PMID- 18158333 TI - Mechanism of botulinum neurotoxin B and G entry into hippocampal neurons. AB - Botulinum neurotoxins (BoNTs) target presynaptic nerve terminals by recognizing specific neuronal surface receptors. Two homologous synaptic vesicle membrane proteins, synaptotagmins (Syts) I and II, bind toxins BoNT/B and G. However, a direct demonstration that Syts I/II mediate toxin binding and entry into neurons is lacking. We report that BoNT/B and G fail to bind and enter hippocampal neurons cultured from Syt I knockout mice. Wild-type Syts I and II (but not Syt I loss-of-function toxin-binding domain mutants) restored binding and entry of BoNT/B and G in Syt I-null neurons, thus demonstrating that Syts I/II are protein receptors for BoNT/B and G. Furthermore, mice lacking complex gangliosides exhibit reduced sensitivity to BoNT/G, and binding and entry of BoNT/A, B, and G into hippocampal neurons lacking gangliosides is diminished. These data suggest that gangliosides are the shared coreceptor for BoNT/A, B, and G, supporting a double-receptor model for these three BoNTs for which the protein receptors are known. PMID- 18158332 TI - Axons of retinal ganglion cells are insulted in the optic nerve early in DBA/2J glaucoma. AB - Here, we use a mouse model (DBA/2J) to readdress the location of insult(s) to retinal ganglion cells (RGCs) in glaucoma. We localize an early sign of axon damage to an astrocyte-rich region of the optic nerve just posterior to the retina, analogous to the lamina cribrosa. In this region, a network of astrocytes associates intimately with RGC axons. Using BAX-deficient DBA/2J mice, which retain all of their RGCs, we provide experimental evidence for an insult within or very close to the lamina in the optic nerve. We show that proximal axon segments attached to their cell bodies survive to the proximity of the lamina. In contrast, axon segments in the lamina and behind the eye degenerate. Finally, the Wld(s) allele, which is known to protect against insults to axons, strongly protects against DBA/2J glaucoma and preserves RGC activity as measured by pattern electroretinography. These experiments provide strong evidence for a local insult to axons in the optic nerve. PMID- 18158334 TI - DNA-activated protein kinase functions in a newly observed S phase checkpoint that links histone mRNA abundance with DNA replication. AB - DNA and histone synthesis are coupled and ongoing replication is required to maintain histone gene expression. Here, we expose S phase-arrested cells to the kinase inhibitors caffeine and LY294002. This uncouples DNA replication from histone messenger RNA (mRNA) abundance, altering the efficiency of replication stress-induced histone mRNA down-regulation. Interference with caffeine-sensitive checkpoint kinases ataxia telangiectasia and Rad3 related (ATR)/ataxia telangiectasia mutated (ATM) does not affect histone mRNA down- regulation, which indicates that ATR/ATM alone cannot account for such coupling. LY294002 potentiates caffeine's ability to uncouple histone mRNA stabilization from replication only in cells containing functional DNA-activated protein kinase (DNA PK), which indicates that DNA-PK is the target of LY294002. DNA-PK is activated during replication stress and DNA-PK signaling is enhanced when ATR/ATM signaling is abrogated. Histone mRNA decay does not require Chk1/Chk2. Replication stress induces phosphorylation of UPF1 but not hairpin[corrected]-binding protein/stem loop binding protein at S/TQ sites, which are preferred substrate recognition motifs of phosphatidylinositol 3-kinase-like kinases, which indicates that histone mRNA stability may be directly controlled by ATR/ATM- and DNA-PK-mediated phosphorylation of UPF1. PMID- 18158335 TI - Huntingtin-interacting protein 14, a palmitoyl transferase required for exocytosis and targeting of CSP to synaptic vesicles. AB - Posttranslational modification through palmitoylation regulates protein localization and function. In this study, we identify a role for the Drosophila melanogaster palmitoyl transferase Huntingtin-interacting protein 14 (HIP14) in neurotransmitter release. hip14 mutants show exocytic defects at low frequency stimulation and a nearly complete loss of synaptic transmission at higher temperature. Interestingly, two exocytic components known to be palmitoylated, cysteine string protein (CSP) and SNAP25, are severely mislocalized at hip14 mutant synapses. Complementary DNA rescue and localization experiments indicate that HIP14 is required solely in the nervous system and is essential for presynaptic function. Biochemical studies indicate that HIP14 palmitoylates CSP and that CSP is not palmitoylated in hip14 mutants. Furthermore, the hip14 exocytic defects can be suppressed by targeting CSP to synaptic vesicles using a chimeric protein approach. Our data indicate that HIP14 controls neurotransmitter release by regulating the trafficking of CSP to synapses. PMID- 18158336 TI - Low concentrations of sphingosylphosphorylcholine enhance pulmonary artery vasoreactivity: the role of protein kinase C delta and Ca2+ entry. AB - Sphingosylphosphorylcholine (SPC) is a powerful vasoconstrictor, but in vitro its EC(50) is approximately 100-fold more than plasma concentrations. We examined whether subcontractile concentrations of SPC (100 nmol/L of SPC, and independent of the endothelium, 2 aminoethoxydiphenylborane-sensitive Ca(2+) entry, and Rho kinase. It was abolished by the phospholipase C inhibitor U73122, the broad spectrum protein kinase C (PKC) inhibitor Ro31-8220, and the PKC delta inhibitor rottlerin, but not by Go6976, which is ineffective against PKC delta. The potentiation could be attributed to enhancement of Ca(2+) entry. SPC also potentiated the responses to prostaglandin F(2 alpha) and U436619, which activate a 2 aminoethoxydiphenylborane sensitive nonselective cation channel in intrapulmonary arteries. In this case, potentiation was partially inhibited by diltiazem but abolished by 2-aminoethoxydiphenylborane, Ro31-8220, and rottlerin. SPC (1 micromol/L) caused translocation of PKC delta to the perinuclear region and cytoskeleton of cultured intrapulmonary artery smooth muscle cells. We present the novel finding that low, subcontractile concentrations of SPC potentiate Ca(2+) entry in intrapulmonary arteries through both voltage-dependent and independent pathways via a receptor-dependent mechanism involving PKC delta. This has implications for the physiological role of SPC, especially in cardiovascular disease, where SPC is reported to be elevated. PMID- 18158337 TI - Lysyl oxidase inhibition is responsible for the vascular elastic fiber phenotype. PMID- 18158338 TI - Conversion of renal angiotensin II to angiotensin III is critical for AT2 receptor-mediated natriuresis in rats. AB - In the kidney, angiotensin II (Ang II) is metabolized to angiotensin III (Ang III) by aminopeptidase A (APA). In turn, Ang III is metabolized to angiotensin IV by aminopeptidase N (APN). Renal interstitial (RI) infusion of Ang III, but not Ang II, results in angiotensin type-2 receptor (AT(2)R)-mediated natriuresis. This response is augmented by coinfusion of PC-18, a specific inhibitor of APN. The present study addresses the hypotheses that Ang II conversion to Ang III is critical for the natriuretic response. Sprague-Dawley rats received systemic angiotensin type-1 receptor (AT(1)R) blockade with candesartan (CAND; 0.01 mg/kg/min) for 24 hours before and during the experiment. After a control period, rats received either RI infusion of Ang II or Ang II+PC-18. The contralateral kidney received a RI infusion of vehicle in all rats. Mean arterial pressure (MAP) was monitored, and urinary sodium excretion rate (U(Na)V) was calculated separately from experimental and control kidneys for each period. In contrast to Ang II-infused kidneys, U(Na)V from Ang II+PC-18-infused kidneys increased from a baseline of 0.03+/-0.01 to 0.09+/-0.02 micromol/min (P<0.05). MAP was unchanged by either infusion. RI addition of PD-123319, an AT(2)R antagonist, inhibited the natriuretic response. Furthermore, RI addition of EC-33, a selective APA inhibitor, abolished the natriuretic response to Ang II+PC-18. These data demonstrate that RI addition of PC-18 to Ang II enables natriuresis mediated by the AT(2)R, and that conversion of Ang II to Ang III is critical for this response. PMID- 18158339 TI - Dietary n-3 polyunsaturated fatty acids and direct renin inhibition improve electrical remodeling in a model of high human renin hypertension. AB - We compared the effect n-3 polyunsaturated fatty acids (PUFAs) with direct renin inhibition on electrophysiological remodeling in angiotensin II-induced cardiac injury. We treated double-transgenic rats expressing the human renin and angiotensinogen genes (dTGRs) from week 4 to 7 with n-3 PUFA ethyl-esters (Omacor; 25-g/kg diet) or a direct renin inhibitor (aliskiren; 3 mg/kg per day). Sprague-Dawley rats were controls. We performed electrocardiographic, magnetocardiographic, and programmed electrical stimulation. Dietary n-3 PUFAs increased the cardiac content of eicosapentaenoic and docosahexaenoic acid. At week 7, mortality in dTGRs was 31%, whereas none of the n-3 PUFA- or aliskiren treated dTGRs died. Systolic blood pressure was modestly reduced in n-3 PUFA treated (180+/-3 mm Hg) compared with dTGRs (208+/-5 mm Hg). Aliskiren-treated dTGRs and Sprague-Dawley rats were normotensive (110+/-3 and 119+/-6 mm Hg, respectively). Both n-3 PUFA-treated and untreated dTGRs showed cardiac hypertrophy and increased atrial natriuretic peptide levels. Prolonged QRS and QT(c) intervals and increased T-wave dispersion in dTGRs were reduced by n-3 PUFAs or aliskiren. Both treatments reduced arrhythmia induction from 75% in dTGRs to 17% versus 0% in Sprague-Dawley rats. Macrophage infiltration and fibrosis were reduced by n-3 PUFAs and aliskiren. Connexin 43, a mediator of intermyocyte conduction, was redistributed to the lateral cell membranes in dTGRs. n-3 PUFAs and aliskiren restored normal localization to the intercalated disks. Thus, n-3 PUFAs and aliskiren improved electrical remodeling, arrhythmia induction, and connexin 43 expression, despite a 70-mm Hg difference in blood pressure and the development of cardiac hypertrophy. PMID- 18158340 TI - Metabolic syndrome and vascular alterations in normotensive subjects at risk of diabetes mellitus. AB - We evaluated the possible association between early vascular abnormalities and the metabolic syndrome (MS) in 77 normotensive subjects (mean age: 50 years) at risk of developing diabetes for family history of diabetes, obesity, or impaired fasting glucose. Fifty healthy subjects were recruited as controls. MS was defined according to the ATP III criteria. Brachial artery endothelium-dependent and -independent vasodilation were assessed as flow-mediated dilation (FMD) and response to glyceryl trinitrate (GTN, 25 microg sublingual), respectively, by automatic computerized edge detection system. Carotid-femoral pulse wave velocity (PWV) and radial augmentation index (AIx) were assessed by applanation tonometry. PWV was significantly (P<0.01) higher in subjects with MS (n=29, 9.0+/-1.9 m/s) as compared with those without MS (n=48, 7.7+/-1.2 m/s) and controls (7.2+/-1.5 m/s). FMD was significantly (P<0.05) reduced in both subjects with (5.8+/-2.7%) and without MS (6.1+/-3.7%) as compared with controls (6.9+/-2.5%). No significant differences were found for response to GTN and AIx. PWV and FMD were significantly (P<0.05) affected by increasing number of MS components. Among the components of the MS, only blood pressure significantly affected PWV, whereas blood pressure and fasting glucose influenced FMD. Logistic regression analysis showed that MS was associated with increased risk of altered PVW (odd ratio: 7.95, confidence limits: 1.06 to 69.11), whereas only blood pressure component was significantly related with increased risk of impaired FMD (odd ratio: 3.60, confidence limits: 1.01 to 12.78). In conclusion, in normotensive subjects at risk of developing diabetes mellitus, the presence of MS is associated with a selective alteration of central PWV. PMID- 18158341 TI - Osteopontin in rat renal fibroblasts: functional properties and transcriptional regulation by aldosterone. AB - Osteopontin (OPN), a proinflammatory cytokine, plays an important role in renal fibrosis. We reported that plasma OPN levels were higher in patients with primary aldosteronism than with essential hypertension. However, the regulatory mechanism of OPN by aldosterone remains unclear. Here, we report the transcriptional regulation of OPN expression by aldosterone and the functional effects of aldosterone-mediated OPN transcription in renal fibroblasts. Aldosterone induced OPN expression in a dose-dependent manner with significant responses at 10 nmol/L (1.6+/-0.2-fold of controls, P<0.05, n=5) and elicited maximal effects at 10 micromol/L (3.5+/-0.4-fold of controls, P<0.01, n=5). Aldosterone increased OPN expression in a time-dependent manner with a maximal effect after 48 hours (2.7+/ 0.3-fold of controls, P<0.01, n=5). This effect was abolished by the mineralocorticoid receptor (MR) antagonist spironolactone. Luciferase promoter deletion assays identified a novel cis regulatory element (-2153 to -1758) in the OPN promoter that is responsive to aldosterone. This element contains an activator protein-1 (AP-1) and nuclear factor kappa B (NF kappaB) site. Electrophoretic mobility shift assays, supershift assays, and chromatin immunoprecipitation assays identified both AP-1 and NF kappaB as the DNA binding proteins induced by aldosterone with spironolactone inhibiting aldosterone induced AP-1 or NF kappaB activity. OPN-siRNA inhibited completely the induction of cell proliferation, type I, III, and IV collagen synthesis by aldosterone. These results indicate that aldosterone induced MR-mediated OPN expression through AP-1 and NF kappaB activation and suggest that aldosterone plays an important role in renal fibrosis through the induction of OPN. PMID- 18158342 TI - Insulin in the brain increases gain of baroreflex control of heart rate and lumbar sympathetic nerve activity. AB - Chronic central administration of insulin increases the gain of baroreflex control of heart rate, but whether baroreflex control of the sympathetic nervous system is similarly affected is unknown. The sites and mechanisms by which brain insulin influences the baroreflex are also unclear. Therefore, the present study tested the hypothesis that acute infusion of insulin into the brain ventricles of urethane-anesthetized rats increases gain of baroreflex control of heart rate and lumbar sympathetic nerve activity and that this action is gender specific. Furthermore, to identify the location within the brain that mediates these effects, insulin was infused into either the lateral ventricle or the fourth ventricle. Lateral ventricular insulin infusion increased the gain of baroreflex control of heart rate (2.1+/-0.3 to 4.0+/-0.6 bpm/mm Hg; P<0.05) and sympathetic activity (2.3+/-0.3% to 4.8+/-1.1% control/mm Hg; P<0.05) within 60 to 90 minutes; however, the increase in heart rate gain was similar in males and females. Increases in the maximum of baroreflex control of heart rate (395+/-10 to 452+/-13 bpm; P<0.05) and of sympathetic activity (156+/-13% to 253+/-22% control; P<0.05) were also observed. In contrast, fourth ventricular insulin infusion failed to alter baroreflex function. In conclusion, increases in brain insulin act acutely in the forebrain to enhance gain of baroreflex control of heart rate and lumbar sympathetic nerve activity. PMID- 18158343 TI - Angiotensin II: one driving force behind atherogenesis. PMID- 18158344 TI - Cellular stretch increases superoxide production in the thick ascending limb. AB - Superoxide (O(2)(-)) is an important regulator of kidney function. We have recently shown that luminal flow stimulates O(2)(-) production in the thick ascending limb (TAL), attributable in part to mechanical factors. Stretch, pressure and shear stress all change when flow increases in the TAL. We hypothesized that stretch rather than shear stress or pressure per se stimulates O(2)(-) production by TALs. We measured O(2)(-) production in isolated perfused rat TALs using fluorescence microscopy and dihydroethidium. Tubules were perfused with a Na-free solution to eliminate the confounding effect of Na transport. Flow induced an increase in O(2)(-) production from 29+/-4 to 90+/-8 AU/s (P<0.002; n=5). The response to flow is rapidly reversible. O(2)(-) production by TALs perfused at 10 nL/min decreased from 113+/-6 to 25+/-10 AU/s (P<0.003; n=4) 15 minutes after flow was stopped. Increasing pressure and stretch in the absence of shear stress caused a significant increase in O(2)(-) production (40+/-6 to 118+/ 17 AU/s; P<0.02; n=5). In contrast, eliminating shear stress had no effect (107+/ 9 versus 108+/-10 AU/s; n=5). Increasing stretch by 27+/-2% in the presence of flow while reducing pressure stimulated O(2)(-) production from 66+/-7 to 84+/-9 AU/s (29+/-8%; P<0.02; n=5). Tempol inhibited this increase (n=5). We conclude that increasing stretch rather than pressure or shear stress accounts for the mechanical aspect of flow-induced O(2)(-) production in the TAL. Stretch of the TAL during hypertension, diabetes, and salt loading may contribute to renal damage. PMID- 18158345 TI - Oxidative stress causes renal dopamine D1 receptor dysfunction and salt-sensitive hypertension in Sprague-Dawley rats. AB - Renal dopamine plays an important role in maintaining sodium homeostasis and blood pressure (BP) during increased sodium intake. The present study was carried out to determine whether renal dopamine D1 receptor (D1R) dysfunction contributes to increase in salt sensitivity during oxidative stress. Male Sprague-Dawley rats, divided into various groups, received tap water (vehicle); 1% NaCl (high salt [HS]); L-buthionine sulfoximine (BSO), an oxidant; and HS plus BSO with or without Tempol, an antioxidant, for 12 days. Compared with vehicle, HS intake increased urinary dopamine production and decreased basal renal Na/K-ATPase activity but did not affect BP. BSO-treated rats exhibited oxidative stress and a mild increase in BP. In these rats, D1R expression and G protein coupling were reduced, and SKF38393, a D1R agonist, failed to inhibit Na/K-ATPase activity and promote sodium excretion. Concomitant administration of BSO and HS caused oxidative stress, D1R dysfunction, and a marked increase in BP. Although renal dopamine production was increased, it failed to reduce the basal Na/K-ATPase activity in these animals. Treatment of BSO plus HS rats with Tempol decreased oxidative stress and restored endogenous, as well as exogenous, D1R agonist mediated Na/K-ATPase inhibition and normalized BP. In conclusion, during HS intake, the increased dopamine production via Na/K-ATPase inhibition prevents an increase in BP. During oxidative stress, D1R function is defective, and there is mild hypertension. However, in the presence of oxidative stress, HS intake causes marked elevation in BP, which results from a defective renal D1R function leading to the failure of dopamine to inhibit Na/K-ATPase and promote sodium excretion. PMID- 18158346 TI - Resistin, glomerular filtration rate, and insulin resistance. PMID- 18158347 TI - Assessing left ventricular performance: a rashomon effect. PMID- 18158348 TI - Pulse pressure is inversely related to aortic root diameter implications for the pathogenesis of systolic hypertension. AB - Hypertension accelerates the age-associated increase in aortic root diameter (AoD), likely because of chronically elevated distending pressures. However, the pulsatile component of blood pressure may have a different relationship with AoD. We sought to assess the relationship between AoD and pulse pressure (PP) while accounting for left ventricular and central arterial structural and functional properties, which are known to influence PP. The study population was composed of 1256 individuals, aged 30 to 79 years (48% women and 48% hypertensive), none of whom were on antihypertensive medications. Blood pressure was measured in the sitting position with conventional sphygmomanometry. PP was calculated as the difference between systolic and diastolic blood pressures. AoD was measured at end diastole at the level of the sinuses of Valsalva with echocardiography. The relationship between AoD and PP was evaluated with multiple regression analyses. PP was 50+/-14 mm Hg in men and 54+/-18 mm Hg in women, and AoD was 31.9+/-3.5 mm in men and 28.9+/-3.5 mm in women. After adjusting for age, age(2), height, weight, and mean arterial pressure, AoD was independently and inversely associated with PP in both sexes. After further adjustments for central arterial stiffness and wall thickness, reflected waves, and left ventricular geometry, AoD remained inversely associated with PP in both men (coefficient=-0.48; P=0.0003; model R(2)=0.51) and women (coefficient=-0.40; P=0.01; model R(2)=0.61). Thus, AoD is inversely associated with PP, suggesting that a small AoD may contribute to the pathogenesis of systolic hypertension. Longitudinal studies are needed to examine this possibility. PMID- 18158349 TI - Endothelial dysfunction and the development of renal injury in spontaneously hypertensive rats fed a high-fat diet. AB - Obesity and hypertension have been identified as cardiovascular risk factors that contribute to the progression of end-stage renal disease. To examine the mechanisms by which a high-fat diet and hypertension contribute to endothelial dysfunction and renal injury, 8-week-old male spontaneously hypertensive rats and Wistar rats were fed a high-fat (36% fat) or a normal-fat (7% fat) diet for 10 weeks. The high-fat diet increased body weight in Wistar and hypertensive rats by 25 and 31 g, respectively. Systolic blood pressure was higher in the hypertensive rats compared with Wistar rats; however, blood pressure was unaltered by the high fat diet. Afferent arteriole response to acetylcholine was impaired in the high fat groups after just 3 weeks. Renal macrophage infiltration was increased in the hypertensive high-fat group compared with others, and monocyte chemoattractant protein-1 excretion was increased in both of the high-fat-fed groups. Renal PCR arrays displayed significant increases in 2 inflammatory genes in hypertensive rats fed a normal diet, 1 gene was increased in high-fat-fed Wistar rats, whereas 12 genes were increased in high-fat-fed hypertensive rats. Urinary albumin excretion was increased in the hypertensive rats compared with the Wistar rats, which was further exacerbated by the high-fat diet. Glomerular nephrin expression was reduced and desmin was increased by the high-fat diet in the hypertensive rats. Our results indicate that endothelial dysfunction precedes renal injury in normotensive and spontaneously hypertensive rats fed a high-fat diet, and hypertension with obesity induces a powerful inflammatory response and disruption of the renal filtration barrier. PMID- 18158350 TI - Beneficial effects of pioglitazone on hypertensive cardiovascular injury are enhanced by combination with candesartan. AB - The effect of pioglitazone, a peroxisome proliferator-activated receptor gamma agonist, on hypertensive cardiovascular injury is unknown. We examined the effect of pioglitazone on hypertensive cardiovascular injury and the significance of combination of pioglitazone with angiotensin type 1 receptor blocker. Stroke prone spontaneously hypertensive rats (SHRSP) were orally given pioglitazone, candesartan, or combined pioglitazone and candesartan for 4 weeks to compare their effects on cardiovascular injury. Pioglitazone, without lowering blood pressure, significantly suppressed cardiac inflammation and fibrosis and reduced vascular endothelial dysfunction, and these beneficial effects were associated with the reduction of superoxide by inhibition of cardiovascular NADPH oxidase. Thus, pioglitazone protects against hypertensive cardiovascular injury, by inhibiting reactive oxygen species (ROS). Combination of pioglitazone and candesartan suppressed cardiac hypertrophy, inflammation, and interstitial fibrosis of SHRSP to a greater extent than either monotherapy, and reduced vascular endothelial dysfunction of SHRSP more than either monotherapy. Furthermore, more beneficial effects of their combination on cardiovascular injury were associated with more reduction of NADPH oxidase-mediated cardiovascular ROS. To elucidate the underlying molecular mechanism, we examined cardiovascular NADPH oxidase subunits. Pioglitazone monotherapy significantly attenuated cardiovascular p22(phox) and Rac1 in SHRSP, whereas pioglitazone combined with candesartan more attenuated p22(phox) and significantly reduced Nox1. Thus, additive suppression of cardiovascular NADPH oxidase by the combination was attributed to its additive attenuation of p22(phox) and Nox1 protein levels. In conclusion, we showed that pioglitazone protected against hypertensive cardiovascular damage, and the combination of pioglitazone and candesartan exerted more beneficial effects on hypertensive cardiovascular injury by more suppressing ROS. PMID- 18158351 TI - Telmisartan inhibits CD4-positive lymphocyte migration independent of the angiotensin type 1 receptor via peroxisome proliferator-activated receptor-gamma. AB - Migration of CD4-positive lymphocytes into the vessel wall represents an important step in early atherogenesis. Telmisartan is an angiotensin type 1 receptor (AT1R) blocker with peroxisome proliferator-activated receptor (PPAR) gamma-activating properties. The present study examined the effect of telmisartan on CD4-positive cell migration and the role of PPARgamma in this context. CD4 positive lymphocytes express both the AT1R and PPARgamma. Stimulation of CD4 positive lymphocytes with stromal cell-derived factor (SDF)-1 leads to a 4.1+/ 3.1-fold increase in cell migration. Pretreatment of cells with telmisartan reduces this effect in a concentration-dependent manner to a maximal 1.6+/-0.7 fold induction at 10 mumol/L of telmisartan (P<0.01 compared with SDF-1-treated cells; n=22). Three different PPARgamma activators, rosiglitazone, pioglitazone, and GW1929, had similar effects, whereas eprosartan, a non-PPARgamma-activating AT1R blocker, did not affect chemokine-induced lymphocyte migration. Telmisartan's effect on CD4-positive lymphocyte migration was mediated through an early inhibition of chemokine-induced phosphatidylinositol 3-kinase activity. Downstream, telmisartan inhibited F-actin formation, as well as intercellular adhesion molecule-3 translocation. Transfection of CD4-positive lymphocytes with PPARgamma small interfering RNA abolished telmisartan's effect on migration, whereas blockade of the AT1R had no such effect. Telmisartan inhibits chemokine induced CD4-positive cell migration independent of the AT1R via PPARgamma. These data provide a novel mechanism to explain how telmisartan modulates lymphocyte activation by its PPARgamma-activating properties. PMID- 18158352 TI - Protective effect of dietary potassium against vascular injury in salt-sensitive hypertension. AB - Hypertensive cardiovascular damage is accelerated by salt loading but counteracted by dietary potassium supplementation. We suggested recently that antioxidant actions of potassium contribute to protection against salt-induced cardiac dysfunction. Therefore, we examined whether potassium supplementation ameliorated cuff-induced vascular injury in salt-sensitive hypertension via suppression of oxidative stress. Four-week-old Dahl salt-sensitive rats were fed a normal-salt (0.3% NaCl), high-salt (8% NaCl), or high-salt plus high-potassium (8% KCl) diet for 5 weeks, and some of the rats fed a high-salt diet were also given antioxidants. One week after the start of the treatments, a silicone cuff was implanted around the femoral artery. Examination revealed increased cuff induced neointimal proliferation with adventitial macrophage infiltration in arteries from salt-loaded Dahl salt-sensitive rats compared with that in arteries from non-salt-loaded animals (intima/media ratio: 0.471+/-0.070 versus 0.302+/ 0.037; P<0.05), associated with regional superoxide overproduction and reduced nicotinamide-adenine dinucleotide phosphate oxidase activation and mRNA overexpression. On the other hand, simultaneous potassium supplementation attenuated salt-induced neointimal hyperplasia (intima/media ratio: 0.205+/ 0.012; P<0.001), adventitial macrophage infiltration, superoxide overproduction, and reduced nicotinamide-adenine dinucleotide phosphate oxidase activation and overexpression. Antioxidants, which decrease vascular oxidative stress, also reduced neointima formation induced by salt excess. In conclusion, high-potassium diets seems to have a protective effect against the development of vascular damage induced by salt loading mediated, at least in part, through suppression of the production of reactive oxygen species probably generated by reduced nicotinamide-adenine dinucleotide phosphate oxidase. PMID- 18158353 TI - Gene expression pattern in biomechanically stretched cardiomyocytes: evidence for a stretch-specific gene program. AB - Biomechanical stress ie, attributable to pressure overload, leads to cardiac hypertrophy and may ultimately cause heart failure. Yet, it is still unclear how mechanical stress is sensed and transduced on the molecular level. To systematically elucidate the underlying signal transduction pathways, we analyzed the gene expression profile of stretched cardiomyocytes on a genome-wide scale in comparison with other inducers of hypertrophy such as pharmacological stimulation. Neonatal rat ventricular cardiomyocytes were either stretched biaxially or stimulated with phenylephrine (PE), both resulting in a similar degree of hypertrophy. Microarray analyses revealed 164 genes >2.0-fold up- and 21 genes <0.5-fold downregulated (P<0.01). Differential expression was confirmed by real-time polymerase chain reaction. Genes of the "fetal gene program" such as BNP were induced by both stretch (4.2x) and PE (2.9x). We also verified upregulation of known stretch-responsive genes, including HSP70 (20.9x) and c-myc (3.0x). Moreover, several genes were found to be preferentially induced by stretch, such as the cardioprotective cytokine GDF15 (24.8x) and heme oxygenase 1 (Hmox1, 10.8x; both confirmed on protein level). Neither PE nor endothelin-1 upregulated GDF15 and Hmox1, whereas angiotensin II significantly induced both genes. Conversely, the AT(1) receptor blocker irbesartan markedly blunted stretch mediated GDF15 and Hmox1 upregulation, suggesting that the angiotensin receptor transduces the biomechanical induction of these genes. In conclusion, we report a comprehensive gene expression profile of cardiomyocytes subjected to biomechanical stress in comparison with pharmacologically induced hypertrophy. Our data imply that a stretch-specific gene program exists, which is mediated, at least in part, by angiotensin II-dependent signaling. PMID- 18158354 TI - Arterioventricular coupling and ventricular efficiency after antihypertensive therapy: a noninvasive prospective study. AB - Patients with hypertension exhibit impaired energetic coupling between the ventricle and the arterial system, leading to reduced cardiac mechanic efficiency and exercise capacity. We tested whether blood pressure normalization with current antihypertensive therapy can improve arterioventricular coupling. Eighteen hypertensive patients without other cardiovascular disease were examined before and after antihypertensive therapy. Transthoracic echocardiography was performed. Central aortic pressure waveforms, including end-systolic pressure, were derived from radial artery applanation tonometry. Afterload was increased with isometric handgrip exercise. Central aortic end-systolic pressure and ventricular volumes at rest and handgrip were used to calculate ventricular elastance, effective arterial elastance, arterioventricular coupling (effective arterial elastance/ventricular elastance), and mechanical efficiency. After 142+/ 67 days, systolic blood pressure decreased from 150.9+/-14.6 to 119.8+/-9.2 mm Hg (P<0.00001), diastolic blood pressure from 85.9+/-14.8 to 68.8+/-8.4 mm Hg (P=0.00002), and cardiac output from 5.8+/-1.7 to 4.9+/-1.8 L/min (P=0.03). Resting left ventricular end-systolic volume, ejection fraction, and septal thickness did not change. Ventricular elastance increased from 1.7+/-1.0 to 3.2+/ 1.4 mm Hg/mL (P=0.00002), whereas effective arterial elastance decreased from 1.4+/-0.5 to 1.2+/-0.4 mm Hg/mL (P=0.02). Effective arterial elastance/ventricular elastance decreased in all patients, from 1.1+/-0.8 to 0.4+/-0.2 (P=0.0002). Efficiency improved at rest (72.9+/-5.8% versus 83.5+/ 5.7%; P<0.00001) and during handgrip (63.5+/-7.8% versus 78.9+/-7.1%; P<0.00001). In hypertensive patients, optimal brachial and central blood pressure reduction shifts arterioventricular coupling from cardiac output maximization to ventricular mechanical efficiency optimization. This occurs before significant changes in ventricular geometry and may be responsible for early clinical improvements. PMID- 18158355 TI - Angiotensin-converting enzyme C-terminal catalytic domain is the main site of angiotensin I cleavage in vivo. AB - Angiotensin-converting enzyme (ACE) plays a central role in the production of the vasoconstrictor angiotensin II. ACE is a single polypeptide, but it contains 2 homologous and independent catalytic domains, each of which binds zinc. To understand the in vivo role of these 2 domains, we used gene targeting to create mice with point mutations in the ACE C-domain zinc-binding motif. Such mice, termed ACE13/13, produce a full-length ACE protein with tissue expression identical to wild-type mice. Analysis of ACE13/13 mice showed that they produce ACE having only N-domain catalytic activity, as determined by the hydrolysis of domain specific substrates and by chloride sensitivity. ACE13/13 mice have blood pressure and blood angiotensin II levels similar to wild-type mice. However, plasma renin concentration is increased 2.6-fold and blood angiotensin I levels are increased 7.5-fold. Bradykinin peptide levels are not different from wild type levels. ACE13/13 mice have a reduced increase of blood pressure after intravenous infusion of angiotensin I. ACE13/13 mice have a normal renal structure, but they are not able to concentrate urine after dehydration as effectively as wild-type mice. This study shows that the C-domain of ACE is the predominant site of angiotensin I cleavage in vivo. Although mice lacking C domain activity have normal physiology under laboratory conditions, they respond less well to the stress of dehydration. PMID- 18158356 TI - Angiotensin II type 2 receptors and nitric oxide sustain oxygenation in the clipped kidney of early Goldblatt hypertensive rats. AB - Angiotensin-converting enzyme inhibitors (ACEIs) decrease the glomerular filtration rate and renal blood flow in the clipped kidneys of early 2-kidney, 1 clip Goldblatt hypertensive rats, but the consequences for oxygenation are unclear. We investigated the hypothesis that angiotensin II type 1 or angiotensin II type 2 receptors or NO synthase mediate renal oxygenation responses to ACEI. Three weeks after left renal artery clipping, kidney function, oxygen (O(2)) use, renal blood flow, renal cortical blood flow, and renal cortical oxygen tension (Po(2)) were measured after acute administration of an ACEI (enalaprilat) and after acute administration of ACEI following acute administration of an angiotensin II type 1 or angiotensin II type 2 receptor blocker (candesartan or PD-123,319) or an NO synthase blocker (N(G)-nitro-L-arginine methyl ester with control of renal perfusion pressure) and compared with mechanical reduction in renal perfusion pressure to the levels after ACEI. The basal renal cortical Po(2) of clipped kidneys was significantly lower than contralateral kidneys (35+/-1 versus 51+/-1 mm Hg; n=40 each). ACEI lowered renal venous Po(2), cortical Po(2), renal blood flow, glomerular filtration rate, and cortical blood flow and increased the renal vascular resistance in the clipped kidney, whereas mechanical reduction in renal perfusion pressure was ineffective. PD-123,319 and N(G)-nitro L-arginine methyl ester, but not candesartan, reduced the Po(2) of clipped kidneys and blocked the fall in Po(2) with acute ACEI administration. In conclusion, oxygen availability in the clipped kidney is maintained by angiotensin II generation, angiotensin II type 2 receptors, and NO synthase. This discloses a novel mechanism whereby angiotensin can prevent hypoxia in a kidney challenged with a reduced perfusion pressure. PMID- 18158357 TI - Deoxycorticosterone acetate salt hypertension in apolipoprotein E-/- mice results in accelerated atherosclerosis: the role of angiotensin II. AB - Previous studies have shown that administration of angiotensin II to atherosclerosis-prone animal models results in an increase in the extent of atherosclerosis and that this effect may be independent of changes in blood pressure. We sought to determine whether atherosclerosis was increased in the setting of a low renin model of hypertension. Apolipoprotein E-deficient mice were made hypertensive using the deoxycorticosterone acetate salt model. We found that this resulted in a dramatic increase in the atherosclerotic lesion area in the setting of either a low- or high-fat diet. In the hypertensive animals, we observed an increase in angiotensin II staining that was localized to the adventitial macrophages. The increase in atherosclerosis was inhibited by administration of an angiotensin receptor antagonist, an angiotensin-converting enzyme inhibitor, or a renin inhibitor. In addition, blood pressure reduction, with either a calcium channel blocker or hydralazine, reduced the extent of atherosclerosis indicating an important contribution of the mechanical effects of elevated blood pressure. These data suggest that, even in the setting of hypertension that is not associated with activation of the systemic renin angiotensin system, local generation of angiotensin II within the arterial wall may be of pathophysiological relevance to the development of atherosclerosis. PMID- 18158358 TI - Nanoparticle PET-CT imaging of macrophages in inflammatory atherosclerosis. AB - BACKGROUND: Macrophages participate centrally in atherosclerosis, and macrophage markers (eg, CD68, MAC-3) correlate well with lesion severity and therapeutic modulation. On the basis of the avidity of lesional macrophages for polysaccharide-containing supramolecular structures such as nanoparticles, we have developed a new positron emission tomography (PET) agent with optimized pharmacokinetics to allow in vivo imaging at tracer concentrations. METHODS AND RESULTS: A dextranated and DTPA-modified magnetofluorescent 20-nm nanoparticle was labeled with the PET tracer 64Cu (1 mCi/0.1 mg nanoparticles) to yield a PET, magnetic resonance, and optically detectable imaging agent. Peak PET activity 24 hours after intravenous injection into mice deficient in apolipoprotein E with experimental atherosclerosis mapped to areas of high plaque load identified by computed tomography such as the aortic root and arch and correlated with magnetic resonance and optical imaging. Accumulated dose in apolipoprotein E-deficient aortas determined by gamma counting was 260% and in carotids 392% of respective wild-type organs (P<0.05 both). Autoradiography of aortas demonstrated uptake of the agent into macrophage-rich atheromata identified by Oil Red O staining of lipid deposits. The novel nanoagent accumulated predominantly in macrophages as determined by fluorescence microscopy and flow cytometry of cells dissociated from aortas. CONCLUSIONS: This report establishes the capability of a novel trimodality nanoparticle to directly detect macrophages in atherosclerotic plaques. Advantages include improved sensitivity; direct correlation of PET signal with an established biomarker (CD68); ability to readily quantify the PET signal, perform whole-body vascular surveys, and spatially localize and follow the trireporter by microscopy; and clinical translatability of the agent given similarities to magnetic resonance imaging probes in clinical trials. PMID- 18158359 TI - Reversibility of PRKAG2 glycogen-storage cardiomyopathy and electrophysiological manifestations. AB - BACKGROUND: PRKAG2 mutations cause glycogen-storage cardiomyopathy, ventricular preexcitation, and conduction system degeneration. A genetic approach that utilizes a binary inducible transgenic system was used to investigate the disease mechanism and to assess preventability and reversibility of disease features in a mouse model of glycogen-storage cardiomyopathy. METHODS AND RESULTS: Transgenic (Tg) mice expressing a human N488I PRKAG2 cDNA under control of the tetracycline repressible alpha-myosin heavy chain promoter underwent echocardiography, ECG, and in vivo electrophysiology studies. Transgene suppression by tetracycline administration caused a reduction in cardiac glycogen content and was initiated either prenatally (Tg(OFF(E-8 weeks))) or at different time points during life (Tg(OFF(4-16 weeks)), Tg(OFF(8-20 weeks)), and Tg(OFF(>20 weeks))). One group never received tetracycline, expressing transgene throughout life (Tg(ON)). Tg(ON) mice developed cardiac hypertrophy followed by dilatation, ventricular preexcitation involving multiple accessory pathways, and conduction system disease, including sinus and atrioventricular node dysfunction. CONCLUSIONS: Using an externally modifiable transgenic system, cardiomyopathy, cardiac dysfunction, and electrophysiological disorders were demonstrated to be reversible processes in PRKAG2 disease. Transgene suppression during early postnatal development prevented the development of accessory electrical pathways but not cardiomyopathy or conduction system degeneration. Taken together, these data provide insight into mechanisms of cardiac PRKAG2 disease and suggest that glycogen-storage cardiomyopathy can be modulated by lowering glycogen content in the heart. PMID- 18158360 TI - Dietary alpha-lipoic acid supplementation inhibits atherosclerotic lesion development in apolipoprotein E-deficient and apolipoprotein E/low-density lipoprotein receptor-deficient mice. AB - BACKGROUND: Vascular inflammation and lipid deposition are prominent features of atherosclerotic lesion formation. We have shown previously that the dithiol compound alpha-lipoic acid (LA) exerts antiinflammatory effects by inhibiting tumor necrosis factor-alpha- and lipopolysaccharide-induced endothelial and monocyte activation in vitro and lipopolysaccharide-induced acute inflammatory responses in vivo. Here, we investigated whether LA inhibits atherosclerosis in apolipoprotein E-deficient (apoE-/-) and apoE/low-density lipoprotein receptor deficient mice, 2 well-established animal models of human atherosclerosis. METHODS AND RESULTS: Four-week-old female apoE-/- mice (n=20 per group) or apoE/low-density lipoprotein receptor-deficient mice (n=21 per group) were fed for 10 weeks a Western-type chow diet containing 15% fat and 0.125% cholesterol without or with 0.2% (wt/wt) R,S-LA or a normal chow diet containing 4% fat without or with 0.2% (wt/wt) R-LA, respectively. Supplementation with LA significantly reduced atherosclerotic lesion formation in the aortic sinus of both mouse models by approximately 20% and in the aortic arch and thoracic aorta of apoE-/- and apoE/low-density lipoprotein receptor-deficient mice by approximately 55% and 40%, respectively. This strong antiatherogenic effect of LA was associated with almost 40% less body weight gain and lower serum and very low density lipoprotein levels of triglycerides but not cholesterol. In addition, LA supplementation reduced aortic expression of adhesion molecules and proinflammatory cytokines and aortic macrophage accumulation. These antiinflammatory effects of LA were more pronounced in the aortic arch and the thoracic aorta than in the aortic sinus, reflecting the corresponding reductions in atherosclerosis. CONCLUSIONS: Our study shows that dietary LA supplementation inhibits atherosclerotic lesion formation in 2 mouse models of human atherosclerosis, an inhibition that appears to be due to the "antiobesity," antihypertriglyceridemic, and antiinflammatory effects of LA. LA may be a useful adjunct in the prevention and treatment of atherosclerotic vascular diseases. PMID- 18158361 TI - Adiponectin prevents cerebral ischemic injury through endothelial nitric oxide synthase dependent mechanisms. AB - BACKGROUND: Adiponectin is a fat-derived plasma protein that has beneficial actions on cardiovascular disorders. A low level of plasma adiponectin is associated with increased mortality after ischemic stroke; however, the causal role of adiponectin in ischemic stroke is unknown. METHODS AND RESULTS: To explore the role of adiponectin in the development of acute cerebral injury, we subjected adiponectin-deficient (APN-KO) and wild-type (WT) mice to 1 hour of middle cerebral artery occlusion followed by 23 hours of reperfusion. APN-KO mice exhibited enlarged brain infarction and increased neurological deficits after ischemia-reperfusion compared with WT mice. Conversely, adenovirus-mediated supplementation of adiponectin significantly reduced cerebral infarct size in WT and APN-KO mice. APN-KO mice showed decreased cerebral blood flow during ischemia by laser speckle flowmetry methods. Adiponectin colocalized within the cerebral vascular endothelium under transient ischemic conditions by immunohistochemical analysis. Phosphorylation of endothelial nitric oxide synthase in ischemic brain tissues and the production of nitric oxide metabolites in plasma were attenuated in APN-KO mice compared with WT mice. Adenovirus-mediated administration of adiponectin stimulated endothelial nitric oxide synthase phosphorylation and nitric oxide metabolites during cerebral ischemia in both WT and APN-KO mice. Neuronal nitric oxide synthase expression during ischemia did not differ between WT and APN-KO mice. Adenovirus-mediated delivery of adiponectin did not affect brain infarction in mice deficient in endothelial nitric oxide synthase. CONCLUSIONS: These data provide causal evidence that adiponectin exerts a cerebroprotective action through an endothelial nitric oxide synthase-dependent mechanism. Adiponectin could represent a molecular target for the prevention of ischemic stroke. PMID- 18158362 TI - High absolute risks and predictors of venous and arterial thromboembolic events in patients with nephrotic syndrome: results from a large retrospective cohort study. AB - BACKGROUND: No data are available on the absolute risk of either venous thromboembolism (VTE) or arterial thromboembolism (ATE) in patients with nephrotic syndrome. Reported risks are based on multiple case reports and small studies with mostly short-term follow-up. We assessed the absolute risk of VTE and ATE in a large, single-center, retrospective cohort study and attempted to identify predictive factors in these patients. METHODS AND RESULTS: A total of 298 consecutive patients with nephrotic syndrome (59% men; mean age, 42+/-18 years) were enrolled. Mean follow-up was 10+/-9 years. Nephrotic syndrome was defined by proteinuria > or =3.5 g/d, and patients were classified according to underlying histological lesions accounting for nephrotic syndrome. Objectively verified symptomatic thromboembolic events were the primary study outcome. Annual incidences of VTE and ATE were 1.02% (95% confidence interval, 0.68 to 1.46) and 1.48% (95% confidence interval, 1.07 to 1.99), respectively. Over the first 6 months of follow-up, these rates were 9.85% and 5.52%, respectively. Proteinuria and serum albumin levels tended to be related to VTE; however, only the predictive value of the ratio of proteinuria to serum albumin was significant (hazard ratio, 5.6; 95% confidence interval, 1.2 to 26.2; P=0.03). In contrast, neither the degree of proteinuria nor serum albumin levels were related to ATE. Sex, age, hypertension, diabetes, smoking, prior ATE, and estimated glomerular filtration rate predicted ATE (P< or =0.02). CONCLUSIONS: This study verifies high absolute risks of symptomatic VTE and ATE that were remarkably elevated within the first 6 months. Whereas the ratio of proteinuria to serum albumin predicted VTE, estimated glomerular filtration rate and multiple classic risk factors for atherosclerosis were predictors of ATE. PMID- 18158363 TI - Proarrhythmic aspects of atrial fibrillation surgery: mechanisms of postoperative macroreentrant tachycardias. AB - BACKGROUND: Surgery for the treatment of atrial fibrillation may be associated with early or late postoperative atrial arrhythmias. In many cases, the arrhythmias that occur early in the postoperative course may be related to pericardial inflammation or increased catecholamine levels and tend to resolve without further therapy. In contrast, late postoperative arrhythmias often are persistent, highly symptomatic, and refractory to medical therapy. The incidence and mechanisms of these late atrial arrhythmias have not been thoroughly described. METHODS AND RESULTS: In this study, a total of 143 consecutive patients undergoing atrial fibrillation surgery from 1996 to 2005 were prospectively followed up. Sustained atrial tachycardia developed in 22 of 143 patients (15%) >8 weeks after the surgical procedure. These patients underwent electrophysiological study. A total of 25 separate tachycardias were mapped in these 22 patients, including 15 located in right atrium and 10 in the left atrium. Right atrial tachycardias included cavotricuspid isthmus-dependent atrial flutter (n=7), non-isthmus-dependent right atrial reentry (n=7), and 1 focal atrial tachycardia. Left atrial tachycardias included reentry around the mitral valve (n=3) and in the roof of the left atrium (n=7). All tachycardias were successfully treated with radiofrequency catheter ablation with no recurrences in long-term follow-up. CONCLUSIONS: Postoperative arrhythmias are not uncommon late after surgery for atrial fibrillation and typically are reentrant and related to surgically created boundaries. Radiofrequency catheter ablation is highly effective in the management of these late arrhythmias. PMID- 18158364 TI - Negative affect and chemical intolerance as risk factors for building-related symptoms: a controlled exposure study. AB - OBJECTIVE: To assess whether differences in negative affect (NA) and chemical intolerance (CI) affect responses to chemical mixtures and stress in a controlled experimental model. METHODS: Participants were 130 nonsmoking, healthy women, recruited from a university community. Participants completed the Positive and Negative Affect Scale and the Chemical Odor Intolerance Index. In separate sessions 1 week apart, they were exposed to volatile organic compounds (VOCs), VOCs with ozone (VOCs+O3), and ambient or filtered air with a 1-minute spike of VOCs (masked clean air). During each session, half of the participants performed a videotaped speech stressor and half performed simple arithmetic. Before, during, and after each session, salivary cortisol samples were collected, and subjects completed neurobehavioral tests and used a ratio scale to rate physical, cognitive, and anxiety symptoms. RESULTS: Relative to low NA or low CI, neither the high NA nor the high CI groups reported significantly more symptoms in response to any exposure condition. High NA subjects reported more anxiety symptoms in response to the speech stressor but did not have higher cortisol than low NA subjects. High NA subjects, however, were more distressed by the experimental conditions than were low NA subjects. Low NA subjects reported more severe anxiety in the VOCs+O3 with psychological stress condition. CONCLUSIONS: Subjects high in NA were more anxious after a stressor but were not more physically symptomatic in response to increasing chemical exposures. A disposition toward high or low CI did not result in a differential symptomatic response to controlled chemical exposures. PMID- 18158365 TI - Violence exposure, a chronic psychosocial stressor, and childhood lung function. AB - OBJECTIVE: Chronic psychosocial stressors, including violence, and neuropsychological and behavioral development in children as well as physiologic alterations that may lead to broader health effects. METHODS: We studied the relationship between violence and childhood lung function in a prospective birth cohort of 313 urban children (age range = 6-7 years). Mothers reported on their child's lifetime exposure to community violence (ETV) and interparental conflict in the home (Conflict Tactics Scale (CTS)) within 1 year of the lung function assessment. RESULTS: In linear regression analyses, adjusting for maternal education, child's age, race, birthweight, tobacco smoke exposure, and medical history, girls in the highest CTS verbal aggression tertile had a 5.5% (95% confidence interval (CI) = -9.6, -1.5) decrease in percent predicted forced expiratory volume (FEV(1)) and a 5.4% (95% CI = -9.7, -1.1) decrease in forced vital capacity (FVC) compared with girls in the lowest tertile. The CTS verbal aggression subscale was associated with lung function among boys in the same direction, albeit this was not statistically significant. Boys in the highest ETV tertile had a 3.4% (95% CI = -8.0, 1.1) lower FEV(1) and 5.3% lower FVC (95% CI = -10.2, -0.4) compared with boys in the lowest tertile. The ETV score was not a significant predictor of girls' lung function. CONCLUSIONS: Interparental conflict, specifically verbal aggression, and ETV were associated with decreased childhood lung function independent of socioeconomic status, tobacco smoke exposure, birthweight, and respiratory illness history. Gender differences were noted based on the type of violence exposure, which may warrant further exploration. PMID- 18158366 TI - Depression and anxiety in adults with sickle cell disease: the PiSCES project. AB - OBJECTIVE: Depression and anxiety are common in sickle cell disease (SCD) but relatively little is known about their impact on SCD adults. This study measured prevalence of depression and anxiety in SCD adults, and their effects on crisis and noncrisis pain, quality-of-life, opioid usage, and healthcare utilization. METHODS: The Pain in Sickle Cell Epidemiology Study is a prospective cohort study in 308 SCD adults. Baseline variables included demographics, genotype, laboratory data, health-related quality-of-life, depression, and anxiety. Subjects completed daily diaries for up to 6 months, reporting sickle cell pain intensity, distress, interference, whether they were in a sickle cell crisis, as well as health care and opioid utilization. RESULTS: Two hundred thirty-two subjects who completed at least 1 month of diaries were studied; 27.6% were depressed and 6.5% had any anxiety disorder. Depressed subjects had pain on significantly more days than nondepressed subjects (mean pain days 71.1% versus 49.6%, p < .001). When in pain on noncrisis days, depressed subjects had higher mean pain, distress from pain, and interference from pain. Both depressed and anxious subjects had poorer functioning on all eight SF-36 subscales, even after controlling for demographics, hemoglobin type, and pain. The anxious subjects had more pain, distress from pain, and interference from pain, both on noncrisis pain days and on crisis days, and used opioids more often. CONCLUSIONS: Depression and anxiety predicted more daily pain and poorer physical and mental quality-of-life in adults with SCD, and accounted for more of the variance in all domains of quality of-life than hemoglobin type. PMID- 18158367 TI - HPA axis reactivity and lymphocyte glucocorticoid sensitivity in fibromyalgia syndrome and chronic pelvic pain. AB - OBJECTIVE: Chronic pelvic pain (CPP) and fibromyalgia syndrome (FMS) have been associated with hypothalamic-pituitary-adrenal (HPA) axis alterations, i.e., mild hypocortisolism and enhanced feedback sensitivity. We tested the hypothesis of reduced cortisol release in response to a psychosocial stressor and pharmacological stimulation. Furthermore, glucocorticoid (GC) sensitivity was evaluated. METHODS: Plasma total and salivary-free cortisol concentrations were measured in response to a standardized social laboratory stressor, the Trier Social Stress Test, and to adrenocorticotropin (ACTH)(1-24) stimulation. In the Trier Social Stress Test, we additionally measured ACTH. GC sensitivity was measured by dexamethasone inhibition of lipopolysaccharide-induced interleukin-6 and tumor necrosis factor-alpha production in whole blood. RESULTS: There were no HPA axis alterations in women with CPP (N = 18) in these tests. Patients with FMS (N = 17) showed lower total cortisol release in response to the social stressor and exogenous ACTH, but normal free cortisol and ACTH levels compared with controls (N = 24). GC sensitivity was similar in all groups. CONCLUSIONS: Our results suggest normal HPA responses to stress and ACTH stimulation in patients with CPP but reduced adrenal reactivity in patients with FMS, namely in total cortisol release. Free cortisol on the other hand was unaltered, possibly reflecting an adaptation to reduced circulating total cortisol. PMID- 18158368 TI - Racism and ambulatory blood pressure in a community sample. AB - OBJECTIVE: Racism has been identified as a psychosocial stressor that may contribute to disparities in the prevalence of cardiovascular disease. The goal of the present article was to investigate the relationship of perceived racism to ambulatory blood pressure (ABP) in a sample of American-born Blacks and Latinos. METHODS: Participants included English-speaking Black or Latino(a) adults between the ages of 24 and 65. They completed daily mood diaries and measures of perceived racism, socioeconomic status, and hostility. Participants were outfitted with ABP monitors; 357 provided data on waking hours only, and 245 provided data on both waking and nocturnal ABP. RESULTS: Perceived racism was positively associated with nocturnal ABP even when controlling for personality factors and socioeconomic status. CONCLUSIONS: The results suggest that racism may influence cardiovascular disease risk through its effects on nocturnal BP recovery. PMID- 18158369 TI - Race/ethnicity, income, chronic asthma, and mental health: a cross-sectional study using the behavioral risk factor surveillance system. AB - OBJECTIVE: To examine the relationships among race/ethnicity, income, and asthma on mental health outcomes in individuals surveyed as part of the Centers for Disease Control and Prevention 2004 Behavioral Risk Factor Surveillance System (BRFSS). Racial and ethnic disparities in asthma prevalence exist, which may be explained in part by socioeconomic status. Individuals with asthma often have comorbid mental health conditions, the rates of which are also marked by significant racial and ethnic disparities. METHODS: We obtained 2004 BRFSS demographic, asthma, and mental health data on Hispanics, non-Hispanic Whites, and non-Hispanic Blacks. Linear regression analysis was used to examine the main and interaction effects of race/ethnicity, income, and history of asthma on poor mental health (n = 282,011), as well as on depression (n = 14,907) and anxiety (n = 14,871) specifically. RESULTS: A significant three-way interaction emerged among race/ethnicity, income, and history of chronic asthma on number of days of poor mental health. Among the most impoverished (income <$15,000/yr), Hispanics with asthma reported greater number of days of poor mental health than non Hispanic Whites with asthma. However, among those with slightly greater economic resources, Hispanics with asthma reported fewer number of days of poor mental health than non-Hispanic Whites. CONCLUSIONS: The results of this study highlight the complex interactions among race/ethnicity, income, and asthma on mental health outcomes. PMID- 18158370 TI - Neural correlates of levels of emotional awareness during trauma script-imagery in posttraumatic stress disorder. AB - OBJECTIVE: To examine individual differences in levels of emotional awareness as a predictor of the blood oxygenation level dependent (BOLD) response to trauma script-driven imagery in trauma-exposed individuals with (n = 25) and without (n = 16) posttraumatic stress disorder (PTSD). METHODS: Participants completed the Levels of Emotional Awareness Scale (LEAS) and a functional magnetic resonance imaging trauma script-driven imagery paradigm. RESULTS: Patients with PTSD exhibited lower LEAS scores in comparison with the control group. LEAS scores correlated positively with BOLD activity during trauma script-imagery in the ventral anterior cingulate cortex (vACC) in healthy controls, whereas LEAS scores correlated negatively with activation of vACC in individuals with PTSD. CONCLUSION: Patients with PTSD exhibit lower than average levels of emotional awareness. Levels of emotional awareness are differentially associated with vACC response during trauma script-driven imagery in healthy controls versus individuals with PTSD. PMID- 18158371 TI - Childhood socioeconomic status and serotonin transporter gene polymorphism enhance cardiovascular reactivity to mental stress. AB - OBJECTIVE: To test the hypothesis that low socioeconomic status (SES) and the 5HTTLPR L allele are associated with increased cardiovascular reactivity (CVR) to stress in a larger sample and that SES and 5HTTLPR genotypes interact to enhance CVR to stress. CVR to mental stress has been proposed as one mechanism linking stress to the pathogenesis of cardiovascular disease. The more transcriptionally efficient long (L) allele of a polymorphism of the serotonin transporter gene promoter (5HTTLPR) has been found associated with increased risk of myocardial infarction. We found the long allele associated with larger CVR to mental stress in a preliminary study of 54 normal volunteers. METHODS: Subjects included 165 normal community volunteers stratified for race, gender, and SES, who underwent mental stress testing. RESULTS: Childhood SES as indexed by Father's Education Level was associated with larger systolic blood pressure (SBP) (p < .05) and diastolic blood pressure (DBP) (p = .01) responses to mental stress. The L allele was associated with larger SBP (p = .04), DBP (p < .0001), and heart rate (p = .04) responses to mental stress compared with the short (S) allele. Subjects with the SS genotype and high Father's Education exhibited smaller SBP (5.2 mm Hg) and DBP (2.9 mm Hg) responses than subjects with LL genotype and low Father's Education (SBP = 13.3 mm Hg, p = .002; DBP = 9.7 mm Hg, p < .0001). CONCLUSIONS: Both the 5HTTLPR long allele and low SES, particularly during childhood, are associated with increased CVR to mental stress, which could account, at least in part, for the increased cardiovascular disease risk associated with these characteristics. If confirmed in further research, these characteristics could be used to identify persons who might benefit from preventive interventions. PMID- 18158372 TI - Muscular reactivity and specificity in chronic back pain patients. AB - OBJECTIVE: To compare the muscular reactivity of patients with chronic back pain (CBP) to different psychological stressors with the reactions of healthy controls. We also investigated the specificity of muscular reaction near the site of pain in comparison to distal sites. The symptom-specificity model of chronic pain postulates that increased muscle tension in CBP patients may be responsible for the development and maintenance of chronic pain. METHOD: We studied a total of 54 CBP patients with musculoskeletal pain of the lower back, midback, or neck and 62 healthy controls, matched with CPB patients. Muscle tension and skin conductance level (SCL) were assessed. The four experimental conditions included back focusing, a personally relevant stressor, a cognitive stressor, and a social stressor. RESULTS: CBP patients showed patterns of higher muscular reactivity in the lower back region for chronic low back pain (CLBP) patients during the exposure to a personally relevant stressor, a cognitive stressor, and a social stressor. Additionally, CLBP patients showed specific muscular responses in the lower back. CONCLUSION: The results support the assumptions made by the symptom specificity model of CBP, but only for lower back, not for midback and neck. Treatment programs of CLBP should include specific combined relaxation and stress management components. PMID- 18158373 TI - Stress and blood pressure during pregnancy: racial differences and associations with birthweight. AB - OBJECTIVE: To extend findings that African American women report greater stress during pregnancy, have higher blood pressure (BP), and are twice as likely to have low birthweight infants relative to white women. This study examines a) racial differences in associations between stress and BP during pregnancy, and b) the combined effects of stress and BP on infant birthweight in a sample of 170 African American and white women. METHODS: A prospective, longitudinal study of pregnant women was conducted in which measures of BP, stress, and other relevant variables were collected. Multiple measures of systolic and diastolic BP were taken at each of three points during pregnancy (18-20, 24-26, and 30-32 weeks gestation). RESULTS: Both systolic blood pressure (SBP) and diastolic blood pressure (DBP) were positively associated with stress in pregnant African American women and not in pregnant white women. In analyses of birthweight, there were no main effects of BP or stress. However, a significant interaction demonstrated that, when stress was high, DBP was negatively associated with birthweight and a combination of high stress and high DBP predicted the lowest birthweight in the sample. Furthermore, African American women were twice as likely as white women to have a combination of high stress and high DBP. CONCLUSIONS: Racial differences in relationships between stress and BP, and the interactive effect of stress and DBP on birthweight together suggest that a high stress-high BP profile may pose a risk for lower birthweight among African American women, in particular, and possibly for all pregnant women. PMID- 18158374 TI - Do changes in job control predict differences in health status? Results from a longitudinal national survey of Canadians. AB - OBJECTIVE: To examine the effect of changes in job control on health behaviors, psychological distress and health status. METHODS: Using a path analysis model, we examined the effects of change in job control over a 4-year period on levels of physical activity, smoking, and psychological distress; and on self-rated health over an additional 2 years, among a representative sample of 2221 Canadians. RESULTS: Over the 4-year period, 280 respondents reported decreases in job control, and 256 reported increases in job control. Health at baseline was not associated with the likelihood of changes in job control. We found a graded relationship between change in job control and levels of physical activity and psychological distress over a 4-year period; and levels of self-rated health over a 6-year period, with positive change in job control associated in higher levels of physical activity and self-rated health and lower levels of distress. CONCLUSIONS: The results of this study suggest that both level of job control and changes in job control have direct and indirect effects on health status over time. Future research should focus on developing precise measures of work exposures, and examine differences between changes in job control due to only changes in perceptions and changes due to work redesign. PMID- 18158375 TI - Shared temperament risk factors for anorexia nervosa: a twin study. AB - OBJECTIVE: To answer two questions about the nature of the relationship between anorexia nervosa (AN) and dimensional temperament traits: Which traits are comorbid with AN? Which traits share transmitted liabilities with AN? METHODS: A community sample of 1002 same-gender female twins was selected with respect to participation in two earlier waves of data collection. Measures of eating disorder diagnoses and features were ascertained through interview and continuous measures of temperament were ascertained from self-report measures. RESULTS: Four temperaments were comorbid with AN, namely, higher levels of perfectionism (concern over mistakes, personal standards, doubt about actions), and higher need for organization. Comparison between the female co-twins of AN probands and controls (who had never had an eating disorder) showed that the former group reported higher levels of personal standards, organization, and reward dependence. The association between personal standards and reward dependence remained when controlling for the temperament of the proband or control in monozygotic twins. CONCLUSIONS: The evidence overall supports the suggestion that AN may represent the expression of a common underlying familial liability to a temperament style that reflects a striving for perfectionism, a need for order, and a sensitivity to praise and reward. The nature of the shared risk factors is likely to be, in part, genetic. PMID- 18158376 TI - Close relationships and emotional processing predict decreased mortality in women with breast cancer: preliminary evidence. AB - OBJECTIVE: To examine close relationships and emotional processing as predictors of breast cancer mortality. METHODS: Ninety women were enrolled at 14 +/- 5 months after diagnosis of Stage II/III breast cancer. The Nottingham Prognostic Index (NPI) quantified disease severity. Cox proportional hazards analyses were used to predict mortality using standardized variables. RESULTS: Twenty-one subjects developed recurrent disease and 16 died during an 8-year follow-up. NPI predicted increased mortality: risk ratio (RR) = 1.60 (CI = 1.05-2.41). Decreased mortality was predicted by confiding marriage (CONF): RR = 0.31 (CI = 0.10-0.99), and number of dependable, nonhousehold supports (SUPP): RR = 0.41 (CI = 0.21 0.80). A composite measure of close relationships (standardized CONF + SUPP = SUPPCONF) had a strong protective effect: RR = 0.30 (CI = 0.13-0.69). Two emotion processing variables, acceptance of emotion and emotional distress (POMS-TOT) were found to be negatively correlated (r = -.49). Acceptance of emotion predicted decreased mortality (RR = 0.46 (CI = 0.24-0.86)) when analyzed together with emotional distress, but not separately. There was a trend for a protective effect of emotional distress: RR = 0.37 (CI = 0.12-1.09) in the same analysis. RRs for mortality in a multivariable analysis were: SUPPCONF: RR = 0.55 (CI = 0.30-1.00); acceptance of emotion: RR = 0.48 (CI = 0.25-0.91); and emotional distress: RR = 0.40 (CI = 0.14-1.19). CONCLUSIONS: Two aspects of close relationships--marital confiding and dependable, nonhousehold supports--were protective against breast cancer progression. Acceptance of emotion, after controlling for emotional distress, also predicted decreased mortality. Analysis of close relationships together with emotion processing variables suggested unique protective effects against mortality, but a larger study is necessary to determine whether this is the case. PMID- 18158377 TI - No higher risk of myocardial infarction among bipolar patients in a 6-year follow up of acute mood episodes. AB - OBJECTIVE: The aim of this study is to estimate the risk of acute myocardial infarction (AMI) among bipolar disorder patients during a 6-year follow-up after acute mood episodes. The risk is compared with that of a cohort of patients who underwent appendectomy operations during the same period. METHODS: We used administrative claims data from the Taiwan National Health Insurance Research Database covering the years 1997-2002, with the two study cohorts comprising patients hospitalized for bipolar disorder (n = 1429) or appendectomies (n = 4993) in 1997. Multiple logistic regression analyses were performed to compare the crude odds ratio of patients in these cohorts developing AMI following the index discharge by gender. RESULTS: A total of 2.24% of the bipolar disorder patients developed AMI during the 6-year follow-up period, when compared with 1.72% of the appendectomy patients. The multiple logistic regression analyses revealed that there were no significant relationships between the patients in the two cohorts developing AMI, regardless of gender. CONCLUSIONS: There were no significant differences in the risk of developing AMI between patients with bipolar disorder and patients undergoing appendectomy operations, when compared either by gender or as whole groups. PMID- 18158379 TI - A bidirectional relationship between psychosocial factors and atopic disorders: a systematic review and meta-analysis. AB - OBJECTIVE: There is growing epidemiological literature focusing on the bidirectional association between psychosocial factors and atopic disorders, but no efforts to quantify the relationship systematically have been published. METHODS: We searched Medline, PsycINFO, Web of Science, and PubMed up to June 2007. The studies included were prospective cohort studies investigating the influence of psychosocial factors on atopic disorders and the effect of atopic disorders on mental health. Two investigators independently extracted data and determined study quality. RESULTS: There were 43 studies (in 22 articles), of which 34 evaluated the effect of psychosocial factors on atopic disorders and 9 evaluated the effect of atopic disorders on mental health. The major atopic disease assessed in these studies was asthma (90.7%) with allergic rhinitis, 4.7%; atopic dermatitis, 2.3%; and food allergies, 2.3%. The overall meta analysis exhibited a positive association between psychosocial factors and future atopic disorder (correlation coefficient (r) as combined size effect .024; 95% confidence interval, 0.014-0.035; p < .001) as well as between atopic disorders and future poor mental health (r = .044, 95% confidence interval, 0.021-0.067, p < .001). More notably, the subgroup meta-analysis on the healthy and atopic disorder populations showed psychosocial factors had both an etiological and prognostic effect on atopic disorders. CONCLUSIONS: The current review revealed a robust relationship between psychosocial factors and atopic disorders. This supports the use of psychological in addition to conventional physical and pharmacological interventions, in the successful prevention and management of atopic disorders. PMID- 18158378 TI - Depression, the metabolic syndrome and cardiovascular risk. AB - BACKGROUND: The relationship between depression and the metabolic syndrome is unclear, and whether metabolic syndrome explains the association between depression and cardiovascular disease (CVD) risk is unknown. METHODS: We studied 652 women who received coronary angiography as part of the Women's Ischemia Syndrome Evaluation (WISE) study and completed the Beck Depression Inventory (BDI). Women who had both elevated depressive symptoms (BDI > or =10) and a previous diagnosis of depression were considered at highest risk, whereas those with one of the two conditions represented an intermediate group. The metabolic syndrome was defined according to the ATP-III criteria. The main outcome was incidence of adverse CVD events (hospitalizations for myocardial infarction, stroke, congestive heart failure, and CVD-related mortality) over a median follow up of 5.9 years. RESULTS: After adjusting for demographic factors, lifestyle and functional status, both depression categories were associated with about 60% increased odds for metabolic syndrome compared with no depression (p = .03). The number of metabolic syndrome risk factors increased gradually across the three depression categories (p = .003). During follow-up, 104 women (15.9%) experienced CVD events. In multivariable analysis, women with both elevated symptoms and a previous diagnosis of depression had 2.6 times higher risk of CVD. When metabolic syndrome was added to the model, the risk associated with depression only decreased by 7%, and both depression and metabolic syndrome remained significant predictors of CVD. CONCLUSIONS: In women with suspected coronary artery disease, the metabolic syndrome is independently associated with depression but explains only a small portion of the association between depression and incident CVD. PMID- 18158380 TI - Effects of total sleep deprivation in major depression: overnight improvement of mood is accompanied by increased pain sensitivity and augmented pain complaints. AB - OBJECTIVE: Major depressive disorder (MDD) is associated with more pain complaints and an altered pain perception. Studies regarding the longitudinal relationship between depressive symptoms and pain processing have rarely been performed and have produced inconsistent results. To clarify how short-term alleviation of depressive mood is linked to changes in pain processing, the effect of sleep deprivation (SD) on pain and somatosensory thresholds, pain complaints, and mood was investigated in MDD patients. METHODS: Nineteen drug free inpatients with Diagnostic and Statistical Manual of Mental Disorders, fourth edition, diagnosis of MDD were investigated for 3 weeks. All patients received cognitive-behavioral therapy and were randomized to obtain either additional SD therapy (six nights of total SD, separated by recovery sleep) or no SD therapy (control group). Heat/cold pain thresholds, warmth/cold thresholds, measures of current pain complaints, and mood were assessed the evening before and the morning after SD as well as before and after a normal night sleep in the control group. Long-term changes of depressive symptomatology were assessed by weekly mood ratings. RESULTS: Both treatment groups improved markedly in mood over the 3-week treatment period. SD regularly induced a moderate but statistically nonsignificant overnight improvement of mood, which was abolished by recovery sleep. Compared with the control condition, SD significantly decreased heat pain thresholds and nearly significantly cold pain thresholds; SD significantly augmented pain complaints the next morning. No such effects were observed for somatosensory thresholds. CONCLUSIONS: SD induced differential short term effects on mood and pain, with the patients being less depressed but more pain vulnerable. PMID- 18158381 TI - Could multicenter trials optimize the cost of prolonged mechanical ventilation? PMID- 18158382 TI - Collaboration on the rise? PMID- 18158383 TI - Nursing in the face of constant terrorism threat is more stressful. PMID- 18158385 TI - Symptoms across the continuum of acute coronary syndromes: differences between women and men. AB - BACKGROUND: The urgency and level of care provided for acute coronary syndromes partially depends on the symptoms manifested. OBJECTIVES: To detect differences between women and men in the type, severity, location, and quality of symptoms across the 3 clinical diagnostic categories of acute coronary syndromes (unstable angina, myocardial infarction without ST-segment elevation, and myocardial infarction with ST-segment elevation) while controlling for age, diabetes, functional status, anxiety, and depression. METHODS: A convenience sample of 112 women and 144 men admitted through the emergency department and hospitalized for acute coronary syndromes participated. Recruitment took place at 2 urban teaching hospitals in the Midwest. Data were collected during structured interviews in each patient's hospital room. Forty-eight symptom descriptors were assessed. Demographic characteristics, health history, functional status, anxiety, and depression levels also were measured. RESULTS: Regardless of clinical diagnostic category, women reported significantly more indigestion (beta = 0.25; confidence interval [CI] = 0.01-0.49), palpitations (beta = 0.31; CI = 0.06-0.56), nausea (beta = 0.37; CI = 0.10-0.65), numbness in the hands (beta = 0.29; CI = 0.02 0.57), and unusual fatigue (beta = 0.60; CI = 0.27-0.93) than men reported. Differences between men and women in dizziness, weakness, and new-onset cough did differ by diagnosis. Reports of chest pain did not differ between men and women. CONCLUSIONS: Women with acute coronary syndromes reported a higher intensity of 5 symptoms (but not chest pain) than men reported. Whether differences between the sexes in less typical symptoms are clinically significant remains unclear. PMID- 18158386 TI - Response to signs and symptoms of acute coronary syndrome: differences between Lebanese men and women. AB - BACKGROUND: Signs and symptoms of acute coronary syndromes differ between men and women, but whether men and women respond differently to these indications is not well understood. Such responses influence health outcomes because success of treatment depends on how quickly healthcare is sought. OBJECTIVE: To explore differences between Lebanese men and women in cognitive, emotional, and behavioral responses to signs and symptoms of acute coronary syndromes. METHODS: A convenience sample of 149 men and 63 women with unstable angina or acute myocardial infarction were interviewed within 72 hours of admission to coronary care in a tertiary center by using the Response to Symptoms Questionnaire. Demographic and clinical data were obtained from medical records. RESULTS: Women were older, less educated, and more often widowed than men. More women had hypertension but more men were current smokers. Women had shoulder pain, dyspnea, nausea and vomiting, and palpitations more often than men did. Women's signs and symptoms were rated more severe by the women than men's were by the men. Women were less likely to know signs and symptoms of myocardial infarction than were men and delayed coming to the hospital longer than men did. Delay correlated with the characteristics of the signs and symptoms and not realizing their importance in men and with dyspnea and taking the "wait and see" approach in women. CONCLUSION: Factors related to promptness in seeking care for acute coronary syndromes differ between Lebanese men and women. PMID- 18158387 TI - A national online survey on the effectiveness of clinical alarms. AB - PURPOSE: To develop a national online survey to be administered by the American College of Clinical Engineers Healthcare Technology Foundation to hospitals and healthcare workers to determine the problems associated with alarms in hospitals. METHODS: An online survey was developed by a 16-member task force representing professionals from clinical engineering, nursing, and technology to evaluate the reasons health-care workers do not respond to clinical alarms. RESULTS: A total of 1327 persons responded to the survey; most (94%) worked in acute care hospitals. About half of the respondents were registered nurses (51%), and one third of respondents (31%) worked in a critical care unit. Most respondents (>90%) agreed or strongly agreed with the statements covering the purpose of clinical alarms and the need for prioritized and easily differentiated audible and visual alarms. Likewise, many respondents identified nuisance alarms as problematic; most agreed or strongly agreed that the alarms occur frequently (81%), disrupt patient care (77%), and can reduce trust in alarms and cause caregivers to disable them (78%). CONCLUSIONS: Effective clinical alarm management relies on (1) equipment designs that promote appropriate use, (2) clinicians who take an active role in learning how to use equipment safely over its full range of capabilities, and (3) hospitals that recognize the complexities of managing clinical alarms and devote the necessary resources to develop effective management schemes. PMID- 18158389 TI - Characteristics associated with unplanned extubations in a surgical intensive care unit. AB - BACKGROUND: Unplanned extubations can result in serious complications. OBJECTIVES: To determine characteristics of patients and nurses and risk factors that affect extubations. Methods A retrospective exploratory design was used. Ram say Sedation Scale scores, need for reintubation, time between intubation and extubation, use of sedation and analgesia in the preceding 48 hours, and use of restraints were collected on 31 patients in a surgical intensive care unit who had unplanned extubations. For nurses, data collected included years' experience in nursing and as an intensive care nurse, professional credentials, and location at the time of extubation. RESULTS: All unplanned extubations were self extubations; 15 required reintubation. Most patients had low levels of sedation in the hour preceding the extubation (mean Ramsay score, 2.42; SD, 1.06). Patients who needed reintubation had higher mean Ramsay scores (2.85; SD, 1.14) than patients who did not (2.00; SD, 0.86; P = .04). Ramsay scores correlated with need for reintubation (r=0.423; P=.03). Of the 31 patients, 27 (87%) were restrained at the time of extubation (chi2 = 17.06; df=1; P<.001). Among the nurses, 32.3% had less than 5 years' experience in nursing, and 51.6% had less than 5 years' experience in intensive care; 89% of extubations occurred when the nurse was away from the bedside. Sedative and analgesic doses in the 24 hours before extubation did not differ significantly from those in the 2 hours before extubation. CONCLUSIONS: Levels of sedation and use of restraints are associated with unplanned extubations and need for reintubation. PMID- 18158390 TI - Evaluation of delivery of enteral nutrition in critically ill patients receiving mechanical ventilation. AB - BACKGROUND: Published reports consistently describe incomplete delivery of prescribed enteral nutrition. Which specific step in the process delays or interferes with the administration of a full dose of nutrients is unclear. OBJECTIVES: To assess factors associated with interruptions in enteral nutrition in critically ill patients receiving mechanical ventilation. METHODS: An observational prospective study of 59 consecutive patients who required mechanical ventilation and were receiving enteral nutrition was done in an 18-bed medical intensive care unit of an academic center. Data were collected prospectively on standardized forms. Steps involved in the feeding process from admission to discharge were recorded, each step was timed, and delivery of nutrition was quantified. RESULTS: Patients received approximately 50% (mean, 1106.3; SD, 885.9 Cal) of the prescribed caloric needs. Enteral nutrition was interrupted 27.3% of the available time. A mean of 1.13 interruptions occurred per patient per day; enteral nutrition was interrupted a mean of 6 (SD, 0.9) hours per patient each day. Prolonged interruptions were mainly associated with problems related to small-bore feeding tubes (25.5%), increased residual volumes (13.3%), weaning (11.7%), and other reasons (22.8%). Placement and confirmation of placement of the small-bore feeding tube were significant causes of incomplete delivery of nutrients on the day of admission. CONCLUSIONS: Delivery of enteral nutrition in critically ill patients receiving mechanical ventilation is interrupted by practices embedded in the care of these patients. Evaluation of the process reveals areas to improve the delivery of enteral nutrition. PMID- 18158391 TI - Residual volume measurement should be retained in enteral feeding protocols. PMID- 18158392 TI - Critical care nurses' knowledge of evidence-based guidelines for preventing infections associated with central venous catheters: an evaluation questionnaire. AB - BACKGROUND: Lack of adherence to recommended evidence-based guidelines for preventing infections associated with use of central venous catheters may be due to nurses' lack of knowledge of the guidelines. OBJECTIVE: To develop a reliable and valid questionnaire for evaluating critical care nurses' knowledge of evidence-based guidelines for preventing infections associated with central venous catheters. METHODS: A total of 10 nursing-related strategies were identified from current evidence-based guidelines for preventing infections associated with use of central venous catheters. Face and content validation were determined for selected interventions and multiple-choice questions (1 question per intervention). The test results of 762 critical care nurses were evaluated for item difficulty, item discrimination, and quality of the response alternatives or options for answers (possible responses). RESULTS: All 10 items had face and content validity. Values for item difficulty ranged from 0.1 to 0.9. Values for item discrimination ranged from 0.05 to 0.41. The quality of the response alternatives (0.0-0.8) indicated widespread misconceptions among the critical care nurses in the sample. CONCLUSION: The questionnaire is reliable and has face and content validity. Findings from surveys in which this questionnaire is used can lead to better educational programs for critical care nurses on infections associated with use of central venous catheters. PMID- 18158393 TI - Flushing hemodynamic catheters: what does the science tell us? PMID- 18158394 TI - Torsade. PMID- 18158395 TI - Rapid ventricular response. PMID- 18158396 TI - Postgraduate ophthalmic education in India: are we on the right track? PMID- 18158397 TI - Orbital endoscopic surgery. AB - Minimally invasive "keyhole" surgery performed using endoscopic visualization is increasing in popularity and is being used by almost all surgical subspecialties. Within ophthalmology, however, endoscopic surgery is not commonly performed and there is little literature on the use of the endoscope in orbital surgery. Transorbital use of the endoscope can greatly aid in visualizing orbital roof lesions and minimizing the need for bone removal. The endoscope is also useful during decompression procedures and as a teaching aid to train orbital surgeons. In this article, we review the history of endoscopic orbital surgery and provide an overview of the technique and describe situations where the endoscope can act as a useful adjunct to orbital surgery. PMID- 18158398 TI - An evaluation of medical college departments of ophthalmology in India and change following provision of modern instrumentation and training. AB - AIM: To evaluate teaching and practice in medical college ophthalmology departments in a representative Indian state and changes following provision of modern instrumentation and training. STUDY TYPE: Prospective qualitative study. MATERIALS AND METHODS: Teaching and practice in all medical colleges in the state assessed on two separate occasions by external evaluators. Preferred criteria for training and care were pre-specified. Methodology included site visits to document functioning and conduct interviews. Assessments included resident teaching, use of instrumentation provided specifically for training and standard of eye care. The first evaluation (1998) was followed by provision of modern instrumentation and training on two separate occasions, estimated at Rupees 34 crores. The follow-up evaluation in 2006 used the same methodology as the first. RESULTS: Eight departments were evaluated on the first occasion; there were 11 at the second. On the first assessment, none of the programs met the criteria for training or care. Following the provision of modern instrumentation and training, intraocular lens usage increased dramatically; but the overall situation remained essentially unchanged in the 8 departments evaluated 8 years later. Routine comprehensive eye examination was neither taught nor practiced. Individually supervised surgical training using beam splitters was not practiced in any program; neither was modern management of complications or its teaching. Phacoemulsification was not taught, and residents were not confident of setting up practice. Instruments provided specifically for training were not used for that purpose. Students reported that theoretical teaching was good. CONCLUSIONS: Drastic changes in training, patient care and accountability are needed in most medical college ophthalmology departments. PMID- 18158399 TI - Assessment of anterior segment parameters under photopic and scotopic conditions in Indian eyes using anterior segment optical coherence tomography. AB - PURPOSE: To compare the anterior segment parameters in photopic and scotopic conditions using anterior segment optical coherence tomography (AS OCT) in Indian eyes. MATERIALS AND METHODS: One hundred eyes of 100 normal subjects of both sexes, aged 19 to 76 years, underwent anterior segment evaluation by AS OCT (Visante OCT). Central corneal thickness (CCT), central anterior chamber depth (ACD), pupil diameter (PD) and the temporal and nasal peripheral irido-corneal angles were assessed in photopic and scotopic conditions. These anterior segment parameters were stratified for age, sex and refractive error. RESULTS: Mean values of the parameters measured in photopic and scotopic conditions respectively were as follows: ACD (mm) 2.88 +/- 0.32, 2.89 +/- 0.32 (P = 0.10); nasal angle (degrees) 28.80 +/- 5.91, 22.28 +/- 7.50 (P < 0.001); temporal angle (degrees) 29.95 +/- 6.74, 22.82 +/- 8.43 (P < 0.001); pupil diameter (mm) 4.08 +/ 0.91, 4.68 +/- 0.92 (P < 0.001); CCT (microm) 519 +/- 33.88, 519 +/- 33.88. CONCLUSIONS: There was no significant difference in the ACD in photopic and scotopic conditions. While the nasal and temporal angles showed a significant decrease, the pupil diameter showed a significant increase in scotopic conditions. Mean central ACD decreased with age and was shallower in females than in males. It was highest in myopes and lowest in hypermetropes. CCT was not influenced by photopic and scotopic conditions. PMID- 18158400 TI - A proposed new classification for diabetic retinopathy: the concept of primary and secondary vitreopathy. AB - BACKGROUND: Many eyes with proliferative diabetic retinopathy (PDR) require vitreous surgery despite complete regression of new vessels with pan retinal laser photocoagulation (PRP). Changes in the vitreous caused by diabetes mellitus and diabetic retinopathy may continue to progress independent of laser regressed status of retinopathy. Diabetic vitreopathy can be an independent manifestation of the disease process. AIM: To examine this concept by studying the long-term behavior of the vitreous in cases of PDR regressed with PRP. MATERIALS AND METHODS: Seventy-four eyes with pure PDR (without clinically evident vitreous traction) showing fundus fluorescein angiography (FFA) proven regression of new vessels following PRP were retrospectively studied out of a total of 1380 eyes photocoagulated between March 2001 and September 2006 for PDR of varying severity. Follow-up was available from one to four years. RESULTS: Twenty-three percent of eyes showing FFA-proven regression of new vessels with laser required to undergo surgery for indications produced by vitreous traction such as recurrent vitreous hemorrhage, tractional retinal detachment, secondary rhegmatogenous retinal detachment and tractional macular edema within one to four years. CONCLUSION: Vitreous changes continued to progress despite regression of PDR in many diabetics. We identifies this as "clinical diabetic vitreopathy" and propose an expanded classification for diabetic retinopathy to signify these changes and to redefine the indications for surgery. PMID- 18158401 TI - Helical computed tomographic dacryocystography and its role in the diagnosis and management of lacrimal drainage system blocks and medial canthal masses. AB - AIM: To study the indications, technique and diagnostic utility of helical computed tomographic dacryocystography (CTDCG). MATERIALS AND METHODS: Retrospective analysis of 13 patients who underwent CTDCG with subsequent surgical intervention, during the period January 2003 to December 2005, was done. Axial plain computed tomography (CT) scan was performed, followed by administration of water-soluble contrast in the conjunctival cul de sac or by cannulation of the lacrimal passages. Thin-slice helical CT with two-dimensional (2D) and three-dimensional (3D) coronal and sagittal reformation was done. RESULTS: Four patients were males and 9 were females. Age range was 5 to 62 years. Seven patients presented with watering and 6 patients with a medial canthal mass. Three patients had history of trauma. CTDCG was performed by instillation technique in 10 patients and by cannulation in 3 patients. CTDCG showed mass lesion displacing the sac in 5 cases, nasolacrimal duct obstruction in 6 cases and mucocele in 2 cases. Based on the findings on CTDCG, 5 patients underwent mass excision, 7 underwent dacryocystorhinostomy and 1 patient underwent primary silicone tube intubation. CONCLUSION: Helical CTDCG is a safe and useful diagnostic tool for the lacrimal surgeon. Instillation technique is a physiological and convenient method, and cannulation is needed only in cases where adequate visualization is not achieved. PMID- 18158402 TI - Evaluation of subjective and objective cyclodeviation following oblique muscle weakening procedures. AB - PURPOSE: To evaluate the subjective and objective cyclodeviational changes following different weakening procedures on superior and inferior oblique muscles. DESIGN: Comparative case series. MATERIALS AND METHODS: In a prospective institution based study, 16 cases of A pattern horizontal strabismus having superior oblique overaction were randomized to superior oblique weakening procedures: either silicon expander or translational-recession. Similarly, 20 cases of V pattern horizontal strabismus with inferior oblique overaction were randomized for inferior oblique weakening procedures: either 10 mm Fink's recession or modified Elliot and Nankin's anteropositioning. Cyclodeviation was assessed subjectively with the synoptophore and objectively using the fundus photograph before surgery and 3 months postoperatively. Change in cyclodeviation was measured by subjective and objective methods. The index of surgical effect (ISE) was defined as the net torsional change postoperatively. RESULTS: The difference between the extorsional change induced by the two superior oblique procedures, silicone expander (-6 degrees ) and translational recession (-11.3 degrees), was statistically significant (P=0.001). Translational recession caused more extorsional change (ISE=296%) than silicone expander surgery (ISE=107%). The two inferior oblique weakening procedures, Fink's recession (+2.5 degrees) and modified Elliot and Nankin's anteropositioning (+4.7 degrees) produced equitable amount of intorsional shift with no statistical difference (P=0.93). Objective measurements were significantly more than the subjective measurements. CONCLUSIONS: Different weakening procedures on oblique muscles produce different changes in cyclodeviation, which persists even up to 3 months. Subjective cyclodeviation is less than the objective measurements indicating partial compensation by sensorial adaptations. PMID- 18158403 TI - Understanding and using sensitivity, specificity and predictive values. AB - In this article, we have discussed the basic knowledge to calculate sensitivity, specificity, positive predictive value and negative predictive value. We have discussed the advantage and limitations of these measures and have provided how we should use these measures in our day-to-day clinical practice. We also have illustrated how to calculate sensitivity and specificity while combining two tests and how to use these results for our patients in day-to-day practice. PMID- 18158404 TI - A community-based study of asthenopia in computer operators. AB - CONTEXT: There is growing body of evidence that use of computers can adversely affect the visual health. Considering the rising number of computer users in India, computer-related asthenopia might take an epidemic form. In view of that, this study was undertaken to find out the magnitude of asthenopia in computer operators and its relationship with various personal and workplace factors. AIMS: To study the prevalence of asthenopia among computer operators and its association with various epidemiological factors. SETTINGS AND DESIGN: Community based cross-sectional study of 419 subjects who work on computer for varying period of time. MATERIALS AND METHODS: Four hundred forty computer operators working in different institutes were selected randomly. Twenty-one did not participate in the study, making the nonresponse rate 4.8%. Rest of the subjects (n = 419) were asked to fill a pre-tested questionnaire, after obtaining their verbal consent. Other relevant information was obtained by personal interview and inspection of workstation. STATISTICAL ANALYSIS USED: Simple proportions and Chi square test. RESULTS: Among the 419 subjects studied, 194 (46.3%) suffered from asthenopia during or after work on computer. Marginally higher proportion of asthenopia was noted in females compared to males. Occurrence of asthenopia was significantly associated with age of starting use of computer, presence of refractive error, viewing distance, level of top of the computer screen with respect to eyes, use of antiglare screen and adjustment of contrast and brightness of monitor screen. CONCLUSIONS: Prevalence of asthenopia was noted to be quite high among computer operators, particularly in those who started its use at an early age. Individual as well as work-related factors were found to be predictive of asthenopia. PMID- 18158405 TI - Prevalence and distribution of glaucoma in central India (Glaucoma Survey 2001). AB - PURPOSE: A community-based survey was conducted in Rajnandangaon district of Chhattisgarh state of central India in 2001 to assess the prevalence of glaucoma in the age group of =35 years. DESIGN: Community-based cross-sectional survey. MATERIALS AND METHODS: Ophthalmologists measured ocular pressure using Perkins applanation tonometer. Best corrected visual acuity was checked by ETDRS chart. After dilating the pupil the fundus was examined. A sketch diagram was drawn to note glaucomatous changes in optic disc and the surrounding retina. The field of vision was tested on Bjerrum screen. Gonioscopy was performed to determine type of glaucoma. Persons and their relatives were interviewed to find out risk factors and glaucoma treatment in the past. RESULTS: Seven thousand four hundred and thirty-eight (87.3%) persons were examined. The age-sex standardized prevalence of glaucoma was 3.68% (95% CI 3.27 to 4.07). Gender variation of glaucoma was not significant. [OR = 1.13 (CI 95% 0.88 to 1.44)] Glaucoma varied significantly by age groups. (chi2 = 48.2, degree of freedom = 3 P < 0.001) Among those patients diagnosed to suffer from glaucoma, the proportion of open angle, closed angle, secondary glaucoma, ocular hypertension and glaucoma suspects was 13.1%, 21.2%, 21.2%, 14.5% and 30% respectively. Different types of visual disabilities were associated with glaucoma. However, unilateral blindness in glaucoma was unusual. Twenty-five per cent of the glaucoma cases were detected for the first time during the survey. CONCLUSIONS: The prevalence of glaucoma was high and the angle closure type was more compared to the open angle glaucoma. PMID- 18158406 TI - Pupil dilation with intracameral lidocaine during phacoemulsification: Benefits for the patient and surgeon. AB - Topical and/or intracameral administration of anticholinergic and/or sympathomimetic mydriatic agents which are usually used for pupillary dilation during cataract surgery, have some disadvantages such as slow onset of dilation and adverse ocular and systemic effects. We evaluated intracameral injection of preservative-free 1% lidocaine without using any preoperative or intraoperative mydriatics to induce pupil dilation in 31 consecutive eyes scheduled for phacoemulsification cataract extraction and intraocular lens implantation. Pupil diameter was measured before and 90 sec after intracameral lidocaine injection. After intracameral lidocaine injection, the mean pupil diameter was significantly greater than the baseline measurement (P< 0.001). No additional mydriatics were needed up to the end of the operations. Intracameral preservative-free lidocaine 1% has a rapid and effective mydriasis that could be a safe alternative to topical and intracameral mydriatics in phacoemulsification. PMID- 18158407 TI - Diagnosis of Aspergillus fumigatus endophthalmitis from formalin fixed paraffin embedded tissue by polymerase chain reaction-based restriction fragment length polymorphism. AB - New molecular biological technique of Polymerase Chain Reaction (PCR) based Restriction Fragment Length Polymorphism (RFLP) can identify the species from paraffin-embedded tissue section. We demonstrated Aspergillus fumigatus fungus by PCR-based RFLP technique from paraffin section of an eyeball of an eight-month old child removed for endogenous endophthalmitis. PMID- 18158408 TI - Single lens to lens duplication: the missing link. AB - Congenital anomalies of the lens include a wide range from lens coloboma to primary aphakia and doubling of lens. There have been few case reports of double lens; the etiology suggested is metaplastic changes in the surface ectoderm that leads to formation of two lens vesicles and hence resulting in double lens. We report a case with bilobed lens, which raises the possibility of explaining the etiology of double lens. PMID- 18158409 TI - Atypical presentation and diagnostic pitfalls: a case of rapidly progressive bilateral proptosis in a child aged 18 months. AB - We report an atypical presentation of non-Hodgkin lymphoma (NHL) in a child aged 18 months who presented with rapidly progressive bilateral proptosis. Computerized search using Medline did not reveal a similar presentation of NHL in such a young child. It stresses the need for an early histopathological study including immunohistochemistry and demonstrates the dramatic local response to combined radiotherapy and chemotherapy even in advanced stages without any ocular side-effects. PMID- 18158410 TI - Bilateral conjunctival retention cysts in the aftermath of Stevens-Johnson syndrome. AB - In this case report, we describe the rare occurrence of bilateral conjunctival retention cysts in a child with Stevens-Johnson syndrome. The case was managed conservatively as there were no functional or cosmetic problems. PMID- 18158411 TI - An unusual presentation of lupus retinopathy. AB - We have described an unusual presentation of lupus retinopathy in the form of macular arterio-arterial and arterio-venular shunts with extensive macular ischemia as a presenting sign. PMID- 18158412 TI - Branch retinal artery occlusion secondary to dengue fever. AB - Dengue is known to affect the posterior segment of the eye, with a range of hemorrhagic and inflammatory sequelae. A 28-year-old lady convalescing from dengue fever complained of unilateral blurring of inferior visual field. She was evaluated clinically and with fluorescein angiography. Her best-corrected visual acuity was 20/20 bilaterally. Fundus examination revealed a branch retinal artery occlusion in the right eye. Fluorescein angiogram confirmed the clinical diagnosis; and also revealed a late staining and leakage from the affected arterial segment. The patient maintained status quo over a follow-up of six months. We report a major vascular occlusion complicating classic dengue fever even in the absence of severe systemic manifestations. PMID- 18158413 TI - Bilateral choroidal metastasis from carcinoma of the submandibular gland. AB - Metastatic tumor is the most common uveal malignancy. However, choroidal metastasis from a salivary gland neoplasm is extremely rare. We report a case of bilateral, multifocal choroidal metastasis from carcinoma of the submandibular gland. PMID- 18158414 TI - Intravitreal live adult Brugian filariasis. AB - Human ocular infestation by live filarial worm is a rare occurrence and has been reported mostly form South-East Asia. It involves the eyelids, conjunctiva, cornea, anterior chamber and uvea. No case of intravitreal Brugian microfilaria / adult worm has been found by Medline search. Here we report a case of live intravitreal adult Brugian filaria, where the parasite was successfully removed by pars plana vitrectomy. Identification of the worm was done by light microscopy and confirmed by immuno chromatographic test. PMID- 18158415 TI - Anterior dislocation of a sulcus fixated posterior chamber intraocular lens in a high myope. AB - A 31-year-old man with high axial myopia and strabismus fixus convergens underwent bilateral refractive lens exchange followed by a squint surgery (bilateral superior partial Jensen's procedure and medial rectus recession). After one year he presented with traumatic anterior dislocation of the sulcus fixated posterior chamber polymethyl methacrylate lens. The lens was dialed back into the ciliary sulcus without any complications. This case highlights the importance of implanting an intraocular lens (IOL) in-the-bag. If the IOL needs to be implanted in the sulcus, a larger diameter of the IOL with larger optic size and overall length is desirable, especially in highly myopic eyes. PMID- 18158416 TI - Oculosporidiosis. PMID- 18158417 TI - Transcorneal extrusion of a posterior chamber intraocular lens: an unusual presentation of intraocular lens dislocation. PMID- 18158418 TI - Rapidly growing pilomatrixoma on eyebrow. PMID- 18158419 TI - Trans-pupillary thermotherapy for circumscribed choroidal hemangioma. PMID- 18158420 TI - Surgery of basal cell carcinoma around the lacrimal canaliculus can be necessary after primary treatment with intralesional interferon alpha 2b. PMID- 18158421 TI - Chronic bilateral lacrimal gland pseudotumor in an adult. PMID- 18158422 TI - The role of botulinum toxin in correcting frontalis-induced eyelid pseudo retraction post ptosis surgery. PMID- 18158423 TI - Cortical blindness with absent visually evoked potential in non-ketotic hyperglycemia. PMID- 18158424 TI - Inverse Duane's retraction syndrome following myocysticercosis. PMID- 18158425 TI - Spontaneous late-onset comitant acute nonaccommodative esotropia in children. PMID- 18158426 TI - The future of physical & rehabilitation medicine as a medical specialty in the era of evidence-based medicine. AB - Ebenbichler G, Kerschan-Schindl K, Brockow T, Resch KL: The future of physical & rehabilitation medicine as a medical specialty in the era of evidence-based medicine. PMID- 18158427 TI - Effects of a new radio frequency-controlled neuroprosthesis on gait symmetry and rhythmicity in patients with chronic hemiparesis. AB - OBJECTIVE: To assess the effects of a new neuroprosthesis (NESS L300) designed to ameliorate foot drop on gait symmetry and rhythmicity during walking. DESIGN: Twenty-four patients (mean age: 54.0 +/- 13.5 yrs) with chronic hemiparesis (5.8 +/- 5.2 yrs) whose walking was impaired by foot drop. Subjects walked for 6 mins while wearing force-sensitive insoles, once with and once without the neuroprosthesis, in randomized order. Additional assessments with the neuroprosthesis were conducted after using the device for 4 and 8 wks. Walking speed, swing, and stride time were determined, along with a gait asymmetry index and stride time variability-both markers of gait stability and fall risk. RESULTS: While wearing the neuroprosthesis, the gait asymmetry index instantly improved by 28% (from 0.58 +/- 0.30 to 0.42 +/- 0.22) and by 45% (to 0.32 +/- 0.20; P < 0.001) after 8 wks. Stride time variability decreased by 23% immediately (from 5.7 +/- 2.9% to 4.4 +/- 1.3%) and by 33% (to 3.8 +/- 1.4%; P = 0.002) after 8 wks. Walking speed improved initially by 17% (from 0.53 +/- 0.24 to 0.62 +/- 0.22 m/sec) and after 8 wks by 34% (to 0.71 +/- 0.25 m/sec; P < 0.001). CONCLUSIONS: The studied neuroprosthesis enhances gait and improves dynamic stability in chronic hemiparetic patients, supporting the idea that this is a viable treatment option in the rehabilitation of patients with foot drop. PMID- 18158428 TI - The value of ultrasonography in the detection of meniscal tears diagnosed by magnetic resonance imaging. AB - OBJECTIVE: To evaluate the diagnostic value of ultrasonography in meniscal tears that were diagnosed by magnetic resonance imaging (MRI). DESIGN: Twenty-seven knees with meniscal tears and 14 knees without tears on MRI were prospectively evaluated. A radiologist performed the ultrasonography and evaluated the presence and locations of the meniscal tears. MRI was used as the reference standard. RESULTS: Twenty-nine menisci with tears and 53 menisci without tears were identified by MRI. Twenty-two tears were in the medial menisci, and seven tears were in the lateral menisci. In the 29 meniscal tears, the ultrasonographic diagnosis was correct in 25 (86.2%) and incorrect in 4 (13.8%) menisci. In the 53 menisci without tears, the ultrasonographic diagnosis was correct in 45 (84.9%) and incorrect in 8 (15.1%) menisci. Ultrasonography showed a sensitivity, specificity, accuracy, and positive and negative predictive values for meniscal tears of 86.2%, 84.9%, 85.4%, 75.8%, and 91.8%, respectively. Of the four intracapsular injuries observed by MRI in injured knees, an osteochondritis dessecans was only observed by ultrasonography. CONCLUSIONS: Ultrasonography is an accurate imaging study for diagnosing meniscal tears. The results correlated with those obtained by MRI; this suggests that ultrasonography can be a useful imaging modality in uninjured knees. PMID- 18158429 TI - Association between body mass index and functional independence measure in patients with deconditioning. AB - OBJECTIVE: To assess the association of body mass index (BMI) with functional independence measure (FIM) score in patients with deconditioning. We also examined whether the association was different for motor and cognitive subscales of the FIM instrument. DESIGN: A retrospective study of 1077 inpatients admitted to the general medicine service for deconditioning at an acute rehabilitation hospital. Patients were classified into underweight (BMI < 18.5), normal range (BMI = 18.5-24.9), overweight (BMI = 25.0-29.9), obese class I (BMI = 30.0-34.9), obese class II (BMI = 35.0-39.9), and obese class III (BMI > or = 40). RESULTS: Median gain in FIM scores from admission to discharge was highest in obese class I patients (27 points), followed by obese class II patients (26 points). The most gain in FIM scores was accounted for by the motor subscale. Adjusting for age, gender, and length of in-hospital stay, obese class I patients had a 5.8-point (95% confidence limits = 1.2, 7.0) higher gain in FIM score compared with patients with BMI in the normal range. CONCLUSIONS: In an acute rehabilitation setting, obese patients had higher gains in FIM scores as compared with normal range-BMI patients. Most of the improvements in FIM scores were accounted for by the motor subscale, with little or no improvement on the cognitive scale. PMID- 18158430 TI - Left-ventricular function and physical performance on the 6-min walk test in older patients after inpatient cardiac rehabilitation. AB - OBJECTIVE: The 6-min walk test (6mWT) is widely used to assess physical performance in cardiac rehabilitation settings. Factors affecting the walked distance before starting physical training have been described, whereas information on factors affecting the increase of the walked distance after physical training is still scant. The aim of this study was to verify, in a large sample of elderly patients soon after cardiac surgery, the role of left ventricular function (LVF) in increases in distances walked after an intensive rehabilitation program. DESIGN: We enrolled 459 patients (300 males and 159 females, mean [+/-SD] age 70 +/- 11 yrs). According to the echographic ejection fraction, patients were classed into two categories, LVF > or = 40% and LVF < 40%. All patients performed the 6mWT at the beginning and end of the rehabilitation program. RESULTS: Longer walked distances before and after the rehabilitation program were significantly associated with preserved or moderately depressed LVF, whereas greater relative increases of the distance walked after the rehabilitation program were significantly associated with poor LVF (P < 0.001 for all). CONCLUSIONS: Among elderly patients admitted as inpatients to an intensive rehabilitation program soon after cardiac surgery, those with poor LVF are most likely to respond more favorably to physical training. Therefore, instead of considering poor LVF a risk for starting physical training in these patients, it should be considered a strong indication, to avoid further physical deconditioning and disability. PMID- 18158431 TI - Bilateral sciatic nerve entrapment due to heterotopic ossification in a traumatic brain-injured patient. AB - We report a 22-yr-old man who was admitted to our inpatient rehabilitation center 17 mos after a traumatic brain injury (TBI). He was also diagnosed to have bilateral sciatic nerve entrapment attributable to hip heterotopic ossification (HO). Presenting the first report of a sciatic nerve entrapment secondary to a HO in TBI, we herein discuss the concurrence of a lower-motor nerve problem in a patient with an upper-motor nerve injury. PMID- 18158432 TI - Repetitive transcranial magnetic stimulation of the unaffected hemisphere in a patient who was forced to use the affected hand. AB - We present a case report of a 56-yr-old chronic stroke patient with right hemiparesis who was treated with repetitive transcranial magnetic stimulation (rTMS) therapy. Before stroke, the patient had suffered an accident that led to paralysis and contracture of the left upper limb, and, subsequently, he was forced to use only his right upper limb for routine activities, despite right hemiparesis. We performed subthreshold rTMS (1 Hz, 25 mins) and sham stimulation of the contralesional primary motor cortex (M1) at different times. Immediately after rTMS, the patient was able to write characters with increased speed and accuracy, and this effect continued for more than 7 days; however, this was not the case after sham stimulation. Moreover, the writing practice after rTMS improved the patient's pinch force. PMID- 18158433 TI - Chronic pain: second, do no harm. AB - Pain may be undertreated--contributing to anguish, as reported by the World Health Organization. Pain may be overtreated--inadvertently contributing to drug addiction, drug diversion, and even death. Pain may be misunderstood-contributing to illness propagation, as reported in somatization literature. Pain words may even be presented as a tool of manipulation, where report of pain is verbiage in pursuit of utilitarian social consequence. Thus, primum non nocere--first, do no harm--is not easily achieved in the pharmacological treatment of pain, particularly in pain reported chronically. Herein, we examine the pharmacological treatment of chronic pain, and we suggest strategies for improved management that are based on solid principles derived from extensive experience which may protect against the problems derived from the vague and subjective nature of pain symptoms. Optimal treatment of chronic pain may be assisted by three paradigms: (1) an adequate model of appraisal, (2) treatment focused on pathophysiology (whether physical, psychosocial, or some combination of these), and (3) frequent reassessment of total social function. By these approaches, contribution to drug abuse, diversion, and life deterioration can be largely avoided. Whereas the emphasis here is pharmacological management, the principles may be more widely applied to other therapies of chronic pain. PMID- 18158434 TI - Unilateral predominant toe walking gait in a patient with ankylosing spondylitis. PMID- 18158435 TI - A retrospective observational study of drotrecogin alfa (activated) in adults with severe sepsis: comparison with a controlled clinical trial. AB - OBJECTIVE: To compare characteristics and outcomes of patients treated with drotrecogin alfa (activated) (DrotAA) in clinical practice to those treated in a phase III randomized controlled trial (PROWESS). DESIGN: Observational data were collected retrospectively from patients who received DrotAA as part of physician directed treatment. SETTING: Intensive care units of five teaching institutions. PATIENTS: Patients were > or = 18 yrs old, had severe sepsis (confirmed/suspected infection with one or more sepsis-induced organ dysfunctions), and received DrotAA. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Baseline demographics, severity of illness, time from organ dysfunction onset to DrotAA treatment, daily assessment of organ dysfunction, serious bleeding events, and in hospital mortality were reported. Timing from severe sepsis documentation to start of DrotAA infusion was categorized: day 0 (same calendar day); day 1 (next calendar day); and day > or = 2 (second calendar day or later). Clinical practice patients (n = 274) were younger, had more comorbidities, had higher severity of illness (as measured by organ dysfunction or greater vasopressor/ventilator use), and received DrotAA later than PROWESS patients (all p < .05). Overall hospital mortality for clinical practice patients was 42%, compared with 37% for DrotAA treated PROWESS patients with Acute Physiology and Chronic Health Evaluation II score > or = 25. Mortality for day 0, day 1, and day > or = 2 groups was 33%, 40%, and 52%, respectively. In PROWESS, the vast majority were treated on day 0 or day 1. Serious bleeding events during infusion were noted in 4.0% of clinical practice patients compared with 2.2% of PROWESS DrotAA-treated patients with Acute Physiology and Chronic Health Evaluation II score > or = 25. CONCLUSIONS: Patients treated in clinical practice differed from those in PROWESS. Patients were younger, had more comorbidities, had greater severity of illness, and had longer mean time from severe sepsis onset to the start of DrotAA. Hospital mortality for patients treated within 1 day of severe sepsis onset was similar to DrotAA-treated PROWESS patients. While the low number of serious bleeding events precludes a definitive assessment, the observed incidence of serious bleeding events in clinical practice patients was numerically higher than in DrotAA treated PROWESS patients. PMID- 18158438 TI - Meta-analysis: convenient assumptions and inconvenient truth. PMID- 18158437 TI - Surviving Sepsis Campaign: international guidelines for management of severe sepsis and septic shock: 2008. AB - OBJECTIVE: To provide an update to the original Surviving Sepsis Campaign clinical management guidelines, "Surviving Sepsis Campaign Guidelines for Management of Severe Sepsis and Septic Shock," published in 2004. DESIGN: Modified Delphi method with a consensus conference of 55 international experts, several subsequent meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. This process was conducted independently of any industry funding. METHODS: We used the Grades of Recommendation, Assessment, Development and Evaluation (GRADE) system to guide assessment of quality of evidence from high (A) to very low (D) and to determine the strength of recommendations. A strong recommendation (1) indicates that an intervention's desirable effects clearly outweigh its undesirable effects (risk, burden, cost) or clearly do not. Weak recommendations (2) indicate that the tradeoff between desirable and undesirable effects is less clear. The grade of strong or weak is considered of greater clinical importance than a difference in letter level of quality of evidence. In areas without complete agreement, a formal process of resolution was developed and applied. Recommendations are grouped into those directly targeting severe sepsis, recommendations targeting general care of the critically ill patient that are considered high priority in severe sepsis, and pediatric considerations. RESULTS: Key recommendations, listed by category, include early goal-directed resuscitation of the septic patient during the first 6 hrs after recognition (1C); blood cultures before antibiotic therapy (1C); imaging studies performed promptly to confirm potential source of infection (1C); administration of broad spectrum antibiotic therapy within 1 hr of diagnosis of septic shock (1B) and severe sepsis without septic shock (1D); reassessment of antibiotic therapy with microbiology and clinical data to narrow coverage, when appropriate (1C); a usual 7-10 days of antibiotic therapy guided by clinical response (1D); source control with attention to the balance of risks and benefits of the chosen method (1C); administration of either crystalloid or colloid fluid resuscitation (1B); fluid challenge to restore mean circulating filling pressure (1C); reduction in rate of fluid administration with rising filing pressures and no improvement in tissue perfusion (1D); vasopressor preference for norepinephrine or dopamine to maintain an initial target of mean arterial pressure > or = 65 mm Hg (1C); dobutamine inotropic therapy when cardiac output remains low despite fluid resuscitation and combined inotropic/vasopressor therapy (1C); stress-dose steroid therapy given only in septic shock after blood pressure is identified to be poorly responsive to fluid and vasopressor therapy (2C); recombinant activated protein C in patients with severe sepsis and clinical assessment of high risk for death (2B except 2C for postoperative patients). In the absence of tissue hypoperfusion, coronary artery disease, or acute hemorrhage, target a hemoglobin of 7-9 g/dL (1B); a low tidal volume (1B) and limitation of inspiratory plateau pressure strategy (1C) for acute lung injury (ALI)/acute respiratory distress syndrome (ARDS); application of at least a minimal amount of positive end-expiratory pressure in acute lung injury (1C); head of bed elevation in mechanically ventilated patients unless contraindicated (1B); avoiding routine use of pulmonary artery catheters in ALI/ARDS (1A); to decrease days of mechanical ventilation and ICU length of stay, a conservative fluid strategy for patients with established ALI/ARDS who are not in shock (1C); protocols for weaning and sedation/analgesia (1B); using either intermittent bolus sedation or continuous infusion sedation with daily interruptions or lightening (1B); avoidance of neuromuscular blockers, if at all possible (1B); institution of glycemic control (1B), targeting a blood glucose < 150 mg/dL after initial stabilization (2C); equivalency of continuous veno-veno hemofiltration or intermittent hemodialysis (2B); prophylaxis for deep vein thrombosis (1A); use of stress ulcer prophylaxis to prevent upper gastrointestinal bleeding using H2 blockers (1A) or proton pump inhibitors (1B); and consideration of limitation of support where appropriate (1D). Recommendations specific to pediatric severe sepsis include greater use of physical examination therapeutic end points (2C); dopamine as the first drug of choice for hypotension (2C); steroids only in children with suspected or proven adrenal insufficiency (2C); and a recommendation against the use of recombinant activated protein C in children (1B). CONCLUSIONS: There was strong agreement among a large cohort of international experts regarding many level 1 recommendations for the best current care of patients with severe sepsis. Evidenced-based recommendations regarding the acute management of sepsis and septic shock are the first step toward improved outcomes for this important group of critically ill patients. PMID- 18158439 TI - Ventilator discontinuation process: evidence and guidelines. PMID- 18158440 TI - Euthanasia, eye of the beholder? PMID- 18158441 TI - Effectiveness of drotrecogin alfa (activated) in clinical practice: a stitch in time. PMID- 18158442 TI - Melatonin, shedding new light on darkness. PMID- 18158443 TI - Visiting hours in the intensive care unit: more evidence that open visitation is beneficial. PMID- 18158444 TI - From national to global outcome research in the intensive care unit: a challenge to win. PMID- 18158445 TI - Sizing up (or down) extravascular lung water as a predictor of outcome in acute lung injury/acute respiratory distress syndrome. PMID- 18158446 TI - N-acetylcysteine: multiple organ saver during cardiac surgery? PMID- 18158447 TI - How the qualitative method comes to the rescue of caregivers for their comprehension of the complex management of pain and mental disorders in the intensive care unit setting. PMID- 18158448 TI - Decisional incapacity and delirium in the critically ill: what is the real value of the informed consent form? PMID- 18158449 TI - Diagnosis of pneumonia in the critically ill patient: is it time to abandon bronchoscopy? PMID- 18158450 TI - Suicidal intention and self-immolation: what is the outcome? PMID- 18158451 TI - Separating pharmaconutrition from classic nutrition goals: a necessary step. PMID- 18158452 TI - Pathophysiology and diagnostic criteria for disseminated intravascular coagulation associated with sepsis. PMID- 18158453 TI - Assessing the benefits of noninvasive ventilation: the tissue is the issue. PMID- 18158454 TI - Post-cardiac arrest myocardial dysfunction: adding insult to injury. PMID- 18158455 TI - Cyclooxygenase-2 inhibition and increased arterial vasoconstriction to vasopressin: what is the link? PMID- 18158456 TI - Implementation of new technologies in automatic external defibrillators using guidelines for cardiopulmonary resuscitation. PMID- 18158457 TI - Treating lung infections with anticoagulation: mechanisms matter. PMID- 18158458 TI - Cardiopulmonary resuscitation: from flying blind to flying right. PMID- 18158459 TI - Novel insights into the effects of inducible nitric oxide synthase inhibition during sepsis. PMID- 18158460 TI - Stress proteins and acute lung injury: dreams can come true... eventually. PMID- 18158461 TI - Can cytokine removal in brain-dead patients improve transplant organ survival? PMID- 18158462 TI - A breath of fresh air: the potential use for hyperoxia in traumatic brain injury. PMID- 18158463 TI - Mediators of cellular stress response in bacterial meningitis. PMID- 18158464 TI - 24-hour intensivist staffing: balancing benefits and costs. PMID- 18158465 TI - Obesity and intensive care unit survival: pass the potatoes? PMID- 18158467 TI - Ethical challenges in living organ donation. PMID- 18158468 TI - Acute heart failure: emerging from the shadows. PMID- 18158469 TI - Inotropes in the management of acute heart failure. AB - Impaired cardiac contractility is a fundamental component of the heart failure syndrome, initiating the cycle of vasoconstriction, neurohormonal and inflammatory activation, and adverse ventricular remodeling that leads to heart failure progression. Based on this core paradigm, drugs that increase cardiac contractility (positive inotropes) are theoretically appealing as a heart failure therapy, and such agents have been extensively investigated in both acute and chronic heart failure. Although these agents clearly improve cardiac output, their use in heart failure has consistently been associated with increased myocardial oxygen demand, cardiac arrhythmias, and mortality in a variety of clinical settings. Based on these data, the routine use of inotropes as heart failure therapy is not indicated in either the acute or chronic setting. Inotropes may be a necessary evil in a subset of acute heart failure patients, such as those with acute heart failure decompensation in the setting of clinically evident hypoperfusion or shock, or as a bridge to more definitive treatment, such as revascularization or cardiac transplantation. Currently available inotropes, such as dobutamine and milrinone, act (directly or indirectly) by increasing cyclic adenylate monophosphate and therefore intracellular calcium flux. Whether newer inotropes with differing mechanisms of action will realize the potential clinical benefits of inotropic therapy without the risk remains a subject of ongoing investigation. PMID- 18158470 TI - New pharmacologic therapies for acute heart failure. AB - Given the limitations of high-dose diuretics and vasodilators and the increasing literature showing that inotropes, regardless of the dose used, have a detrimental effect on mortality, a variety of new agents are under investigation for the treatment of pulmonary and systemic congestion and restoration of cardiac output in the setting of acute heart failure syndromes. The new therapeutic approach is based on two goals: short-term improvement in symptoms together with long-term improvement of cardiac function. This review describes new agents that are in preclinical and in clinical phases with realistic prospects: anti endothelin, natriuretic peptides, istaroxime, levosimendan, myosin activators, and vasopressin antagonists. Those new therapeutic strategies aim to act at the cellular level to improve vessel and heart functions, with minimal side effects, together with improved sodium and water balance. PMID- 18158471 TI - Devices in acute heart failure. AB - The optimal management strategy for patients with acutely decompensated heart failure has been limited to the use of various pharmacologic agents. However, with technological advances in mechanical devices, nonpharmacologic approaches now are available to supplement pharmacologic management. These devices range in sophistication and expertise and target various features of the acute heart failure patient, such as circulatory failure, volume overload, renal insufficiency, and respiratory failure. As more innovations and technologies emerge, these new devices may become a cornerstone of the management strategy in cardiac patients. PMID- 18158472 TI - Practical recommendations for prehospital and early in-hospital management of patients presenting with acute heart failure syndromes. AB - Guideline recommendations for the prehospital and early in-hospital (first 6-12 hrs after presentation) management of acute heart failure syndromes are lacking. The American College of Cardiology/American Heart Association and European Society of Cardiology guidelines direct the management of these acute heart failure patients, but specific consensus on early management has not been published, primarily because few early management trials have been conducted. This article summarizes practical recommendations for the prehospital and early management of patients with acute heart failure syndromes; the recommendations were developed from a meeting of experts in cardiology, emergency medicine, and intensive care medicine from Europe and the United States. The recommendations are based on a unique clinical classification system considering the initial systolic blood pressure and other symptoms: 1) dyspnea and/or congestion with systolic blood pressure >140 mm Hg; 2) dyspnea and/or congestion with systolic blood pressure 100-140 mm Hg; 3) dyspnea and/or congestion with systolic blood pressure <100 mm Hg; 4) dyspnea and/or congestion with signs of acute coronary syndrome; and 5) isolated right ventricular failure. These practical recommendations are not intended to replace existing guidelines. Rather, they are meant to serve as a tool to facilitate guideline implementation where data are available and to provide suggested treatment approaches where formal guidelines and definitive evidence are lacking. PMID- 18158473 TI - Advances in congestive heart failure management in the intensive care unit: B type natriuretic peptides in evaluation of acute heart failure. AB - BACKGROUND: Circulating concentrations of B-type natriuretic peptide (BNP) and the aminoterminal fragment (NT-proBNP) of its prohormone (proBNP) are increased in congestive heart failure in proportion to the severity of symptoms, the degree of left ventricular dysfunction, and cardiac filling pressures. Following the introduction of rapid, automated assays for determination of BNP and NT-proBNP, these peptides are increasingly used for diagnostic and prognostic purposes. OBJECTIVE: To review studies evaluating the diagnostic and prognostic value of BNP and NT-proBNP, with special emphasis on their performance as indicators of acute heart failure in the intensive care unit. RESULTS: In patients presenting with acute dyspnea, both BNP and NT-proBNP are accurate indicators of acute heart failure and provide prognostic information above and beyond conventional risk markers. Increased plasma levels of BNP and NT-proBNP are not specific for heart failure and may be influenced by a variety of cardiac and noncardiac conditions commonly seen in the intensive care unit, including myocardial ischemia, cardiac arrhythmias, sepsis, shock, anemia, renal failure, hypoxia, acute pulmonary embolism, pulmonary hypertension, and acute respiratory distress syndrome. CONCLUSIONS: The diagnostic performance of BNP and NT-proBNP as indicators of acute heart failure depends on the clinical setting. In the intensive care unit, particular caution should be used in the interpretation of elevated BNP and NT proBNP levels. PMID- 18158474 TI - Imaging studies in patients with heart failure: current and evolving technologies. AB - Technological advances continue to expand the clinical role of echocardiography in the intensive care unit, particularly in patients with heart failure. It has many advantages over tomographic techniques such as echo cardiac magnetic resonance imaging and cardiac computed tomography, can provide rapid bedside cardiac assessment, and facilitate emergent decision-making for critically ill patients. Image quality problems in the intensive care setting have largely been overcome by the use of harmonic imaging, contrast opacification, and when indicated, transesophageal echocardiography. Newer techniques promise to advance the scope and prognostic power of echocardiography, and to expand the portability and availability of this tool. PMID- 18158475 TI - Acute heart failure in the intensive care unit: epidemiology. AB - More than a million patients are admitted annually to U.S. hospitals with acute heart failure. Multicentered hospital-based registries and surveys in the United States and Europe have shown that the typical patient is >70 yrs of age, with a history of heart failure, coronary artery disease, and hypertension. There are an equal number of men and women. Patients typically spend several days on the intensive care unit, with longer admissions in Europe than the United States. The in-hospital mortality rate is around 4% to 7%. The risk of subsequent hospital readmission is high. The elderly, those with comorbidities, and those with cardiogenic shock or renal failure do particularly badly. Better treatment by those with expertise in the management of this syndrome and good follow-up care are likely to improve the outcome for this large group of patients. PMID- 18158476 TI - Hemodynamic monitoring in acute heart failure. AB - Hemodynamic monitoring has moved in the last few years from being the holy grail of evaluating patients with acute heart failure to being all but extinct. Recent studies have not demonstrated any sustained benefits from right heart catheterization, and some studies have even suggested harm due to adverse events related to this invasive procedure. It is possible that this lack of efficacy is related to multiple inherent deficiencies in the design of these studies, including the inclusion of patients with chronic heart failure or mild acute heart failure, use of the reduction in pulmonary artery occlusion pressure as the main hemodynamic target for intervention, choice of treatment algorithms, and selection of ambitious long-term efficacy and safety end points. This review discusses the role of hemodynamic monitoring in patients with acute heart failure. We suggest that right heart catheterization should be reserved for patients with acute heart failure and impending respiratory or circulatory failure especially in the presence of a diagnostic or therapeutic dilemma or when encountering acute heart failure or hemodynamic lability refractory to conventional therapy. Therapeutic algorithms emphasizing modern variables for cardiovascular performance and using safer and more efficacious individualized therapies and possibly noninvasive measurement of certain hemodynamic variables may enhance the likelihood of a beneficial effect for hemodynamic guided therapy. PMID- 18158477 TI - Systolic heart failure: chronic and acute syndromes. AB - Systolic heart failure is characterized by ventricular dilation and reduced ejection fraction, and this syndrome may be either chronic or acute. Left ventricular remodeling is the principal cause of progression of systolic heart failure. Acute heart failure resulting from cardiomyopathy has similar functional and morphologic abnormalities. This review discusses remodeling, initial therapy based on neurohormonal modulation, and treatment of decompensated and refractory heart failure. Diagnosis, prognosis, and management of acute myocarditis are also discussed. PMID- 18158478 TI - Acute heart failure with preserved systolic function. AB - Many patients with acute heart failure have marked hypertension and preserved left ventricular ejection fraction. In these patients, the heart failure usually does not result from transient systolic dysfunction or valvular abnormalities but rather results from diastolic dysfunction. Treatment of this condition includes control of hypertension, cautious diuresis, and, if necessary, ventilatory support. Further workup after the acute phase should be directed by the overall clinical picture. Other potential contributing factors, such as renal artery stenosis, valvular heart disease, and ischemia, should be strongly considered. Unfortunately, chronic therapy for diastolic heart failure has not yet been standardized due to the paucity of clinical trial data. Strict control of hypertension appears to be of paramount importance. Angiotensin-converting enzyme inhibitors or receptor blockers may be of benefit in preventing repeat hospitalizations. PMID- 18158479 TI - Pathophysiology of right ventricular failure. AB - Right ventricular failure may be defined as the inability of the right ventricle of the heart to provide adequate blood flow through the pulmonary circulation at a normal central venous pressure. Critical care specialists encounter right ventricular failure routinely in their practice, but until recently right ventricular failure as a primary clinical entity received scant consideration. Indeed, there is still not a single published practice guideline focused on right ventricular failure. Right ventricular failure is usually due to a combination of right ventricular pressure overload and contractile abnormalities of the right ventricular free wall. Decompensation may occur abruptly and catastrophically because of unique aspects of right ventricular physiology. This review will focus on the pathophysiology of acute right ventricular failure in the critical care setting and summarize the limited management options available. PMID- 18158480 TI - Cardiogenic shock. AB - Cardiogenic shock is the most common cause of death in patients hospitalized with acute myocardial infarction and is associated with a poor prognosis. More than 75% of cases are due to extensive left ventricular infarction and ventricular failure. Other causes include right ventricular infarction and papillary muscle rupture with acute severe mitral regurgitation. Activation of neurohormonal systems and the systemic inflammatory response worsens shock. To improve outcomes, cardiogenic shock needs to be recognized early in its course and its cause needs to be diagnosed rapidly. Treatment strategies using intra-aortic balloon counterpulsation and emergency revascularization by percutaneous coronary interventions or coronary bypass surgery have been shown to improve outcomes. To decrease the incidence of cardiogenic shock, public education regarding early presentation to hospital in the course of acute chest pain is important. Emergency medical transport systems may need to take patients with complicated acute myocardial infarction to hospitals with the capability to perform urgent revascularization. PMID- 18158481 TI - Acute decompensated heart failure and the cardiorenal syndrome. AB - Heart failure is one of the leading causes of hospitalizations in the United States. Concomitant and significant renal dysfunction is common in patients with heart failure. Increasingly, the syndrome of heart failure is one of cardiorenal failure, in which concomitant cardiac and renal dysfunctions exist, with each accelerating the progression of the other. One fourth of patients hospitalized for the treatment of acute decompensated heart failure will experience significant worsening of renal function, which is associated with worse outcomes. It remains unclear whether worsening renal function specifically contributes to poor outcomes or whether it is merely a marker of advanced cardiac and renal dysfunction. Diuretic resistance, with or without worsening renal function, is also common in acute decompensated heart failure, although the definition of diuretic resistance, its prevalence, and prognostic implications are less well defined. The term cardiorenal syndrome has been variably associated with cardiorenal failure, worsening renal function, and diuretic resistance but is more comprehensively defined as a state of advanced cardiorenal dysregulation manifest by one or all of these specific features. The pathophysiology of the cardiorenal syndrome is poorly understood and likely involves interrelated hemodynamic and neurohormonal mechanisms. When conventional therapy for acute decompensated heart failure fails, mechanical fluid removal via ultrafiltration, hemofiltration, or hemodialysis may be needed for refractory volume overload. While ultrafiltration can address diuretic resistance, whether ultrafiltration prevents worsening renal function or improves outcomes in patients with cardiorenal syndrome remains unclear. Evidence regarding the potential renal preserving effects of nesiritide is mixed, and further studies on the efficacy and safety of different doses of nesiritide in heart failure therapy are warranted. Newer therapeutic agents, including vasopressin antagonists and adenosine antagonists, hold promise for the future, and clinical trials of these agents are underway. PMID- 18158482 TI - Diuretics: still the mainstay of treatment. AB - The mainstay of treatment of acute decompensated heart failure is diuretic therapy. While there are no data showing a morbidity or mortality benefit from the use of chronic diuretic therapy, diuretics rapidly improve symptoms associated with volume overload. Thus, despite concerns that some diuretics may cause harm by neurohormonal activation, these agents continue to be the first line treatment for patients with heart failure. There is no conclusive evidence that one means of diuresis is better than another. When administration of moderate doses of loop diuretics is not sufficient, patients can be treated with higher doses, continuous infusions, or the addition of a thiazide diuretic or aldosterone antagonist. Diuretics improve symptoms but should be used in addition to other agents that improve the long-term outcome of patients with heart failure. PMID- 18158483 TI - Cytokines and acute heart failure. AB - In patients with chronic heart failure, ongoing myocardial injury partially results from activation of the inflammatory system, with production and release of proinflammatory cytokines, activation of the complement system, production of autoantibodies, overexpression of major histocompatibility complex molecules, and expression of adhesion molecules that may perpetuate the inflammatory state. Acute decompensated heart failure modifies the course of chronic heart failure and worsens outcomes via a combination of potential mechanisms, including neurohormonal activation, apoptosis, and the inflammatory cascade. Proinflammatory cytokines, including tumor necrosis factor-alpha and interleukin 6, play a pathogenetic role in chronic heart failure, and anti-inflammatory immune therapy is currently under investigation. In acute decompensation of chronic heart failure, the change in the inflammatory cytokine activation cascade is less clear. Larger investigational studies are needed to assess the exact roles of circulating and intracardiac cytokines in this particular patient population. PMID- 18158484 TI - Vasodilators in the management of acute heart failure. AB - Recent guidelines by the Heart Failure Society of America have recommended consideration for use of nitroprusside, nitroglycerin, or nesiritide in addition to diuretics to achieve hemodynamic and symptomatic improvement. This article reviews the results of previous studies evaluating the pharmacologic and clinical effects and safety profiles of these drugs in patients with heart failure. PMID- 18158485 TI - Preparing for the challenges of an aging population: the graying of America. PMID- 18158486 TI - My father's experience with macular degeneration: implications for the home healthcare nurse. AB - Macular degeneration has become a problem of substantial proportions for older adults. This article can help the home health nurse gain a greater understanding of macular degeneration and therefore be more effective in the home setting. The addition of vision rehabilitation also can prolong older adults' independent years and enhance their satisfaction with life. Using Parse's human-becoming theory as a theoretical framework, the home health nurse can coparticipate in the older adult's experience of living and thus enhance the probability that the last years will be a rewarding time of life. PMID- 18158488 TI - Partnering for prevention: a Pressure Ulcer Prevention Collaborative project. AB - In a statewide initiative, coordinated by the New Jersey Hospital Association (NJHA) Quality Institute, hospitals together with nursing home and home care agencies were asked to participate in a Pressure Ulcer Prevention Collaborative. The goal of this collaborative was to decrease the incidence and prevalence of pressure ulcers across the state by 25% within a 12-month period. This article discusses the rationale for the Collaborative as well as the requirements and implementation of the initiative within Community Medical Center's Home Health Program. PMID- 18158489 TI - Artificial nutrition and hydration for the terminally ill: a reasoned approach. AB - Knowledge about evidence-based research findings on the physiologic effects of artificial nutrition and hydration (ANH) can be used as a framework for assisting patients and families with decision making at the end of life. This article provides a review of the credible evidence related to ANH obtained through scientifically sound research, supplies a framework for discussion about ANH with patients and families, and lists basic educational recommendations and resources that both home health and hospice nurses can use to assist with patient and family supportive efforts. The charge to nurses caring for dying patients and their families is consistently to honor patient beliefs about life--their goals and values--in a supportive, unbiased, and informational manner, which will improve the quality of life--at the end of life. PMID- 18158490 TI - Elders at risk during disasters. AB - Recent natural and manmade disasters such as the September 11, 2001 terrorist attacks, the hurricanes of 2005, and Chicago heat waves demonstrate the vulnerability of older adults to such events. In this article, the specific physical, psychosocial, and cultural characteristics of older adults that place them at greater risk during disasters and emergencies are discussed. Unique concerns of older adults and their families in disasters and emergencies are addressed. In addition, the impact that these characteristics have on the ability of older adults to respond to such events and recover from them is discussed. Finally, strategies that home health providers can use in working with vulnerable older adults are explored. PMID- 18158492 TI - Managing the patient with dysphagia. AB - Dysphagia, or difficulty swallowing, is a common medical condition among today's patients. It is estimated the approximately 15% of individuals older than 60 years experience dysphagia. Approximately 40% of institutionalized patients (i.e., patients residing in nursing homes, group homes, or assistive care facilities) have swallowing difficulties (Zagaria 2005). The swallowing mechanism is quite complex, involving more than 50 pairs of muscles. Impairment of any one or more of these muscles may result in a serious health risk to the patient. The patient may be at risk of aspirating food or saliva into the lungs, creating a potentially life-threatening medical condition. PMID- 18158494 TI - VNAA brings the full force of its advocacy to Washington. PMID- 18158495 TI - The emotional needs of care recipients and the psychological well-being of informal caregivers: implications for home care clinicians. AB - In this exploratory study, 77 informal caregivers of older persons in Ohio completed telephone interviews that included questions regarding their perceived difficulty providing emotional and physical care, perceived quality of care, demographic items including caregivers' health status, and a measure of their psychological well-being. Findings suggest that psychological well-being of informal caregivers is diminished when they experience greater difficulty meeting the care recipients' emotional care needs, are in poorer health, or are older. Suggestions for clinical implications and future research based on these findings are discussed. PMID- 18158496 TI - Research challenges and lessons learned from a heart failure telehomecare study. AB - Examining the benefits of nursing interventions through research significantly contributes to both professional nursing and quality healthcare, yet few benefits of home healthcare have been rigorously studied. Research challenges and lessons learned during a clinical trial of heart failure patients receiving home health nursing services are presented with recommendations. PMID- 18158497 TI - Infection prevention and control: achieving a culture of zero tolerance. PMID- 18158498 TI - PCNA: Preventive Cardiovascular Nurses Association. PMID- 18158499 TI - Foreword: American Heart Association's top 10 research advances in 2006: what makes a top 10? PMID- 18158500 TI - High-dose statin therapy for secondary prevention of stroke: stroke prevention by aggressive reduction in cholesterol levels study review. AB - It has been shown that HMG-CoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase inhibitors (statins) lower the incidence of a first stroke in patients with coronary heart disease, diabetes, or risk factors for cardiovascular disease. However, it is unknown whether statin therapy could reduce the incidence of a second stroke in patients without evidence of heart disease. This article reviews the results of the Stroke Prevention by Aggressive Reduction in Cholesterol Levels trial, a prospective, randomized, multicentered, double-blind, placebo controlled, international trial designed to examine the effect of high-dose atorvastatin on secondary stroke prevention. Trial participants (4,731) had experienced a stroke or transient ischemic attack within 1 to 6 months before randomization into the study. Over the 5-year follow-up period, incidence of second stroke or transient ischemic attack was significantly reduced in the atorvastatin treatment group compared with the placebo group. In addition, high dose atorvastatin therapy significantly decreased major coronary artery and other negative cardiovascular events. The reduction in incidence of secondary stroke was specific to ischemic stroke as opposed to hemorrhagic stroke. Results of the trial are clinically significant and support extension of the latest secondary stroke prevention guidelines to include statin therapy for those patients without coronary heart disease. PMID- 18158501 TI - Use of selective cyclooxygenase-2 inhibitors and nonselective nonsteroidal antiinflammatory drugs in high doses increases mortality and risk of reinfarction in patients with prior myocardial infarction. AB - The selective cyclooxygenase-2 (COX-2) inhibitors and other nonselective nonsteroidal antiinflammatory drugs (NSAIDs) have been associated with increased cardiovascular risk, but the risk in patients with established cardiovascular disease is unknown. In the present study, we analyzed the risk of rehospitalization for acute myocardial infarction (re-MI) and death related to the use of NSAIDs including selective COX-2 inhibitors in patients with a prior myocardial infarction (MI). We included 58,432 patients discharged alive after a first MI, and subsequent use of all NSAIDs was identified from a nationwide register of drug dispensing from pharmacies. We found a dose-dependent increase in risk of death for both the selective COX-2 inhibitors and the nonselective NSAIDs (all of the drugs tested). There were trends for increased risk of re-MI associated with the use of both the selective COX-2 inhibitors and the nonselective NSAIDs (high dosages). Selective COX-2 inhibitors in all dosages and nonselective NSAIDs in high dosages should be used with particular caution in patients with a prior MI. PMID- 18158502 TI - Clinical implications of increased congenital malformations after first trimester exposures to angiotensin-converting enzyme inhibitors. AB - Angiotensin-converting enzyme (ACE) inhibitors are some of the most frequently prescribed antihypertensives in the United States. These agents are contraindicated during the second half of pregnancy because prior studies have demonstrated that use of ACE inhibitors late in pregnancy can cause oligohydramnios, fetal growth restriction, skull defects, infant anuria and renal failure, and death. Interestingly, little was known about the effects of ACE inhibitors when taken early during pregnancy. The purpose of the ACE Inhibitors in Early Pregnancy study was to clarify the safety of the use of ACE inhibitors during pregnancy by conducting an epidemiologic study using a large Medicaid database in which medications prescribed for pregnant women as a part of routine care and infant outcomes were studied. In the study, among 209 infants with first trimester exposure to ACE inhibitors, 7.1% had any major congenital malformation. Compared with 29,096 infants with no exposure to any antihypertensive medication, among whom 2.6% had any major congenital malformation, the adjusted risk of major congenital malformations was increased more than 2-fold. The risks of any congenital malformations and risks of specific organ system malformations, including cardiac malformations, were not increased in 202 infants with first trimester exposure to other antihypertensives when compared with infants with no antihypertensive exposure. Although this was an exploratory study whose findings should be confirmed, there are some important clinical insights that can be drawn from the study's conclusions. Further information on the pregnancy risks of ACE inhibitors and almost every other medication potentially used by pregnant women is needed. Thus, well-controlled studies to identify these risks should be undertaken. Until such information is available, alternative medications to ACE inhibitors should be considered in women of child-bearing age who are pregnant or who are likely to become pregnant while taking the medication. PMID- 18158503 TI - Ventricular assist devices as a bridge to heart transplantation in children. AB - The increase in time waiting for appropriate pediatric allografts for heart transplantation has mandated the use of long-term mechanical assistance in the pediatric population. Extracorporeal membrane oxygenation support has been routinely used but is limited by both its inability to provide support without life-threatening complications for longer than 2 to 3 weeks as well as the inability of patients to achieve mobility. For the past 10 years, pediatric programs have increasing experience with the use of ventricular assist devices (VADs) to bridge patients to heart transplant. This retrospective study analyzed the clinical features and outcomes of 99 pediatric patients who underwent VAD implant as a bridge to heart transplant. METHODS: Between 1993 and 2003, the Pediatric Heart Transplant Study Group enrolled 2,375 patients (age 1 day-17.9 years) listed for heart transplant from 23 participating centers. Four percent (99 patients) of those listed received VAD support as a bridge to transplantation. Seventy-seven (77%) patients survived to transplant with a mean time on support of 57 days. There were 17 deaths on support and 5 bridged to recovery. Overall incidence of adverse events was similar to the adult data with a 19% risk of stroke. There was no difference in 5-year survival after transplant for patients on VAD at time of transplant compared with those (n = 2,293) not requiring VAD (77% vs 73%, P = .8). These data suggest that despite the lack of pediatric specific devices and relatively high adverse event rate, VADs may be used as a bridge to transplant therapy in appropriate-sized children with the expectation of a successful outcome in most patients. PMID- 18158504 TI - Lack of benefit with percutaneous intervention for late persistent occlusion after myocardial infarction: summary of the occluded artery trial. AB - The clinical utility of establishing late patency of the persistently occluded infarct-related artery with percutaneous coronary intervention (PCI) was uncertain. The Occluded Artery Trial was a National Heart, Lung, and Blood Institute-supported, international, multicenter, randomized controlled trial comparing a test strategy of late PCI (3-28 days) of the occluded infarct-related artery and optimal medical therapy to optimal medical therapy alone. The primary end point of the trial was a centrally adjudicated composite of death, reinfarction, and New York Heart Association class IV heart failure over 4 years (mean follow-up, 1,059 +/- 11 days). The final study population of 2,166 patients gave the trial 94% power to detect the anticipated 25% reduction in event rate with PCI. The combined primary outcome occurred in 161 patients in the PCI group and in 140 subjects receiving medical therapy alone. The 4-year cumulative primary event rate was 17.2% in the PCI group and 15.6% in the medical therapy group (hazard ratio, 1.16; 95% confidence interval, 0.92-1.45; P = .20; covariate adjusted hazard ratio, 1.17; 95% confidence interval, 0.93 to 1.47; P = .18). Rates of New York Heart Association class IV heart failure and death were similar in both groups. A trend toward increased nonfatal myocardial infarction in the PCI group (hazard ratio, 1.44; 95% confidence interval, 0.96-2.16; P = .08) unrelated to periprocedural events was apparent. No significant interaction between treatment effect and prespecified subgroups was observed. This lack of clinical benefit supports optimal medical therapy alone for Occluded Artery Trial eligible patients in current clinical practice. PMID- 18158505 TI - Secondary stroke prevention. AB - Stroke is the most common life-threatening neurological disorder. Based on limited acute therapies, clinicians have opted to focus on preventive strategies to limit its recurrence. Targets for prevention include modifiable risk factors such as hypertension, diabetes mellitus, dyslipidemia, cigarette smoking, obesity, alcohol use, and physical inactivity among others. The American Stroke Association and American Heart Association guideline for the secondary prevention of stroke published in 2006 provides comprehensive and timely evidence-based recommendations on the prevention of ischemic stroke among survivors of stroke or transient ischemic attack. This guideline helps healthcare providers who have arrived at a potential explanation of the cause of stroke in an individual patient to select therapies that reduce the risk of recurrent events and other vascular events. The purpose of this review is to highlight the recently published American Stroke Association/American Heart Association guidelines for the secondary prevention of stroke. PMID- 18158507 TI - Summary of the American Heart Association scientific statement: promoting physical activity in children and youth: a leadership role for schools. AB - Schools have long played an important role in providing students with healthful physical activity. However, the decline in population-level physical activity suggests that schools should play an even greater role in providing and promoting physical activity. Recently, the American Heart Association issued a set of recommendations that, if implemented, would position schools as leaders in helping children and youth become more physically active. This article summarizes an American Heart Association scientific statement on physical activity and the schools that was recently developed by the Association's Council on Nutrition, Physical Activity, and Metabolism in collaboration with the Council on Cardiovascular Disease in the Young and the Council on Cardiovascular Nursing. PMID- 18158508 TI - Association of evidence-based care processes and outcomes among patients with acute coronary syndromes: performance matters. AB - The American College of Cardiology and American Heart Association publishes evidence-based guideline recommendations, yet the degree to which these guidelines are followed and the association between hospital guideline adherence and patient outcomes are unknown. Using data from 350 US centers participating in the "Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes With Early Implementation of the American College of Cardiology/American Heart Association Guidelines" Quality Improvement Initiative, we evaluated the in hospital treatment and outcomes from 64,775 patients with non-ST-segment elevation acute coronary syndromes. Overall, guideline-recommended treatments were followed in 74% of eligible instances. However, hospitals varied considerably in their composite adherence rates (median lowest to highest adherence hospital quartiles 63% to 82%). We also noted significant age, gender, and racial disparities in the use of guideline-recommended therapies as well as significant facility type and regional variability in care. Composite guideline adherence rates were significantly associated with in-hospital mortality. After risk adjustment, every 10% increase in composite adherence at a hospital was associated with a 10% reduction in its patients' likelihood of in-hospital mortality. Our findings support the use of broad, guideline-based performance metrics as a means of assessing and helping improve hospital quality. PMID- 18158509 TI - Age-specific lipid and lipoprotein thresholds for adolescents. AB - Plasma lipid and lipoprotein values that are relatively stable in adulthood undergo continuous changes during the developmental years, with most of these changes reflecting natural fluctuations that occur with growth and maturation. Therefore, a clinical classification system appropriate for youth should not consist of uniform lipoprotein thresholds that can be used at all ages to define healthy and increased risk values. This is, however, the approach used in the National Cholesterol Education Program pediatric guidelines. A recent study created the first age-specific lipid and lipoprotein thresholds for adolescents based on a large representative sample of American adolescents aged 12 to 19 years. The adolescent thresholds were linked to the health-based thresholds used in adults using growth curve modeling. Although studies are needed to validate the newly defined adolescent lipid and lipoprotein thresholds, this classification system should provide a more accurate diagnosis of dyslipidemia and associated cardiovascular health risks in adolescents. PMID- 18158510 TI - "In vivo tissue-engineered" valved conduit with designed molds and laser processed scaffold. AB - BACKGROUND: "In body tissue architecture" technology is a practical concept of regenerative medicine that uses the living recipient body's reaction to a foreign object as a reactor for autologous tissue organization. A novel autologous valved conduit was produced by creating a specially designed conduit-mold composite and elastomeric scaffold for this unique in vivo tissue engineering. METHODS: Convex and concave plastic molds assembled with a small aperture of 500-800 microm were inserted into a microporous elastomeric conduit scaffold. The assembly was placed in a subcutaneous pocket of Japanese white rabbits for 1 month. The molds were pulled out from the edge of the harvested implant to obtain the valved conduit. RESULTS: Homogenous and well-balanced trileaflet of membranous tissue was developed in the optimized aperture of molds. The valve leaflet closed and opened rapidly in synchronization with the backward and forward flow of the pulsatile flow circuit in vitro. CONCLUSIONS: A tissue-engineered conduit incorporated with a functional autologous trileaflet valve was developed in an in vivo reactor by optimizing the microstructures of conduit scaffolds and newly designing the composite molds. The method holds promise for a safe, biocompatible, and economical heart valve prosthesis. PMID- 18158511 TI - Progress in prevention: preventing cardiovascular disease: a global challenge and a call to action. PMID- 18158512 TI - Effects of acculturation on smoking behavior in Asian Americans: a meta-analysis. AB - BACKGROUND: Cigarette smoking is the most preventable risk factor for many negative health consequences, such as cancer, heart disease, and lung disease. In the United States, the prevalence rate in Asian immigrants is high (26%-70%), with Southeast Asian men having the highest rate. Acculturation has been associated with smoking behavior in this ethnic group. OBJECTIVE: The purposes of this meta-analysis are to describe the extent to which acculturation affects smoking behavior in Asian immigrants and to compare the direction and magnitude of the effect between subgroups by gender and age. METHODS: Databases within PubMed, CINAHL, The Cochrane Library, and PsycINFO were searched. Twenty-one studies published in English or Korean from 1994 through 2005 met criteria, and 9 of these studies contained sufficient data. Among the 9 studies, 3 presented gender-specific data; thus, these studies were entered separately for men and women, making a total of 12 entries for final analysis. The odds ratio was used as an effect size statistic. The values of odds ratios were calculated from the data in the studies. RESULTS: The average effect size for men was 0.53 (95% confidence interval, 0.28-0.99), indicating that acculturated men are 53% less likely to smoke than nonacculturated or "traditional" men. The average effect size for women was 5.26 (2.75-10.05), suggesting that acculturated women are 5 times more likely to smoke than traditional women. In adolescents, the average effect size was 1.92 (1.22-3.01), indicating that acculturated adolescents are almost 2 times more likely to smoke than traditional adolescents. CONCLUSIONS: Acculturation may have a protective effect on smoking behavior in Asian men and a harmful effect in Asian women and adolescents. The magnitude of effect is larger in women and adolescents than in men. Smoking cessation programs should target acculturated women, adolescents, and traditional men. PMID- 18158513 TI - Witnessed resuscitation: beneficial or detrimental? AB - This article explores the existing literature and discusses the benefits and disadvantages of witnessed resuscitation for health professionals, relatives, and patients themselves. Keywords "witnessed resuscitation," "patient perspective," "health professionals," and "resuscitation room" were entered into MEDLINE, Medscape, and Science Direct databases. The issue of witnessed resuscitation, along with the benefits and disadvantages of its implementation, is discussed widely with increasing controversy among health professionals. Many authors accept the existence of benefits of witnessed resuscitation, but they each have reservations on certain aspects of the practice. Although witnessed resuscitation has demonstrable benefits, the dearth of research literature on the subject makes it difficult to come to a concrete conclusion about its value in practice. More studies are needed focusing on the impact of witnessed resuscitation on staff, family members, and patients. Larger sample sizes are needed in future studies, and studies are needed in which geographical, cultural, religious, and sociological factors are taken into consideration. PMID- 18158514 TI - Retrospective screening of patients for age of initial diagnosis of coronary artery disease: shouldn't we just ask? PMID- 18158516 TI - The shadow of your smile. PMID- 18158517 TI - New York University College of Nursing takes on the challenge of innovation for the healthcare system. PMID- 18158518 TI - Should elective dental procedures be done during pregnancy? PMID- 18158519 TI - Should elective dental procedures be done during pregnancy? PMID- 18158520 TI - Improving oral health in women: nurses' call to action. AB - The purpose of this article is to discuss the most significant oral health and related problems experienced by women, and to provide a Nurse's Plan of Action to respond to these largely preventable diseases. Oral health is integral to women's overall health and well-being, with poor oral health being associated with cancer, heart disease, diabetes, depression, and the birth of preterm, low birthweight babies. Poor nutrition and lifestyle, principally tobacco and heavy alcohol use, can further increase the risk for oral diseases. Disparities are evident in women's reported poor access of regular dental care related to lack of dental insurance and low income. These facts are disturbing because most oral diseases are preventable. The Surgeon General's report on oral health in America (U.S. Department of Health and Human Services, 2000) and, more recently, the "National Call to Action to Promote Oral Health" (U.S. Department of Health and Human Services, 2003) emphasized the need for partnerships of key stakeholders, including nurses, to get involved in oral disease prevention. Nurses are in an ideal position to provide health promotion education and screening across the multitude of settings in which they work regarding oral health and risk factors for oral disease. Nursing interventions aimed at promoting healthy outcomes and preventing disease should include a focus on oral health. PMID- 18158522 TI - Promoting oral health from birth through childhood: prevention of early childhood caries. AB - Early childhood caries (ECC) is the single most common chronic childhood disease in the United States. Associated with a characteristic clinical presentation, ECC results from a culmination of risk factors, including cariogenic bacteria, inappropriate feeding practices, and social variables. It is essential that nurses become knowledgeable about ECC health promotion and prevention interventions to facilitate their efforts in everyday patient encounters and improve the overall oral health status of infants and children. This article provides an overview of ECC along with suggestions for actions that nurses can take to improve the oral health of children. PMID- 18158524 TI - Oral health of adolescents--it's more than dental caries. AB - Of all the health issues common to adolescents, oral health has not seemed to receive much attention. However, just as "eyes are the windows to the soul," the oral health of an adolescent can be a reliable and expedient indicator of general health. Lesions discovered during an oral exam can indicate systemic problems that may not otherwise be disclosed during the health history. Tobacco use, sexually transmitted infections, and type II diabetes can manifest in the mouth, and many of these lesions mimic other illnesses. It is essential that nurses who work with adolescents be aware of possible differential diagnoses beyond the typical canker sores, herpes simplex type I, and periodontal disease so commonly seen. Adolescents rarely enter the healthcare system, but school and camp nurses and nurse practitioners have access to teens on a regular basis. They can screen adolescents for oral health problems and teach them practices that may prevent health problems with lifetime consequences. Capitalizing on this opportunity to educate adolescents can be critical to their long-term health. PMID- 18158525 TI - Pregnancy and oral health: a review and recommendations to reduce gaps in practice and research. AB - This article presents a review of the research relevant to oral health during pregnancy and includes nursing practice recommendations for referral of women to a dentist for safe and effective dental care during pregnancy. In recent years, research linking periodontitis to the risk for adverse birth outcomes has resulted in increased interest in the topic of oral health during pregnancy. The achievement of optimal oral health in pregnant women as its own benefit, however, has in the past been hampered by myths surrounding the safety of dental care during pregnancy. Many women also lack access to dental care and dental insurance, which interferes with their ability to receive adequate oral care during pregnancy. Intraoral changes that occur with pregnancy because of hormonal changes, combined with lack of routine exams and delays in treatment for oral disease, place pregnant women at higher risk for dental infections. PMID- 18158526 TI - Nursing care & management of pathological oral conditions among women and children. AB - Oral health is receiving increased recognition as an important factor for the health of women and children. This article describes pathological oral conditions and the physiological mechanisms involved in the maintenance of oral health during illness and hospitalization, including the importance of the production and secretion of adequate saliva. Alterations in physiology of saliva induced by hormonal changes that occur during pregnancy are described along with possible roles in pathophysiological conditions associated with pregnancy. The roles of xerostomia and periodontal disease as sources of pathological bacteria involved in aspiration pneumonia, ventilator-associated pneumonia, and chronic inflammation are described. The bidirectional interaction between oral health and general overall health is described, with special emphasis on cardiovascular disease. Specific evidence-based nursing measures to reduce the complications associated with poor oral health in at-risk patients are provided. PMID- 18158527 TI - Maternal periodontal disease, pregnancy, and neonatal outcomes. AB - The purpose of this article is to review the current literature on the association between maternal periodontal disease and poor pregnancy and neonatal outcomes and outline the role nurses can play in improving the oral health of pregnant women. Maternal periodontal disease is linked to preterm birth, low birthweight, and preterm low birthweight, but treatment of periodontal disease during pregnancy has been shown to be safe and effective. Nurses, nurse practitioners, and nurse-midwives are in a position to educate pregnant women on the benefits of good oral health and identify and refer women who are in need of dental care for treatment. PMID- 18158528 TI - Oral health & HIV/AIDS. AB - Oral lesions are common in women and children with HIV/AIDS and may decrease the overall quality of life in these patients because of pain, dry mouth, and difficulty in eating. An oral cavity screening is an easy, noninvasive, quick, and inexpensive procedure that provides nurses with invaluable information about the need for referral, treatment, and health education. Nurses can use the information obtained from a careful oral screening to decrease the symptoms experienced with oral lesions and optimize a patient's ability to chew and enjoy food. Common oral manifestations of HIV infection include fungal, viral, and bacterial infections, although neoplasms, periodontal disease, salivary gland disease, and lesions of uncertain origin are also seen. Oral lesions such as candidiasis, oral hairy leukoplakia, herpetic ulcers, and Kaposi's sarcoma are often among the first symptoms of HIV infection. PMID- 18158529 TI - Oral health getting off to a good start. PMID- 18158530 TI - Go online for oral health! PMID- 18158531 TI - Global oral health in women and children. PMID- 18158538 TI - Taxonomies. PMID- 18158539 TI - Behaviour change guidance. National Institute for Health and Clinical Excellence. PMID- 18158540 TI - Little evidence of any difference between surgical or non-surgical approaches for retreatment of periapical lesions. Is surgical treatment more effective than nonsurgical treatment when patients need retreatment of periapical disease? PMID- 18158541 TI - Using a cold test to assess pulpal anaesthesia. Is a cold test effective in determining whether a patient will experience pain during root canal therapy? PMID- 18158542 TI - Clinical significance of early treatment of overjet is questionable. At what age should orthodontic treatment for prominent upper teeth be carried out? PMID- 18158543 TI - Bond or band? Is there a difference in clinical failure rates between bonded and banded first molar attachments during fixed appliance therapy? PMID- 18158544 TI - More caries in primary teeth of children who have cleft lip and palate. Do children who have cleft lip and palate have elevated caries levels in the primary and permanent dentition? PMID- 18158545 TI - Mineral trioxide aggregate a reliable alternative material for pulpotomy in primary molar teeth. Is mineral trioxide aggregate more effective than formocresol for pulpotomy in primary molars? PMID- 18158546 TI - Home visits providing diet advice may reduce early childhood caries. Do home visits that provide new mothers with advice about breastfeeding and weaning reduce the odds of infants developing early childhood caries? PMID- 18158547 TI - Limited evidence to support the use of physical therapy for temporomandibular disorder. How effective are physical therapy interventions in the management of temporomandibular disorder? PMID- 18158548 TI - Non-surgical care should be the primary treatment for TMJ closed lock. Does medical management improve pain and jaw function as much and as quickly as rehabilitation with/ without surgery in the initial management of temporomandibular joint closed lock? PMID- 18158549 TI - Edentulous people prefer lingualised or anatomic posterior occlusal tooth forms. What is the best posterior tooth form for complete dentures? PMID- 18158550 TI - Is periodontal disease associated with poor pregnancy outcomes? What is the strength of association between periodontal disease in mothers and birth of their infants preterm or with low birthweights? PMID- 18158551 TI - Oral health and respiratory diseases. Is there an association between oral health and pneumonia or other respiratory diseases? PMID- 18158552 TI - The EQUATOR network and website. PMID- 18158556 TI - F2-isoprostane receptors on hepatic stellate cells. AB - F2-isoprostanes are considered as the most reliable markers of oxidative stress and can be used to evaluate the oxidative status in a number of human pathologies. Besides being markers of oxidative stress, F2-isoprostanes proved to be mediators of important biological effects and would act through the activation of receptors analogous to those for thromboxane A2. In a previous work, we provided evidence that F(2)-isoprostanes, generated during carbon tetrachloride induced hepatic fibrosis, mediate hepatic stellate cell (HSC) proliferation and collagen hyperproduction. To investigate whether TxA2 receptor (TxA2r or TPr) is involved in the effects of F2-isoprostanes on HSC, experiments on DNA synthesis were carried out in the presence of 8-epi-prostaglandin F(2alpha) (8-epi PGF(2alpha)) or the TxA2r-specific agonist I-BOP ([1S [1alpha,2alpha(Z),3beta(1E,3S*), 4alpha]]-7-[3-[3-hydroxy-4-(4-iodophenoxy)-1 butenyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5-heptenoic acid). Both agonists significantly stimulated DNA synthesis, which was almost completely inhibited by the TxA2r-specific antagonist SQ29548 ([1S-[1alpha,2alpha(Z),3alpha,4alpha]]-7-[3 [[2-[(phenylamino)carbonyl] hydrazino] methyl]-7-oxabicyclo[2.2.1]hept-2-yl]-5 heptanoic acid), suggesting that much of the effect of 8-epi-PGF(2alpha) is mediated by the TxA2r. Further studies showed that increasing concentrations of SQ29548 progressively inhibit DNA synthesis, suggesting a possible competitive antagonism between the two molecules. In addition, we demonstrated that the stimulatory effect of 8-epi-PGF(2alpha) on collagen synthesis could be mediated by TxA2r. The occurrence of TxA2r on HSC was also investigated using western blotting analysis and immunocytochemistry, which reveals that TP is distributed both on plasma membranes and within the cells. Moreover, binding studies indicated the presence of a specific binding site for 3H-SQ29548 on HSC. Competition binding studies indicated that 8-epi-PGF(2alpha) and I-BOP were both able to displace 3H-SQ29548 binding with a very different affinity (K(i)=4.14+/ 1.9 x 10(-6) M and K(i)=1.15+/-0.3 x 10(-9) M, respectively), suggesting the involvement of a modified form of isoprostane receptor, homologous to the classic thromboxane A2-binding site in F2-isoprostanes-evoked responses on HSC. PMID- 18158557 TI - High telomerase activity and long telomeres in advanced hepatocellular carcinomas with poor prognosis. AB - Telomerase reactivation and telomere maintenance are crucial in carcinogenesis and tumor progression. In this study, the relationships between telomere parameters, chromosomal instability and clinicopathological features were evaluated in hepatocellular carcinomas (HCCs). Telomere length (TL), telomerase activity (TA) and human telomerase reverse transcriptase (hTERT) mRNA levels were measured in 49 hepatitis B virus (HBV)-related HCCs and corresponding non tumorous tissues. The results were compared with clinicopathological data, including differentiation, multipolar mitosis (MM), anaphase bridge, immunohistochemical stain results for cytokeratin 19 (CK19) and patient outcome. TL of HCCs ranged from 4.7 to 13.1 kb, and 44.4% of HCCs showed telomere lengthening. hTERT mRNA levels and TA were closely related (P=0.008), and were significantly higher in HCCs than non-tumorous tissues. TL was significantly higher in HCCs with strong TA (P=0.048), high hTERT mRNA levels (P=0.001) and poor differentiation (P=0.041). Frequent MM was associated with poor differentiation (P=0.007) and advanced stage (P<0.001). TA was positively correlated with MM, anaphase bridges and advanced stage (P=0.019, P=0.017 and P=0.029). Thirteen (28.3%) HCCs were CK19+ and demonstrated longer telomeres than CK19- HCCs (P=0.046). Overall survival was poor in HCCs with MM >0.4 per field (P=0.016), high TA (P=0.009) and high TL ratio (HCC/non-HCC) >0.8 (P=0.044). Our results show that long telomeres, high TA and high mitotic instability are poor prognostic markers for HBV-related HCCs and their close association suggests that telomere maintenance may be important for the progression of HCCs with high chromosomal instability to more aggressive ones. PMID- 18158558 TI - Elevated hydrostatic pressure promotes protein recovery from formalin-fixed, paraffin-embedded tissue surrogates. AB - High-throughput proteomic studies on formalin-fixed, paraffin-embedded (FFPE) tissues have been hampered by inefficient methods to extract proteins from archival tissue and by an incomplete knowledge of formaldehyde-induced modifications to proteins. We previously reported a method for the formation of 'tissue surrogates' as a model to study formalin fixation, histochemical processing, and protein retrieval from FFPE tissues. In this study, we demonstrate the use of high hydrostatic pressure as a method for efficient protein recovery from FFPE tissue surrogates. Reversal of formaldehyde-induced protein adducts and crosslinks was observed when lysozyme tissue surrogates were extracted at 45 000 psi and 80-100 degrees C in Tris buffers containing 2% sodium dodecyl sulfate and 0.2 M glycine at pH 4. These conditions also produced peptides resulting from acid-catalyzed aspartic acid cleavage. Additives such as trimethylamine N-oxide or copper (II) chloride decreased the total percentage of these aspartic acid cleavage products, while maintaining efficient reversal of intermolecular crosslinks in the FFPE tissue surrogates. Mass spectrometry analysis of the recovered lysozyme yielded 70% sequence coverage, correctly identified all formaldehyde-reactive amino acids, and demonstrated hydrolysis at all of the expected trypsin cleavage sites. This study demonstrates that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues for proteomic analysis. PMID- 18158559 TI - DNA methylation profiles of gastric carcinoma characterized by quantitative DNA methylation analysis. AB - Transcriptional silencing by CpG island hypermethylation is a potential mechanism for the inactivation of tumor-related genes. Virtually, all types of human cancers show CpG island hypermethylation, and gastric carcinoma (GC) is one of the tumors with a high frequency of aberrant CpG island hypermethylation. In this study, we prescreened DNA methylation of 170 CpG island loci in a training set of 8 paired GC and GC-associated non-neoplastic mucosae (GCN) using MethyLight technology and selected 27 DNA methylation markers showing higher methylation frequency or level in GC than in GCN. These markers were then analyzed in a tester set of 25 paired GC and GCN and 27 chronic gastritis (CG) from non-cancer patients to generate their DNA methylation profiles. We identified 17 novel methylation markers in GC, including SFRP4, SEZ6L, TWIST1, BCL2, KL, TERT, SCGB3A1, IGF2, GRIN2B, SFRP5, DLEC1, HOXA1, CYP1B1, SMAD9, MT1G, NR3C1, and HOXA10. Of the 27 selected CpG island loci, 23 were methylated in GC, GCN, and CG and the remainder four loci (DLEC1, CHFR, CYP1B1, and NR3C1) were only methylated in GC. We found that the number of methylated loci was significantly higher in GC than in GCN or CG and that Helicobacter pylori infection was strongly associated with aberrant CpG island hypermethylation in CG. Hypermethylation was more prevalent in Epstein-Barr virus (EBV)-positive GC than in EBV-negative GC and in diffuse-type GC than in intestinal-type GC. Through our large-scale screening of 170 CpG island loci, we found 17 new DNA methylation markers of GC, which may serve as useful markers that may identify a distinct subset of GC. PMID- 18158560 TI - Pathology of peripheral neuroblastic tumors: significance of prominent nucleoli in undifferentiated/poorly differentiated neuroblastoma. AB - The presence of large cells having simultaneously increased cytoplasmic and nuclear volume accompanied by prominent nucleoli; i.e., differentiating neuroblasts and ganglion cells, is well documented in peripheral neuroblastic tumors (pNTs), and considered as one of the signs of tumor maturation and an indication of a better prognosis of the patients. On the other hand, in 2004 it was reported that large-cell neuroblastoma composed of neuroblastic cells with only nuclear enlargement without recognizable cytoplasmic maturation behaved poorly clinically. Here we are proposing a new pNT subtype in the neuroblastoma category, in addition to the undifferentiated, poorly differentiated and differentiating subtypes: that is large nucleolar neuroblastoma (LNN) characterized by large prominent nucleoli and no or very little amount of discernible cytoplasm. LNN, whose neuroblastic cells are often large in size due to nuclear enlargement, includes those tumors previously categorized into the large-cell neuroblastoma group. LNN tumors, regardless of the size of nuclei, seem to behave aggressively with a very poor prognosis of the patients. It is speculated that nucleolar enlargement without cytoplasmic maturation in LNN tumor cells can be a sign of MYCN amplification. PMID- 18158561 TI - Elevated levels of somatic mutation in a manifesting BRCA1 mutation carrier. AB - Homozygous loss of activity at the breast cancerpredisposing genes BRCA1 and BRCA2 (FANCD1) confers increased susceptibility to DNA double strand breaks, but this genotype occurs only in the tumor itself, following loss of heterozygosity at one of these loci. Thus, if these genes play a role in tumor etiology as opposed to tumor progression, they must manifest a heterozygous phenotype at the cellular level. To investigate the potential consequences of somatic heterozygosity for a BRCA1 mutation demonstrably associated with breast carcinogenesis on background somatic mutational burden, we applied the two standard assays of in vivo human somatic mutation to blood samples from a manifesting carrier of the Q1200X mutation in BRCA1 whose tumor was uniquely ascertained through an MRI screening study. The patient had an allele-loss mutation frequency of 19.4 x 10(-6) at the autosomal GPA locus in erythrocytes and 17.1 x 10(-6) at the X-linked HPRT locus in lymphocytes. Both of these mutation frequencies are significantly higher than expected from age-matched disease-free controls (P < 0.05). Mutation at the HPRT locus was similarly elevated in lymphoblastoid cell lines established from three other BRCA1 mutation carriers with breast cancer. Our patient's GPA mutation frequency is below the level established for diagnosis of homozygous Fanconi anemia patients, but consistent with data from obligate heterozygotes. The increased HPRT mutation frequency is more reminiscent of data from patients with xeroderma pigmentosum, a disease characterized by UV sensitivity and deficiency in the nucleotide excision pathway of DNA repair. Therefore, this BRCA1-associated breast cancer patient manifests a unique phenotype of increased background mutagenesis that likely contributed to the development of her disease independent of loss of heterozygosity at the susceptibility locus. PMID- 18158562 TI - Prognostic significance of loss of c-fos protein in gastric carcinoma. AB - c-fos was first identified as a viral oncoprotein, and has been studied in terms of its oncogenic function in tumorigenesis. Many experimental and clinical data indicated that c-fos expression plays a role in the progression of several types of carcinomas. However, some recent studies challenge this view as they indicate that c-fos has tumor suppressor activity. In the present study, we assessed c-fos protein expression in 625 consecutive gastric cancers immunohistochemically, and analyzed its relationship with clinicopathologic factors and survival. We found that a loss of c-fos expression is correlated with a more advanced stage, lymph node metastasis, lymphatic invasion and shorter survival, indicating that c-fos expression in gastric cancer cells is lost during progression and that this loss is associated with a poor prognosis. The above findings suggest that loss of c fos expression has tumor suppressor activity in gastric cancer and we suspect that this suppressor activity might be related to the pro-apoptotic function of c fos. PMID- 18158563 TI - Expression of the TRAIL receptors in blood mononuclear cells in leukemia. AB - TRAIL receptors are differentially expressed on restricted subpopulations of normal blood cells. In the present study, we investigated the utility of individual TRAIL receptors in evaluating the presence of circulating tumor cells in blood. Patients with chronic myeloid leukemia (CML) carrying the t(9;22) translocation were compared with patients in whom no translocation was detected, with patients with multiple myeloma and with a group of healthy individuals. TRAIL receptor expression was analyzed by RT-PCR in blood mononuclear cells. Blood mononuclear cells of healthy subjects expressed the TRAIL-R1 and TRAIL-R2 death receptors and the TRAIL-R4 decoy receptor while the other decoy receptor TRAIL-R3 was not detectable. This normal expression pattern was also observed in all cases with multiple myeloma and in almost all patients without translocation (42/43; 97.7%). However, in 24/56 (42.9%) of the translocation-positive patients, the expression pattern was completely different. In this group the TRAIL-R4 receptor alone or in combination with TRAIL-R1 disappeared from blood mononuclear cells, while the TRAIL-R2 was expressed at normal level, indicating that the loss of expression is specific for the TRAIL-R4 and TRAIL-R1. This expression pattern was also confirmed by real-time PCR. The differences between the translocation positive and -negative groups for the TRAIL-R4 and TRAIL-R1 expression were highly significant (p=0.0001 and p=0.0004, respectively). However, the differential expression pattern did not correlate with the number of leukemic cells. Our results suggest a correlation between the presence of leukemic cells in circulation and the differential expression pattern of TRAIL receptors in blood mononuclear cells. PMID- 18158564 TI - Inhibition of novel protein kinase C-epsilon augments TRAIL-induced cell death in A549 lung cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has great potential for cancer treatment since it provokes cell death in most tumor cells while leaving most normal cells unscathed. Some cancers, however, show resistance to TRAIL, indicating that TRAIL alone may be insufficient for cancer therapy. Here we studied whether the apoptotic susceptibility of A549 non-small cell lung cancer cells could be modulated by inhibiting protein kinase C (PKC). We show that an inhibitor with preference for novel PKC isozymes, NPC 15437, significantly augmented TRAIL sensitivity of A549 cells, as judged by assessing cell death and mitochondrial membrane potential. Likewise, NPC 15437 also significantly potentiated the responsiveness of DAOY medulloblastoma cells to TRAIL. In contrast, an inhibitor with preference for conventional PKC isozymes, Go6976, did not augment TRAIL sensitivity of A549 cells. To further specify the PKC isozyme responsible for TRAIL sensitization, we used a peptide inhibitor with selectivity for the novel PKC isozyme epsilon, myr-PKC-epsilon V1-2. The inhibition of PKC-epsilon resulted in a significant amplification of the cytotoxic activity of TRAIL in A549 cells. Altogether, our study provides evidence for a considerable role of PKC-epsilon in the apoptotic responsiveness of A549 lung cancer cells, and possibly other malignancies, to TRAIL. PMID- 18158565 TI - Transcriptional characterization of the Notch signaling pathway in rodent multipotent adult progenitor cells. AB - The Notch signaling pathway is a multifunctional, evolutionarily conserved pathway, which plays an important role in development as well as stem cell biology. Multipotent adult progenitor cells (MAPCs) represent a unique stem cell population, which is capable of differentiating into cell types of the ectodermal, mesodermal and endodermal lineages in vitro, and contribute to most somatic cell types in vivo. Our aim was to characterize the gene expression of Notch signaling elements in rodent MAPCs. We show that transcripts for Notch receptors, ligands, regulatory molecules of the pathway and the Hairy/Enhancer of Split-1 (HES-1) target gene are present in mouse and rat low-Oct4 MAPCs. We found that mouse Notch3 and rat Notch1 transcripts increased when cells were cultured at high density for 48 to 96 hours. HES-1 and HES-related transcription factor-1 (HERP-1), transcriptional targets of Notch-signaling, were both elicited by immobilized Delta1 ligand. In addition, mRNA for Notch1 and Notch3 was also induced by Notch-signaling, suggesting the presence of regulatory feedback loops. Slight differences between mouse and rat derived MAPCs suggest that the exact function, transcriptional regulation and the fine-tuning of the signal may be species specific. Taken together, we characterized the gene expression profile of the Notch pathway in rodent low-Oct4-MAPCs, and showed that the pathway is functional and can be modulated. Our results provide an additional tool and a further basis for a better understanding of stem cell biology. PMID- 18158566 TI - Inappropriate Notch activity and limited mesenchymal stem cell plasticity in the bone marrow of patients with myelodysplastic syndromes. AB - Myelodysplastic syndromes (MDSs) are a heterogeneous group of hematological disorders characterized by ineffective hematopoiesis, enhanced bone marrow apoptosis and frequent progression to acute myeloid leukemia. Several recent studies suggested that, besides the abnormal development of stem cells, microenvironmental alterations are also present in the MDS bone marrow. In this study, we have examined the relative frequencies of stem and progenitor cell subsets of MDS and normal hematopoietic cells growing on stromal cell layers established from MDS patients and from normal donors. When hematopoietic cells from MDS patients were co-cultured with normal stromal cells, the frequency of either early or late cobblestone area-forming cells (CAFC) was significantly lower compared to the corresponding normal control values in 4 out of 8 patients. In the opposite situation, when normal hematopoietic cells were incubated on MDS stromal cells, the CAFC frequencies were decreased in 5 out of 6 patients, compared to normal stromal layer-containing control cultures. Moreover, a soluble Notch ligand (Jagged-1 protein) was an inhibitor of day-35-42 CAFC when normal hematopoietic cells were cultured with normal or MDS stromal cells, but was unable to inhibit MDS stem and early progenitor cell growth (day-35-42 CAFC) on pre-established stromal layers. These findings suggest that in early hematopoietic cells isolated from MDS patients the Notch signal transduction pathway is disrupted. Furthermore, there was a marked reduction in the plasticity of mesenchymal stem cells of MDS patients compared with those of normal marrow donors, in neurogenic and adipogenic differentiation ability and hematopoiesis supporting capacity in vitro. These results are consistent with the hypothesis that when alterations are present in the myelodysplastic stroma environment along with intrinsic changes in a hematopoietic stem/progenitor cell clone, both factors might equally contribute to the abnormal hematopoiesis in MDS. PMID- 18158567 TI - Chromogranin A-positive tumor cells in human esophageal squamous cell carcinomas. AB - Gastrointestinal cancers have frequently shown neuroendocrine (NE) differentiation, but whether NE differentiation occurs in esophageal squamous cell carcinoma (ESCC) remains unclear. In this study, tissue sections obtained from 43 patients with ESCC from a high-incidence area of Northern China were used for the assessing of NE differentiation by immunohistochemistry using antibody against chromogranin A (CGA). In addition, the malignant characteristics and proliferation capacity of CGA-positive cells were also examined by immunohistochemistry. The clinicopathological significance of these CGA-positive tumor cells in ESCC was assessed. Of 43 ESCC samples, CGAimmunoreactive tumor cells were detected in 10 cases (23.26%). However, the CGA-positive tumor cells were scattered at a very low number among non-immunoreactive tumor cells and were rarely constituted a major part of cancer cell nests. Only 4.65% (2/43) cases showed a high density (>10 cells but <1% of total tumor cell mass) of CGA positive tumor cells. P53 immunoreactivity was frequently shown, while Ki67 was hard to detect in these CGApositive cells. In addition, no relationship between CGA positivity rate and clinicopathological parameters was found. Thus, we concluded that lowdensity CGA-positive tumor cells can be detected in ESCC, supporting the notion that heterogeneous NE differentiation also exists in tumors that lack neuroendocrine cells in their normal epithelial counterparts. PMID- 18158568 TI - Significance of PML and p53 protein as molecular prognostic markers of gallbladder carcinomas. AB - Molecular markers for cancers are not only useful for cancer detection and prognostic prediction, but may also serve as potential therapeutic targets. In order to identify reliable molecular markers for prognostic prediction in gallbladder carcinoma (GBC), we evaluated the immunohistochemical expression of 15 proteins, namely p53, p27, p16, RB, Smad4, PTEN, FHIT, GSTP1, MGMT, E cadherin, nm23, CD44, TIMP3, S100A4, and promyelocytic leukemia (PML) in 138 cases of GBC using the tissue microarray method. The prognostic significance was analyzed for each protein. Overexpression of p53 and S100A4, and loss of p27, p16, RB, Smad4, FHIT, E-cadherin and PML expression were associated with poor survival. In particular, PML and p53 showed considerable potential as independent prognostic markers. Patients with normal PML and p53 expression displayed favorable outcomes, compared to those showing abnormal expression of either or both proteins (49% vs. 23% in a 5-year survival rate; 60 months vs. 11 months in median survival, respectively; P=0.009). Thus, PML and p53 are potential candidates for development as clinically applicable molecular prognostic markers of GBC, and may be effective therapeutic targets for the disease in the future. PMID- 18158569 TI - Expression of connective tissue growth factor (CTGF/CCN2) in a mouse model of rhabdomyosarcomagenesis. AB - Connective tissue growth factor (CTGF/CCN2) is a cysteine-rich matricellular protein that belongs to the CCN (CYR61, CTGF, NOV) protein family. It is highly expressed by human rhabdomyosarcoma cells and sustains their survival. In this study we investigated CCN2 expression in a mouse model of spontaneous rhabdomyosarcomagenesis that combines HER-2/neu oncogene activation and p53 oncosuppressor gene inactivation (BALB-p53neu mice). Murine rhabdomyosarcoma cells showed a 4-26 fold increase in CCN2 mRNA expression regarding to normal thigh muscle. Moreover, they expressed CCN2 protein at levels comparable to human rhabdomyosarcoma cells. Therefore BALBp53neu mice might be useful for the evaluation of the role played by CCN2 in rhabdomyosarcoma in vivo. PMID- 18158570 TI - Increase of hypophyseal hormone levels in male head and neck cancer patients. AB - Head and neck squamous cell carcinoma (HNSCC) develops in at least 80% of cases in men with a history of smoking and heavy alcohol consumption, still it is only diagnosed in a small proportion of alcoholics. Endocrine milieu is an important factor in carcinogenesis and prognosis of several cancer types. The aim of our study was to investigate sex steroid and hypophyseal hormone status of male HNSCC patients in comparison to healthy volunteers and to patients with alcoholic liver disease, to determine possible hormonal alterations characteristic of cancer. Liver function (GGT level), and serum levels of gonadotropic hormones (FSH, LH, prolactin), sex steroids (estradiol, progesterone, testosterone) and sex hormone binding globulin (SHBG) were compared in 130 male HNSCC patients, 54 patients with alcoholic liver disease but no known cancer, and 56 healthy controls. We found abnormal values of liver function in both HNSCC patients and alcoholics compared to healthy controls, suggesting the presence of alcoholic liver disease in the former group as well. On the other hand, a significant elevation in the level of DHEA, FSH and LH was observed in cancer patients exclusively. As a conclusion, abnormal alterations in sex steroid hormone levels can frequently be found in HNSCC patients, which may be caused in part by the alcoholic liver damage accompanying the disease. The significant increase in FSH and LH serum levels, observed only in the cancer patients, indicates that these hormones may play a role in the development and/or progression of HNSCC. PMID- 18158571 TI - Histopathological variation of primary mucosa-associated lymphoid tissue lymphoma of the oral cavity. AB - Primary mucosa-associated lymphoid tissue (MALT) type lymphoma arising in the oral cavity is rare. We examined histopathologic, immunohistological and genotypic findings of seven cases of intraoral MALT lymphoma using formalin-fixed paraffin-embedded tissues. Histologically, two variants have been delineated. (i) In four cases of minor salivary gland type, the lymphoid follicles were surrounded by centrocyte-like (CCL) cells with occasional follicular colonization. The CCL cells invaded the residual salivary gland duct resulting in a lymphoepithelial lesion. CCL cells frequently showed plasmacytic differentiation. (ii) In three cases of follicular growth type, the lesion was characterized by follicular growth pattern resulting from prominent follicular colonization. CCL cells showed minimal plasma cell differentiation. There was no residual epithelial component detected even by cytokeratin immunostaining. There were no Epstein-Barr virus-encoded small RNA-positive cells detected by in situ hybridization. API2-MALT1 fusion transcript does not appear to be associated with either histological variant of primary intraoral MALT lymphoma. PMID- 18158572 TI - Effect of hormone replacement therapy on postmenopausal endometrial bleeding. AB - The aim of the study was to determine the effect of postmenopausal hormone replacement therapy (HRT) (treatment using estrogen only and sequential and continuous combined estrogen-progestogen treatment) on endometrial bleeding and histological changes of the endometrium. In a six-year period (2000-2005), 5893 patients were given care and the incidence of postmenopausal uterine bleeding was detected in groups of patients having and not having received hormonal treatment at the Menopause Outpatient Unit of the authors' department. In the case of bleeding, fractioned abrasion was performed and the samples were analyzed histologically. Among the postmenopausal patients who had not been given hormonal treatment, the incidence of bleeding episodes was significantly higher as among those having received hormonal treatment. In the samples, findings of proliferative endometrium occurred significantly more often in case of non treated patients and those treated with sequential combined hormone therapy compared to patients receiving continuous combined hormone therapy. Although it was statistically not significant, hyperplasia simplex and complex together showed a tendency of reduced incidence in patients medicated by continuous combined treatment. These findings suggest that continuous combined hormonal treatment started at the right time (even before the menopause) may reduce the chances of the development of hyperplasia. A significantly higher incidence of hyperplasia was noted in patients using estrogen treatment only. It is possible that unopposed estrogen treatment further engraves an already diagnosed endometrial hyperplasia. In the group having received hormonal treatment, no complex hyperplasia accompanied by atypia occurred, only hyperplasia simplex was diagnosed in these cases. As a result of continuous reliance on combined preparations, the endometrium had become atrophied, therefore the chance of hyperplasia-related changes and of bleeding as a side effect decreased significantly. According to the authors' experience, hormonal treatment does not pose a risk to the development of endometrial carcinoma; on the contrary, continuous combined preparations appear to reduce the risk of hyperplasia and, indirectly, the chances of the development of adenocarcinoma. PMID- 18158573 TI - Factors influencing serum concentration of CA125 and CA15-3 in Iranian healthy postmenopausal women. AB - Screening for breast and ovarian cancers are required due to the late stage at diagnosis and poor survival. Serum CA125 and CA15-3 are important cancerdetecting agents in patients with ovarian and breast cancers, respectively. Elevation of CA125 and CA15-3 level correlates with malignant and non-malignant conditions. Moreover, a series of individual characteristics affect the serum level of these markers. The objective of the present study was to evaluate CA125 and CA15-3 levels in cancer-free postmenopausal women to investigate the impacts of patient parameters on the serum level of these markers. 203 subjects were studied prospectively. Serum CA125 and CA15-3 assessment was done subsequent to the direct interview. The associations between marker levels and presenting features were examined. CA125 and CA15-3 levels were elevated in 35 (17.2%) and 12 (5.9%) of persons, respectively. A higher CA125 level was associated with advanced age (p = 0.046), while a lower level was correlated with hormone replacement therapy (HRT) and having smoking habits (p = 0.000 and p = 0.01, respectively). CA15-3 level was remarkably lower amongst oral contraceptive (OCP) users (p = 0.03). Serum marker levels were not significantly related to menarche age, age at menopause, height, weight, BMI and parity. Serum CA125 is imperative indicator for malignancies of the ovary; however, personal and medical factors influence its serum level. A fair interpretation of results must be due to an accurate attention to the individual characteristics. PMID- 18158574 TI - Anti-tumor effect of non-steroidal anti-inflammatory drugs on human ovarian cancers. AB - Many reports have demonstrated that non-steroidal anti-inflammatory drugs (NSAIDs) suppress malignant transformation and tumor growth, and some NSAIDs are expected to be new anti-cancer agents. In this study, we examined the anti-tumor effects of the non-specific cyclooxygenase (COX) inhibitors aspirin and piroxicam, and the selective COX-2 inhibitor meloxicam on xenotransplanted ovarian cancer. Tumor growth and survival were compared in female nu/nu mice, xenografted with subcutaneous OVCAR-3 tumors or with intraperitoneal DISS tumors and treated with aspirin (200 ppm in diet, everyday), piroxicam (150 ppm in diet, everyday) or meloxicam (162 ppm in diet, everyday). Al, of the agents tested significantly suppressed the growth of OVCAR-3 tumors xenotransplanted subcutaneously as compared to the control. There was a significant difference in inhibition of OVCAR-3 tumor growth between meloxicam and aspirin treatment. Meloxicam and piroxicam treatment significantly prolonged survival of mice with malignant ascites derived from DISS cells as compared to control and aspirin treatment. Mice treated with meloxicam survived significantly longer than those treated with piroxicam. There was no significant difference in survival between control and aspirin treatment. Necropsy revealed that one of the 6 cancer-bearing mice treated with piroxicam suffered from stomach perforation. These results indicate that a selective COX-2 inhibitor produces greater anti-tumor effect against ovarian cancer than a nonselective COX inhibitor and that meloxicam may have a potential of leading to a novel therapeutic strategy against ovarian cancer. PMID- 18158575 TI - Genetic alterations of E-cadherin and beta-catenin in germinoma and teratoma: report of two central nervous system cases. AB - The genetic basis as well as mechanisms of development of germ cell tumors of the CNS are still unexplained. In the present article changes of Ecadherin (CDH1) and beta-catenin (CTNNB1) genes in two CNS germ cell tumors are reported. Both gene products are components of adherens junctions, but are also involved in the Wnt signaling pathway. A case of germinoma of the central nervous system and a case of spinal channel teratoma were tested for loss of heterozygosity (LOH) of E cadherin gene by PCR amplification of tetranucleotide polymorphism (D16S752). Changes of beta-catenin were tested by heteroduplex method. Both germ cell tumors analyzed demonstrated LOH of the CDH1 gene. Analysis of exon 3 of the CTNNB1 gene showed additional band in the germinoma, suggesting that this sample harbors mutation in the beta-catenin gene. Immunostaining showed that LOHs in our samples were accompanied with the absence of E-cadherin protein. We also investigated E cadherin expression in four other germinomas, of which three were negative and one was mildly positive. Our findings may contribute to better understanding of the genetic profile of germ cell tumors. PMID- 18158576 TI - Rhabdomyomatous mesenchymal hamartoma presenting as a skin tag in the sternoclavicular area. AB - Rhabdomyomatous mesenchymal hamartoma (RMH) is a rare congenital lesion of the dermis and subdermis. It has been described predominantly in newborns, with 30 cases reported in the English literature. Typically, it appears as a skin tag, papule, nodule or a mass involving the face or sternal notch. A 28-day-old girl presented with a 1.4 x 0.8 cm soft skin tag in the right sternoclavicular area. Physical examination revealed no congenital anomalies. A shaved biopsy showed that the core of the lesion contained striated muscle fibers mixed with hair follicles and sebaceous and eccrine glands. Thin epidermis lined the outside of the tag. We report a patient with a RMH in a site not previously reported and discuss the differential diagnosis. PMID- 18158577 TI - Unusual case of subcutaneous angiosarcoma metastatic to the ovary. AB - The ovaries are common sites for metastatic disease, however, the most frequent ones are carcinomas. Metastatic sarcomas are very rare in ovary and most of them arise from genital tract. We present the case of a 33-year-old woman with subcutaneous angiosarcoma who had metastatic disease to the ovary resulting in acute abdominal pain. Discussion of the case and a review of the literature are presented. PMID- 18158578 TI - Hepatitis C virus infection and locally advanced splenic marginal zone lymphoma. AB - Splenic marginal zone lymphoma (SMZL) is a rare malignant B-cell neoplasm, usually with an indolent clinical course and favorable prognosis. Treatment options include chemotherapy, surgery, radiation and immunotherapy. In some recent studies an increased incidence of hepatitis C virus (HCV) infection in patients with SMZL was reported and its possible role in lymphomagenesis was emphasized. A 66-year-old woman with twelve-year history of HCV infection was admitted due to locally advanced abdominal tumor involving the spleen and the left part of the diaphragm. Transaminase serum levels were not elevated. Neither peripheral lymphadenopathy nor bone marrow pathology was found. Absolute blood lymphocyte, erythrocyte and platelet counts were normal. A splenectomy with partial diaphragm resection in one block was performed. Recovery was uneventful. Pathologic examination with immunohistochemistry revealed SMZL and confirmed a neoplastic infiltration of the resected diaphragm. Following surgery, chemotherapy (CHOP regimen) and immunotherapy (anti-CD20 antibody) were given. At the last follow-up 15 months after surgery, the patient was free of any symptoms of lymphoma. Surgical resection of even locally advanced SMZL with involvement of adjacent tissues can be performed as a diagnostic and therapeutic procedure. Splenectomy is especially indicated in symptomatic patients without other sites of the disease. HCV infection may result in increased risk of SMZL due to the induction of B-cell lymphoproliferation. Because of possible lymphoma regression following anti-viral therapy, a systematic screening for HCV in patients with SMZL seems to be valuable and helpful for treatment planning. PMID- 18158579 TI - Detection of CD55- and CD59-deficient granulocytic populations in patients with myelodysplastic syndrome. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by absence of CD55 and CD59 from the surface of affected cells. PNH has been associated with myelodysplastic syndromes (MDS). The aim of our study was to estimate the prevalence of the PNH clone in MDS patients by detecting CD55 and CD59 deficiency. We studied 90 MDS patients: 19 patients with RA, 15 with refractory anemia with ringed sideroblasts (RARS), 18 with refractory anemia with excess of blasts (RAEB), 17 with refractory anemia with excess of blasts in transformation (RAEB-t), and 21 with chronic myelomonocytic leukemia (CMML). Twenty healthy individuals were also studied as the control group. We studied the PNH clone on granulocytes of these patients with the aid of flow cytometry. CD55- and CD59 deficient granulocytic populations were detected in 15.5% of MDS patients compared to 2.8% of normal individuals. Among the subgroups of the study, significant difference was present in three cases: (1) between CMML and control, (2) between CMML and RA, and (3) between CMML and RARS. These data indicate a possible association between PNH phenotype and MDS. MDS patients of worse prognosis (CMML) express more strongly the PNH clone compared to those of better prognosis (RA and RARS). Perhaps, the examination of MDS patients for the PNH clone by flow cytometry could provide us with a valuable prognostic tool. PMID- 18158580 TI - The diagnostic yield from implantable loop recorders in children and young adults. AB - BACKGROUND: Syncope and palpitations occur frequently in young patients. Noninvasive diagnostic testing may be inconclusive. AIM: To assess the diagnostic yield of implantable loop recorders in young patients. PATIENTS AND METHODS: Thirty-three young patients underwent implantation of a loop recorder for long term monitoring of cardiac rhythm, to establish symptom-rhythm correlation. They belonged to one of three subgroups: those with structurally normal heart, normal electrocardiogram at rest, and negative family history (n = 16); patients with structural heart disease and previous surgical repair (n = 11), and patients with proven or suspected primary electrical disease (n = 6). A combination of automatic and patient-activated recordings was used to monitor cardiac rhythm during symptomatic episodes. RESULTS: There were no procedural complications. Diagnostic electrograms could be obtained in all patients. A high degree of symptom-rhythm correlation was established. In 8/33 patients, no recurrence of symptoms was observed either until end of battery life of the device (n = 4) or until last follow-up (n = 2). Specific cardiac therapy was required, based on rhythms recorded by the device in 15 patients (until last follow-up). This consisted of catheter ablation of a tachyarrhythmia (n = 7), pacemaker implantation or upgrade (n = 5) or ICD implantation (n = 5). In the remaining patients (n = 10), recurrence of symptoms was associated with a normal electrocardiogram, and in two of these patients a non-cardiac diagnosis was made. CONCLUSIONS: In selected patients, the implantable loop recorder provides valuable diagnostic information to guide further therapy. PMID- 18158582 TI - Filoviruses: Interactions with the host cell. AB - The highly pathogenic filoviruses, Marburg and Ebola virus, are difficult to handle and knowledge of the interactions between filoviruses and their host cells remained enigmatic for many years. Two developments were crucial for the presented advances in our understanding of the cell biology of filoviruses, which is still fragmentary. On the one hand, the number of high containment laboratories increased where handling of the highly pathogenic filoviruses is possible. On the other hand, molecular biological tools have been developed that allow investigation of certain aspects of filoviral replication under normal laboratory conditions which considerably accelerated research on filoviruses. This review describes advances in understanding the interactions between host cells and filoviruses during viral attachment, entry, transcription, assembly and budding. PMID- 18158583 TI - Towards understanding Pseudomonas aeruginosa burn wound infections by profiling gene expression. AB - Pseudomonas aeruginosa is a key opportunistic pathogen causing severe acute and chronic nosocomial infections in immunocompromised or catheterized patients. It is prevalent in burn wound infections and it is generally multi-drug resistant. Understanding the genetic programs underlying infection is essential to develop highly needed new strategies for prevention and therapy. This work reviews expression profiling efforts conducted worldwide towards gaining insights into pathogenesis by P. aeruginosa, in particular in burn wounds. Work on various infection models, including the burned mouse model, has identified several direct virulence factors and elucidated their mode of action. In vivo gene expression experiments using In vivo Expression Technology (IVET) ascertained distinct regulatory circuits and traits that have helped explain P. aeruginosa s success as a general pathogen. The sequencing of the whole genome from a number of P. aeruginosa strains and the construction of genome-wide microarrays have paved the road to the several insightful studies on the (interacting) traits underlying infection. A series of in vitro and initial in vivo gene expression studies revealed specific traits pivotal for infection, such as quorum sensing systems, iron acquisition and oxidative stress responses, and toxin production among others. The data sets obtained from global transcriptional profiling provide insights that will be essential for the development of new targets and options for prevention and intervention. PMID- 18158581 TI - Protein factors in pre-mRNA 3'-end processing. AB - Most eukaryotic mRNA precursors (premRNAs) must undergo extensive processing, including cleavage and polyadenylation at the 3'-end. Processing at the 3'-end is controlled by sequence elements in the pre-mRNA (cis elements) as well as protein factors. Despite the seeming biochemical simplicity of the processing reactions, more than 14 proteins have been identified for the mammalian complex, and more than 20 proteins have been identified for the yeast complex. The 3'-end processing machinery also has important roles in transcription and splicing. The mammalian machinery contains several sub-complexes, including cleavage and polyadenylation specificity factor, cleavage stimulation factor, cleavage factor I, and cleavage factor II. Additional protein factors include poly(A) polymerase, poly(A)-binding protein, symplekin, and the C-terminal domain of RNA polymerase II largest subunit. The yeast machinery includes cleavage factor IA, cleavage factor IB, and cleavage and polyadenylation factor. PMID- 18158584 TI - Generation of superoxide anion in chloroplasts of Arabidopsis thaliana during active photosynthesis: a focus on rapidly induced genes. AB - The antioxidant defense system involves complex functional coordination of multiple components in different organelles within the plant cell. Here, we have studied the Arabidopsis thaliana early response to the generation of superoxide anion in chloroplasts during active photosynthesis. We exposed plants to methyl viologen (MV), a superoxide anion propagator in the light, and performed biochemical and expression profiling experiments using Affymetrix ATH1 GeneChip microarrays under conditions in which photosynthesis and antioxidant enzymes were active. Data analysis identified superoxide-responsive genes that were compared with available microarray results. Examples include genes encoding proteins with unknown function, transcription factors and signal transduction components. A common GAAAAGTCAAAC motif containing the W-box consensus sequence of WRKY transcription factors, was found in the promoters of genes highly up-regulated by superoxide. Band shift assays showed that oxidative treatments enhanced the specific binding of leaf protein extracts to this motif. In addition, GUS reporter gene fused to WRKY30 promoter, which contains this binding motif, was induced by MV and H(2)O(2). Overall, our study suggests that genes involved in signalling pathways and with unknown functions are rapidly activated by superoxide anion generated in photosynthetically active chloroplasts, as part of the early antioxidant response of Arabidopsis leaves. PMID- 18158585 TI - Histology in neuroscience: from the symposia: homage of the histology chairs to Santiago Ramon y Cajal, 18-19 December, 2006, Albacete, Spain. PMID- 18158586 TI - OPG and RANKL mRNA and protein expressions in the primary and secondary metaphyseal trabecular bone of PTH-treated rats are independent of that of SOST. AB - Sclerostin, encoded by the SOST gene, is a recently identified protein which seems to affect bone remodeling by inhibiting bone formation via Wnt pathways. A previous study on OPG and RANKL, two cytokines involved in the control of osteoclastogenesis, showed that the anabolic effect produced by intermittent treatment with parathyroid hormone was characterized by an increase in OPG/RANKL mRNA ratio in the primary spongiosa of metaphyseal bone of rat femur, and by its falling in the secondary spongiosa, in comparison to controls (Silvestrini et al. (2007a)). Considering that Wnt pathway components seem to regulate osteoclast formation and bone resorption by repression of RANKL transcription and by positive regulation of OPG gene in osteoblastic cells, we have evaluated, in the same rats, whether and how SOST mRNA and protein in the primary and secondary metaphyseal bone are affected by PTH. SOST mRNA and protein significantly fell in both primary and secondary spongiosa where only a few osteocytes were positive to sclerostin. These data show that in the two metaphyseal areas no relationship does exist between the trends of OPG and RANKL mRNA and that of SOST, suggesting that there are no direct links between the effects induced by PTH on these molecules, at least in terms of gene expression. PMID- 18158587 TI - Role of the specifically targeted lysine residues in the glycation dependent loss of chaperone activity of alpha A- and alpha B-crystallins. AB - Earlier studies have shown significant loss of chaperone activity in alpha crystallin from diabetic lenses. In vitro glycation studies have suggested that glycation of alpha-crystallin could be the major cause of chaperone activity loss. The following lysine (K) residues in alpha-crystallin have been identified as the major glycation sites: K11, K78, and K166 in alpha A-crystallin and K90, K92, and K166 in alpha B-crystallin. The present study was aimed to assess the contribution of each of the above glycation site in the overall glycation and loss of chaperone activity by mutating them to threonine followed by in vitro glycation with fructose. Level of glycated protein (GP) was determined by phenylboronate affinity chromatography, advanced glycation end products (AGEs) by direct ELISA using anti-AGE polyclonal antibody, and chaperone activity by using alcohol dehydrogenase as the target protein. K11T, K78, and K166T mutants of alpha A showed 33, 17, and 27% decrease in GP and 32, 18, and 21% decrease in AGEs, respectively, as compared to alpha A-wt. Likewise, K90T, K92T, K90T/K92T, and K166T mutants of alpha B showed 18, 21, 29, and 12% decrease in GP and 22, 24, 32, and 16% decrease in AGEs, respectively. Chaperone activity also showed concomitant increase with decreasing glycation and AGEs formation. alpha A-K11T and alpha B-K90T/K92T mutants showed the largest decrease in glycation and increase in chaperone activity. PMID- 18158588 TI - Atorvastatin accelerates extracellular nucleotide degradation in human endothelial cells. AB - HMG-CoA reductase inhibitors (statins) exert pleiotropic effects in the cardiovascular system beyond its cholesterol-lowering action. We aimed to investigate how atorvastatin affects extracellular nucleotide degradation in human endothelial cells, as increased activity of this pathway would facilitate conversion of pro-inflammatory nucleotides into anti-inflammatory adenosine. Primary cultures of human endothelial cells were treated with 1 microM, 10 microM and 100 microM atorvastatin for 24 h. Enzyme assays were performed as well as intact cell studies, to evaluate capacity of cells to degrade ATP to adenosine. Atorvastatin significantly increased ATP breakdown and adenosine formation in the medium of intact cells in a dose-dependent manner. The activities of ATPase, ADPase and ecto-5'-nucleotidase (eN) in cell homogenates following Atorvastatin treatment were also increased while no change was observed in the lactate dehydrogenase activity. We suggest a new mechanism of protective effect of atorvastatin by activation of endothelial enzymes involved in extracellular nucleotide degradation in human endothelial cells. PMID- 18158589 TI - Expression and distribution of junctional adhesion molecule-1 in the human cornea. AB - PURPOSE: To investigate the expression and distribution of junctional adhesion molecule 1 (JAM-1) in human corneal tissue and cells. METHODS: Reverse transcriptase-polymerase chain reaction was used to detect the expression of JAM 1, ZO-1, and occludin mRNAs in corneal cells, while the presence of JAM-1 protein was analyzed by flow cytometry (FACS). Double immunofluorescence staining was used to determine the tissue distribution of JAM-1 and occludin in human corneas. RESULTS: Strong expression of JAM-1, ZO-1, and occludin mRNAs was observed in primary cultured corneal epithelial and endothelial cells, but not in primary cultured keratocytes. The expression of JAM-1 protein in cultured epithelial and endothelial cells was confirmed by FACS. When keratocytes were cultured in medium with 10% fetal calf serum for several passages, differentiation into corneal myofibroblasts occurred. The expression of JAM-1 was detected in these corneal myofibroblasts at both RNA and protein levels. JAM-1 immunoreactivity was seen at cell borders throughout the entire epithelium, but not in keratocytes from normal corneal tissue. On the other hand, JAM-1 immunoreactivity was detected in the cytoplasm of corneal endothelial cells. CONCLUSIONS: JAM-1 is expressed by human corneal epithelial and endothelial cells, but not by keratocytes, although its expression is induced in corneal myofibroblasts. PMID- 18158590 TI - Proteomic analysis of human lens epithelial cells exposed to microwaves. AB - PURPOSE: To study proteomic changes in human lens epithelial cells (HLECs) exposed to 1800-MHz Global System for Mobile Communication (GSM)-like microwaves. METHODS: In three separate experiments, HLECs were exposed and sham-exposed (six dishes each) to 1800-MHz GSM-like radiation for 2 h. The specific absorption rates were 1.0, 2.0, or 3.5 W/kg. Immediately after radiation, the proteome was extracted from the HLECs. Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis(2-DE; silver staining) and PDQuest 2-DE analysis software were used to separate and analyze the proteome of exposed and sham-exposed HLECs. Four differentially expressed protein spots were selected and identified by using electrospray ionization tandem mass spectrometry (ESI-MS-MS). RESULTS: When the protein profiles of exposed cells were compared with those of sham-exposed cells, four proteins were detected as upregulated. After analysis by ESI-MS-MS and through a database search, heat-shock protein (HSP) 70 and heterogeneous nuclear ribonucleoprotein K (hnRNP K) were determined to be upregulated in the exposed cells. CONCLUSIONS: Two-dimensional polyacrylamide gel electrophoresis combined with mass spectrometry may be a powerful tool for screening potential electromagnetic-reaction protein markers. HSP70 and hnRNP K are involved in the stress reaction of HLECs exposed to microwaves. These cell responses are nonthermal effects of the electromagnetic field. PMID- 18158591 TI - Association between open-angle glaucoma and gene polymorphism for heat-shock protein 70-1. AB - PURPOSE: Heat-shock proteins (HSPs) or antibodies against them may contribute to glaucomatous optic neuropathy. We investigated the associations of HSP70-1 polymorphisms with open-angle glaucoma (OAG) in a Japanese population. METHODS: In 241 normal Japanese controls and 501 Japanese OAG patients, including 211 with primary open-angle glaucoma (POAG) and 290 with normal-tension glaucoma (NTG), two single-nucleotide polymorphisms, A-110C and G+190C, of HSP70-1 were identified by using an Invader assay and polymerase chain reaction-restriction fragment length polymorphism, respectively. Genotype distributions were compared between controls and OAG patients. Age at diagnosis, untreated maximum intraocular pressure, and visual field defects at diagnosis were examined for associations with the polymorphisms. RESULTS: Distribution of the A-110C genotype (AA versus AC+CC) differed significantly between controls and OAG patients (P = 0.007), POAG patients (P = 0.007), or NTG patients (P = 0.032). The genotype distribution of the G+190C polymorphism did not differ significantly between the controls and any patient group. No significant differences in the clinical characteristics of the patients were detected between genotype-defined groups by logistic regression analysis. CONCLUSION: The A-110C polymorphism of HSP70-1 may be associated with OAG pathogenesis in Japanese patients. PMID- 18158592 TI - Involvement of the Rho/Rho kinase signaling pathway in platelet-derived growth factor BB-induced vascular endothelial growth factor expression in diabetic rat retina. AB - PURPOSE: To examine the role played by the Rho/Rho kinase pathway in the platelet derived growth factor BB (PDGF-BB)-induced expression of vascular endothelial growth factor (VEGF) in the diabetic rat retina. METHODS: Male Sprague-Dawley rats were made diabetic by a single intraperitoneal injection of streptozotocin. Diabetic rats and PDGF-BB-exposed primary cultured porcine retinal pericyte cells (PRPCs) were treated with fluvastatin, an HMG-CoA reductase inhibitor, and fasudil, a selective Rho kinase inhibitor. The retinal expression of VEGF was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The mRNA expression of VEGF and the activation of Rho A were studied by Northern and Western blot analyses. RESULTS: RT-PCR showed that in rats that were diabetic for 4 weeks VEGF mRNA expression levels were 1.8-fold those in control rats. This enhanced expression was blocked by treatment with fluvastatin or fasudil. Exposure of PRPCs to PDGF-BB increased their VEGF mRNA expression threefold, and fluvastatin suppressed this effect. Fluvastatin also suppressed the PDGF-BB induced activation of Rho GTPase in PRPCs. CONCLUSIONS: The Rho/Rho kinase pathway may be involved in the upregulation of VEGF expression in the diabetic retina, suggesting that fasudil might be useful for the prevention of diabetic retinopathy. PMID- 18158593 TI - Association of Major Histocompatibility Complex Class I Chain-Related Gene A and HLA-B Alleles with Behcet's Disease in Turkey. AB - PURPOSE: Behcet's disease (BD) is known to be associated with HLA-B*51 in many different ethnic groups. Recently, the major histocompatibility complex class I chain-related gene A (MICA), located near the HLA-B gene, has been proposed as a candidate gene for BD susceptibility in several ethnic groups. To compare the relative contribution of MICA polymorphisms and HLA-B*51 to BD in different ethnic groups, we studied MICA polymorphisms in Turkish BD patients. METHODS: Thirty-three Turkish BD patients and 65 healthy controls were enrolled for analysis of polymorphisms in the extracellular domains of MICA. RESULTS: The phenotype frequencies of MICA*009 were significantly higher in BD patients (75.8%) than in controls (29.2%) (P = 0.000015). HLA-B*51 was also significantly more frequent in BD patients (81.8%) than in controls (29.2%) (P = 0.0000007). A strong association existed between MICA*009 and HLA-B*51. To assess the confounding effect of MICA*009 on HLA-B*51, we performed a stratification analysis that showed that BD was distinctly associated only with HLA-B*51. CONCLUSION: Our results indicate that the major susceptibility gene for BD is HLA B*51 and that the association between MICA*009 and BD arises from a strong linkage disequilibrium with HLA-B*51. However, we suggest that MICA*009 likely elicits an immune effect secondary to BD. PMID- 18158594 TI - Acquired nasolacrimal duct obstruction in children. AB - PURPOSE: To study the clinical characteristics of acquired nasolacrimal duct (NLD) obstruction in Korean children. METHODS: The records of 38 patients under the age of 15 years who had undergone silicone intubation or dacryocystorhinostomy (DCR) for acquired NLD obstruction were evaluated. Patients who had congenital NLD obstruction or a history of NLD obstruction due to trauma were excluded. RESULTS: The mean age of the patients was 3 years (range, 1-8 years). The causes of the NLD obstruction were epidemic keratoconjunctivitis (EKC) in 32 patients, herpetic blepharitis in one, and idiopathic in five. In patients with a history of EKC, symptoms were observed at a mean age of 12.7 months (range, 3-51 months). These patients demonstrated multifocal features of obstruction, unlike those in the idiopathic group. Silicone intubation was performed in 32 patients, external DCR in three, and endonasal DCR in three. The lacrimal procedures were beneficial in 36 of the 38 patients. The mean follow-up period was 7.6 months. CONCLUSIONS: Acquired NLD obstruction in children was mostly caused by EKC in our study. It is important to suspect NLD obstruction in children with persistent epiphora that develops after EKC. PMID- 18158595 TI - Persistent epithelial defects due to neurotrophic keratopathy treated with a substance p-derived peptide and insulin-like growth factor 1. AB - PURPOSE: The loss of corneal sensation results in the development of persistent corneal epithelial defects. The combination of a substance P-derived peptide, phenylalanine -glycine-leucine-methionine (FGLM)-amide, and insulin-like growth factor 1 (IGF-1) stimulates corneal epithelial migration in vitro and corneal epithelial wound closure in vivo. The clinical efficacy of eye drops containing FGLM-amide and IGF-1 for the treatment of persistent epithelial defects in individuals with neurotrophic keratopathy was examined in a prospective open study. METHODS: Eleven patients with persistent corneal epithelial defects associated with neurotrophic keratopathy were treated for up to 28 days by the administration of eye drops containing FGLM-amide and IGF-1. The course of epithelial healing was monitored by slit-lamp examination, and visual acuity was measured before and after treatment. RESULTS: Complete epithelial resurfacing was achieved in eight of the nine (89%) cases of persistent epithelial defects in the nine patients who received the full course of treatment. Epithelial defects had completely resurfaced in two of nine (22%) and five of nine (56%) cases within 1 and 2 weeks, respectively, after treatment initiation. No adverse effects of treatment were observed in any of the 11 patients. CONCLUSIONS: Eye drops containing FGLM-amide and IGF-1 induced the rapid resurfacing of persistent epithelial defects in individuals with neurotrophic keratopathy. PMID- 18158596 TI - Frequency doubling technology threshold visual field results vary with software version. AB - PURPOSE: To determine the comparability of test results obtained using two versions of commercially available frequency doubling technology (FDT) software for visual field testing. METHODS: One hundred glaucoma patients underwent duplicate N-30 threshold FDT visual field examinations of one eye; one test was performed with the recently released software version 3.0 and the other test with the earlier version 2.6. Results were compared in terms of test duration, mean deviation (MD), pointwise sensitivity, pointwise deviations from the age-normal threshold, and average sensitivity. RESULTS: Results obtained with version 3.0 differed significantly from those obtained with version 2.6. Version 3.0 tests took on average 14.2% less time (P < 0.0001), and the average MD was 0.53 dB more negative (P < 0.01). Version 3.0 threshold estimates derived from a mixed-model analysis were significantly lower at 15 of 19 test points (P = 0.0074). At 14 of the 19 test point locations, the mixed-model analysis also showed that threshold measurements were more frequently outside the normal range with version 3.0 (P = 0.022). CONCLUSIONS: We found statistically significant differences between results obtained using FDT versions 3.0 and 2.6, which might affect clinical management. New age-corrected normal values for version 3.0 should be obtained and made available to users, and new baseline field data should be established whenever a patient is switched from version 2.6 to version 3.0. PMID- 18158597 TI - Abnormal retinal blood vessels in Ehlers-Danlos syndrome type VI. AB - BACKGROUND: Ehlers-Danlos syndrome (EDS) is a hereditary connective tissue disorder caused by defective collagen synthesis and categorized into 11 types based on genetic mutations and clinical features.(1) Its principal clinical characteristics include hyperelasticity and vulnerability of the skin and joints to laxity, and fragility of blood vessels. CASE: We recently performed vitreous surgery with introduction of an artificial cornea and corneal allograft transplantation in a boy with EDS type VI, whose visual acuity had decreased rapidly because of vitreous hemorrhage. OBSERVATIONS: An ultrasound examination revealed posterior vitreous detachment with fresh vitreous hemorrhage. After vitrectomy with introduction of an artificial cornea and cornea allograft transplant were carried out, visual acuity improved to 0.05 and was maintained for at least 22 months. CONCLUSIONS: Abnormal collagen synthesis is thought to lead to systemic disorders such as hyperelasticity, vulnerability to skin and joint laxity, and fragile blood vessels in patients with Ehlers-Danlos syndrome. PMID- 18158598 TI - In vivo measurements of cone photoreceptor spacing in myopic eyes from images obtained by an adaptive optics fundus camera. AB - PURPOSE: To determine the cone spacing in normal and myopic eyes from the images obtained by an adaptive optics (AO) fundus camera. METHODS: Nineteen eyes of 19 healthy volunteers with a mean +/- SD spherical equivalent refractive error of 3.7 +/- 3.3 diopters (D) (range, -0.3 to -11.1 D) and a mean axial length of 25.4 +/- 1.61 mm (range, 23.4-28.0 mm) were investigated in a prospective cross sectional study. An AO fundus camera equipped with a liquid crystal phase modulator was used to obtain the images of the photoreceptor mosaic. The spacing between the cones was calculated manually at a retinal locus 2 degrees temporal from the center of the fovea. The magnification of the image was calculated by the axial length measured with an IOL Master. RESULTS: The axial length was correlated with the refractive error (Pearson, r = -0.869; P < 0.001). The average cone spacing in the moderate- to high-myopia group (-6.5 +/- 2.3 D, n = 9) was 4.71 +/- 0.44 microm, which was significantly greater (P = 0.002) than the 3.90 +/- 0.47 microm in the normal and low-myopia groups (-1.1 +/- 0.9 D, n = 10). The cone spacing was significantly correlated with the axial length (r = 0.77, P < 0.001). CONCLUSIONS: The AO fundus camera is capable of acquiring images of the photoreceptors in normal and myopic eyes. The greater spacing between cones in the myopia group is consistent with histological findings. These results suggest that retinal expansion should be considered in addition to Knapp's law when aniseikonia is evaluated in axial myopia. PMID- 18158599 TI - Relationship between intensity of reflected light and temperature increase: assessment of fundus pigmentation for transpupillary thermotherapy. AB - PURPOSE: To develop a new device for measuring reflected light during diode-laser irradiation in transpupillary thermotherapy (TTT) and to assess the correlation between fundus pigmentation, the measured intensity of reflected light, and the increase in fundus temperature. METHODS: We developed a device to measure reflected light by modifying a slit-lamp-mounted 810-nm diode-laser delivery system used clinically for TTT. The diode laser was used to irradiate test charts with various degrees of reflectance in in vitro experiments, and the fundus of nonpigmented or pigmented rabbits in in vivo experiments; then, the intensity of the reflected light and the temperature increase at the target were measured. The retinal sections were also examined histologically. RESULTS: There was a significant negative correlation between the intensity of the reflected light and the temperature increase in the target that depended on the degree of reflectance of the charts or the pigmentation of the rabbit eyes. On histopathologic examination, the extent of the changes in the irradiated retina after TTT was clearly different between pigmented and nonpigmented rabbits. CONCLUSIONS: Correlations between fundus pigmentation, the intensity of reflected light during diode-laser irradiation, and the temperature increase in TTT were demonstrated in vitro and in vivo. Our results suggest that measurement of the intensity of reflected light should be helpful for modulating the laser power in TTT according to the degree of fundus pigmentation. PMID- 18158600 TI - Incomplete oculomotor palsy with pupil sparing caused by compression of the oculomotor nerve by a posterior communicating posterior cerebral aneurysm. AB - PURPOSE: To report a patient with agenesis of both internal carotid canals who presented with incomplete oculomotor palsy with pupil sparing. METHODS: The incomplete oculomotor palsy was followed clinically, and the precise anatomical relation of the aneurysm to the subarachnoid oculomotor nerve was investigated during clipping surgery for the aneurysm. RESULTS: A 39-year-old woman with agenesis of both internal carotid arteries was admitted because of diplopia and left blepharoptosis. The left superior palpebral and the superior rectus muscles were severely palsied. The paralysis of the medial rectus muscle was milder than that of the former two muscles, and the inferior rectus was the least affected muscle. The papillary reflexes were normal. Examination during clipping surgery showed that the aneurysm was located below the oculomotor nerve in the subarachnoid space about 6.5 mm from its exit from the midbrain. The differences in severity and resolution time of the palsies of the extraocular muscles suggested that the fibers destined for the superior levator and the superior rectus were concentrated on the caudomedial portion of the subarachnoid oculomotor nerve. The fibers innervating the medial rectus muscle were located within the core of the nerve, and the fibers innervating the pupils and the inferior rectus muscle occupied a more rostral part. CONCLUSIONS: The functional distribution of fibers within the subarachnoid oculomotor nerve about 6.5 mm from its exit from the midbrain succeeds to that of the intraparenchymal oculomotor nerve. PMID- 18158601 TI - Stem cell factor in the tear fluid of vernal keratoconjunctivitis patients. AB - PURPOSE: To evaluate the contribution of stem cell factor (SCF) to the pathogenesis of vernal keratoconjunctivitis (VKC), we determined levels of soluble SCF in the tear fluid of VKC patients. METHODS: Samples of tear fluid were obtained from 38 VKC patients and 13 normal controls after we received informed consent from each participant. In the 38 VKC patients, disease severity was mild in 13, moderate in 17, and severe in 8. The levels of soluble SCF in tear fluid were determined by enzyme immunoassay. RESULTS: Mean +/- SD SCF levels in tears were 90.3 +/- 23.4 pg/ml in patients with mild VKC, 225.3 +/- 101.2 pg/ml in those with moderate VKC, and 872.0 +/- 313.2 pg/ml in those with severe VKC, and 68.3 +/- 15.3 pg/ml in healthy controls. The increase in SCF levels in patients with moderate or severe VKC was statistically significant compared with the level in healthy controls. CONCLUSION: Soluble SCF may make a crucial contribution to the pathogenesis of VKC. PMID- 18158602 TI - Alterations of retinal pigment epithelium in central serous chorioretinopathy treated by laser photocoagulation. PMID- 18158603 TI - Lobular structure of the choriocapillaris in a patient with ophthalmic artery occlusion. PMID- 18158604 TI - Compressive optic neuropathy caused by a paranasal sinus cyst of Wegener's granulomatosis. PMID- 18158605 TI - Successful heart transplantation following neonatal necrotic enterovirus myocarditis. AB - Enterovirus myocarditis is a potentially devastating diagnosis in the neonatal setting, with an associated high mortality and risk for chronically impaired cardiac function. Transplantation may be a possible therapeutic option, but, due to the comparative scarcity of the presentation, the limited donor pool, and fears of viral persistence and recurrence of myocarditis (especially in the immunocompromised, post-transplant setting), there are no reported cases in the literature. This case report illustrates an encouraging one-year outcome following heart transplantation for necrotic enterovirus myocarditis in a neonate. PMID- 18158606 TI - Patient-activated antitachycardia pacing to terminate atrial tachycardias with 1:1 atrioventricular conduction in congenital heart disease. AB - A young girl with complex congenital heart disease and a Fontan procedure presented with frequent episodes of atrial tachycardias with 1:1 atrioventricular conduction. The patient underwent placement of a dual-chamber AT500 pacemaker with epicardial leads but because of the detection algorithm requiring > or = 2:1 AV relation, she received no antitachycardia therapy. Installation of special software AT500 TPARx v2.1 allowed use of patient-activated antitachycardia pacing for 1:1 atrial tachycardias. Since the installation of this application, the patient has successfully pace-terminated her tachycardia multiple times. PMID- 18158607 TI - Genioplasty and chin augmentation with Medpore implants: a report of 650 cases. AB - BACKGROUND: Genioplasty is an old and useful technique. Chin augmentation with Medpore has become popular in recent years. The authors briefly examine their experience with the two techniques. METHODS: From 2000 to date, 500 genioplasties and 150 chin augmentations with Medpore have been performed. The versatility and benefits of the two techniques are discussed. RESULTS: Most of the patients achieved satisfactory results with few complications. Genioplasty is recommended for severe developmental or complicated retrogenia and for deviated chin. Both techniques can be used for mild to modest retrogenia. Medpore is especially useful for patients who need a meticulous revision of the chin shape or contour. CONCLUSIONS: Chin shape and contour are as important as the chin position. Although genioplasty has the widest range of indications, chin augmentation with Medpore has the advantage of minimal risk for some patients who need to change the chin shape or contour. Medpore is totally different from other alloplastic chin implants such as silicone. PMID- 18158609 TI - Productivity of Thai Brahman and Simmental-Brahman crossbred (Kabinburi) cattle in central Thailand. AB - The productivity of the new crossbred cattle Kabinburi (K) was compared to that of Thai Brahman (TB) using 756 production records from K cattle and 1,316 production records from TB cattle kept at three locations in Thailand. The data were analyzed for the effect of breeds and locations. The ambient temperature, the humidity, the Temperature-Humidity Index (THI) and the rainfall of the three locations were different. Lamphayaklang Livestock Research and Breeding Center (LP) had the highest rainfall/year followed by Nongkwang Livestock Research and Breeding Center (NK), and Prachinburi Livestock Breeding Station (PC). Kabinburi cattle had a higher bodyweight at birth as well as at 200, 400 and 600 days of age than TB cattle. Furthermore, K heifers gave birth to their first calf at a younger age and had a shorter calving interval than TB cows. Thai Brahman cattle kept at LP had significantly higher bodyweight at 400 and 600 days than the animals kept at NK, but bodyweight at birth and 600 days of age were not significantly different. Thai Brahman cattle kept at LP were younger at first calving and had a shorter calving interval than the animals kept at NK. K cattle kept at NK were heavier at birth and at 200, 400 and 600 days of age than the animals kept at PC. Furthermore, Kabinburi cows kept at NK were younger at first calving (P<0.01), but the calving interval was not different between the two groups kept at NK or PC. PMID- 18158608 TI - RpoS regulation of gene expression during exponential growth of Escherichia coli K12. AB - RpoS is a major regulator of genes required for adaptation to stationary phase in E. coli. However, the exponential phase expression of some genes is affected by rpoS mutation, suggesting RpoS may also have an important physiological role in growing cells. To test this hypothesis, we examined the regulatory role of RpoS in exponential phase using both genomic and biochemical approaches. Microarray expression data revealed that, in the rpoS mutant, the expression of 268 genes was attenuated while the expression of 24 genes was enhanced. Genes responsible for carbon source transport (the mal operon for maltose), protein folding (dnaK and mopAB), and iron acquisition (fepBD, entCBA, fecI, and exbBD) were positively controlled by RpoS. The importance of RpoS-mediated control of iron acquisition was confirmed by cellular metal analysis which revealed that the intracellular iron content of wild type cells was two-fold higher than in rpoS mutant cells. Surprisingly, many previously identified RpoS stationary-phase dependent genes were not controlled by RpoS in exponential phase and several genes were RpoS regulated only in exponential phase, suggesting the involvement of other regulators. The expression of RpoS-dependent genes osmY, tnaA and malK was controlled by Crl, a transcriptional regulator that modulates RpoS activity. In summary, the identification of a group of exponential phase genes controlled by RpoS reveals a novel aspect of RpoS function. PMID- 18158610 TI - Cheyne stokes breathing at high altitude: a helpful response or a troublemaker? AB - Sleep disorders at high altitude are common and well-known for centuries. One symptom of the complex is periodic breathing (PB). PB occurs from a disbalance of the negative feedback loop of ventilation control, and at high altitude, it is increased by a phase shift of 180 degrees between hyperventilation and hypoxia. This paper explains the mechanisms that trigger the problem and discusses whether PB may be of advantage or disadvantage for the person going to high altitude. Up to about 3,000-3,500 m, PB may be of advantage because it stabilizes oxygen saturation at a relatively high level. At higher altitudes, disadvantages predominate because frequent arousals cause total sleep deprivation and mental and physical impairment of the victim. Correct acclimatization and "defensive" altitude profiles are gold standard, which minimize PB and optimizes recreative sleep, although they cannot mask PB completely, especially at extreme altitude. PMID- 18158611 TI - Studies on synchronous egress of coccidian parasites (Neospora caninum, Toxoplasma gondii, Eimeria bovis) from bovine endothelial host cells mediated by calcium ionophore A23187. AB - Neospora caninum, Toxoplasma gondii and Eimeria bovis are coccidian parasites of veterinary importance. Tachyzoites of N. caninum and T. gondii and sporozoites of E. bovis are able to invade and replicate in endothelial cells in vivo and in vitro. As it holds true for all eukaryotic cells, the survival of parasitized host cells and the parasites themselves should be dependent on ion balances, especially on extra- and intracellular calcium concentrations. Addition of the calcium ionophore A23187 reliably did release merozoites from mature N. caninum and T. gondii meronts grown in cultured primary bovine umbilical vein endothelial cells (BUVEC). Extent and time course of merozoite release depended on both, maturity of the meronts and concentration of the calcium ionophore. Attempts, however, to achieve synchronous release of merozoites from E. bovis first generation meronts by ionophore treatment failed, suggesting a different biological behaviour of this parasite. According to microscopical observations, the quite variable time of E. bovis macromeront maturation and a hampered merozoite exit owing to dense parasite-induced cytoskeleton elements surrounding the meront may be a reason for the lack of inducible synchronous release. PMID- 18158612 TI - Diagnostic accuracy of myocardial perfusion imaging with single photon emission computed tomography and positron emission tomography: a comparison with coronary angiography. AB - OBJECTIVE: The aim of this study was to compare the diagnostic accuracy of myocardial perfusion imaging (MPI) by positron emission tomography (PET) with the diagnostic accuracy of MPI by single photon emission computed tomography (SPECT) in two comparable patient cohorts, using coronary angiography (CA) as the standard of reference. METHODS: A "SPECT-group" of 80 patients (15 female, 65 male; mean age 60+/-9 years) and a "PET-group" of 70 patients (14 female, 56 male; mean age 57+/-10 years) underwent a one day stress/rest examination either with attenuation-corrected (13)N-ammonia PET or attenuation-corrected (201)TlCl SPECT or (99m)Tc-hexakis-methoxy-isobutyl-isonitril (MIBI) SPECT. PET and SPECT results were semiquantitatively graded using a 6-segment heart model. All patients underwent CA, and stenoses were graded as a diameter reduction >or=50%. RESULTS: Coronary findings between both groups did not significantly differ at CA. For the SPECT-group overall sensitivity and specificity for localisation of stenoses was 77% and 84%. Respective values for the PET-group were 97% and 84%. The specificity of MPI by SPECT in the detection of ischemia was 74% and 91% for MPI by PET. The diagnostic accuracy of MPI improves when the individual coronary dominance and previous coronary revascularisations are taken into account. CONCLUSION: MPI by (13)N-ammonia PET is more sensitive in the detection and localisation of coronary stenoses, and more specific in the detection of ischemia than MPI by (201)TlCl/(99m)MIBI SPECT. PMID- 18158613 TI - Analysis of OPLA scaffolds for bone engineering constructs using human jaw periosteal cells. AB - For bone regeneration constructs using human jaw periosteal cells (JPC) the extent of osteoinductive ability of different three-dimensional scaffolds is not yet established. We analyzed open-cell polylactic acid (OPLA) scaffolds for their suitability as bone engineering constructs using human JPC. Cell adhesion and spreading was visualized on the surface of scaffolds by scanning electron microscopy. JPC proliferation within OPLA scaffolds was compared with proliferation within collagen and calcium phosphate scaffolds. We found a significant increase of proliferation rates in OPLA scaffolds versus Coll/CaP scaffolds at three time points. Live-measurements of oxygen consumption within the cell-seeded scaffolds indicate that the in vitro culturing time should not exceed 12-15 days. OPLA scaffolds, which were turned out to be the most beneficial for JPC growth, were chosen for osteogenic differentiation experiments with or without BMP-2. Gene expression analyses demonstrated induction of several osteogenic genes (alkaline phosphatase, osterix, Runx-2 and insulin-like growth factor) within the 3D-scaffolds after 12 days of in vitro culturing. Element analysis by EDX spectrometry of arising nodules during osteogenesis demonstrated that JPC growing within OPLA scaffolds are able to form CaP particles. We conclude that OPLA scaffolds provide a promising environment for bone substitutes using human JPC. PMID- 18158614 TI - Biocompatible zwitterionic copolymer networks with controllable swelling and mechanical characteristics of their hydrogels. AB - The equilibrium swelling ratio in both water and physiological solution of the biocompatible copolymer networks of 3-dimethyl(methacryloyloxyethyl)ammonium propane sulfonate (DMAPS) and N-vinyl-2-pyrrolidone (NVP) is determined as a function of copolymer composition. It is established that equilibrium swelling ratio of the polymer networks in physiological solution increase with raise of zwitterionic monomer unit fraction. A sharp decrease of this ratio in water with increase of zwitterionic monomer unit fraction is related to the formation of thermolabile physical junctions produced by dipole-dipole interactions between the zwitterionic side groups. The same fact affects considerably the storage and loss moduli of the copolymer hydrogels as well as the morphology of the dried networks. Scanning electron microscopy images provided evidence of the occurrence of a lamellar structure forming the morphology of the polymers. This was corroborated by differential scanning calorimetry experiments. In this way a possibility for effective control on swelling ratio in different solutions and the mechanical properties of these novel biocompatible networks are established. PMID- 18158615 TI - Repair of goat tibial defects with bone marrow stromal cells and beta-tricalcium phosphate. AB - Tissue engineering techniques have been proven effective in bone regeneration and repairing load-bearing bone defects. Previous studies, however, have heretofore been limited to the use of slowdegradable or natural biomaterials as scaffolds. There are, however, no reports on using biodegradable, synthetic beta-tricalcium phosphate (beta-TCP) as scaffolds to repair weight-bearing bone defects in large animals. In the present study, highly porous beta-TCP scaffolds prepared by the polymeric sponge method were used to repair goat tibial defects. Fifteen goats were randomly assigned to one of three groups, and a 26 mm-long defect at the middle part of the right tibia in each goat was created. In Group A (six goats), a porous beta-TCP ceramic cylinder that had been loaded with osteogenically induced autologous bone marrow stromal cells (BMSCs) was implanted in the defect of each animal. In Group B (six goats), the same beta-TCP ceramic cylinder without any cells loaded was placed in the defect. In Group C (three goats), the defect was left untreated. In Group A, bony union can be observed by gross view, X-ray and micro-computed tomography (Micro-CT) detection, and histological observation at 32 weeks post-implantation. The implanted beta-TCP scaffolds were almost completely replaced by tissue-engineered bone. Bone mineral density in the repaired area of Group A was significantly higher (p < 0.05) than that of Group B, in which scant new bone was formed in each defect and the beta-TCP hadn't been completely resorbed at 32 weeks. Moreover, the tissue-engineered bone of Group A had similar biomechanical properties as that of the normal left tibia in terms of bending strength and Young's modulus (p > 0.05). In Group C, little or no new bone was formed, and non-union occurred, showing that the 26 mm segmental defect of the goat tibia was critical sized at 32 weeks. Thus, it can be concluded that the mechanical properties of the BMSCs/beta-TCP composites could be much improved via tissue engineering approach and beta-TCP might be used to repair the weight bearing segmental defects of goat tibias. PMID- 18158616 TI - Effects of carbon nanotubes (CNTs) on the processing and in-vitro degradation of poly(DL-lactide-co-glycolide)/CNT films. AB - Nanocomposite films based on single wall carbon nanotubes (SWNTs) and poly(DL lactide-co-glycolide) copolymer (50:50 PLGA) were processed and analyzed. The purpose of this study was to investigate the effect of different functionalization systems on the physical stability and morphology of PLGA films. Both covalent and non covalent functionalization of carbon nanotubes were considered in order to control the interactions between PLGA and SWNTs and to understand the role of the filler in the biodegradation properties. Using a solvent casting process, different PLGA/SWNT nanocomposites were prepared and incubated using organic solution under physiological conditions. In-vitro degradation studies were conducted by measurements of weight loss, infrared spectroscopy, glass transition temperature and SEM observations as a function of the incubation time, over a 9-week period. All PLGA films were degraded by hydrolitical degradation. However, a different degradation mechanism was observed in the case of functionalized SWNTs with respect to pristine material. It has been observed that system composition and SWNT functionalization may play a crucial role on the autocatalytic effect of the degradation process. These studies suggest that the degradation kinetics of the films can be engineered by varying carbon nanotube (CNT) content and functionalization. The combination of biodegradable polymers and CNTs opens a new perspective in the self-assembly of nanomaterials and nanodevices. PMID- 18158617 TI - Why the human penis is larger than in the great apes. PMID- 18158618 TI - Pain catastrophizing, physiological indexes, and chronic pain severity: tests of mediation and moderation models. AB - Catastrophizing about pain is related to elevated pain severity and poor adjustment among chronic pain patients, but few physiological mechanisms by which pain catastrophizing maintains and exacerbates pain have been explored. We hypothesized that resting levels of lower paraspinal muscle tension and/or lower paraspinal and cardiovascular reactivity to emotional arousal may: (a) mediate links between pain catastrophizing and chronic pain intensity; (b) moderate these links such that only patients described by certain combinations of pain catastrophizing and physiological indexes would report pronounced chronic pain. Chronic low back pain patients (N = 97) participated in anger recall and sadness recall interviews while lower paraspinal and trapezius EMG and systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate (HR) were recorded. Mediation models were not supported. However, pain catastrophizing significantly interacted with resting lower paraspinal muscle tension to predict pain severity such that high catastrophizers with high resting lower paraspinal tension reported the greatest pain. Pain catastrophizing also interacted with SBP, DBP and HR reactivity to affect pain such that high catastrophizers who showed low cardiovascular reactivity to the interviews reported the greatest pain. Results support a multi-variable profile approach to identifying pain catastrophizers at greatest risk for pain severity by virtue of resting muscle tension and cardiovascular stress function. PMID- 18158619 TI - Id-1 regulates Bcl-2 and Bax expression through p53 and NF-kappaB in MCF-7 breast cancer cells. AB - Although increasing evidence supports the protective role of inhibitor of differentiation and DNA binding-1 (Id-1) against anticancer drug-induced apoptosis, the underlying molecular mechanisms seem to vary depending on the tumor system. Here, we examined the direct role of Id-1 in MCF-7 breast cancer cells by ectopically overexpressing Id-1 under serum-free condition, where the endogenous Id-1 expression was suppressed. Id-1 expression resulted in increased number of viable cells, reduced Bax expression, enhanced Bcl-2 expression, but no change in Bcl-xL expression. The expression of nuclear factor-kappaB (NF-kappaB) was augmented, while those of p53 and IkappaB were reduced. Such changes in p53 and NF-kappaB pathways were also functional, as assessed by real-time polymerase chain reactions and reporter assays of their known downstream targets, p21 and Il 6, as well as Bax and Bcl-2 genes. Finally, Id-1 played a protective role against taxol-induced apoptosis in breast cancer cells as assessed by MTT assay and apoptotic cell count upon taxol treatment (0-200 nM). Reduced Bax expression and enhanced Bcl-2 expression by Id-1 were also noted in the presence of taxol. Taken together, we present a molecular mechanism where Id-1 regulates p53 and NF-kappaB pathways, which in turn regulates Bax and Bcl-2 genes, thus providing a survival advantage under exogenous stress such as serum-free or taxol treatment in MCF-7 breast cancer cells. In this regard, inactivation of Id-1 may provide a potential therapeutic strategy leading to inhibition of breast cancer progression and anti cancer drug resistance. PMID- 18158620 TI - Exemestane as primary systemic treatment for hormone receptor positive post menopausal breast cancer patients: a phase II trial of the Austrian Breast and Colorectal Cancer Study Group (ABCSG-17). AB - BACKGROUND: A multicenter phase II study was conducted to analyze the clinical activity of the steroidal aromatase inhibitor exemestane in the neoadjuvant treatment of post-menopausal women with strongly ER- and/or PgR- positive operable breast cancer. PATIENTS AND METHODS: From September 2000 to December 2003, 80 women were recruited for treatment with exemestane 25 mg once daily for 4 months. The primary end-point was the clinical response rate according the WHO criteria; the secondary end-points included toxicity and the number of patients who qualified for breast conserving surgery at the end of treatment, comparability of evaluation methods for response, potential alterations of hormone receptor and Her2/neu status during treatment. RESULTS: On an intention to evaluate analysis, according to the prespecified criteria the overall clinical objective response rate was 34%, the pCR rate was 3% and the rate of breast conserving surgery was 76%. When sonographic and mammographic longitudinal measurements were included in patients with missing palpation data, response rates were 38% and 41%, respectively. The tumor response was independent of the Her2/neu status which remained unchanged during treatment. In contrast, while the ER expression remained unaltered, downregulation of the PgR was observed. The treatment was well tolerated with no grade 3 and 4 toxicities except gastrointestinal (one grade 3 case) and hot flushes (two grade 3 cases). CONCLUSION: This study shows that exemestane is effective and safe as a preoperative therapy in post-menopausal patients with strongly hormone receptor positive breast cancer. PMID- 18158621 TI - Tissue and serum alpha 2-3- and alpha 2-6-linkage specific sialylation changes in oral carcinogenesis. AB - Increased sialylation of cell surface glycoconjugates is among the key molecular changes associated with malignant transformation and cancer progression. We investigated significance of linkage-specific sialylation changes in oral carcinogenesis. Tissue and serum levels of total sialic acid (TSA), linkage specific sialyltransferases (ST) and sialoproteins were analyzed from patients with oral precancerous conditions (OPC) and oral cancer as well as the post treatment follow-up blood samples of oral cancer patients. TSA levels were measured using a spectrophotometric method. The linkage-specific lectins, Sambusus nigra (SNA) and Maackia amurensis (MAM) detects alpha 2-6- and alpha 2-3 linked sialic acid, respectively, were used to analyze ST activity and sialoproteins. Malignant tissues showed significantly higher levels of TSA, reactivity of SNA and MAM, and alpha 2,3-ST activity compared to the adjacent normal tissues. alpha 2,6-ST was also higher in malignant tissues. Similarly, the marker levels were higher in precancerous tissues than their adjacent normal tissues. Serum levels of TSA, TSA/ total proteins, alpha 2-6-sialoproteins and alpha 2,6-ST were markedly increased in untreated oral cancer patients compared to the controls and OPC as well as responder (CR) patients. Serum levels of the markers were higher or comparable between untreated oral cancer patients and non responders (NR). Serum levels of alpha 2-3-sialylation were elevated in non responders compared with the responders. Further, the observed sialylation changes in tissue and serum were found to be associated with various clinicopathological features and disease progression. Thus, the data suggest potential utility of sialylation markers in early detection, prognostication and treatment monitoring of oral cancer. PMID- 18158622 TI - Introduction: aminopeptidases and hypertension-mechanisms of action and therapeutic strategies. PMID- 18158623 TI - Fibronectin-mediated activation of Akt2 protects human ovarian and breast cancer cells from docetaxel-induced apoptosis via inhibition of the p38 pathway. AB - Although multiple mechanisms have been implicated in chemoresistance, recent evidence has suggested that the attachment of cells to extracellular matrix proteins such as fibronectin (FN) may mediate the signals that participate in cell survival and resistance to apoptosis. We established previously that human ovarian cancer cells and breast cancer cells adhering to FN acquire a survival advantage through activation of the PI3-kinase/Akt2 pathway. However, the mechanism by which Akt2 regulates chemoresistance in adherent cells is unknown. In the present study, we have investigated the role of the interaction between the Akt2/survivin survivial pathway and the ASK1/p38 apoptotic pathway in the phenomenon of resistance to docetaxel. We show here that the resistance of FN adhered A2780 or MDA-MB-231 cells to docetaxel requires survivin, and we present evidence that attenuation of the antiapoptotic activity of survivin is p38 dependent. The activation of p38 kinase in response to docetaxel, on the other hand, is abolished by FN adhesion. We further demonstrate that FN adhesion mediated inhibition of p38 activation was governed by Akt2 via the promotion of direct protein association of ASK1 with p38. Our results indicate for the first time that p38 plays a critical role in FN adhesion-mediated resistance to docetaxel. The present findings may help us to understand the formation of FN adhesion-mediated chemoresistance and facilitate development of novel antineoplastic strategies. PMID- 18158624 TI - Depression in public community long-term care: implications for intervention development. AB - The objective of this paper is to increase understanding of geriatric depression in the public community long-term care system to guide intervention development. Protocols included screening 1,170 new clients of a public community long-term care agency and interviewing all clients with major, dysthymia, or subthreshold depression (n = 299) and a randomly selected subset of nondepressed older adults (n = 315) at baseline, 6-month, and 1 year. Six percent had major depression, one half of a percent had dysthymia only, and another 19% had subthreshold depression. Over the year observation period, 40% were persistently depressed; 32% were assessed as depressed only at the first observation; and the remainder was intermittently depressed. There were high levels of comorbid medical, functional, and psychosocial conditions. Mental health service use was low, and clients reported attitudinal and other barriers to depression treatment. Findings suggest the need for universal screening for depression with some strategies for triaging the most severely and persistently depressed for treatment. Although there will be challenges to the development of depression interventions, the public community long-term care system has high potential to assist vulnerable older adults receive help with depression. PMID- 18158625 TI - Oral contraceptives: 40 years after release. PMID- 18158626 TI - Factors influencing pregnancy outcome in women with type 2 versus type 1 diabetes mellitus. AB - AIM: To compare pregnancy outcome and factors related to adverse perinatal outcome in women with type 1 versus type 2 diabetes mellitus (DM). MATERIAL AND METHODS: Multicentre retrospective study. Some 404 women were studied, 257 with type 1 DM and 147 with type 2 DM. Main outcome measures were rates of prematurity, macrosomia, instrumental deliveries, congenital malformations, need for neonatal intensive care unit (NICU), and perinatal mortality. RESULTS: There were no significant differences in pregnancy outcome between women with type 1 and type 2 DM, except for an increased rate of instrumental deliveries in women with type 1 DM. In these women, duration of diabetes was a significant predictor of caesarean delivery (OR =1.06 (1.01-1.12); p=0.02). Chronic hypertension was positively related to prematurity (p=0.02), and proved to be a significant predictor of birth weight lower than 2,500 g (OR =3.54 (1.4-12.49); p=0.043) and perinatal mortality (OR =10.6 (1.15-117.6); p=0.04). In women with type 2 DM, third trimester higher glycosylated haemoglobin was related to both prematurity (OR 4.9 (1.7-14.4; p=0.004) and low birth weight. Macrosomia was a significant risk factor for caesarean section. First trimester glycosylated haemoglobin was related to congenital malformations and proved to be a significant predictor of perinatal mortality (OR =2.4 (1.02-5.74); p=0.04). CONCLUSIONS: Duration of DM and chronic hypertension were the most influential factors related to adverse perinatal outcomes in women with type 1 DM, and poor metabolic control and macrosomia in women with type 2 DM. PMID- 18158627 TI - Group B streptococcal carriage in Sweden: a national study on risk factors for mother and infant colonisation. AB - BACKGROUND: To study group B streptococcus (GBS) colonisation in parturients and infants in relation to obstetric outcome and to define serotypes and antibiotic resistance in GBS isolates acquired. METHODS: A population-based, national cohort of parturients and their infants was investigated. During 1 calendar week in 2005 all women giving birth (n=1,754) were requested to participate in the study. RESULTS: A total of 1,569 mother/infant pairs with obstetric and bacteriological data were obtained. Maternal carriage rate was 25.4% (95% confidence interval (CI): 23.3-27.6). In GBS-positive mothers with vaginal delivery and no intrapartum antibiotics, the infant colonisation rate was 68%. Some 30% of infants were colonised after acute caesarean section, and 0% were colonised after an elective procedure. Duration of transport of maternal recto/vaginal swabs of more than 1 day impeded culture sensitivity. Infant mMales were more frequently colonised than females (76.9 versus 59.8%, odds ratio (OR): 2.16; 95% CI: 1.27 3.70), as were infants born after rupture of membranes > or =24 h (p =0.039). Gestational age, birth weight and duration of labor did not significantly influence infant colonisation. Some 30% of parturients with at least one risk factor for neonatal disease received intrapartum antibiotics. The most common GBS serotypes were type III and V. Some 5% of the isolates were resistant to clindamycin and erythromycin, respectively. CONCLUSIONS: Maternal GBS prevalence and infant transfer rate were high in Sweden. Males were more frequently colonised than females. The sensitivity of maternal cultures decreased with the duration of sample transport. Clindamycin resistance was scarce. The use of intrapartum antibiotics was limited in parturients with obstetric risk factors for early onset group B streptococcal disease. PMID- 18158628 TI - Adverse pregnancy outcome in women with mild glucose intolerance: is there a clinically meaningful threshold value for glucose? AB - BACKGROUND: The diagnostic criteria of gestational diabetes mellitus (GDM) have been based on the risk of future maternal diabetes rather than the short-term risk of mother and infant. Our aim was to illustrate the relation between various adverse pregnancy outcomes and maternal glucose levels in women with mild glucose intolerance using a graphical approach. METHODS: Observational study of 2,885 pregnant women examined with a 2-h, 75-g oral glucose tolerance test (OGTT) based on risk indicators. Only women with 2-h capillary blood glucose <9.0 mmol/l were included, as women with 2-h values > or =9.0 mmol/l were treated for GDM. Empirical frequencies of adverse outcomes were related to 2-h values by linear and quadratic logistic models. Adjustments for well-known confounders were performed by a multiple logistic model. RESULTS: Linear trends were demonstrated for the outcomes: shoulder dystocia, caesarean section rate (univariate analysis only), spontaneous preterm delivery, and macrosomia (large-for-gestational age infants). None of the outcomes deviated significantly from linearity. No significant trend was found for hypertension or neonatal hypoglycaemia and jaundice. CONCLUSIONS: A gradually increasing risk for a number of adverse pregnancy outcomes was found with increasing glucose levels. No obvious threshold value for GDM was demonstrated for 2-h values up to 9.0 mmol/l. PMID- 18158629 TI - Preinduction cervical ripening with prostaglandin E2 at preterm. AB - OBJECTIVE: To evaluate the efficacy of prostaglandin E2 (PGE2) for cervical ripening at preterm and to identify factors predicting ripening failure. DESIGN: A retrospective study of women admitted for preinduction cervical ripening with PGE2 vaginal tablets at preterm (n=444). PGE2 vaginal tablets were administered at 6-8 h intervals until cervical ripening was achieved (Bishop score > or =7) or ripening failed (Bishop score <7 after administration of 4 PGE2 tablets). Data were compared with a control group of women admitted for preinduction cervical ripening with PGE2 vaginal tablets at term (n=1,029). A multivariate logistic regression analysis was performed to detect factors predicting ripening failure. RESULTS: Women at preterm required a significantly higher number of PGE2 tablets compared to women at term (2.0+/-1.1 versus 1.6+/-0.9, p<0.001). Overall rates of ripening failure and caesarean section due to failed induction were significantly higher in preterm than term gestations (11.4 versus 1.1% and 9.7 versus 0.4%, p <0.001), and were inversely related to gestational age. Nulliparity, lower gestational age at delivery, lack of cervical dilatation, and higher maternal age were significant predictors of ripening failure at preterm. CONCLUSION: The use of PGE2 for preinduction cervical ripening at preterm may be associated with an increased risk of ripening failure and caesarean section compared with term gestations. This information may be useful when consulting women regarding the available options when premature delivery is necessary. PMID- 18158630 TI - Early maternal feeding following caesarean delivery: a prospective randomised study. AB - OBJECTIVE: To evaluate the effect of early maternal feeding on maternal satisfaction and gastrointestinal complications on patients following simple and complicated caesarean delivery (CD). METHODS: A prospective, randomised study was designed including 179 women undergoing first or repeated CD. Women who were assigned to the early feeding group received clear fluids and solid food within 8 h of surgery, in accordance with their request. The traditionally fed women received clear fluids 8-12 h after the surgery subsequent to physician examination confirming bowel sounds. RESULTS: Some 82 women were assigned to the traditionally fed group and 97 women were assigned to the early fed group. Maternal satisfaction was significantly higher among the early fed women. No significant differences were noted between the groups regarding post-operative complications. CONCLUSIONS: Early maternal feeding following CD is associated with higher maternal satisfaction. However, it is not associated with higher rates of post-operative complications. PMID- 18158631 TI - Analysis of 127 peripartum hypoxic brain injuries from closed claims registered by the Danish Patient Insurance Association. AB - BACKGROUND: One of the most feared complications in medicine is hypoxic brain damage to a newborn. The authors investigated the circumstances of registered peripartum hypoxic brain injuries in order to identify potential opportunities to improve patient safety and prevent injuries. METHODS: The authors retrospectively investigated peripartum hypoxic brain injuries registered by the Danish Patient Insurance Association. RESULTS: From 1992 to 2004, 127 approved claims concerning peripartum hypoxic brain injuries were registered and subsequently analysed. Thirty-eight newborns died, and a majority of the 89 surviving children suffered from major handicaps, primarily cerebral palsy. In 69 of the cases, misinterpretation of or late action on an abnormal cardiotocography (CTG) were the reasons for the majority of the hypoxic brain injuries. CONCLUSIONS: All injuries could potentially have been avoided using established obstetric practice. CTGs are often misinterpreted. In the authors' opinion, education and training in CTG interpretation is essential. The use of ST-analysis of the fetus ECG (STAN) could probably reduce the number of these injuries. PMID- 18158632 TI - What is the recurrence rate of postmenopausal bleeding in women who have a thin endometrium during a first episode of postmenopausal bleeding? AB - OBJECTIVE: To determine the incidence and significance of recurrent postmenopausal bleeding among women diagnosed with an endometrial thickness < or =4 mm after a first episode of postmenopausal bleeding. METHODS: Consecutive patients not using hormone replacement therapy (HRT) presenting with a first episode of postmenopausal bleeding and an endometrial thickness < or =4 mm at transvaginal ultrasonography (TVU) were managed expectantly. In case of recurrent bleeding, the patient was evaluated according to the hospital's local policy with TVU, office endometrial sampling, hysteroscopy or dilatation and curettage (D&C) or a combination of these tests. We evaluated the incidence of recurrent bleeding, potential risk factors for recurrent bleeding, and the diagnosis made after recurrent bleeding. RESULTS: A total of 607 patients were registered with a first episode of postmenopausal bleeding, of whom 249 had an endometrial thickness < or =4 mm. Follow-up took place with a median of 174 weeks (range: 4 250 weeks). During follow-up, 25 of the 249 patients (10%; 95% CI: 6.6-14%) had recurrent bleeding. Median time until recurrence of bleeding was 49 weeks (range: 9-186 weeks). Two patients with recurrent bleeding turned out to have an endometrial carcinoma (8%; 95% CI: 2.2-25%), and 1 patient had a malignant melanoma. Time since menopause, age, body mass index, hypertension, diabetes and anticoagulants were not predictive for recurrent bleeding. CONCLUSION: The recurrence rate after a first episode of postmenopausal bleeding managed expectantly is low and cannot be predicted by patient characteristics. Patients with recurrent bleeding should be re-evaluated, as they bear a considerable risk of carcinoma. PMID- 18158633 TI - New attempt using labio-vestibular flap technique to manage circumcised women with Rokitansky syndrome. AB - OBJECTIVE: To assess the efficacy of the labio-vestibular flap technique in managing circumcised women with Rokitansky syndrome and in correcting the sexuality defects caused by the anomaly present and by female circumcision. DESIGN: A prospective, comparative, clinical study. SETTING: Kasr El Aini School of Medicine, Cairo University, Egypt. PATIENTS: Thirty-four circumcised patients with Rokitansky syndrome were divided into 2 groups; Group 1 consisted of 22 cases treated with the labio-vestibular flap technique, and Group 2 consisted of 12 cases treated with McIndoe's technique. MAIN OUTCOME MEASURES: Pre and postoperative clinical assessment of the newly formed vagina and sexuality. The new flap is formed of the remaining parts of the labia minora and the adjoining parts of the vestibule, and is used to cover the posterior and lateral walls. Meanwhile, the anterior wall is covered by the epithelium of the original blind vaginal pouch. The modified Kasr El Aini sexual assessment sheet assesses sexuality. RESULTS: Rokitansky syndrome could be classified into 4 clinical types according to the degree of development of the uterus. The labio-vestibular technique was simple and not associated with any graft rejection, hair growth or contraction of the reformed tract. Dyspareunia and marital failure were less significantly recorded in the labio-vestibular technique than in McIndoe's technique. But, the most significant results were the improvement in sexuality, in addition to restoration of genital continuation, menstruation and fertility in some cases. CONCLUSION: The labio-vestibular flab technique is the most simple and most suitable line of treatment for circumcised cases with Rokitansky syndrome. In these cases, the technique achieved good results in correcting sexual defects after circumcision. PMID- 18158634 TI - Does box model training improve surgical dexterity and economy of movement during virtual reality laparoscopy? A randomised trial. AB - OBJECTIVE: Laparoscopic box model trainers have been used in training curricula for a long time, however data on their impact on skills acquisition is still limited. Our aim was to validate a low cost box model trainer as a tool for the training of skills relevant to laparoscopic surgery. DESIGN: Randomised, controlled trial (Canadian Task Force Classification I). SETTING: University Hospital. MEASUREMENTS AND MAIN RESULTS: Sixteen gynaecologic residents with limited laparoscopic experience were randomised to a group that received a structured box model training curriculum, and a control group. Performance before and after the training was assessed in a virtual reality laparoscopic trainer (LapSim and was based on objective parameters, registered by the computer system (time, error, and economy of motion scores). Group A showed significantly greater improvement in all performance parameters compared with the control group: economy of movement (p=0.001), time (p=0.001) and tissue damage (p=0.036), confirming the positive impact of box-trainer curriculum on laparoscopic skills acquisition. CONCLUSIONS: Structured laparoscopic skill training on a low cost box model trainer improves performance as assessed using the VR system. Trainees who used the box model trainer showed significant improvement compared to the control group. Box model trainers are valid tools for laparoscopic skills training and should be implemented in the comprehensive training curricula in gynaecology. PMID- 18158635 TI - Endometrial thermal balloon ablation has a beneficial long-term effect on menorrhagia. AB - BACKGROUND: More data on the long-term results of thermal balloon endometrial ablation are needed. METHODS: A retrospective observational study of 190 women who underwent endometrial thermal balloon ablation in the treatment of menorrhagia. The mean follow-up period was 6 years. RESULTS: Of the 172 women analysed after exclusion, 28 (16%) had a hysterectomy during follow-up. Women with regular menstrual periods had the best outcome. Some 152 (89%) women filled in a questionnaire concerning their satisfaction with the procedure, and 76% were satisfied. Amenorrhoea was reported by 14% and eumenorrhoea by 54% of the women who completed the questionnaire. CONCLUSION: Endometrial thermal balloon ablation has good long-term efficacy and can be considered an applicable alternative in the treatment of menorrhagia, especially for women who have regular periods and are over 40 years of age. PMID- 18158636 TI - Decreased endopelvic fascia elastin content in uterine prolapse. AB - BACKGROUND: Genital prolapse is a debilitating manifestation of pelvic floor dysfunction. The cause of this condition has not been elucidated. The purpose of this study was to determine elastin content and RNA expression of related enzymes of elastin synthesis in uterosacral ligament biopsies from women with severe prolapse, and controls with normal pelvic support. METHODS: Biopsies were taken from the uterosacral ligament tissue of 31 women with Grade III or greater prolapse and 29 women with normal pelvic support. Elastin content was assessed by measuring desmosine using radioimmunoassay, and quantitative real time PCR was performed to quantify mRNA levels of lysyl oxidase (LOX), lysyl oxidase like-1 (LOXL1), LOXL2 and fibulin-5 (FIB-5). RESULTS: The mean desmosine concentration found in uterosacral ligaments of women with prolapse (n =26) was 103.3+/-59.3 pmolD/mgP compared to controls (n =29) 120.5+/-47.4 pmolD/mgP (p =0.1943). In the subgroup of subjects with complete procidentia (n =8), mean desmosine concentration was 50.6+/-25.8 and 127.1+/-42.2 pmolD/mgP in age-matched controls (n =12) (p <0.05). In tissue from subjects with more than 2 vaginal deliveries (n =18), the mean desmosine concentration was 99.9+/-60.7 and 133.0+/-44.0 pmolD/mgP in controls (n =17) (p <0.05). Expression of LOX, LOXL1 and LOXL2 decreased 8.2 fold+/-3.4, 5.0-fold+/-1.7 and 15.2-fold+/-5.2, respectively (mean+/-SD) in cases versus controls (p<0.05). Expression of FIB-5 was increased 3.1-fold+/-0.7 compared to controls (p<0.05). CONCLUSIONS: Significantly decreased desmosine content was measured in the uterosacral ligament tissue from women with prolapse versus controls in women with parity >2 and in women with complete procidentia. Suppression of mRNA for LOX and two LOX isoenzymes was correspondingly present. These results suggest that altered elastin metabolism is present in women with uterine prolapse. PMID- 18158637 TI - Ultrasonographic assessment of tape location following tension-free vaginal tape and transobturator tape procedure. AB - BACKGROUND: The purpose of this study was to perform an ultrasonographic comparison of tension-free vaginal tape (TVT) and the TVT-obturator (TVT-O) procedure in women with stress urinary incontinence (SUI). METHODS: We retrospectively reviewed 159 women with SUI undergoing either TVT (n=91) or TVT-O (n=68). All subjects underwent perineal ultrasonographies, pelvic examination, urinalyses, 1-h pad tests, multichannel urodynamic studies and a personal interview using the Bristol Female Lower Urinary Tract Symptoms Questionnaire before and 1 year after surgery. RESULTS: At rest or during Valsalva, the middle of the TVT-O tape localised more distally than the TVT on ultrasound (p<0.01). A higher rate of urethral kinking during straining was observed in the TVT group compared with the TVT-O group after surgery (86.9 versus 23.9%, p<0.01). Mean operative time was significantly shorter in the TVT-O group (16.2 versus 28.6 min, p<0.01). The rate of objective cure was comparable for the TVT and TVT-O groups (94.5 versus 88.2%, p<0.05). However, subjective cure rate was significantly lower for the TVT-O than the TVT group (82.4 versus 93.4%, p=0.042). There was no statistical difference between the 2 groups regarding patient characteristics and complication rates. CONCLUSIONS: TVT-O tape results in a less acute angle and localises to a more distal part of the urethra, resulting in less urethral compression and a lower rate of urethral dynamic kinking. PMID- 18158641 TI - Middle ear gas exchange via the mucosa: estimation by hyperventilation. AB - CONCLUSION: It is thought that gas exchange via the mucosa occurred in relation to the partial pressure gradient, and it was impaired mainly by inflammatory changes in the mastoid mucosa. It was verified that gas exchange via the mucosa is less likely to be impaired than gas exchange via the eustachian tube. OBJECTIVES: To evaluate the capacity of middle ear gas exchange via the mucosa by examining the effect of hyperventilation on middle ear pressure. SUBJECTS AND METHODS: A total of 55 patients, 40 patients with a type A tympanogram and 15 with type C, were selected. Tympanometry was performed in one ear every 2 min while hyperventilation was forcibly continued for 44-6 min in the supine position. The middle ear pressure and the pressure of end-tidal carbon dioxide (PETCO2) was measured, and sonotubometry was performed. RESULTS: PETCO2 decreased gradually as hyperventilation continued in all cases. Although middle ear pressure decreased by hyperventilation in 49 of 55 patients, in 6 patients it hardly decreased despite the decrease in PETCO2. These six patients were treated for otitis media with effusion within 1 month before this examination. PMID- 18158642 TI - Regulation of NADPH oxidase subunit p22(phox) by NF-kB in human aortic smooth muscle cells. AB - Accumulating evidence demonstrates the involvement of oxidative stress in the pathophysiology of cardiovascular diseases. The molecular mechanisms accountable for the increased production of reactive oxygen species remain uncertain. Among others, NADPH oxidase is one of the most important sources of superoxide in vascular cells. Here we investigate the role of NF-kB in the regulation of p22(phox) subunit and NADPH oxidase activity, in human aortic smooth muscle cells. Overexpression of p65/RelA or IKKbeta up-regulated p22(phox) gene promoter activity. Transcription factor pull-down assays demonstrated the physical interaction of p65/RelA protein with predicted NF-kB binding sites. Real time PCR and Western blotting analysis showed that p22(phox) mRNA and protein expression are significantly down-regulated by NF-kB decoy oligodeoxynucleotides and N-alpha tosyl-l-phenylalanine chloromethyl ketone (TPCK). Lucigenin-enhanced chemiluminescence assay revealed that NF-kB inhibitors reduce the NADPH-dependent superoxide production. Regulation of NADPH oxidase by NF-kB may represent a possible mechanism whereby pro-inflammatory factors induce oxidative stress in atherosclerosis, hypertension, diabetes, stroke or heart failure. PMID- 18158643 TI - Dynamics of insulin signalling in liver during hyperinsulinemic euglycaemic clamp conditions in vivo and the effects of high-fat feeding in male mice. AB - Insulin is an important regulator of hepatic carbohydrate, lipid, and protein metabolism, and the regulation of these processes by insulin is disturbed under conditions of insulin resistance and type 2 diabetes. Despite these alterations, the impact of insulin resistance on insulin signalling in the liver is not well defined. Variations in time and dose of insulin stimulation as well as plasma glucose levels may underlie this. The present study aimed at determining the dynamics of activation of hepatic insulin signalling in vivo at insulin concentrations resembling those achieved after a meal, and addressing the effects of high-fat feeding. An unexpected finding of this study was the biphasic activation pattern of the IRS-PI3K-PKB/Akt pathway. Our findings indicate that the first burst of activation contributes to regulation of glucose metabolism. The physiological function of the second peak is still unknown, but may involve regulation of protein synthesis. Finally, high-fat feeding caused hepatic insulin resistance, as illustrated by a reduced suppression of hepatic glucose production. A sustained increased phosphorylation of the serine/threonine kinases p70S6kinase and Jun N-terminal kinase in the absence of insulin may underlie the abrogated phosphorylation of the IRS proteins and their downstream targets. PMID- 18158644 TI - Rosiglitazone treatment improves cardiac efficiency in hearts from diabetic mice. AB - Isolated perfused hearts from type 2 diabetic (db/db) mice show impaired ventricular function, as well as altered cardiac metabolism. Assessment of the relationship between myocardial oxygen consumption (MVO(2)) and ventricular pressure-volume area (PVA) has also demonstrated reduced cardiac efficiency in db/db hearts. We hypothesized that lowering the plasma fatty acid supply and subsequent normalization of altered cardiac metabolism by chronic treatment with a peroxisome proliferator-activated receptor-gamma (PPARgamma) agonist will improve cardiac efficiency in db/db hearts. Rosiglitazone (23 mg/kg body weight/day) was administered as a food admixture to db/db mice for five weeks. Ventricular function and PVA were assessed using a miniaturized (1.4 Fr) pressure volume catheter; MVO(2) was measured using a fibre-optic oxygen sensor. Chronic rosiglitazone treatment of db/db mice normalized plasma glucose and lipid concentrations, restored rates of cardiac glucose and fatty acid oxidation, and improved cardiac efficiency. The improved cardiac efficiency was due to a significant decrease in unloaded MVO(2), while contractile efficiency was unchanged. Rosiglitazone treatment also improved functional recovery after low flow ischemia. In conclusion, the present study demonstrates that in vivo PPARgamma-treatment restores cardiac efficiency and improves ventricular function in perfused hearts from type 2 diabetic mice. PMID- 18158645 TI - Essential role of p38 MAPK for activation of skeletal muscle glucose transport by lithium. AB - Lithium increases glucose transport and glycogen synthesis in insulin-sensitive cell lines and rat skeletal muscle, and has been used as a non-selective inhibitor of glycogen synthase kinase-3 (GSK-3). However, the molecular mechanisms underlying lithium action on glucose transport in mammalian skeletal muscle are unknown. Therefore, we examined the effects of lithium on glucose transport activity, glycogen synthesis, insulin signaling elements (insulin receptor (IR), Akt, and GSK-3beta), and the stress-activated p38 mitogen activated protein kinase (p38 MAPK) in the absence or presence of insulin in isolated soleus muscle from lean Zucker rats. Lithium (10 mM LiCl) enhanced basal glucose transport by 62% (p < 0.05) and augmented net glycogen synthesis by 112% (p < 0.05). Whereas lithium did not affect basal IR tyrosine phosphorylation or Akt ser(473) phosphorylation, it did enhance (41%, p < 0.05) basal GSK-3beta ser(9) phosphorylation. Lithium further enhanced (p < 0.05) the stimulatory effects of insulin on glucose transport (43%), glycogen synthesis (44%), and GSK 3beta ser(9) phosphorylation (13%). Lithium increased (p < 0.05) p38 MAPK phosphorylation both in the absence (37%) and presence (41%) of insulin. Importantly, selective inhibition of p38 MAPK (using 10 microM A304000) completely prevented the basal activation of glucose transport by lithium, and also significantly reduced (52%, p < 0.05) the lithium-induced enhancement of insulin-stimulated glucose transport. Theses results demonstrate that lithium enhances basal and insulin-stimulated glucose transport activity and glycogen synthesis in insulin-sensitive rat skeletal muscle, and that these effects are associated with a significant enhancement of GSK-3beta phosphorylation. Importantly, we have documented an essential role of p38 MAPK phosphorylation in the action lithium on the glucose transport system in isolated mammalian skeletal muscle. PMID- 18158646 TI - The central role of glutathione in the pathophysiology of human diseases. AB - Reduced glutathione (L-gamma-glutamyl-L-cysteinyl-glycine, GSH) is the prevalent low-molecular-weight thiol in mammalian cells. It is formed in a two-step enzymatic process including, first, the formation of gamma-glutamylcysteine from glutamate and cysteine, by the activity of the gamma-glutamylcysteine synthetase; and second, the formation of GSH by the activity of GSH synthetase which uses gamma-glutamylcysteine and glycine as substrates. While its synthesis and metabolism occur intracellularly, its catabolism occurs extracellularly by a series of enzymatic and plasma membrane transport steps. Glutathione metabolism and transport participates in many cellular reactions including: antioxidant defense of the cell, drug detoxification and cell signaling (involved in the regulation of gene expression, apoptosis and cell proliferation). Alterations in its concentration have also been demonstrated to be a common feature of many pathological conditions including diabetes, cancer, AIDS, neurodegenerative and liver diseases. Additionally, GSH catabolism has been recently reported to modulate redox-sensitive components of signal transduction cascades. In this manuscript, we review the current state of knowledge on the role of GSH in the pathogenesis of human diseases with the aim to underscore its relevance in translational research for future therapeutic treatment design. PMID- 18158647 TI - The roles of hyperglycaemia and oxidative stress in the rise and collapse of the natural protective mechanism against vascular endothelial cell dysfunction in diabetes. AB - Vascular endothelial cell (VEC) dysfunction in diabetes has been associated with hyperglycaemia-induced intra- and extracellular glycation of proteins and to overproduction of glucose-derived free radicals. VEC protect their intracellular environment against an increased influx of glucose in face of hyperglycaemia by reducing the expression and plasma membrane abundance of their glucose transporter-1 (GLUT-1). We investigated the hypothesis that glucose-derived free radicals induce this down-regulatory mechanism in VEC, but proved the contrary. In fact, pro-oxidants significantly increased the expression and plasma membrane abundance of GLUT-1 and the rate of glucose transport in VEC while abolishing high-glucose-induced down-regulation of the hexose transport system. The resulting uncontrolled influx of glucose followed by overproduction of glucose derived ROS further up-regulates the rate of glucose transport, and vice versa. This perpetuating glycoxidative stress finally leads to the collapse of the auto regulatory protective mechanism and accelerates the development of dysfunctional endothelium in blood vessels. PMID- 18158648 TI - 2007 Donald E. Cummings Memorial Award Lecture. The industrial hygiene profession: current and future issues. PMID- 18158649 TI - Games as teaching tools in a surgical residency. AB - BACKGROUND: Didactic lectures have been the mainstay of core teaching in the surgical residency program at our school. Our concerns about the educational impact of these passive activities led us to consider more interactive teaching approaches. METHODS: We developed an interactive games-based approach to learning. One set of games was labeled "Who wants to be a Surgeon" (WS) and the other was called "Senior Face-off" (SF). We evaluated the impact of this innovation using an end-of-year questionnaire. RESULTS: Enjoyment, teaching quality and preference over lectures were high for both games. However, the WS sparked interest significantly more in junior residents (4.3 +/- 0.21 vs 3.3 +/- 0.31, p = 0.015) and senior residents found both games more stressful than did junior residents (WS: 2.88 +/- 0.32 vs 2.00 +/- 0.21, p = 0.038, and SF: 3.54 +/- 0.29 vs 1.80 +/- 0.33, p = 0.001). CONCLUSIONS: This innovative teaching technique promoted learner interest and was regarded as a worthwhile educational activity. Games with a competitive emphasis may unduly stress senior residents. PMID- 18158650 TI - Computer-based testing of the modified essay question: the Singapore experience. AB - BACKGROUND: The modified essay question (MEQ), featuring an evolving case scenario, tests a candidate's problem-solving and reasoning ability, rather than mere factual recall. Although it is traditionally conducted as a pen-and-paper examination, our university has run the MEQ using computer-based testing (CBT) since 2003. AIMS: We describe our experience with running the MEQ examination using the IVLE, or integrated virtual learning environment (https://ivle.nus.edu.sg), provide a blueprint for universities intending to conduct computer-based testing of the MEQ, and detail how our MEQ examination has evolved since its inception. METHODS: An MEQ committee, comprising specialists in key disciplines from the departments of Medicine and Paediatrics, was formed. We utilized the IVLE, developed for our university in 1998, as the online platform on which we ran the MEQ. We calculated the number of man-hours (academic and support staff) required to run the MEQ examination, using either a computer-based or pen-and-paper format. RESULTS: With the support of our university's information technology (IT) specialists, we have successfully run the MEQ examination online, twice a year, since 2003. Initially, we conducted the examination with short-answer questions only, but have since expanded the MEQ examination to include multiple-choice and extended matching questions. A total of 1268 man-hours was spent in preparing for, and running, the MEQ examination using CBT, compared to 236.5 man-hours to run it using a pen-and-paper format. Despite being more labour-intensive, our students and staff prefer CBT to the pen and-paper format. CONCLUSIONS: The MEQ can be conducted using a computer-based testing scenario, which offers several advantages over a pen-and-paper format. We hope to increase the number of questions and incorporate audio and video files, featuring clinical vignettes, to the MEQ examination in the near future. PMID- 18158651 TI - The profession of medicine: a joint US-German collaborative project in medical education. AB - BACKGROUND: International collaborations between medical institutions occur frequently in research and clinical training, but less often in undergraduate medical education. Collaborative programs in psychosocial, ethical and cultural topics are rare. "The Profession of Medicine", an elective undergraduate course based on the "Patient, Physician and Society" curriculum at Northwestern University's Feinberg School of Medicine in Chicago, USA, was introduced in 2005 at the Charite-Universitatsmedizin Berlin, Germany, in order to provide students with a learning opportunity in this field and to introduce an international context to student education. METHODS: The five-day course is offered to preclinical medical students twice a year and includes topics such as cultural diversity, end of life issues, mistakes in medicine, vulnerable populations, and interactions with the pharmaceutical industry. The course language is English, in a format consisting of both plenary and small group sessions. To date, one-fifth of the instructors have been from the U.S. institution. Educational methods include small group discussions, presentations by guest speakers, movie and video clips, role plays, and reading and writing assignments. RESULTS: The participants have evaluated the course very favourably, with average scores ranging from 1.2 to 1.5 (1 = best/agree the most and 5 = worst/agree the least). Pre- and post course self-assessment with regard to knowledge, interest and professional attitude revealed a statistically significant increase for all course topics. CONCLUSIONS: In sum, the integration of Northwestern University's Feinberg School of Medicine's "Patient, Physician and Society" course concept into the Charite - Universitatsmedizin Berlin has been an effective method to develop students' professional skills and to stimulate international educational collaboration. PMID- 18158652 TI - Development of a leadership skills workshop in paediatric advanced resuscitation. AB - BACKGROUND: Paediatric residency programs rarely prepare trainees to assume resuscitation team leadership roles despite the recognized need for these skills by specialty accreditation organizations. We conducted a needs-assessment survey of all residents in the McGill Pediatric Residency Program, which demonstrated that most residents had minimal or no experience at leading resuscitation events and felt unprepared to assume this role in the future. AIMS: We developed an educational intervention (workshop) and evaluated immediate and long term learning outcomes in order to determine whether residents could acquire and retain team leadership skills in pediatric advanced resuscitation. METHODS: Fifteen paediatric residents participated in a workshop that we developed to fulfill the learning needs highlighted with the needs assessment, as well as the Objectives of Training in Pediatrics from the Royal College of Physicians and Surgeons of Canada. It consisted of a plenary session followed by 2 simulated resuscitation scenarios. Team performance was evaluated by checklist. Residents were evaluated again 6 months later without prior interactive lecture. Learning was also assessed by self-reported retrospective pre/post questionnaire. RESULTS: Checklist score (assigning roles, limitations of team, communication, overall team atmosphere) expressed as % correct: initial workshop scenario 1 vs. scenario 2 (63 vs. 82 p < 0.05); 6-month scenario with prior workshop exposure vs. control (74 vs. 50 p < 0.01); initial workshop scenario 2 vs. 6-month scenario control (82 vs. 50 p < 0.001). Retrospective pre/post survey (5 point Likert scale) revealed self-reported learning in knowledge of tasks, impact and components of communication, avoidance of fixation errors and overall leadership performance (p < 0.001). CONCLUSIONS: Residents acquired resuscitation team leadership skills following an educational intervention as shown by both observational checklist scores and self-reported survey. The six-month follow-up evaluation demonstrated skill retention beyond the initial intervention. A control group suggested that these results were due to completion of the first workshop. PMID- 18158653 TI - Teaching professionalism: a South African perspective. AB - BACKGROUND: Incorporating the teaching and assessment of professionalism in an undergraduate medical curriculum is a challenging process. There are two components that require attention: the cognitive and the experiential. This article outlines how the School of Medicine at the University of Pretoria (UP), South Africa is addressing the teaching and assessment of professionalism. AIMS: To embed teaching and assessment of professionalism in a revised six-year undergraduate medical curriculum. METHOD: This is a descriptive paper of a curriculum process that has stretched over a number of years. The eight principles regarding teaching professionalism provided by Cruess & Cruess (2006) are used to reflect on the current strengths and weaknesses of teaching and assessing professionalism in the UP undergraduate medical curriculum. The cognitive component of the curriculum has been addressed by introducing a series of stage appropriate, contextualized assignments in each year of the curriculum. Faculty committee structures supported the development and implementation of this component. Students' responses to an assignment on the influence of role models provided insight into how the experiential component impacts on their professional development. RESULTS: The teaching and assessment of the cognitive component were relatively easy to put in place. The challenge is to maintain this component. The experiential component appears to be much the same as it was prior to revision. CONCLUSIONS: Our way forward is to expand the teaching and assessment of professionalism in the experiential component. We want to facilitate faculty to develop a charter on professionalism. The principles and commitments of the charter will underlie a white coat ceremony for undergraduates. These principles will also provide guidelines for role models in their supervision of experiential learning and can inform criteria for assessment. An assessment strategy to assess professionalism in the experiential component needs to be developed and implemented. PMID- 18158654 TI - Medical education in Africa: not always a level playing field. PMID- 18158655 TI - Workplace-based assessment as an educational tool: AMEE Guide No. 31. AB - BACKGROUND: There has been concern that trainees are seldom observed, assessed, and given feedback during their workplace-based education. This has led to an increasing interest in a variety of formative assessment methods that require observation and offer the opportunity for feedback. AIMS: To review some of the literature on the efficacy and prevalence of formative feedback, describe the common formative assessment methods, characterize the nature of feedback, examine the effect of faculty development on its quality, and summarize the challenges still faced. RESULTS: The research literature on formative assessment and feedback suggests that it is a powerful means for changing the behaviour of trainees. Several methods for assessing it have been developed and there is preliminary evidence of their reliability and validity. A variety of factors enhance the efficacy of workplace-based assessment including the provision of feedback that is consistent with the needs of the learner and focused on important aspects of the performance. Faculty plays a critical role and successful implementation requires that they receive training. CONCLUSIONS: There is a need for formative assessment which offers trainees the opportunity for feedback. Several good methods exist and feedback has been shown to have a major influence on learning. The critical role of faculty is highlighted, as is the need for strategies to enhance their participation and training. PMID- 18158656 TI - Twelve tips for effective international clinical placements. AB - As universities adopt an increasingly international focus, student health professionals are keen to gain clinical experiences in other countries. Such clinical placements provide students with the opportunity to share their knowledge and at the same time acquire new clinical and cultural skills. The experience gained will not only enhance their clinical practice overseas, but will also enhance it in their home country where they are likely to work with people from culturally, linguistically and clinically diverse backgrounds. Careful preparation and adequate supports are critical if students and the host institutions are to gain maximum benefit from cross-cultural clinical placements. The tips below are based on an ongoing collaboration between the Indian Institute of Cerebral Palsy, Kolkata, India and The University of Sydney, as well as recommendations from the burgeoning literature on international clinical placements. The authors are from both the sending and the receiving institutions, and have found that close collaboration between the home and host institutions along with student reflection, evaluation and the opportunity to integrate new knowledge with other clinical experiences are keys to a satisfactory outcome for all concerned. PMID- 18158657 TI - Challenges in training the ideal Doctor for Africa: lessons learned from Zimbabwe. AB - BACKGROUND: Limited competencies among doctors and reduced numbers from medical migration in Africa could be corrected through innovative curricula and retention of trained manpower. The University of Zimbabwe Medical School simultaneously decided to increase the quality and quantity of doctors to address shortages. AIMS: To evaluate the outcome of innovative medical education at the University of Zimbabwe Medical School. METHOD: A structured questionnaire was administered to a broad group of staff and student representatives. In addition, a desk review of academic documents and policies and procedures was carried out. RESULTS: Early patient contact and community attachment which were introduced to the traditional curriculum remained but other teaching methodologies were not sustained with traditional didactic training still taking centre stage with limited staff development and retention. Whilst the annual student enrolment increased from less than 80 to 200 per year the vacancy rate of academic staff increased to 50%. CONCLUSION: Innovative curricula were partially implemented. The annual student intake increased but the staff complement declined. There is an urgent need to monitor and evaluate outcomes of medical education in Africa to arrest further decline in the quality of health care services. PMID- 18158658 TI - Challenges and issues in health professions education in Africa. AB - The challenges facing health professions education in Africa focus on physical infrastructure, accreditation systems, student selection and faculty recruitment, retention, and development. Higher education in the health professions must be closely aligned with community health needs, and with the training and support of community health workers. A key ingredient in changing institutions in this way is implementation of effective strategies for strengthening the faculty. In addition, information systems, and their prerequisite stable electric power supply, must be supported by internal public resources combined with external aid. These infrastructure investments will facilitate better quality education, improved diffusion of information among schools, and more useful data for self study by institutions. PMID- 18158659 TI - Global paradigm shift in medical education: issues of concern for Africa. AB - INTRODUCTION: When medical education became established in Africa, many curricula were adopted from the West so as to achieve comparable standards in training. Over the last half a century however, major global pedagogical shifts have occurred in medical education without African keeping pace. METHODS: This article reviews key pedagogical changes and other innovations in medical education that have occurred over the last half a century as reported in the literature and identifies some of the issues that need to be addressed in Africa. DISCUSSION AND CONCLUSION: Socioeconomic and political instability, failure to rapidly overcome the inertia for change by substituting the old curriculum with a more problem, system and student-based one and redefining the goals of medical education are some of the issues of concern for Africa, and its ability to keep up in the dynamic world of medical education. There are only few faculty and school managers with effective medical education backgrounds to initiate, evaluate and sustain these changes. African medical academics, national governments and the international community need to come together to assist Africa to rise up to these challenges to ensure attainment and sustenance of global standards in medical training. PMID- 18158660 TI - Medical migration and Africa: an unwanted legacy of educational change. AB - The opportunities given for medical staff to travel, work and remain in countries other than that of their domicile or graduation have led to the phenomenon of medical migration. This has been supported by ease of travel, improved technology and a drive to share good examples of medical education through improved communication. Whilst these opportunities create positive advantages to the individuals and countries involved, through the transfer of knowledge and medical management, the situation does not always lead to long term benefits, and clear disadvantages begin to emerge. The gulf between the developed and developing countries becomes pronounced, leading to a general drift of resources away from the areas where they are most needed and subsequent profound effects upon the indigenous population. This paper suggests that it is a responsibility of medical educators throughout the world to recognize this effect and create opportunities whereby the specialty of medical education positively effects medical migration to the benefit of the less fortunate areas of the world. PMID- 18158661 TI - Developing family medicine in South africa: a new and important step for medical education. AB - The announcement by the National Ministry of Health on the 17th August 2007 (South African Government 2007), officially recognizing family medicine as a speciality in its own right is an indication of just how far 'general practice' has developed in South Africa. From January 2008 there is to be a compulsory, full-time four- year training programme for registrars in Family Medicine which should result in a well trained cohort of Family Physicians suitable to staff Community Health Centres and Primary Care Hospitals in the future. This article seeks to explore some of the milestones reached in the development of the community of family medicine professionals and teachers, the roles taken by the major protagonists in the development and the way that medical education can promote and sustain the discipline. PMID- 18158662 TI - Medical education in Nigeria. AB - Nigeria is the most populated black African nation, with a population of 140 million from the 2006 population census. Medical education began with the establishment of the University College Hospital, Ibadan as a College branch of the University of London in 1948. Since then four generations of medical schools have evolved. The newer medical schools adopted the curriculum of the older schools with little modification. The subsequent introduction of changes and modification in the curriculum of medical education worldwide did not seem to affect Nigerian medical schools. Teacher training in educational methods for medical educators has not been introduced, nor has there been any curriculum planning or review. Efforts made regionally and nationally to change the medical curriculum and improve quality of medical training has not yielded the desired results. The regulatory bodies, the National Universities Commission and the Medical and Dental Council of Nigeria introduced separate curricula for medical schools, as a guide to minimum standards, but there has been so far a varying degree of compliance by the Universities. There is an urgent need to bring Medical Education and teacher training issues to the fore. Needs assessment of medical school faculty to determine their training would be seen to be a good starting point, as well as curriculum transformation and the introduction of newer assessment methods. It is suggested that the Medical and Dental Council of Nigeria (MDCN) should take a leading role by collaborating with outside bodies concerned with medical education. PMID- 18158663 TI - Medical education in South Africa-exciting times. AB - The dramatic political changes of the first ten years of democracy in South Africa have seen major shifts in vested power. Social change and political will have resulted in a new face of our medical student population. Diversity of colour, gender, religion and previously disadvantaged groups make student profiles as different as they could be from a decade ago. The forces of curriculum change, the devolution of power and resources from tertiary centres to primary care facilities and the financial squeeze on university coffers have resulted in enormous challenges for medical educators. South African doctors have been readily accepted in English-speaking countries around the world as they have been well trained, resourceful and usually extensively experienced in clinical situations. Some of these attributes remain. This paper outlines the new prospects facing those who will be educating the future doctors of South Africa. PMID- 18158664 TI - Medical education in the Sudan: its strengths and weaknesses. AB - The history of medical education in the Sudan is both long and interesting. It began in 1924 and has passed through several different phases and stages. Despite numerous difficulties and constraints along the way, vast experience has been gained and many achievements made, all of which have had positive impacts on the health system in the Sudan and the Region. This paper aims to share the experiences and lessons that have emerged from the journey of medical education in the Sudan, and explores the future need for continuing support and dialogue from international colleagues to maintain momentum. PMID- 18158665 TI - Early clerkships. AB - BACKGROUND: Early clinical experience is being introduced in innovative, vertically integrated undergraduate medical curricula. While in many cases, this early clinical experience is limited to the presence of patients during lectures, in Utrecht students gain 'hands on' experience of daily clinical practice during 6-week clerkships. We studied students' and teachers' opinions of these full clerkships in the third year of a medical course. METHODS: All 238 clinical teachers and 89 students involved in the 2003/2004 third year clerkships of the new Utrecht medical curriculum were asked to fill in a 16-item questionnaire about background knowledge, clinical skills, communication skills, and readiness for clerkships. Answers were given on a 5-point Likert scale. Response was 61%. RESULTS: Both students and clinical teachers answered all statements positively (score > 3.00), although some statements were awarded higher scores by students than teachers. In conclusion, students and teachers appear to appreciate clerkships early in the curriculum. CONCLUSIONS: Less background knowledge and a lower age than is usual for the more traditional (later) clerkships do not appear to hinder successful completion of an early clerkship. Indeed, early clerkships have several advantages, such as early observation of the future profession, increased motivation for further study, contextual learning, and improvement of clinical skills. PMID- 18158666 TI - Changes in self-perceived abilities among male and female medical students after the first year of clinical training. AB - BACKGROUND: It is known that male and female medical students have different experiences in their clinical training. AIMS: To assess whether male and female medical students change in their self-rated work habits and interpersonal habits during the first year of clinical training. METHOD: Longitudinal study of self- and peer-assessment among 224 medical students in 3 consecutive classes at a private US medical school. Students rated themselves on global work habits (WH) and interpersonal attributes (IA). Students also rated and were rated by 6-12 peers on the same scale. RESULTS: In the second year of medical school, there were no differences between men and women in quartiles of self-assessed WH or IA. At the end of the third year, however, women were more likely to be in the lower quartiles of self-assessed WH (X(2) = 6.77; p = 0.03), as well as the highest quartiles of self-assessed IA (X(2) = 11.36; p = 0.003). In both years, women rated their own WH skills significantly lower than they rated their peers, while men rated themselves similarly to peers. There were no sex differences in self assessed IA. CONCLUSIONS: Although second-year male and female medical students appear similar to one another in terms of self-assessed WH and IA, by the end of the third year women rate themselves relatively lower in WH, while men rate themselves relatively lower in IA. PMID- 18158667 TI - Should non-expert clinician examiners be used in objective structured assessment of communication skills among final year medical undergraduates? AB - BACKGROUND: Adoption of the objective structured clinical examination may be hindered by shortages of clinicians within a specialty. Clinicians from other specialties should be considered as alternative, non-expert examiners. AIMS: We assessed the inter-rater agreement between expert and non-expert clinician examiners in an integrated objective structured clinical examination for final year medical undergraduates. METHODS: Pairs of expert and non-expert clinician examiners used a rating checklist to assess students in 8 oral communication stations, representing commonly encountered scenarios from medicine, paediatrics, and surgery. These included breaking bad news, managing an angry relative, taking consent for lumbar puncture; and advising a mother on asthma and febrile fits, and an adult on medication use, lifestyle changes and post-suture care of a wound. 439 students participated in the OSCE (206 in 2005, 233 in 2006). RESULTS: There was good to very good agreement (intraclass coefficient: 0.57-0.79) between expert and non-expert clinician examiners, with 5 out of 8 stations having intraclass coefficients > or =0.70. Variation between paired examiners within stations contributed the lowest variance to student scores. CONCLUSION: These findings support the use of clinicians from other specialties, as 'non-expert' examiners, to assess communication skills, using a standardized checklist, thereby reducing the demand on clinicians' time. PMID- 18158668 TI - A gaming approach to learning medical microbiology: students' experiences of flow. AB - BACKGROUND: There is a growing awareness in medical education of general skills(1) required for lifelong learning. Such skills are best achieved when students experience positive affective states while they are learning, as put forth by the Csikszentmihalyian theory of flow. This study describes how a quiz type board game was used in the School of Medicine of the Faculty of Health Sciences at the University of the Free State to address students' negativity towards medical microbiology. METHODS: The study population consisted of third year medical students who had recently completed the Infections module of the undergraduate Learning Programme for Professional Medicine. RESULTS: Data gathered by means of two questionnaire surveys and direct observation showed that the game impacted positively on students' perceptions of and attitudes towards medical microbiology as a subject. A high perceived probability of the game contributing to the acquisition of general skills was recorded, since the experience of positive affect during the process of informal learning went hand in-hand with heightened team effort and spontaneous communication. CONCLUSIONS: This article may be of value to health educators who wish to supplement formal teaching with informal learning so as to enhance not only the recall of factual knowledge, but also the advancement of general skills. PMID- 18158669 TI - More than mere games: a review of card and board games for medical education. AB - During recent years, attempts have been made to complement more classical concepts of medical teaching by introducing card and board games on medical topics. These teaching tools cover every age and education group, and many different medical topics. In this article we have reviewed all card and board games for medical education purpose listed in NCBI PubMed database and Internet game databases (n = 29). It summarizes games that might be useful to medical teaching staff. To categorize these games, a new schema for medical games categorization, based on the game mechanism and theories on experiential learning circles, is proposed and discussed. Additionally we have a view on card and board games with medical topics for entertainment (n = 22). PMID- 18158670 TI - A qualitative evaluation of medical student learning with concept maps. AB - PURPOSE AND AIMS: The purpose of this study was to investigate the ways in which the use of concept maps influenced the learning processes of third year internal medicine students in the context of medical education. Reported here are the qualitative results of this study. METHODS: One-hundred thirty four medical students were taught to use concept mapping as a learning strategy at the beginning of their internal medicine rotations. Upon completion of the internal medicine rotation students were asked to evaluate how concept maps fostered the process of linking theoretical information to clinical practice. Additionally, students described how concept maps impacted their learning. RESULTS: In this study, concept maps fostered a positive connection between theory and practice. Additionally, students described three major themes impacting their learning: concept mapping as a facilitator of knowledge integration and critical thinking, as a teaching methodology and finally, as a learning method. CONCLUSIONS: This study suggests concept maps may be an instructional method to foster the learning and thinking process of medical students. PMID- 18158671 TI - Integration of role-playing into technical skills training: a randomized controlled trial. AB - INTRODUCTION: Recently, efforts have been undertaken to enhance the face validity of technical skills training by introducing role-plays and standardised patients. Since little is known about the effects of role-playing with respect to the realism of a training situation and students' objective performance, we performed a randomized controlled trial. METHODS: 36 medical students participated in videotaped small group skills-lab sessions on the topics of Doppler sonography and gastric tube insertion. One half of the students participated in role-plays and the other half practised without role-playing. Realism of the training situation was analysed by means of post-intervention self-selected student survey evaluations. Technical performance and patient-physician communication were assessed by independent ratings of the videotaped sessions. RESULTS: The physician's role was regarded to be significantly more realistic when performing role-plays. Assessment of videotaped sessions showed that practising technical skills by performing role-plays resulted in significantly better patient physician communication whereas students' technical performance did not differ between groups. CONCLUSION: Introducing role-plays enhances the realism of technical skills training and leads to better patient-physician communication. Students do not seem to be overstrained by practising clinical technical skills using role-plays. We conclude that role-playing is a valuable method in practising technical skills. PMID- 18158672 TI - IVIMEDS: a short report on an evaluation of the cardiovascular system learning module. AB - BACKGROUND: On-line learning technology can be used to present curricular material in a variety of formats to stimulate and support student learning in both content and potentially skills-based areas in medicine. In 2005, second-year medical students at the University of Queensland evaluated the IVIMEDS' cardiovascular module, and were given access to all learning objects in the module. AIM: The study aimed to determine the value to students of the IVIMEDS' Cardio-vascular System (CVS) module compared with existing CVS learning resources. METHOD: Research and control groups (n = 50 respectively) completed a 16-item questionnaire relating either to the IVIMEDS' CVS module or to the existing CVS resources. Responses were analysed for common themes, and the performance of the control and research groups on the mid-year summative assessment results was compared. RESULTS: Thirty-five students in the research group completed the IVIMEDS evaluation. Thirty-one-percent had difficulty learning to navigate the software, and 17% felt that they would have derived greater benefit with prior training. Students in the research group scored significantly higher on one question in the summative assessment than did students in the control group (F(66) = 2.1, p < 0.5). Qualitative data suggest that for students to fully accept the IVIMEDS' material, it will be essential to ensure that students are fully aware of its place in the medical program so they are confident that by using it, they will be able to achieve the set learning objectives. CONCLUSIONS: The students appreciated the potential of the IVIMEDS' cardiovascular module, but a full evaluation of the package would require that the module be presented at an appropriate time using an easily navigable system, after training in use of the software package. PMID- 18158673 TI - Long-term effect of a course on in-training assessment in postgraduate specialist education. AB - BACKGROUND: In-training assessment has become an important part of clinical teachers' responsibilities. One way to ensure that clinical teachers are qualified for this role is setting up a course. A "Teach the teachers" course focusing on in-training assessment was designed for anaesthesiologists in Denmark. AIMS: To evaluate short and longer term effects of a course on in training assessment for clinical teachers in Anaesthesiology. METHOD: Fifty-one anaesthesiologists attended a 2-day interactive course about in-training assessment. Effects of the course on knowledge were assessed using identical pre- and post- tests. Longer- term effects were measured six months after the course using the same test. Self-reported use of in-training assessment methods was evaluated using supplemental questions in the follow-up test. RESULTS: There were significant increases in knowledge about in-training assessment immediately following the course (effect size, Cohens d = 1, 5). The knowledge was retained six months later. Knowledge about assessment by clinical structured observation and by written assignments showed further increases in the follow-up period. Participants used the various assessment methods in their daily practice during the six-month study period. CONCLUSION: A focused "Teach the teachers" course during the implementation phase of a new assessment programme increased participants' knowledge about in-training assessment. PMID- 18158674 TI - A comparison of teaching methods: interactive lecture versus game playing. AB - BACKGROUND: We compared an interactive lecture with a game based on charades as a means of teaching child development to fifth year medical students. METHODS: Students were randomised to either intervention and the data analyst was blinded to which method of teaching the students had received. Performance of the students was assessed after the teaching with a multiple choice questionnaire. Long term performance was assessed by comparing the students' OSCE scores in the child development station. RESULTS: The students that received an interactive lecture performed significantly better in the MCQ than those who were taught using the game. CONCLUSIONS: Neither group had any difference in long term performance in this subject following the interventions. PMID- 18158675 TI - Integrating physical examination and trunk anatomy; a new course for second year medical students. AB - The aim of this study was to determine the effect of an integrated course of physical examination and trunk anatomy on second year medical students' attitude and knowledge of anatomy. Thirty nine second year medical students attended a physical examination course (5 small group sessions) related to topics in trunk anatomy (2004/05). Students in control group attended routine anatomy lectures and problem based learning discussions relevant to their physiology course. Routine final anatomy exam (65 multiple choices questions) was performed. Attitude toward course impacts in intervention group were assessed by a likert questionnaire (18 items). A T-test was performed to compare anatomy scores. The Intervention group had higher score exam anatomy (46.6 +/- 6.3) than others (41.4 +/- 6.0, p = 0.04). The mean score of attitude questionnaire was 3.48 (SD = 0.8). In conclusion teaching physical examinations integrated with trunk anatomy improves students' knowledge and motivated them in learning anatomy. PMID- 18158676 TI - Evaluation using simulated students for reliability of multiple standardized patients scoring in clinical performance examinations. AB - BACKGROUND: The consistency of rating among different standardized patients (SPs) is very important in clinical performance examinations (CPX). AIMS: We evaluated the usefulness of simulated students to assess the reliability of a group of standardized patients in a CPX. METHODS: Five SPs in each case were trained to simulate the same patient in a CPX. Ten fourth-year medical student volunteers were selected to act as simulated students. After the simulated students encountered the first SP, they reviewed a video of their performance and sequentially repeated the performance with the other 4 SPs. RESULTS: The average maximum difference (Deltamax +/- SD) of multiple SPs' percent scores on a simulated student was 18 +/- 7. In 10 pairs of SPs who gave largest Deltamax on a given student, the median Deltamax was 30 (range, 26-38) while median difference of percent score in the faculty scoring on the same simulated student was 3 (range, 0-7). Five problematic SPs whose ratings were significantly different from 3 or more other SPs' ratings in each case were identified. CONCLUSIONS: Simulated students may be a useful means to check the consistency of rating among different SPs in a CPX. PMID- 18158677 TI - The impact of pre-clinical pharmacology and pharmacotherapy training on students' abilities and perceptions during clinical rotations. AB - BACKGROUND: Due to curricular changes at Leiden University Medical School, students have been exposed to different amounts of pharmacology and pharmacotherapy education. Cohort 1999 received little training; whereas cohort 2000 was provided with self-study materials preclinically and extra attention to pharmacotherapy during rotations. This study compares the cohorts by performance in and perception of pharmacotherapeutic abilities during the internal medicine rotation. RESULTS: Preclinically, both cohorts were unable to adequately complete therapeutic plans. While cohort 1999 did not improve during the rotation, cohort 2000 did. For both cohorts the percentage of students feeling confident about various pharmacotherapeutic abilities increased significantly during rotations, but only 25% felt secure about their overall ability. CONCLUSIONS: The extra education helped cohort 2000 perform better, but did not improve their confidence in their pharmacotherapeutic ability. Thorough preclinical training and compulsory practice with feedback and assessment are necessary for students to develop competence in pharmacotherapy. PMID- 18158678 TI - An innovative web-based peer support system for medical students in Hong Kong. AB - BACKGROUND: Psychological morbidity is commonly found in medical students. AIMS: The Mental Health Support Group (MSG), a student-initiated and student-run web based peer support service aims to provide mental health information, e mail counseling and an online forum for medical students. METHODS: The development process of MSG is described in the paper with presentation of preliminary evaluation results. RESULTS: Preliminary evaluation shows promising results. Student members of MSG acquired valuable skills in counseling, communication, webpage design and maintenance of an online forum. CONCLUSIONS: Future challenges include succession issues, strategies to keep up the momentum, enhancement of publicity and further diversification of service to meet the needs of our students. PMID- 18158679 TI - Online master's degree in education for healthcare professionals: early outcomes of a new program. AB - BACKGROUND: The Online Master's Degree in Education for Healthcare Professionals program, a collaboration of the University of Cincinnati Colleges of Medicine and Education, was launched nationally in 2002. The purpose of this survey-based, descriptive study was to describe the early outcomes of this new program. METHODS: Surveys were sent via email to 32 eligible students (students who have completed at least 6 of 12 required courses in the program) which included all 8 graduates. RESULTS: Based on 21 returns (66%) participants have had 19 journal publications, 99 national presentations, 13 teaching awards, 5 promotions, and acquisition of extramural funding for educational projects totaling over $3 million. CONCLUSIONS: Based on early results, graduates of the program are productive in academic medical education. We anticipate that graduates of the program will have a significant impact on medical education and will achieve promotion and advancement at their institutions. PMID- 18158680 TI - A study of student performance on self-scheduled, computer-based examinations in a medical histology course: is later better? AB - BACKGROUND: Student self-scheduling of computer-based examinations during a defined window of opportunity is a protocol used by some medical school courses and licensing bodies. Students scheduling late have more time to prepare, which may provide them with an unfair advantage. METHODS: ANOVA was used to compare the average scores of three different cohorts of test-takers (early, middle and late) on 13 Microscopic Anatomy exams taken over a 3-year period (2003-2005) at the University of Arkansas for Medical Sciences. RESULTS: On 11 of the 13 exams, the average score of students who took the exam last was lower than that of students who took the exam first. These differences were significant for eight (73%) exams. There were two exams on which the average score of students who took the exam last was higher than that of students who took the exam first, but these differences were not significant. CONCLUSIONS: Scheduling exams late in the window of opportunity does not appear to provide a performance advantage and may even be detrimental since these students are responsible for new content information in the course and in other concurrently running courses while they are preparing for a 'late' exam. PMID- 18158681 TI - Incorporating prescription writing into a therapeutics examination. PMID- 18158682 TI - Do you know? PMID- 18158684 TI - EMedical teacher. PMID- 18158686 TI - New insights in trauma and memory: a special issue of memory. PMID- 18158687 TI - Review of the contemporary literature on how children report sexual abuse to others: findings, methodological issues, and implications for forensic interviewers. AB - Methods used during forensic interviews with children are driven by beliefs about how children recall and report child sexual abuse (CSA) to others. Summit (1983) proposed a theory (Child Sexual Abuse Accommodation Syndrome) contending that, due to the specific traumatic characteristics of CSA, children will often delay disclosing abuse or altogether fail to disclose during childhood, deny abuse when asked, and often recant abuse allegations. His theory has had a tremendous impact on the field of CSA forensic evaluations, despite its dearth of empirical support. In this paper, we review and critique the contemporary literature from two main sources: retrospective accounts from adults reporting CSA experiences and studies of children undergoing forensic evaluation for CSA. We conclude that data support the notion that children often delay abuse disclosure, but that among valid abuse cases undergoing forensic evaluation, denial and recantation are not common. Methodological issues and implications for forensic interviewers are discussed. PMID- 18158688 TI - Familiarity breeds distortion: the effects of media exposure on false reports concerning media coverage of the terrorist attacks in london on 7 July 2005. AB - The present experiment investigated whether increased media exposure could lead to an increase in memory distortions regarding a traumatic public event: the explosion of the No. 30 bus in Tavistock Square, London on 7 July 2005. A total of 150 Swedish and 150 UK participants completed a series of questionnaires about their memory of either (i) the aftermath of the explosion, (ii) a non-existent computerised reconstruction of the moment of the explosion, or (iii) non-existent closed circuit television footage of the moment of the explosion. In line with the availability heuristic, U.K. participants were more likely than Swedish participants to claim to have seen all three types of footage. Furthermore, a subsample of U.K. participants who appeared to have developed false "memories" of seeing the No. 30 bus explode scored significantly higher on measures of dissociation and fantasy proneness than participants who did not develop false "memories". This experiment provides further support for the role of imaginative processes in the development of false memories. PMID- 18158689 TI - Further mechanical considerations between polar and SRM mobile ergometer systems during laboratory-based high-intensity, intermittent cycling activity. AB - The purpose of this article is to outline mechanical issues related to the use of the Polar S710 heart monitor with Power Unit when compared with the SRM Powercrank system. There are issues outlined in this article that refer to the suitability of the Polar S710 for the quantification of performance during downhill cycling that relate to chain vibration, chain tension, and time interval sampling rates. PMID- 18158690 TI - The effects of vitamin E and vitamin C supplementation on bioenergetics index. AB - In order to study the effects of vitamin E and vitamin C supplementation on the bioenergetic index, 36 male physical education students were selected nonrandomly and assigned to a different supplementation protocol. The average age, weight, height, and fat percentage were 22.48 +/- 1.84 years, 64.93 +/- 7.84 kg, 175.4 +/ 5.66 cm, and 10.94 +/- 5.29%, respectively. The period of supplementation lasted 3 weeks. The subjects from group 1 consumed a daily dose of 400 mg of vitamin E, subjects from group 2 ingested 1000 mg of vitamin C, subjects from group 3 ingested 400 mg of vitamin E along with 1000 mg of vitamin C, and subjects from group 4 (control group) consumed a placebo. The tests applied were the running anaerobic sprint test (RAST) and the Cooper 12-min run test. The results indicate that there were no significant differences between groups during the study in anaerobic power assessed by RAST. We found a significant difference between group's, however, in aerobic power (p < 0.05). We concluded that daily consumption of vitamin E, vitamin C, and a combination of vitamin E and vitamin C for a period of 3 week significantly improved aerobic power. PMID- 18158691 TI - Cycling and running performance, not anthropometric factors, are associated with race performance in a Triple Iron Triathlon. AB - Sixteen well-trained and well-experienced male Caucasian triathletes participated in World Championship Triple Iron Triathlon 2006 in Moosburg, Karnten, Austria, which required athletes to perform 11.6 km swimming, 540 km cycling, and 126.6 km running within a time limit of 58 hours. Body mass, body height, skinfold thicknesses, circumferences of extremities, skeletal muscle mass, and body fat were measured. The results showed that race time was not significantly influenced by the anthropometric properties (p > 0.05). No significant influence was observed between race time and the calculated parameters of body mass index (BMI), body fat, and skeletal muscle mass (p > 0.05). In contrast, cycling performance (r(2) = 0.49, p < 0.01) and running performance (r(2) = 0.73, p < 0.01) are associated with total race performance. It summery, in an ultratriathlon, BMI, body height, skinfold thicknesses, circumference of extremities, skeletal muscle mass, and body fat have no influence on race performance in world-class ultratriathletes. Total race performance is associated with performance in cycling and running, not in swimming. PMID- 18158692 TI - Effect of a 15-day pragmatic hamstring stretching program on hamstring flexibility and single hop for distance test performance. AB - The purpose of this study was to determine the effects of a 15-day pragmatic hamstring stretching program on lower extremity performance as measured by the single hop for distance test (SHDT) in subjects with limited hamstring flexibility. Thirteen cadets enrolled at a military academy participated in the study. Subjects randomly were assigned to stretch the hamstrings of one lower extremity for 15 consecutive days (experimental lower extremity group) and not stretch the other (control lower extremity group). Pre- and post-test measurements showed significant improvements in the hamstring flexibility and SHDT scores for the experimental lower extremities. No significant change occurred for the control lower extremity group for the AKET and SHDT scores. The results suggest that in addition to a routine 15-day stretching program, a brief purposeful delay between stretching and performance testing may negate the detrimental effect on performance that may be caused by the acute effects of stretching. Because subjects also stretched on the day of the SHDT posttest, but with a purposeful delay of 10 minutes between stretching and testing, it is believed the design of this study more closely mimics stretching activities as commonly performed by athletes. PMID- 18158693 TI - Gender bias on the effects of instruction on kinematic and kinetic jump parameters of high-level athletes. AB - The purposes of this study were to examine gender differences during drop jumps, to evaluate the effect of a set of simple instructions on kinematic and kinetic parameters during a drop jump, and to determine if there are gender differences in the effects of instruction on those parameters. Twenty-five basketball players, 13 men and 12 women, performed drop jumps from a box (height 30.5 cm) after being asked to perform a land and jump movement as they normally would do when aiming at maximum jump height. The experimental group received a set of instructions designed to make them land softer. The measured parameters were impact force at landing, ground contact time, flight time, frontal plane knee angle, sagittal plane knee angle, and distance between the knees. When the groups were examined by gender, no differences in the parameters were seen in men after instruction, but in women the following significant differences were detected: an increase in contact time, a decrease in landing force, and a decrease in inward movement of the knees after landing. These data indicate that females respond differently to jumping/landing instructions. PMID- 18158694 TI - Effect of an ankle stabilizing orthosis on selected ankle kinematics during walking. AB - This study evaluated the stability effects of an ankle stabilizing orthosis on unstable ankles on selected three-dimensional ankle kinematics during two speeds (2.5 and 4.5 mph) of treadmill walking. Eleven subjects with one unstable ankle walked at these two treadmill speeds with and without the ankle stabilizing orthosis. The angular excursions and velocities of the ankle in the sagittal and frontal planes were recorded with a three-dimensional ProReflex Imaging System and analyzed with two-way repeated-measure ANOVA and paired t test. It was found that the ankle stabilizing orthosis provided significant stability control in the angular excursions of plantarflexion, and it did not affect the maximum velocity of plantarflexion and the walking speed during the treadmill walking. As the speed increased, the angular excursions and the maximum angular velocities in the sagittal and frontal planes increased significantly. PMID- 18158695 TI - MRI of the small bowel with oral contrast or nasojejunal intubation in Crohn's disease: randomized comparison of patient acceptance. AB - OBJECTIVE: Magnetic resonance imaging of the small bowel with an oral contrast (MRI per os) is believed to generate fewer adverse symptoms compared with installation of the contrast in a nasojejunal catheter (MRE). However, there is very little evidence to support this assumption. The aim of this study was to evaluate the compliance of patients examined with both of these MRI methods. MATERIAL AND METHODS: Patients with suspected or known Crohn's disease referred for MRI of the small bowel underwent both MRI per os and MRE within 7 days. A questionnaire was answered immediately and 24 h after the procedure. Adverse symptoms were graded on a visual analogue scale (VAS). Responses were compared with paired and independent sample t-tests and signed-rank tests. Correlation of each symptom to the overall discomfort was evaluated with Spearman's correlation coefficient. RESULTS: Thirty-eight patients (18 F, 20 M) were included in the analysis. Abdominal pain and discomfort were lower with MRI per os than with MRE (mean VAS pain score immediately after: 10 mm and 33 mm, respectively, p < 0.001; mean VAS discomfort score 24 h after: 18 mm and 62 mm, respectively, p < 0.001). Nausea and abdominal pain were correlated with overall discomfort after MRI per os (r = 0.56 for both, p < 0.001). No symptoms were significantly correlated with discomfort experienced with MRE. More patients accepted repeat MRI per os examination (n = 36) than MRE (n = 22, p = 0.001). CONCLUSION: Patients preferred and experienced less abdominal pain and discomfort with MRI per os than with MRE. PMID- 18158696 TI - Incidence of the focal type of autoimmune pancreatitis in chronic pancreatitis suspected to be pancreatic carcinoma: experience of a single tertiary cancer center. AB - OBJECTIVE: With an increase in autoimmune pancreatitis (AIP) being reported, the focal type of AIP, which shows localized narrowing of the main pancreatic duct and focal swelling of the pancreas, has recently been recognized. Therefore, cases of focal-type AIP subjected to surgical intervention for presumptive malignancy might previously have been diagnosed as mass-forming chronic pancreatitis. The aim of this study was to elucidate the incidence of focal-type AIP in resected chronic pancreatitis at a single tertiary cancer center. The clinical and radiological features of focal-type AIP were also evaluated. MATERIAL AND METHODS: We re-evaluated 15 patients who underwent pancreatic resection with a presumed diagnosis of pancreatic ductal adenocarcinoma, and who in the past had been diagnosed pathologically as having chronic pancreatitis. RESULTS: Seven of 15 patients showed AIP, and the other 8 patients were diagnosed as having mass-forming chronic pancreatitis not otherwise specified by pathological retrospective examination. In other words, nearly half of the cases of resected chronic pancreatitis that were suspected to be pancreatic carcinoma preoperatively showed focal-type AIP. Regarding the characteristic findings of focal-type AIP, narrowing of the pancreatic duct on endoscopic retrograde pancreatography (ERP) might be diagnostic. CONCLUSIONS: Focal-type AIP is not a rare clinical entity and might be buried in previously resected pancreatic specimens that in the past were diagnosed simply as mass-forming pancreatitis. PMID- 18158697 TI - Combating childhood type 2 diabetes mellitus: it will take a village. PMID- 18158698 TI - Epidemiology of paediatric metabolic syndrome and type 2 diabetes mellitus. AB - The epidemic in childhood obesity is a driving force behind the increase in paediatric metabolic syndrome, a collection of abnormalities that is associated in adults with increased risk for cardiovascular disease and type 2 diabetes mellitus. Although there is no clear consensus about the paediatric definition for metabolic syndrome, the prevalence of this syndrome is clearly rising. Children with metabolic syndrome are at increased risk for metabolic syndrome in adulthood. A late consequence of metabolic syndrome is type 2 diabetes, which increasingly affects adolescents. The rise in metabolic syndrome and type 2 diabetes in children is almost sure to lead to an increase in associated complications in young adulthood, including early cardiovascular disease. This epidemic will bear fruit in forthcoming decades, putting further stress on the healthcare system and probably leading to increased morbidity and a shorter lifespan for future generations. PMID- 18158699 TI - Vascular disease in paediatric type 2 diabetes: the state of the art. AB - Type 2 diabetes is an emerging problem in the paediatric population. Paediatricians and paediatric endocrinologists are struggling with how to best assess, predict and treat cardiovascular risk factors in these patients. There is a notable lack of consensus in how to proceed, even among experts in the field. There are very limited data from quality trials in established paediatric type 2 diabetes. We address the available information regarding traditional and non traditional indices of cardiovascular risk, including examination findings, biochemical markers and non-invasive imaging modalities. We discuss the utility and pitfalls of applying knowledge gained in adult medicine to the paediatric population. Potential treatment strategies are reviewed, including the currently available pharmaceutical options, with the acknowledgement that there are few drugs formally approved in the paediatric population. PMID- 18158700 TI - Physical activity for the prevention and management of youth-onset type 2 diabetes mellitus: focus on cardiovascular complications. AB - With the growing prevalence of childhood obesity and type 2 diabetes mellitus (T2DM) in youth, the challenge of cardiovascular disease risk management has entered the paediatric realm, affecting specialists, family physicians and allied healthcare professionals alike. Currently, there is little evidence to support optimal strategies for management of T2DM in youth and the associated cardiovascular complications. Physical activity plays a powerful role in the prevention and management of T2DM and cardiovascular disease in adults. This review will focus on the role of physical activity for the prevention of T2DM in youth and its associated cardiovascular complications. The first part describes the prevalence of cardiovascular risk factors in this cohort. The second part focuses on the role of physical activity in the prevention and management of T2DM in youth. Collectively, the limited intervention and observation studies published to date suggest that daily targets of 60-90 minutes of physical activity and less than 60 minutes of screen time (i.e. time spent in front of a television, computer or video games) are required for the prevention and management of T2DM in youth. Large-scale intervention studies are needed to determine the most effective physical activity strategies for the prevention and management of T2DM in youth. PMID- 18158701 TI - Targeting obesity to reduce the risk for type 2 diabetes and other co-morbidities in African American youth: a review of the literature and recommendations for prevention. AB - Over the last decade, the prevalence of obesity has continued to rise within the adolescent population of the US. Data show that African American youth are disproportionately affected by the obesity epidemic due to their higher risk for obesity-related co-morbidities, such as type 2 diabetes. Interventions that target risk factors for obesity at the individual, family and community levels are needed in order to prevent the onset of type 2 diabetes and related complications. This article provides an overview of the prevalence of obesity and type 2 diabetes in African American youth, the pathophysiology of the disease, and the behavioural risk factors that have contributed to its high prevalence within the African American adolescent population. Successful intervention strategies that target modifiable risk factors, such as diet and physical activity, will be identified. Finally, recommendations for programmes to prevent the onset of type 2 diabetes within the African American adolescent population are presented. PMID- 18158702 TI - Impact of metabolic syndrome definitions on prevalence estimates: a study in a Portuguese community. AB - This study compared the prevalence of metabolic syndrome (MS) according to World Health Organization (WHO), National Cholesterol Education Program Third Adult Treatment Panel (NCEP-ATP III), International Diabetes Federation (IDF) and American Heart Association/ National Heart, Lung and Blood Institute (AHA/NHLBI) definitions, to evaluate how well the different classifications agreed. The study also compared their 10-year predicted risk of coronary heart disease (CHD) with the Framingham risk score (FRS). Some 886 women and 547 men aged 18-92 years were included in the study. Demographic and personal medical history data were obtained at interview. Four operational definitions of MS were used (those of the WHO, NCEP-ATP III, AHA/NHLBI and IDF). The prevalence of metabolic syndrome was found to be 26.4% (WHO criteria), 24.0% (NCEP-ATP III criteria), 41.9% (IDF criteria) and 37.2% (AHA/NHLBI criteria). According to the definition used, central obesity ranged from 41.9% to 75.1% and high blood pressure from 52.9% to 65.8%. Agreement between classifications ranged from 75.2% (kappa=0.47) to 90.4% (kappa=0.80) and was lower in males. The 10-year predicted risk of CHD by FRS was similar between the different definitions. IDF and AHA/NHLBI definitions resulted in a higher prevalence of MS than the NCEP-ATP III or WHO definition. Overall, however, good agreement was found between definitions, and the predicted 10-year of CHD risk was similar. PMID- 18158703 TI - Levels of adiponectin, C-reactive protein and interleukin-1 receptor antagonist are associated with the relative change in body mass index between childhood and adulthood. AB - Obesity has been related to subclinical inflammation and decreased levels of adiponectin. We examined the relationship between inflammatory markers and adiponectin and the change in body mass index (BMI) between childhood and adulthood. Our study included 368 subjects (176 men and 192 women) from a population-based cohort whose weight and height had been recorded at the age of seven years. They participated in this study as adults (with a mean age of 46 years); levels of adiponectin, interleukin-1 receptor antagonist (IL-1 Ra) and high-sensitivity C-reactive protein (hs-CRP) were measured. The relative change of BMI from childhood to adulthood was significantly associated with levels of IL 1 Ra (men: r=0.27 [95% CI: 0.12 to 0.40] and women: 0.64 [0.55 to 0.72]), hs-CRP (r=0.15 and 0.52, respectively) and adiponectin (r=-0.13 and -0.29, respectively) in both genders. Decreased levels of adiponectin and elevated levels of IL-1 Ra and hs-CRP at adulthood appear to be related to the change in BMI between childhood and adulthood. PMID- 18158704 TI - Double-edged relationship between adiposity and coronary artery calcification in type 1 diabetes. AB - Coronary artery disease (CAD), a leading cause of death in type 1 diabetes (T1D), often occurs two or more decades earlier in this population compared to the population without diabetes. Although CAD generally increases with adiposity, this association is unclear in T1D. In this study, we examined associations of adiposity with coronary artery calcium (CAC) in 315 individuals with T1D. Mean age and diabetes duration were 42 and 34 years, respectively, at study entry. CAC, visceral adiposity (VAT) and subcutaneous adiposity (SAT) were determined by electron beam tomography; and BMI and waist circumference (WC) were determined. The presence of CAC was positively associated with VAT, SAT and BMI in men (p<0.05) and with all four adiposity measures in women (p<0.05) after adjustment for age and other traditional cardiovascular risk factors. However, after adjustment, the degree of CAC was not associated with any of the four adiposity measures, with the exception of SAT in women. Women in the lowest tertile of SAT had more CAC than those in the second tertile (p<0.016). Adiposity was positively associated with the presence of CAC, but the relationship with its severity was either inverse or non-existent. This double-edged association emphasises the complex relationship between adiposity and cardiovascular risk in diabetes. PMID- 18158705 TI - Effective cut-off values of waist circumference to detect the clustering of cardiovascular risk factors of metabolic syndrome in Japanese men and women. AB - The purpose of this study was to determine effective cut-off values of waist circumference (WC) to detect the clustering of cardiovascular risk factors (CCRF) in a Japanese population. The subjects were 2,476 men and women who participated in a health examination in Tokyo. The CCRF was defined according to the International Diabetes Federation (IDF) and the Japanese Committee of the Criteria for Metabolic Syndrome (JCCMS). The effective (accurate and sensitive) values were tested using the receiver operating characteristics analysis. The accurate (maximised sensitivity plus specificity) values were 81 cm and 80 cm using the IDF and JCCMS criteria for men, and 82 cm for both criteria for women. From the ROC curve, 85 cm was identified as an effective value for men. Thus, the effective cut-off value of WC for the Japanese should be approximately 85 cm for men and approximately 82 cm for women. PMID- 18158706 TI - Admission glycaemia and outcome in patients with acute coronary syndrome. AB - Some studies of patients with acute myocardial infarction have reported that hyperglycaemia at admission may be associated with a worse outcome. This study sought to evaluate the association of blood glucose at admission with the outcome of unselected patients with acute coronary syndrome (ACS). Using the Acute Myocardial Infarction and unstable angina in Switzerland (AMIS Plus) registry, ACS patients were stratified according to their blood glucose on admission: group 1: 2.80-6.99 mmol/L, group 2: 7.00-11.09 mmol/L and group 3: > 11.10 mmol/L. Odds ratios for in-hospital mortality were calculated using logistic regression models. Of 2,786 patients, 73% were male and 21% were known to have diabetes. In hospital mortality increased from 3% in group 1 to 7% in group 2 and to 15% in group 3. Higher glucose levels were associated with larger enzymatic infarct sizes (p<0.001) and had a weak negative correlation with angiographic or echographic left ventricular ejection fraction. High admission glycaemia in ACS patients remains a significant independent predictor of in-hospital mortality (adjusted OR 1.08; 95% confidence intervals [CI] 1.05-1.14, p<0.001) per mmol/L. The OR for in-hospital mortality was 1.04 (95% CI 0.99-1.1; p=0.140) per mmol/L for patients with diabetes but 1.21 (95% CI 112-1.30; p<0.001) per mmol/L for non diabetic patients. In conclusion, elevated glucose level in ACS patients on admission is a significant independent predictor of in-hospital mortality and is even more important for patients who do not have known diabetes. PMID- 18158707 TI - The European perspective on diabetes prevention: development and Implementation of A European Guideline and training standards for diabetes prevention (IMAGE). AB - The prevalence of type 2 diabetes has increased dramatically in Europe, particularly in younger people. Nevertheless, jointly developed prevention strategies are still lacking. The development and Implementation of A European Guideline and training standards for diabetes prevention (IMAGE) project was initiated by the Technical University Dresden in response to this major public health concern of the European Union. Within the IMAGE project, four specific objectives will be developed and implemented into clinical practice: a) joint European guidelines; b) a European training curriculum for prevention managers; c) European standards for quality control of diabetes prevention programmes; and d) a European e-health training portal for prevention managers. This three-year project began in June 2007. At the time of writing, 32 institutions from 16 countries have become involved. The development of competent guidelines and education programmes, as well as the establishment of quality standards for the primary prevention of type 2 diabetes, should significantly enhance the ability of healthcare professionals to respond swiftly to its drastic increase and its burden to society. PMID- 18158708 TI - Beneficial effects of ramipril on myocardial diastolic function in patients with type 2 diabetes mellitus, normal LV systolic function and without coronary artery disease: a prospective study using tissue Doppler. AB - Angiotensin-converting enzyme (ACE) inhibitors can improve cardiovascular outcome in patients with type 2 diabetes mellitus (T2DM). Myocardial diastolic function (Ve) is a known marker of cardiovascular prognosis. It could potentially indicate the effects of preventive therapy if evaluated by tissue Doppler. We tested the hypothesis that treatment with the ACE inhibitor ramipril has beneficial effects on Ve. In this study, 16 subjects on insulin therapy (eight receiving 10 mg ramipril/day compared to eight matched controls who were not treated with an ACE inhibitor) were followed up for a period of nine months. Myocardial and vascular function were assessed by tissue Doppler and ultrasound. In the ramipril group, Ve improved significantly after nine months of treatment (7.8+/-0.9 cm/s to 8.6+/ 0.9 cm/s, p<0.04). Systolic blood pressure and intima media thickness (IMT) demonstrated a trend towards improvement. In controls, Ve remained unchanged and there was a trend towards deterioration in stiffness index beta (p<0.07). In conclusion, the observed improvement of myocardial diastolic function with ramipril in patients with T2DM is an encouraging result. It might contribute to the overall improvement that has been observed with hard cardiovascular end points. PMID- 18158709 TI - Microalbuminuria and the metabolic syndrome in non-diabetic black Africans. AB - It is recognised that the metabolic syndrome promotes the development of cardiovascular disease. Although several studies have shown a relationship between the metabolic syndrome and kidney disease, few of these have used non diabetic subjects, especially in the African population. This was a cross sectional study of subjects of African origin, using the metabolic syndrome (MS) criteria of the National Cholesterol Education Program (NCEP) third Adult Treatment Panel (ATP III). Subjects with impaired fasting glucose, with two-hour glucose >or= 11.1 mmol/L after a glucose tolerance test, were excluded. Spot urine for albumin-to-creatinine ratio (ACR) was measured and the glomerular filtration rate (GFR) was estimated using the Modification of Diet in Renal Disease (MDRD) equation. Microalbuminuria was defined as ACR between 3-30 mg/mmol. There was a significant decline in GFR and a significant increase in ACR with increasing number of MS traits. ACR increased four-fold between subjects with no MS traits and those with four or more traits. In subjects with the metabolic syndrome, there was a significant correlation between ACR and systolic blood pressure (SBP), diastolic blood pressure (DBP) and fasting glucose. Estimated GFR correlated significantly and inversely with body mass index (BMI) and serum leptin. These observations raise major clinical and public health concerns for developing countries, where both the metabolic syndrome and kidney disease are being reported more and more frequently. The potential economic impact is huge. PMID- 18158710 TI - Atherosclerosis in type 2 diabetes: a role for fibrate therapy? AB - Vascular complications associated with type 2 diabetes confer significant morbidity and mortality. Atherosclerosis develops much earlier and progresses more rapidly than in subjects without diabetes. The clustering of cardiovascular risk factors associated with type 2 diabetes is mainly responsible for accelerated atherosclerotic disease. While statins remain the primary lipid modifying therapy, the pharmacological profile of the fibrates suggests potential as an alternative or additional treatment for reducing the risk of atherosclerotic vascular complications in type 2 diabetes. PMID- 18158711 TI - Waist circumference in type 2 diabetes: is routine measurement helpful? PMID- 18158712 TI - A novel quantitative method for analyzing the distributions of nanoparticles between different tissue and intracellular compartments. AB - The penetration, translocation, and distribution of ultrafine and nanoparticles in tissues and cells are challenging issues in aerosol research. This article describes a set of novel quantitative microscopic methods for evaluating particle distributions within sectional images of tissues and cells by addressing the following questions: (1) is the observed distribution of particles between spatial compartments random? (2) Which compartments are preferentially targeted by particles? and (3) Does the observed particle distribution shift between different experimental groups? Each of these questions can be addressed by testing an appropriate null hypothesis. The methods all require observed particle distributions to be estimated by counting the number of particles associated with each defined compartment. For studying preferential labeling of compartments, the size of each of the compartments must also be estimated by counting the number of points of a randomly superimposed test grid that hit the different compartments. The latter provides information about the particle distribution that would be expected if the particles were randomly distributed, that is, the expected number of particles. From these data, we can calculate a relative deposition index (RDI) by dividing the observed number of particles by the expected number of particles. The RDI indicates whether the observed number of particles corresponds to that predicted solely by compartment size (for which RDI = 1). Within one group, the observed and expected particle distributions are compared by chi-squared analysis. The total chi-squared value indicates whether an observed distribution is random. If not, the partial chi-squared values help to identify those compartments that are preferential targets of the particles (RDI > 1). Particle distributions between different groups can be compared in a similar way by contingency table analysis. We first describe the preconditions and the way to implement these methods, then provide three worked examples, and finally discuss the advantages, pitfalls, and limitations of this method. PMID- 18158713 TI - Performance of the vibrating membrane aerosol generation device: Aeroneb Micropump Nebulizer. AB - The output and particle size distribution of several series of aqueous solutions were measured to define quantitatively the practical limits of the solution properties acceptable for aerosol production by the aeroneb micropump nebulizer. Aerosol output measurements were made gravimetrically and the particle size distributions were obtained by laser diffractometry. Solution properties were obtained from the literature by interpolation of the best-fit curve of the property plotted as a function of composition. For nonionic solutes, addition of sodium chloride dramatically increased the output rate and also decreased the droplet size at low solute concentrations. Increasing viscosity also caused a significant decrease in output. Cesium chloride displayed increased output rate with concentration due to the rising density. Based on calculations with the number of apertures and oscillatory frequency, low output rates appeared to be a consequence of apertures failing to produce a droplet with each oscillation. Overall, ionic strength, density, surface tension, and viscosity affected the output rate in a manner that can be now empirically predicted. PMID- 18158714 TI - Preclinical safety evaluation of inhaled cyclosporine in propylene glycol. AB - Cyclosporine inhalation solution has the potential to improve outcomes following lung transplantation by delivering high concentrations of an immunosuppressant directly to the allograft while minimizing systemic drug exposure and associated toxicity. The objective of these studies was to evaluate the potential toxicity of aerosolized cyclosporine formulated in propylene glycol when given by inhalation route to rats and dogs for 28 days. Sprague-Dawley rats received total inhaled doses of 0 (air), 0 (vehicle, propylene glycol), 7.4, 24.3, and 53.9 mg cyclosporine/kg/day. In a separate study, beagle dogs were exposed to 0, 4.4, 7.7, and 9.7 mg cyclosporine/kg/day. Endpoints used to evaluate potential toxicity of inhaled cyclosporine were clinical observations, body weight, food consumption, respiratory functions, toxicokinetics, and clinical/anatomic pathology. Daily administration of aerosolized cyclosporine did not result in observable accumulation of cyclosporine in blood or lung tissue. Toxicokinetic analysis from the rat study showed that the exposure of cyclosporine was approximately 18 times higher in the lung tissue compared to the blood. Systemic effects were consistent with those known for cyclosporine. There was no unexpected systemic toxicity or clinically limiting local respiratory toxicity associated with inhalation exposure to cyclosporine inhalation solution at exposures up to 2.7 times the maximum human exposure in either rats or dogs. There were no respiratory or systemic effects of high doses of propylene glycol relative to air controls. These preclinical studies demonstrate the safety of aerosolized cyclosporine in propylene glycol and support its continued clinical investigation in patients undergoing allogeneic lung transplantation. PMID- 18158715 TI - Reliability in the assessment of videotaped inhalation technique. AB - Inhalation medication is essential in the treatment of asthma and chronic obstructive pulmonary disease (COPD) patients. Incorrect inhalation technique reduces the effects of medication and has been reported to range from 22% to 95% from optimal. The objective of this study was to determine inter- and intraobserver reliability in inhalation technique assessment. For interobserver reliability three observers scored after three times viewing a total of 49 video recorded inhalation demonstrations using device-specific checklists and mutually agreed scoring rules. Intraobserver reliability was assessed for two observers after 8 months by scoring inhalation demonstrations a second time. Both inter- and intraobserver reliability were expressed by mean percent agreement and mean Kappa scores. All inhaler devices revealed a high mean percent agreement and a substantial or almost perfect Kappa scoring for both inter- and intraobserver reliability. Only one item, "exhale to residual volume," showed poor intraobserver reliability. Assessment of video recorded inhalation technique using device-specific checklists, triple viewing, and mutual agreed scoring rules is reliable. This method enables blind observation of inhalation technique. PMID- 18158716 TI - The effects of Heliox on the output and particle-size distribution of salbutamol using jet and vibrating mesh nebulizers. AB - There are theoretical benefits of delivering drug aerosols to patients with asthma and chronic obstructive pulmonary disease (COPD) using Heliox as a carrier gas. The objective of this study was to develop systems to allow bronchodilators nebulized by a breath enhanced jet nebulizer and a vibrating mesh nebulizer to be delivered to patients in Heliox. This was achieved by attaching a reservoir to the nebulizers to ensure inhaled Heliox was not diluted by entrained air. For the vibrating mesh nebulizer, the total output was significantly higher after 5 min of nebulization when Heliox rather than air was used as the delivery gas (p < 0.001). The proportion of drug in particles <5 microm was 58.1% for Heliox and 50.1% when air was entrained. When the breath enhanced nebulizer was used a much higher driving flow of Heliox, compared to air, was required to deliver a similar dose of drug (p < 0.05). The total amount of drug likely to be inhaled was significantly higher when the vibrating mesh nebulizer (Aerogen) was used compared to the breath enhanced jet nebulizer (Pari LC plus) (p < 0.001). The amount of drug likely to be inhaled was also significantly greater for the adult as opposed to pediatric breathing pattern for all nebulizers and flows tested with the exception of the Aeroneb and Heliox entrainment. In this case, total amounts were similar for both patterns but for the pediatric pattern, the time taken to reach this output was longer. Such information is required to allow appropriate interpretation of clinical trials of drug delivery using Heliox. PMID- 18158717 TI - IL-1 receptor antagonist as an aerosol in inflammation. AB - The feasibility of efficient aerosol delivery of the human IL-1 receptor antagonist (IL-1Ra) for reduction of acute lung inflammation was demonstrated in a mouse model study. The therapeutic efficacy of dry powder formulations PM(2), PM(10) of IL-1Ra was studied at nonforced inhalation in an aerosol chamber using the DPI "Spinhaler". Micronized powder formulations for insufflation were produced by air-jet milling. The anti-inflammatory effect of IL-Ra preparation was assessed by differential cell counts and biochemical composition of bronchoalveolar lavage (BAL). Reactive oxygen species (ROS), metabolic parameters of BAL (pH, redox potential, total protein, and lactate), and morphological lung changes were investigated by methods of luminol-dependent chemoluminescence, electrochemistry, microscopy, optical, and NMR spectroscopy. Inhalation of IL-1Ra aerosol ensured the systemic absorption of IL-1Ra in the circulatory system and reduced the acute inflammatory response to intranasal lipopolysaccharide challenge. The inhaled anti-inflammatory dosage in aerosol administration appeared to be comparable with i.p. injection. The mechanism of positive action of pulmonary aerosol delivery of Il-Ra includes normalization of the oxidative activity of bronchoalveolar cells, prevention of neutrophil recruitment to the bronchoalveolar tract, and improving of cell respiration. The results were used to develop mathematical models of the anti-inflammatory effects of IL-Ra as functions of the doses and dispersion grades of IL-Ra preparations. Aerosol application of IL-Ra may be an apparent way for prophylactic treating of respiratory inflammation caused by bacterial antigens. PMID- 18158718 TI - Design optimization of a novel pMDI actuator for systemic drug delivery. AB - Pressurized metered dose inhalers (pMDIs) are the most widely prescribed and economical respiratory drug delivery systems. Conventional pMDI actuators-those based on "two-orifice-and-sump" designs-produce an aerosol with a reasonable respirable fraction, but with high aerosol velocity. The latter is responsible for high oropharyngeal deposition, and consequently low drug delivery efficiency. Kos' pMDI technology is based on a proprietary vortex nozzle actuator (VNA), an innovative actuator configuration that seeks to reduce aerosol plume velocity, thereby promoting deep lung deposition. Using VNA development as a case study, this paper presents a systematic design optimization process to improve the actuator performance through use of advanced optical characterization tools. The optimization effort mainly relied on laser-based optical diagnostics to provide an improved understanding of the fundamentals of aerosol formation and interplay of various geometrical factors. The performance of the optimized VNA design thus evolved was characterized using phase Doppler anemometry and cascade impaction. The aerosol velocities for both standard and optimized VNA designs were found to be comparable, with both notably less than conventional actuators. The optimized VNA design also significantly reduces drug deposition in the actuator as well as USP throat adapter, which in turn, leads to a significantly higher fine particle fraction than the standard design (78 +/- 3% vs. 63 +/- 2% on an ex valve basis). This improved drug delivery efficiency makes VNA technology a practical proposition as a systemic drug delivery platform. Thus, this paper demonstrates how advanced optical diagnostic and characterization tools can be used in the development of high efficiency aerosol drug delivery devices. PMID- 18158719 TI - Comparison of three carbon monoxide monitors for determination of smoking status in smokers and nonsmokers with and without COPD. AB - In this (CAMOXI) study, three carbon monoxide (CO) monitors and salivary cotinine are assessed regarding their ability to distinguish smokers from nonsmokers, both in chronic obstructive pulmonary disease (COPD) and healthy people. Twenty-six healthy smokers, 25 healthy nonsmokers, 25 smoking, and 25 former smoking stable COPD patients (age 40-72 years) were included based on self-report (N = 101). All volunteers were measured following a 12-h abstinence period. Sensitivity, specificity, and predictive values of a positive and negative test result were assessed for a range of cutoff points for both CO and salivary cotinine. The prescribed 9-ppm cutoff point of the Breath CO generates a sensitivity of 68% and 42% for COPD patients and healthy people, respectively. Using the prescribed cutoff point (10 ppm) the Smokelyzer produces 56% sensitivity for COPD patients and 23% for healthy people. Both monitors generate 100% specificity in both groups. The cutoff point for the Micro CO meter (5 ppm) generates 88% sensitivity and 92% specificity for COPD patients, and for healthy people 92% and 88%, respectively. The optimal cutoff points depend upon the goal of the test. Salivary cotinine has a 100% sensitivity, specificity, positive predictive value, and negative predictive value over the range of 15 ng/mL through 40 ng/mL for healthy participants and at 10 ng/mL for COPD patients. The prescribed cutoff points for all three CO monitors generate misleading results concerning the determination of the smoking status in both populations. Salivary cotinine measurement outperforms CO measurements and a combination of the two tools is recommended. PMID- 18158720 TI - The size and concentration of droplets generated by coughing in human subjects. AB - This work investigated the size distribution of the droplet nuclei and coughed droplets by test subjects. The size distributions of droplet nuclei coughed by test subjects were determined with an aerodynamic particle sizer (APS) and scanning mobility particle sizer (SMPS) system (system 1). Coughed droplets were only sampled with the APS system (system 2). Two different schemes were employed in system 2. Furthermore, the size distribution of coughed droplets of different ages and gender was investigated to identify the effects of age and gender on droplet size distribution. Results indicated the total average size distribution of the droplet nuclei was 0.58-5.42 microm, and 82% of droplet nuclei centered in 0.74-2.12 microm. The entire average size distribution of the coughed droplets was 0.62-15.9 microm, and the average mode size was 8.35 microm. The size distribution of the coughed droplets was multimodal. The size distribution of coughed droplets showed three peaks at approximately 1 microm, 2 microm, and 8 microm. These analytical findings indicate that variation for average droplet size among the three age groups was insignificant (p > 0.1). Moreover, the variation in average droplet size between males and females was also insignificant (p > 0.1). Also, the variation in droplet concentration between males and females was significant (p > 0.1). Droplet nuclei concentrations from male subjects were considerably higher than that from females. Comparison of the droplet concentrations for subjects in different age groups demonstrated that subjects in the 30-50-year age group have the largest droplet concentrations. PMID- 18158721 TI - The influence of spray properties on intranasal deposition. AB - While numerous devices, formulations, and spray characteristics have been shown to influence nasal deposition efficiency, few studies have attempted to identify which of these interacting factors plays the greatest role in nasal spray deposition. The deposition patterns of solutions with a wide range of surface tensions and viscosities were measured using an MRI-derived nasal cavity replica. The resulting spray plumes had angles between 29 degrees and 80 degrees and contained droplet sizes (D(v50)) from 37-157 microm. Each formulation contained rhodamine 590 as a fluorescent marker for detection. Administration angles of 30 degrees , 40 degrees , or 50 degrees above horizontal were tested to investigate the role of user technique on nasal deposition. The amount of spray deposited within specific regions of the nasal cavity was determined by disassembling the replica and measuring the amount of rhodamine retained in each section. Most of the spray droplets were deposited onto the anterior region of the model, but sprays with small plume angles were capable of reaching the turbinate region with deposition efficiencies approaching 90%. Minimal dependence on droplet size, viscosity, or device was observed. Changes in inspiratory flow rate (0-60 L/min) had no significant effect on turbinate deposition efficiency. Both plume angle and administration angle were found to be important factors in determining deposition efficiency. For administration angles of 40 degrees or 50 degrees , maximal turbinate deposition efficiency (30-50%) occurred with plume angles of 55 65 degrees , whereas a 30 degrees administration angle gave an approximately 75% deposition efficiency for similar plume angles. Deposition efficiencies of approximately 90% could be achieved with plume angles <30 degrees using 30 degrees administration angles. Both the plume angle and administration angle are critical factors in determining deposition efficiency, while many other spray parameters, including particle size, have relatively minor influences on deposition within the nasal cavity. PMID- 18158722 TI - Bronchial challenge test in asthmatics sensitized to mites: role of particle size in bronchial response. AB - Although major house dust mite allergen (Der p 1) is carried mainly on large particles (>10 microm), standard bronchial challenge tests (BCT) use nebulizers that deliver smaller particles (sizes from 1 to 5 microm) and may therefore not reflect actual domestic exposure. The objective of this study was to evaluate the influence of particle size of Dermatophagoides pteronyssinus extract on bronchial response. Specific BCT were performed with different mass median aerodynamic diameters (MMAD): 1.1, 5.6, and 9.7 microm. Each of the 19 mite-sensitized patients underwent mite BCT three times, once with each nebulizer. IL-5 levels were assessed in induced sputum and blood samples. The PD(20) for Der p 1 differed substantially with particle size, with less Der p 1 (11.2 ng) needed to produce a PD(20) with the largest particles (9.7 microm), compared to 18.1 ng for the 5.6 microm particles and 142.5 ng for the 1.1 microm particles (p < 0.0001). Large particles also induced an early phase response significantly more often than small particles (100% vs. 63%). Although the late phase reaction (LPR) frequency was similar with all three particle sizes, lower mean oral corticosteroid doses were needed to treat LPR with the largest particles (23 mg), compared to the smaller particles, with 34 mg for the 5.6 microm particles and 51 mg for the 1.1 microm. The 1.1 microm particles produced a significantly greater increase in IL-5 concentrations in sputum and blood compared to the larger particles. Large particles clearly play a role in the immediate bronchial response in asthmatics sensitized to mites and, therefore, should be included in pharmacological studies in humans. PMID- 18158723 TI - A simple mechanistic model of deposition of water-soluble aerosol particles in the mouth and throat. AB - Aerosol drugs are usually delivered to the lung by inhalation via the oral route, since aerosol deposition is much lower in the oral than in nasal airways. In the present study a practical, non-CFD-based, mechanistic model is developed, which permits an efficient calculation of deposition along the oral route with simple computational means. A simplified geometrical description of the mouth and throat region is used, based on a sequence of conducting ducts. The numerical model takes into account aerosol dynamics, which enables to express the impact on aerosol transport and deposition of the hygroscopic growth of water-soluble particles. Simulations are made for coarse particles in the range 1-17 microm, and the model predictions are found in good agreement with the available experimental data. The model predicts inertial impaction to be the dominant mechanism, and correctly reproduces the increase in the deposition with an increasing flow rate and particle diameter. Higher deposition is calculated in the oropharyngeal region than the laryngeal region, due to the significant flow direction change and constriction at the end of the oral cavity. According to the model, highly soluble particles may deposit up to 50% more than inert aerosols in the mouth-throat region. The proposed model will be useful for quick, practical calculations of deposition with a full account of aerosol dynamical processes. PMID- 18158725 TI - Role of complement in immune regulation and its exploitation by virus. AB - Complement is activated during the early phase of viral infection and promotes destruction of virus particles as well as the initiation of inflammatory responses. Recently, complement and complement receptors have been reported to play an important role in the regulation of innate as well as adaptive immune responses during infection. The regulation of host immune responses by complement involves modulation of dendritic cell activity in addition to direct effects on T cell function. Intriguingly, many viruses encode homologs of complement regulatory molecules or proteins that interact with complement receptors on antigen-presenting cells and lymphocytes. The evolution of viral mechanisms to alter complement function may augment pathogen persistence and limit immune mediated tissue destruction. These observations suggest that complement may play an important role in both innate and adaptive immune responses to infection as well as virus-mediated modulation of host immunity. PMID- 18158726 TI - Meeting review: options for the control of influenza VI. AB - A pandemic outbreak of viral influenza could occur if highly virulent zoonotic strains (such as avian H5N1) were to acquire the ability to transmit among humans. This possibility has increased concern, interest, and investigation in the influenza field. This heightened interest was reflected in the participation of numerous health officials, clinicians, and scientific investigators in the Sixth Conference on the Options for the Control of Influenza held in Toronto, Ontario, Canada from June 17-23, 2007. A recurring theme was that H5N1 viruses and the now-available reassortant/recombinant viruses from the 1918 pandemic have striking pathological and molecular similarities. Understanding these similarities, and how the viruses differ from currently circulating strains will be critical for timely preparation of the appropriate vaccines and antiviral therapies if a new pandemic should arise. To that end, this international conference was a fully comprehensive exchange of information covering public health, pandemic planning/preparedness, current and developing vaccines, animal ecology/zoonosis, mathematical modeling, virology, and viral immunology. In this review I will focus on basic research of pathogen-host interactions and immune responses to influenza A virus reported at this conference. PMID- 18158727 TI - Type I interferon regulates respiratory virus infected dendritic cell maturation and cytokine production. AB - Activation of dendritic cells (DCs) by viruses is critical for both innate and adaptive immune responses. In this report, we investigated the role of type I interferon (IFN) in the activation of DCs by respiratory syncytial virus (RSV). Using DCs from type I IFNR-/- mice, these studies indicate that maturation, including upregulation of co-stimulatory molecules and optimal cytokine production, by RSV infection was dependent on type I IFN receptor signaling. Subsequently, studies using DCs from wild type mice demonstrate that continued production of type I IFN during later stages of DC maturation could alter their activation profiles. IFN-alpha and IFN-beta were upregulated in DCs grown from bone marrow of wild type mice after infection with RSV. In order to determine their function in competent DCs, blocking antibodies were used to specifically inhibit IFN-alpha/beta . The data demonstrate that production of IFN-beta, but not IFN-alpha, in RSV-infected wild type DCs promotes chemokine production and toll-like receptor (TLR) expression, while limiting IL-12 production. The inhibition of IL-12p70 by IFN-beta correlated with suppressed IL-12p40 expression levels. Furthermore, the addition of recombinant IFN-beta potently inhibited IL 12p40 expression in mature DC subsets during RSV infection, while only the highest dose of IFN-alpha had any inhibitory effect. Together, our studies provide insight into the complex regulation of DC maturation and IL-12 production co-ordinated by type I interferons in RSV-infected dendritic cells, and demonstrate that type I IFN has specific roles depending upon the stage of DC maturation. PMID- 18158728 TI - Lack of in vitro and in vivo effects of lipopolysaccharide on porcine circovirus type 2 infection. AB - Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS). The presence of immunostimulating factors or concurrent infections seems to be crucial for PMWS development. Lipopolysaccharide (LPS) is a potent immunological activator and has recently been suggested to enhance PCV2 replication in vitro. This study was designed to evaluate the effects of different LPS products on PCV2 in vitro replication of pulmonary macrophages (PMs), and on the potential ability to trigger PMWS in cesarean-derived, colostrum-deprived (CDCD) PCV2-inoculated piglets. In vitro studies using two different PCV2 isolates (Stoon-1010 and 1452/3) showed the presence of PCV2 antigen within the cytoplasm to a variable degree; PCV2 Stoon 1010 was barely detectable (<1% of stained cells), and PCV2 1452/3 was seen in the cytoplasm of more than 85% of PMs. However, no differences were found in intracytoplasmic PCV2 signals among different LPS treatments, or between the LPS treated and non-treated PMs. Moreover, almost no intranuclear signals for PCV2 antigen were detected in PMs. The in vivo experiment included twenty 7-day-old CDCD piglets divided into four groups: control (n = 4), control/LPS (n = 4), PCV2 (n = 6), and PCV2/LPS (n = 6). The control and control/LPS groups were inoculated intranasally with a cell culture medium (MEM), and the PCV2 and PCV2/LPS groups were inoculated with a Spanish isolate of PCV2 (Burgos). The control/LPS and PCV2/LPS groups were inoculated intraperitoneally with LPS on PCV2 inoculation day. All pigs remained clinically healthy during the entire experimental period (29 days). Animals inoculated with LPS had significant hyperthermia within the first 24 hours post-inoculation. No differences in gross or histological findings were observed among the PCV2 and PCV2/LPS inoculated pigs. All PCV2-infected piglets developed a subclinical infection with the virus. Our results showed that LPS did not increase in vitro viral replication and did not trigger PMWS in PCV2 inoculated pigs. PMID- 18158729 TI - The functional evaluation of dendritic cell vaccines based on different hepatitis C virus nonstructural genes. AB - Hepatitis C virus (HCV) nonstructural (NS) genes are relatively conserved and play critical roles in cellular immune responses against HCV. The aim of the study was to evaluate the immunogenicity of the different HCV NS genes through transduction of DCs and presentation to T cells. Monocyte-derived DCs from healthy donors were infected with the recombinant adenovirus (Ad) harboring HCV NS3 (AdNS3), NS4 (NS4A and NS4B; AdNS4), NS5 (NS5A and NS5B; AdNS5), NS3/NS4 (AdNS3/NS4), and NS4/NS5 (AdNS4/NS5) genes, and then used to stimulate autologous lymphocytes in vitro. Antigen-specific cellular immune responses were detected by interferon-gamma (IFN-gamma), interleukin 4 (IL-4), and Granzyme B (GrB) enzyme linked immunospot assays (ELISPOT). DCs expressing different HCV NS genes all induced positive immune responses. Furthermore, DCs transfected with AdNS3/NS4 were superior to DCs infected with AdNS3 or AdNS4 in inducing HCV-specific immunity. The same results were obtained when we compared DCs infected with AdNS4/NS5 to AdNS4 or AdNS5. DCs transduced with NS3/NS4 or NS4/NS5 had similar ability to elicit specific immune responses to HCV. PMID- 18158730 TI - Mannan-binding lectin (MBL) serum concentration in relation to propagation of infectious bronchitis virus (IBV) in chickens. AB - Mannan-binding lectin (MBL) is a collectin that mediates activation of the complement system and is of importance for host defenses. In humans low concentrations of MBL in serum have been associated with susceptibility to several viral diseases. To understand the function of MBL in relation to infectious viral diseases two chicken lines were selected for high and low concentrations of MBL in serum for several generations. Offspring from the two sub-lines were subjected to infection with infectious bronchitis virus (IBV) in order to determine their genetic susceptibility to the virus. Results suggested that MBL plays a role in the innate immunity against IBV in the way that it performs an acute phase response, is able to activate complement, and inhibits the propagation of the virus in the trachea. PMID- 18158731 TI - IFN-gamma is the only anti-rotavirus cytokine found after in vitro stimulation of memory CD4+ T cells from mice immunized with a chimeric VP6 protein. AB - CD4+ T cells are the only lymphocytes required for protection of mice against rotavirus shedding after mucosal immunization with chimeric VP6 (MBP::VP6) and the adjuvant LT(R192G). One possible effector of protection is CD4+ T-cell cytokines. To determine if memory CD4+ T cells of immunized mice produce cytokines with direct anti-rotavirus activity, an in vitro infection model was developed using mouse CMT-93 cells and rhesus rotavirus (RRV). Spleen and lamina propria (LP) cells, as well as purified splenic CD4T cells obtained after intranasal immunization of BALB/c mice with MBP::VP6/LT(R192G) released large quantities of two cytokines (IL-17 and IFN-gamma) into cell supernatants when stimulated with MBP::VP6. Production of these same cytokines is rapidly upregulated in intestinal lymphocytes after rotavirus inoculation of immunized mice. IL-17 pretreatment of CMT-93 cells had no effect on subsequent RRV replication, but IFN-gamma was the most potent inhibitor within a panel of nine cytokines tested. Supernatants obtained after in vitro stimulation of splenic CD4+ T cells of immunized mice had high levels of anti-RRV activity and their pretreatment with mAb against IFN-gamma caused essentially complete loss of activity. Thus, IFN-gamma was the only cytokine identified in stimulated CD4+ T cells from immunized mice that directly inhibited rotavirus replication. PMID- 18158732 TI - Cross-priming of a single viral protein from lymphocytic choriomeningitis virus alters immunodominance hierarchies of CD8+ T cells during subsequent viral infections. AB - Immunogenic epitopes that stimulate CD8+ T cells can be organized into an immunodominance hierarchy, based on their ability to induce T-cell priming and subsequent expansion. Cytotoxic CD8+ T cells can be primed through the cross priming pathway, where exogenous viral proteins are acquired by professional antigen-presenting cells (pAPCs). We have previously reported that lymphocytic choriomeningitis nucleoprotein (LCMV-NP) expressed in HEK cells (HEK-NP) induces cross-priming of CD8+ T cells in vivo. In this study, we have used this HEK-NP model to study the effects of LCMV-NP cross-priming on the LCMV immunodominance hierarchy following viral challenge. Our results highlight the contribution of cross-priming to the immune response, since the T-cell hierarchy was significantly altered as a result of exogenous processing of a single virus protein, and this phenomenon was maintained throughout the memory response. Moreover, as a result of cross-priming, in vivo CD8+ T-cell killer activity was enhanced during subsequent virus assaults. These findings have significant implications for immunotherapy because they demonstrate that exogenous delivery of specific T-cell epitopes can be utilized to manipulate the host's CD8+ T-cell memory immunodominance responses. PMID- 18158733 TI - CCR1 deficiency increases susceptibility to fatal coronavirus infection of the central nervous system. AB - The role of CC chemokine receptor 1 (CCR1) in host defense and disease development was determined in a model of viral-induced neurologic disease. Intracerebral (IC) infection of mice with mouse hepatitis virus (MHV) results in an acute encephalitis followed by a chronic demyelinating disease similar in pathology to the disease multiple sclerosis (MS). No increase in mortality was observed during the acute phase of disease following MHV infection of mice lacking CCR1 (CCR1-/-) as compared to wild-type (CCR1+/+) mice. However, by 21 d post-infection, 74% of CCR1-/- mice had succumbed to death compared to only 32% mortality of CCR1+/+ mice, indicating that chemokine signaling through CCR1 significantly (p or=160 to dengue 1. We describe the existence of dengue-specific antibodies in the serum of people infected more than 60 years earlier. PMID- 18158742 TI - Understanding and tackling aging: two fields communicating (a little) at last. AB - Those who have followed this journal's, and this author's, efforts over recent years to stimulate the rational design of interventions to combat aging have good reason for bewilderment that the concerted application of our knowledge of biology to the defeat of humanity's foremost killer needed to be kick-started by a bearded troublemaker whose formal academic training was not even in biology at all. Elsewhere in this issue I bemoan the persistent balkanisation of traditional gerontology, whereby biologists, clinicians, sociologists and psychologists studying the elderly seem almost studiously to avoid each other even when participating in the same conference. In this commentary, however, I have something more positive to report. A string of recent and forthcoming conferences, organized not only by those at the forefront of life-extension research but also by highly influential mainstream groups, have publicly endorsed the Methuselah Foundation's goal of defeating aging. The field of biomedical gerontology-the interface between biogerontology and geriatrics, where biological knowledge is focused on developing the geriatrics of tomorrow-is not a traditional component of gerontology, having been poorly appreciated by biogerontologists and geriatricians alike, but these developments show that it is rapidly taking its place at that table. PMID- 18158743 TI - Dissertations: a survey of selected recent theses relevant to combating aging. PMID- 18158744 TI - The Glenn Foundation for Medical Research, Santa Barbara, CA: interview with Paul F. Glenn and Mark R. Collins. Interview by Vicki Glaser. PMID- 18158746 TI - Editorial: good vibrations: more evidence of reciprocity? PMID- 18158747 TI - Effect of low-level laser therapy on inflammatory reactions during wound healing: comparison with meloxicam. AB - OBJECTIVE: This study evaluated the action of low-level laser therapy (LLLT) on the modulation of inflammatory reactions during wound healing in comparison with meloxicam. BACKGROUND DATA: LLLT has been recommended for the postoperative period because of its ability to speed healing of wounds. However, data in the literature are in disagreement about its anti-inflammatory action. METHODS: Standardized circular wounds were made on the backs of 64 Wistar rats. The animals were divided into four groups according to the selected postoperative therapy: group A-control; group B-administration of meloxicam; and groups C and D irradiation with red (lambda = 685 nm) and infrared (lambda = 830 nm) laser energy, respectively. The animals were killed at 12, 36, and 72 h and 7 days after the procedure. RESULTS: Microscopic analysis revealed significant vascular activation of irradiated sites in the first 36 h. Only group B showed decreases in the intensity of polymorphonuclear infiltrates and edema. Group D showed a higher degree of organization and maturation of collagen fibers than the other groups at 72 h. The animals in group C showed the best healing pattern at 7 days. The anti-inflammatory action of meloxicam was confirmed by the results obtained in this research. The quantification of interleukin-1beta (IL-1beta) mRNA by real time polymerase chain reaction (PCR) did not show any reduction in the inflammatory process in the irradiated groups when compared to the other groups. CONCLUSIONS: LLLT improves the quality of histologic repair and is useful during wound healing. However, with the methods used in this study the laser energy did not minimize tissue inflammatory reactions. PMID- 18158748 TI - Effectiveness of helium-neon laser irradiation on viability and cytotoxicity of diabetic-wounded fibroblast cells. AB - OBJECTIVE: This study investigated the effectiveness of helium-neon (He-Ne) laser irradiation at increasing intervals on diabetic-induced wounded human skin fibroblast cells (WS1) at a morphological, cellular, and molecular level. BACKGROUND DATA: The controversies over light therapy can be explained by the differing exposure regimens and models used. No therapeutic window for dosimetry and mechanism of action has been determined at the level of individual cell types, particularly in diabetic cells in vitro. METHODS: WS1 cells were used to simulate an in vitro wounded diabetic model. The effect of the frequency of He-Ne irradiation (632.8 nm) at a fluence of 5 J/cm(2) was determined by analysis of cell morphology, viability, cytotoxicity, and DNA damage. Cells were irradiated using three different protocols: they were irradiated at 30 min only; irradiated twice, at 30 min and at 24 h; or irradiated twice, at 30 min and at 72 h post wound induction. RESULTS: A single exposure to 5 J/cm(2) 30 min post-wound induction increased cellular damage. Irradiation of cells at 30 min and at 24 h post-wound induction decreased cellular viability, cytotoxicity, and DNA damage. However, complete wound closure as well as an increase in viability and a decrease in cytotoxicity and DNA damage occurs when cells were irradiated at 30 min and at 72 h post-wound induction. CONCLUSIONS: Wounded diabetic WS1 cells irradiated to 5 J/cm(2) showed increased cellular repair when irradiated with adequate time between irradiations, allowing time for cellular response mechanisms to take effect. Therefore, the irradiation interval was shown to play an important role in wound healing in vitro and should be taken into account. PMID- 18158749 TI - Use of near-infrared Raman spectroscopy for identification of atherosclerotic plaques in the carotid artery. AB - OBJECTIVES: The aim of this work was to identify the presence of atherosclerotic plaque in the human carotid artery using near infrared Raman spectroscopy. BACKGROUND DATA: Atherosclerosis is the most common and serious pathology of the cardiovascular system. Raman spectroscopy is an analytical tool that can be used to gather information about both the morphology and chemical composition of tissues. METHODS: A Ti:sapphire laser operating at the near-infrared wavelength of 830 nm pumped by an argon laser was used for excitation of the samples, and the Raman scattering was detected by an optical spectrometer with a liquid nitrogen-cooled CCD detector. Carotid artery samples were classified into five groups: normal, intimal thickening, fatty plaque, fibrous-fatty plaque, and fibrous-calcified plaque. RESULTS: It was observed that the Raman spectrum of atheromatous plaque was different that that of normal tissue. The spectra of atheromatous plaques had bands due to the presence of cholesterol and its esters, with major bands at 1439 and 1663 cm(1), respectively. In normal tissues a peak related to C-H bending appears at 1451 cm(1). Calcified atheromatous plaques had primary bands at 961 and 1071 cm(1), which were due to the presence of phosphate and carbonate in the accumulated calcium. Peaks were seen at 1451 and 1655 cm(1) in the non-atherosclerotic tissue, which were shifted to 1439 and 1663 cm(1) in the atherosclerotic plaque. CONCLUSIONS: Our results indicate that this technique could be used to detect the presence of atherosclerotic plaques in carotid arterial tissue. PMID- 18158750 TI - Effect of lower-level laser therapy on rabbit tibial fracture. AB - OBJECTIVE: The purpose of the study was to demonstrate the biological effects of low-level laser therapy (LLLT) on tibial fractures using radiographic, histological, and bone density examinations. METHODS: Fourteen New Zealand white rabbits with surgically induced mid-tibial osteotomies were included in the study. Seven were assigned to a group receiving LLLT (LLLT-A) and the remaining seven served as a sham-treated control group (LLLT-C). A low-energy laser apparatus with a wavelength of 830 nm, and a sham laser (a similar design without laser diodes) were used for the study. Continuous outflow irradiation with a total energy density of 40 J/cm(2) and a power level of 200 mW/cm(2) was directly delivered to the skin for 50 seconds at four points along the tibial fracture site. Treatment commenced immediately postsurgery and continued once daily for 4 weeks. RESULTS: Radiographic findings revealed no statistically significant fracture callus thickness difference between the LLLT-A and LLLT-C groups (p > 0.05). However, the fractures in the LLLT-A group showed less callus thickness than those in LLLT-C group 3 weeks after treatment. The average tibial volume was 14.5 mL in the LLLT-A group, and 11.25 mL in the LLLT-C group. The average contralateral normal tibial volume was 7.1 mL. Microscopic changes at 4 weeks revealed an average grade of 5.5 and 5.0 for the LLLT-A group and the LLLT-C group, respectively. The bone mineral density (BMD) as ascertained using a grey scale (graded from 0 to 256) showed darker coloration in the LLLT-A group (138) than in the LLLT-C group (125). CONCLUSION: The study suggests that LLLT may accelerate the process of fracture repair or cause increases in callus volume and BMD, especially in the early stages of absorbing the hematoma and bone remodeling. Further study is necessary to quantify these findings. PMID- 18158751 TI - Suitable conditions for sealing of open dentinal tubules using a pulsed Nd:YAG laser. AB - BACKGROUND DATA AND OBJECTIVES: Nd:YAG laser radiation has been applied in treatment of dentine hypersensitivity; dentine melt created after irradiation on a tooth surface can occlude open dentinal tubules. Different conditions (energy of pulses, number of successive irradiations, and type of dye agent) have been reported in the literature. In our study, different conditions are compared and the most suitable ones are suggested. METHODS: One hundred nine dentine samples prepared from extracted human third molars in the form of 2- or 3-mm thick discs and facets from an outer convex surface of a root were irradiated by pulses with duration 0.3 ms and beam diameter 0.3 or 0.6 mm. The laser repetition rate was 15 Hz. Energies per pulse have been changed from 20 mJ to 250 mJ (the corresponding energy densities rho are 22 and 275 J/cm(2) for the beam diameter of 0.3 mm). Dyes including tartrazine, carmine indigo, erythrosin, and methylene blue have been used to cover dentine surfaces and to increase absorption of laser radiation. Irradiated dentine surfaces have been investigated using scanning electron microscopy. RESULTS: Sealing of open dentinal tubules can be accomplished even without dyes, provided that a sufficiently intense laser radiation level is used. To avoid damage to the dentine's surface the use of dyes is necessary. Erythrosin has been found to be the best agent, and closing of dentinal tubules occurred after four doses of irradiation with 30-mJ pulses with a beam diameter of 0.6 mm (energy density rho = 8.25 J/cm(2), total energy density rho(T) = 33 J/cm(2)). The upper limit has been reached using four doses of irradiation by 90-mJ pulses with a beam diameter of 0.3 mm (rho = 99 J/cm(2), rho(T) = 396 J/cm(2)). CONCLUSIONS: Occluding dentinal tubules can safely and effectively be accomplished using pulsed Nd:YAG laser radiation provided that the dentine surface is covered with erythrosin solution. PMID- 18158752 TI - Surface morphology of sound deciduous tooth enamel after application of a photo absorbing cream and infrared low-level laser irradiation: an in vitro scanning electron microscopy study. AB - OBJECTIVES: The purpose of this descriptive scanning electron microscopic study was to characterize surface alterations in deciduous tooth enamel after in vitro infrared diode laser irradiation, using a photo-absorbing agent alone and also combined with fluoride, before and after laser irradiation. BACKGROUND DATA: Previous investigations have demonstrated increased enamel caries resistance after laser irradiation. METHODS: Seven extracted or exfoliated primary molar teeth underwent soft tissue debridement and fluoride-free prophylaxis. Buccal surfaces were determined to be caries free by macroscopic examination. Sample groups were divided into: (1) control (no treatment); (2) infrared diode laser irradiation (lambda = 810 nm, 68 nm, 60 mW/mm(2), 30 W) using the photo-absorbing agent alone (IRDL + PA; 500 J/cm(2)); and (3) infrared diode laser irradiation using a photo-absorbing agent combined with 2% fluoride (IRDL + PFA; 500 J/cm(2)). Buccal surfaces were evaluated following standard scanning electron microscopy preparation techniques. Control samples of enamel surfaces were relatively smooth but presented occasional enamel prism ends. There were no areas with cavitations or surface defects. RESULTS: After the IRDL + PA treatment, irradiated surfaces became rough and mildly to moderately irregular with scarce enamel cavitations and without exposure of enamel prism ends. The surfaces had adherent granules and only occasional fine cracks and porosities in surface coatings were noted. After the IRDL + PFA treatment, there was a homogenous confluent surface that masked typical enamel surface markings. The surfaces had well-defined globules resulting from the IRDL + PFA treatment, that were not seen after IRDL + PA treatment. CONCLUSIONS: Treatment of deciduous tooth enamel with infrared diode laser irradiation using a photo-absorbing agent and a photo absorbing agent combined with 2% fluoride created surface coatings that may act as reservoirs for mineral phases during cariogenic activity on enamel, and also provide a certain degree of protection against cariogenic challenge. PMID- 18158753 TI - The influence of Er:YAG laser conditioning versus self-etching adhesives with acid etching on the shear bond strength of orthodontic brackets. AB - OBJECTIVE: The aim of this study was to compare the shear bond strength (SBS) of orthodontic brackets after acid etching, laser ablation, self-etching primer application, and combined treatments. METHODS: One hundred defect-free premolars were randomly assigned to five groups. The teeth received the following treatments: group 1: phosphoric acid + Transbond XT primer; group 2: Transbond Plus Self-Etching Primer (TPSEP); group 3: Er:YAG laser ablation + Transbond XT primer; group 4: Er:YAG laser ablation + phosphoric acid + Transbond XT primer; and group 5: Er:YAG laser ablation + TPSEP. Transbond XT Adhesive Paste was used in all groups for bracket bonding. Samples were stored in deionized water at 37 degrees C for 24 h. The shear debonding test was performed at a crosshead speed of 5 mm/min. Failed brackets were examined by a stereomicroscope at 16 x magnification to determine the bond failure interface using a modified adhesive remnant index (ARI). SBS values were compared using one-way analysis of variance (ANOVA). The Kruskal-Wallis test was used for ARI scores and pairwise comparisons were done using the Mann-Whitney U test with Bonferroni correction. RESULTS: Mean SBS values (MPa) and standard deviations for the groups were: group 1, 13.18 +/- 3.45; group 2, 13.52 +/- 4.38; group 3, 10.33 +/- 4.74; group 4, 13.00 +/- 3.47; and group 5, 12.46 +/- 6.31. No significant differences were observed in the SBS of all the groups evaluated, but there was a significant difference in ARI scores, with group 3 showing higher ARI scores than groups 1 and 4. CONCLUSIONS: Er:YAG laser ablation and self-etching primer application showed similar SBS values as conventional acidetching for bonding brackets. PMID- 18158754 TI - The impact of photodynamic therapy on the viability of Streptococcus mutans in a planktonic culture. AB - OBJECTIVE: This study investigated the effect of photodynamic therapy (PDT) with toluidine blue O (TBO) and a light-emitting diode (LED) on the viability of Streptococcus mutans cells in a planktonic culture. BACKGROUND DATA: Growth of Streptococcus mutans is the first step in the development of tooth decay. The use of light and dyes promotes cellular death in a noninvasive way, reducing treatment time. METHODS: The LED used in this study had output power of 116 mW, its energy was 21 J, and the fluency was 2.18 J/cm(2). Samples were prepared and divided into five groups: (1) control group (-); (2) control group (+); (3) TBO; (4) LED; and (5) LED + TBO. RESULTS: One hundred percent of the bacteria were killed following irradiation with LED and TBO. The biofilm that formed on the glass surfaces was analyzed by SEM and colony count. CONCLUSIONS: It was demonstrated that PDT was efficient at killing microorganisms and preventing the formation of biofilms. PMID- 18158755 TI - Effect of low-level laser therapy on skin fibroblasts of streptozotocin-diabetic rats. AB - OBJECTIVE: This study explored the effects of low-level laser therapy (LLLT) on cellular changes in cell culture and organ culture of skin from streptozotocin diabetic (STZ-D) rats. BACKGROUND DATA: Growth of skin and its fibroblasts are impaired in diabetes. Therefore the healing of skin wounds is impaired in diabetic patients. The positive effects of LLLT on complications of diabetes in patients and animal models have been shown. METHODS: Diabetes was induced in rats by streptozotocin 30 days after its injection. Two sets of skin samples were extracted from skin under sterile conditions. Fibroblasts that were extruded from the samples were proliferated in vitro, and another set of samples were cultured as organ culture. A 24-well culture medium containing Dulbecco's modified minimum essential medium was supplemented by 12% fetal bovine serum. There were five laser-treated and five sham-exposed groups. A helium-neon laser was used, and 0.9 4 J/cm(2) energy densities were applied four times to each organ culture and cell culture. The organ cultures were analyzed by light microscopy and transmission electron microscopy examinations. Cell proliferation was evaluated by dimethylthiazol-diphenyltetrazolium bromide (MTT) assay. RESULTS: Statistical analysis revealed that 4-J/cm(2) irradiation significantly increases the fibroblast numbers compared to the sham-exposed cultures (p = 0.046). CONCLUSION: It is concluded that LLLT resulted in a significant increase of fibroblast proliferation of STZ-D rats in vitro. PMID- 18158756 TI - Accumulation of protoporphyrin-IX in rat Leydig cells following induction by 5 aminolevulinic acid and tramadol. AB - OBJECTIVES: This study aimed to monitor the accumulation of endogenous protoporphyrin-IX (PpIX) in rat Leydig cells (R(2)C) under the effect of 5 aminolevulinic acid (ALA) and various concentrations of tramadol, an analgesic drug. BACKGROUND DATA: Pain during photodynamic treatment with ALA is one of the adverse effects of this new treatment to eradicate tumor cells. ALA is utilized in photodynamic diagnosis and therapy (PDT) as a compound capable of augmenting the intracellular pool of PpIX, which exhibits properties of a photosensitizer. METHODS: Cellular content of PpIX was determined following incubation of the cells for 1 and 2 h in culture medium that contained ALA and different concentrations of tramadol. The amount of PpIX was determined using fluorescent technique under a confocal microscope (laser wavelength 458 nm and filter LP 585 nm), and evaluated using CytFlu 1.2 software. RESULTS: After 1 h of incubation, no significant alterations were noted in the cellular PpIX concentration. However, 2 h of incubation resulted in a significant increase (p < 0.05) in PpIX fluorescence inside the cells, when the medium contained ALA and tramadol in concentrations ranging from 1-2 mg/1 mL. CONCLUSIONS: The results suggested that in R(2)C cells, exogenous ALA and tramadol induced protoporphyrin accumulation. This information is useful for two reasons. First, it may help to diminish pain after ALA-PDT treatment; and second, it allows the use of lower concentrations of ALA during therapy. PMID- 18158757 TI - Letter to the editor: inadequate statistical analysis hides significant effect of low level laser therapy in carpel tunnel syndrome. PMID- 18158760 TI - Lipoic acid suppression of neutrophil respiratory burst: effect of NADPH. AB - Lipoic acid (LA) and its reduced product dihydrolipoic acid (DHLA) are potent antioxidants with capacity to scavenge reactive oxygen species (ROS) and recycle endogenous antioxidants. LA may increase cellular glutathione (GSH), an antioxidant lacking in the lung's epithelial lining fluid in lung disorders such as idiopathic pulmonary fibrosis (IPF). Neutrophils (PMN) are key innate responders and are pivotal in clearing bacterial infection, therefore it is crucial to understand the impact LA may have on their function. Circulating neutrophils were isolated from healthy volunteers and pretreated with LA or diluent. Cells were subsequently activated with phorbol 12-myristate 13-acetate (PMA, 100 ng/ml) to induce ROS production. SOD-inhibitable reduction of acetylated cytochrome c demonstrated the PMA-dependent respiratory burst was suppressed by LA. Oxygen consumption also was diminished when PMA-stimulated cells were pretreated with LA. PMN respiratory burst was partially restored by addition of NADPH but not other pyridine nucleotides. LA did not inhibit glucose 6-phosphate dehydrogenase activity of PMN. These data together suggest that the reduction of LA to DHLA using cellular NADPH may limit the capacity of the PMN NADPH oxidase to produce superoxide. Further studies will be required to determine if LA can diminish excessive superoxide produced by PMN and/or alveolar macrophages in IPF or relevant disease models in vivo. PMID- 18158761 TI - Cysteine-rich protein reverses weight loss in lung cancer patients receiving chemotherapy or radiotherapy. AB - Oxidative stress plays a role in the tumor-cytotoxic effect of cancer chemotherapy and radiotherapy and also in certain adverse events. In view of these conflicting aspects, a double-blind trial over a 6-month period was performed to determine whether a cysteine-rich protein (IMN1207) may have a positive or negative effect on the clinical outcome if compared with casein, a widely used protein supplement low in cysteine. Sixty-six patients with stage IIIB-IV non-small cell lung cancer were randomly assigned to IMN1207 or casein. Included were patients with a previous involuntary weight loss of > or =3%, Karnofsky status > or =70, and an estimated survival of >3 months. Thirty-five lung cancer patients remained on study at 6 weeks. Overall compliance was not different between treatment arms (42-44% or 13 g/day). The patients treated with the cysteine-rich protein had a mean increase of 2.5% body weight, whereas casein treated patients lost 2.6% (p = 0.049). Differences in secondary endpoints included an increase in survival, hand-grip force, and quality of life. Adverse events were mild or moderate. Further studies will have to show whether the positive clinical effects can be confirmed and related to specific parameters of oxidative stress in the host. PMID- 18158762 TI - The conundrum of personalized cancer management, drug development, and economics. PMID- 18158763 TI - Molecular imaging and personalized medicine: an uncertain future. AB - The Food and Drug Administration has described their view of the role that imaging will play in the approval, and perhaps postapproval, use of new therapeutic drugs. The therapeutic drug industry and regulatory authorities have turned to imaging to help them achieve better efficiency and efficacy. We must extend this initiative by demonstrating that molecular imaging can also improve the efficiency and efficacy of routine treatment with these same drugs. The role of molecular imaging in personalized medicine, using targeted drugs in oncology, is very attractive because of the regional information that it provides (in many cases, with a functional or dynamic component), which cannot be provided by in vitro methods ("regional proteomics"). There is great potential for molecular imaging to play a major role in selecting appropriate patients and providing early proof of response, which is critical to addressing the conflict between the high price of treatment and limited reimbursement budgets. This is a new venture in both molecular imaging and targeted drugs. However, there are various regulatory, financial, and practical barriers that must be overcome to achieve this aim, in addition to the normal scientific challenges of drug discovery. There is an urgent need to reduce the cost (i.e., time and money) of developing imaging agents for routine clinical use. The mismatch between the current regulations and personalized medicine includes molecular imaging and requires the engagement of the regulatory authorities to correct. Therapeutic companies must be engaged early in the development of new targeted drugs and molecular imaging agents to improve the fit between the two drug types. Clinical trials must be performed to generate data that not only shows the efficacy of imaging plus therapy in a medical sense, but also in a financial sense. Molecular imaging must be accepted as not just good science but also as central to routine patient management in the personalized medicine of the future. PMID- 18158764 TI - Role of FDG-PET in the assessment of survival prognosis in melanoma. AB - Positron-emission tomography (PET) employing fluorodeoxyglucose (FDG) has proven to be a highly sensitive and specific diagnostic method in the staging and restaging of various neoplasms, including melanoma, complementing morphologic imaging. FDG uptake has been correlated with proliferation rate, and thus, the degree of malignancy of a given tumor (i.e., grading). Consecutively, a relationship of survival prognosis and the extent of tumor burden as well as degree of FDG accumulation--determined by FDG-PET--has been suggested in various tumors. The aim of this study was to assess the potential of fluor-18-FDG-PET in order to evaluate the survival prognosis in melanoma. Patient data (n=95) were retrospectively analyzed, and the results of functional FDG-PET staging was correlated with survival data. Time of staging (diagnosis of primary versus recurrence) had no statistically significant effect on survival prognosis when patients were matched for pertaining node metastasis (NM) stages. Differences in survival were owing to the presence of metastatic disease rather than time of staging. Tumor (T)-stage (T1-T4) alone had no effect on survival prognosis when patients were matched for NM stages. Differences in survival were also due to higher rates of lymph node (LN) and organ metastases in higher T-stages. Detection of LN metastases (N1M0) had a statistically significant and predominant impact on 5-year survival (N0M0 80% versus N1M0 45%; p<0.01). Additional presence of distant metastases in LN-positive patients (N1M1) had only a statistically insignificant further impact on survival (5-year survival in N1M0 45% versus N1M1 29%; p>0.05). Exclusive presence of organ metastases (N0M1) showed a statistically significant drop of survival with a 5-year survival of 61% in N0M1 versus 80% in N0M0, respectively (p<0.03). Further, the combined presence of LN and distant metastases had the worst prognosis (5-year survival in N1M1 29% versus N0M1 61%; p<0.02). Based on a qualitative 4-point scoring system, patients with malignancy-typical FDG uptake showed an overall 5-year survival of 38%, as compared to patients with malignancy-suspicious lesions (71%; p 3 palpable metastases (sensitivity: 25%), while SPECT was true positive in 23 of 28 cases (sensitivity: 82.1%) and P SPECT in 25 of 28 (sensitivity: 89.3%). SPECT was false negative in 5 patients with nonpalpable 3 metastatic nodes. SPECT should be preferred, significantly improving the sensitivity of planar scintigraphy, especially when using a pinhole collimator. PMID- 18158772 TI - Fractional polynomials in a new metastatic renal carcinoma continuous prognostic index involving histology, laboratory, and clinical predictors. AB - BACKGROUND: We analyzed the effect of histology and various clinical and laboratory predictors in a new continuous prognostic index for metastatic renal cell carcinoma based on fractional polynomials. PATIENTS AND METHODS: We evaluated 322 metastatic renal-cell carcinoma patients treated with subcutaneous recombinant cytokine-based home therapies in consecutive trials. We evaluated papillary histology, along with age, disease localizations, C-reactive protein, and neutrophil count in a new prognostic index, which was based on the multivariable fractional polynomial (MFP) algorithm. RESULTS: The MFP model allowed us to divide patients into three risk groups, using seven selected significant prognostic factors: histology, neutrophils, C-reactive protein, bone metastases, liver metastases, lymph node metastases, and age. Using the multivariable fractional polynomial model, median overall survival for high-, intermediate-, and low-risk patients was 10 months (n=80), 23 months (n=162), and 41 months (n=80) (scheme A; p 95% and the overall radiolabeling time was 160 minutes. The in vitro cell uptake of [ (123)I]2 increased with time. Biodistribution in mice bearing the B16-F10 tumor, after an intravenous injection of [(123)I]2, also showed a time-dependent accumulation of the porphyrin in tumor tissue (10.35 %ID/g at 6 hours). The tumor to-muscle ratio was 3.49, 3.38, 3.07, and 3.13 at 1, 2, 6, and 24 hours, respectively. The gamma-camera images of (123)I-porphyrin demonstrated a high focal accumulation of radioactivity in the B16-F10 melanoma tumor. PMID- 18158779 TI - Prognostic value of bax, cytochrome C, and caspase-8 protein expression in primary osteosarcoma. AB - The prognostic significance of bax, cytochrome c, and caspase-8 in patients with primary osteosarcoma is unknown. We examined the immunohistochemical expression of these genes in 35 surgically treated patients with primary osteosarcoma. Clinicopathological and survival data were correlated with the staining result. Eighteen tissue specimens from non-malignant osseous lesions were used as controls. Bax, cytochrome c, and caspase-8 positive staining was observed in 29 (82.9%), 16 (45.7%), and 0 (0%) patients, respectively, but did not stain any of the 18 benign osseous lesions used as controls. None of the genes studied predicted overall or disease-free survival. Patients, however, bearing bax(+)/cytochrome c(+) or bax(+)/cytochrome c(+ +) tumors had a decreased 4-year disease-free survival rate compared to the rest of the group (p = 0.0489 and p = 0.0208, respectively), identifying two groups of patients where more intensive adjuvant treatment could possibly be applied to prevent high postoperative recurrence rates. PMID- 18158780 TI - Preparation and characterization of new anti-PSMA monoclonal antibodies with potential clinical use. AB - Monoclonal antibodies with high specificity for prostate cancer tissue are of interest for diagnostic and therapeutic applications employing targeted therapy. The prostate-specific membrane antigen (PSMA) is a protein predominantly found in epithelial cells of prostate tissue origin and its expression correlates with tumor aggressiveness. Here, we report the development and characterization of new antibodies against PSMA. Murine monoclonal antibodies (MAb) were obtained by immunizing mice with a peptide corresponding to PSMA extracellular residues 490 500 -- GKSLYESWTKK (PSMA(490-500)). The MAbs react specifically to PSMA and to the prostate cancer cell line LNCaP with an affinity for PSMA in the low nanomolar range. This study also demonstrates the potential use of these antibodies for targeted drug delivery to prostate cancer cells. Nanomolar concentrations of PSMA-specific MAb in association with a molecule with cytotoxic potential were sufficient to allow for binding and uptake by LNCaP cells within minutes, leading to complete cell death within 3 days. These MAbs have potential clinical value in the development of diagnostic and therapeutic applications for prostate cancer. PMID- 18158781 TI - Characterization of a novel monoclonal antibody with restricted specificity to the free beta 2 integrin alpha M CD11b subunit. AB - Leukocyte cell surface expression and function of beta2 integrins require the intracellular association of alpha subunits, CD11a, b, c, d, respectively, with the common CD18 beta2 subunit. We have raised and characterized a murine MAb -- ME-MDF -- directed against the low affinity form of the human integrin alphaM subunit CD11b A-domain. MAb ME-MDF is an IgG2a that has a kDa of 2,45461 +/- 0.12 x 10(-9) M. MAb ME-MDF recognizes both the low and high affinity forms of the CD11b A-domain. Flow cytometry showed that ME-MDF does not recognize the heterodimeric CD11b/CD18 molecule at the surface of polymorphonuclear cells and the human monoblast cell line U937. Western blot analysis of U937 cell line cell surface proteins demonstrated that ME-MDF reacts specifically with the CD11b subunit but does not react with the heterodimeric CD11b/CD18 complex, a feature that differentiates it from other CD11b A-dom-specific MAbs. These observations suggest that ME-MDF recognizes an epitope that is involved in the association of the two subunits and hence is not accessible within the heterodimeric form of the CD11b/CD18 molecule. These data show that the CD11b A-dom engages not only the MIDAS but also the ME-MDF-specific epitope to associate with the CD18 subunit. We have also constructed, and expressed in the yeast Pichia pastoris, the corresponding recombinant scFv form of MAb ME-MDF and characterized the CDRs. MAb ME-MDF is characterized by short VH and VL CDR3. MAb ME-MDF and/or its recombinant scFv form would be very useful to study the structural basis of the association between the alpha and beta2 integrin subunits and to investigate the possibility of modulating CR3 cell surface expression by preventing subunit association. PMID- 18158782 TI - Generation and characterization of novel monoclonal antibodies against murine and human TARSH proteins. AB - TARSH/Abi3bp was originally isolated as a novel target of NESH-SH3 by a two hybrid yeast system. We have already identified murine TARSH (mTARSH) as a cellular senescence-related gene because of its robust induction in the early phase of mouse embryonic fibroblast cellular senescence. We have also revealed that the expression of this gene was dramatically reduced in human lung cancer cell lines and primary lung tumor, while it was predominantly expressed in normal conditions. This evidence suggests that TARSH is involved in both stress-induced senescence and prevention of cancer development; however, little is known about its molecular mechanisms. To reveal the further physiological function of this molecule, we established rat anti-TARSH monoclonal antibodies (MAb). Recombinant His-tagged partial mouse TARSH protein was expressed in Escherichia coli, affinity purified and used as an antigen to immunize rats. Hybridomas were screened by enzyme-linked immunosorbent assay, and we generated six stable hybridoma cell lines that produced antibody against murine TARSH protein, including three clones that represented cross-reactivity with human TARSH. We determined their isotypes and further examined capabilities or limitations in immunoblotting, immunoprecipitation, and immunofluorescence microscopy, realizing the most suitable antibody for each application. These MAbs should therefore be very useful tools for the study of TARSH expression and for following biological function in cellular senescence and tumor suppression. PMID- 18158783 TI - Development of a rabbit monoclonal antibody group against Smads and immunocytochemical study of human and mouse embryonic stem cells. AB - We report here the development of a group of rabbit monoclonal antibodies against Smad1, Smad2, Smad3, and Smad5, and the immunocytochemistry (ICC) staining of human embryonic stem cells (hESC) and mouse embryonic stem cells (mESC). Eight New Zealand rabbits were immunized with synthesized peptides linked to KLH, and splenocytes from these rabbits were fused with rabbit immortal B cell 240E-W2. Resulting hybridomas producing anti-Smad monoclonal antibodies were screened by enzyme-linked immunosorbent assay (ELISA) with BSA-linked peptides. Clones were chosen for antibody production based on their activities in Western blotting and on paraffin-embedded human tissues, and the capacity of the antibodies in immunocytochemistry was demonstrated. Using these antibodies, we performed ICC staining on routinely cultured human and mouse embryonic stem cells, and showed that both cell types strongly express these genes. We propose that both hESCs and mESCs have the ability to transduce signals from both BMPs and TGF-b/Activin. PMID- 18158784 TI - Preparation and characterization of agonistic monoclonal antibodies against Toll like receptor 4-MD-2 complex. AB - Ligands for toll-like receptors (TLR) are known to induce a variety of immune responses. Selective induction of desirable responses would be important for the treatment of individual diseases with various pathogenesis. For this purpose, we established six MAbs against the TLR4/MD-2 complex (UT MAbs) from TLR4(-/-) mice or MD-2(-/-) mice. Three MAbs (UT12, 18, and 22) induced NF-kappaB activation and production of pro-inflammatory cytokines, but the other three (UT15, 41, and 49) did not induce such cell responses. Unlike lipopolysaccharide (LPS), agonistic UT MAbs did not require serum components for the functions. UT41 and UT49 recognized TLR4 in the absence of MD-2. On the other hand, the other four MAbs reacted to the TLR4/MD-2 complex, but not to solo TLR4. Agonistic UT MAbs shared the epitopes, but non-agonistic UT15 reacted to distinct epitope on the complex. UT MAbs appear to be useful analyzing the molecular mechanism of TLR signaling and will contribute to the development of novel immunotherapies. PMID- 18158785 TI - Generation of a monoclonal antibody specific for tissue-nonspecific alkaline phosphatase and its use in a clinical diagnostic study. AB - Tissue-nonspecific alkaline phosphatase (TNSALP) in serum comprises liver alkaline phosphatase (liver-ALP) and bone alkaline phosphatase (bone-ALP). Liver ALP is a marker of liver disease; thus a specific method for its measurement would be useful. Measurement of ALP by electrophoresis is difficult, although all of the isozymes can be assessed simultaneously. Total ALP can also be measured by automated analyzer, but it is difficult to determine the cause of a high ALP value because bone-, intestine-, placenta-, and tumor-ALP are measured together. Thus, anti-TNSALP monoclonal antibodies that can resolve these problems are needed. Here we have generated an anti-TNSALP monoclonal antibody, 3-29-3R. This clone has specificity to liver-ALP rather than to bone-ALP. In electrophoresis, 3 29-3R reacted with TNSALP and shifted the bands. The use of 3-29-3R enabled easy interpretation of the results. Furthermore, we tested 3-29-3R by developing an immunocapture enzymatic assay (IEA). Preliminary results of the IEA show that this method is effective for measurement of liver-ALP. Thus, the monoclonal antibody that we have established may be a useful tool for clinical diagnosis. PMID- 18158786 TI - Generation and characterization of a novel phospho-specific monoclonal antibody to p120-catenin serine 879. AB - To better understand the mechanisms that regulate p120-catenin (p120) and E cadherin function, we are systematically generating phospho-specific monoclonal antibodies (MAb) to the major p120 phosphorylation sites. p120 has emerged recently as a master regulator of E-cadherin stability and an important modulator of RhoGTPase activities. A number of phosphorylation sites have been identified, but none have as yet been linked to specific regulatory roles. Here, we describe a novel phospho-specific monoclonal antibody to the major PKC-induced p120 phosphorylation site, phospho-serine 879 (pS879). With a few exceptions, p120 MAb pS879 is remarkably specific for the phosphorylated S879 epitope and works effectively in common applications such as Western blot analysis, immunoprecipitation, and immunofluorescence. p120 MAb pS879 should facilitate efforts to identify the role of S879 phosphorylation and to map signaling pathways that modify p120 function through activation of PKC. PMID- 18158787 TI - Preparation and characterization of a novel monoclonal antibody specific to human EMILIN-5 protein. AB - We aimed to establish the first monoclonal antibody (MAb) against the human EMILIN-5 protein (elastin microfibril interface located protein-5) and to investigate its distribution in normal human esophageal tissues and esophageal carcinomas. The bacterially expressed 6 His-EMILIN-5 fusion protein was induced and purified. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using either purified 6x His-EMILIN-5 fusion proteins or purified 6x His-ZNRD1 fusion protein as a control. The EMILIN-5 protein-specific MAb was further identified by Western immunoblot analysis. The expression of EMILIN-5 was investigated in 63 cases of esophageal cancer specimens and 60 cases of normal esophageal specimens using immunohistochemical analysis. The expression of 6x His EMILIN-5 fusion proteins was successfully induced. One MAb, H7 (IgG1), effective in detecting the recombinant and the cellular protein, was characterized. H7 bound to native EMILIN-5 protein and should be useful in studies of EMILIN-5 protein function and expression. EMILIN-5 staining was found positive in 37 cases of esophageal cancer tissues (59%) and 7 cases of normal esophageal tissues (12%). The higher-grade frequency of expression of EMILIN-5 in normal esophageal tissues was significantly lower (p < 0.01) than that in tumor tissues. We concluded that EMILIN-5 might play important roles in carcinogenesis of esophageal cancer. PMID- 18158788 TI - Biological activity in vitro of anti-epidermal growth factor receptor monoclonal antibodies with different affinities. AB - Epidermal growth factor receptor (EGFR) is frequently overexpressed in epithelial tumors and is associated with a poor prognosis. An increasing interest in developing anti-EGFR therapies has resulted in the evaluation of monoclonal antibodies with the capacity to bind to the EGFR, inhibiting EGFR-dependent cellular transformation. A differential toxicity and therapeutic effect in vivo are associated with the affinity and isotype of the molecule. In this study, we examined the biological activities of three monoclonal antibodies (MAbs) -- Ior egf/r3 (mouse IgG2a, 10(-9) M), Nimotuzumab (humanized IgG1, 10(-9) M), and Cetuximab (human/mouse chimeric IgG1, 10(-10) M) -- considering inhibition of cell proliferation, apoptosis, and complement-mediated cell death in squamous cell carcinoma A431 in vitro. All the antibodies bound to the EGFR on these cells, inhibiting the receptor phosphorylation, as measured by flow cytometry, inmunocytochemistry, and Western blot. Exposure to the different antibodies inhibited cell proliferation in culture in a range from 50 to 80% compared to controls. Furthermore, similar capabilities to induce either complement-mediated cytotoxicity (ranging between 70 and 90%) or a two-fold increase in the rate of apoptotic cells were found when tumor cells were exposed to the antibodies. These results suggest that the affinity between specific anti-EGFR antibodies and its receptor could affect, but not determine their biological activity at least in those cell lines that exhibit high sensitivity to withheld EGFR. Our findings also confirm previous evidences that blocking EGFR in A431 cells by means of antibodies significantly changes tumor cell biology by promoting apoptosis while decreasing tumor cell proliferation. PMID- 18158789 TI - A simple method for the production of anti-C3d monoclonal antibody. AB - Production of monoclonal antibodies to C3d usually involves the purification of protein. Our method does not require C3 purification; it relies on attachment of C3b to mouse erythrocytes by activation of alternative pathways and further conversion in C3d. We prepared human complement-coated mouse red cells and sensitized mice of the same strain with our own schedule of immunization and applied the classical methods to obtain a mouse monoclonal antibody. We obtained a clone called BMS-11 which produces a monoclonal antibody of IgM class, to C3d with a title of 1:500000. The monoclonal antibody obtained has shown that it is suitable for use as an antiglobulin reagent. PMID- 18158797 TI - Comparison between direct trocar and Veress needle insertion in laparoscopic cholecystectomy. AB - Although the Veress needle (VN) is widely used for insufflation of the peritoneum, direct trocar (DT) insertion is going to be more popular. A lot of complications have been reported because of the use of VN. We designed a prospective, nonrandomized study for the comparison of VN and DT insertion techniques. A pneumoperitoneum was created using the VN in 135 patients and using DT insertion technique in 148 patients during a 3-year period. Although no major complication was seen in the DT group, three major complications were seen in the VN group, but there was no statistically significant difference between both groups. More frequent minor complications were seen in the VN group, but it was statistically insignificant. Surgical skill and experience of the surgeon with the entry technique who is using it is an important factor for the selection of the abdominal insufflation technique. PMID- 18158798 TI - Role of laparoscopy in the management of visceral injuries following abdominal stab wounds. AB - BACKGROUND: Laparoscopy offers several advantages in the treatment of abdominal stab wounds. In this paper, we report our experience during 2004, where hemodynamically stable patients with stab wounds were managed laparoscopically. PATIENTS AND METHODS: Between January and December 2004, 8 hemodynamically stable patients (7 men, 1 woman) underwent laparoscopy for anterior abdominal stab wounds. Median age was 28.5 years (range, 17-55). All patients underwent an abdominal computed tomography (CT) prior to the laparoscopy. RESULTS: Exploration of the wound under aseptic conditions, carried out as a part of the physical examination, confirmed peritoneal penetration in 7 of the 8 cases. Abdominal CT revealed positive findings in 7 (87.5%) cases. Laparoscopy was performed after a median time of 60 minutes (range 30-90). Laparoscopic exploration evidenced peritoneal penetration in 100% of the cases and visceral lesions in 87.5% of the cases. All visceral injuries were managed laparoscopically. Median operative time was 135 minutes (range, 45-200). Operative mortality was 0% and early morbidity was 12.5%. Median hospital stay was 5 days (range, 1-11). After a median follow up of 12 months (range, 1-28), 1 patient complained of persistent chest pain and a ventral hernia at the site of the abdominal stab wound was diagnosed in another patient. CONCLUSIONS: Laparoscopy should be included in management algorithms in patients with anterior abdominal stab wounds who are hemodynamically stable. In addition to its diagnostic ability, this study demonstrates that laparoscopy can be an effective management modality with minimal morbidity and no mortality. PMID- 18158799 TI - Laparoscopic sigmoid and rectal resection using an electrothermal bipolar vessel sealing device. AB - BACKGROUND: Laparoscopic colonic resection has gained popularity as a method to treat colonic diseases. The electrothermal bipolar vessel sealer (EBVS; LigaSure Atlas) is a modern device that allows the secure sealing of vessels with a diameter of up to 7 mm. The aim of the present study was to evaluate the suitability of the device for laparoscopic colonic surgery. METHODS: The immediate outcome of 114 consecutive patients who underwent a sigmoid or rectal resection was prospectively analyzed. The intention was to perform all operations with the EBVS for dissection and ligation of the mesenterial vessels. Details on patient characteristics, peroperative and postoperative complications, and postoperative recovery were recorded prospectively and analyzed. RESULTS: One hundred and fourteen patients were scheduled for elective left-sided colonic or rectal resection. Massive intra-abdominal adhesions in 1 patient required a conversion of the laparoscopic procedure to an open one; In total, 113 laparoscopic operations were thus performed. The mean operative time was 87.7 +/- 2.8 minutes, and the mean time for patients to tolerate solid food was 3.4 +/- 0.1 days and the time to discharge from hospital was 4.6 +/- 0.2 days. There were nine (8.0%) general complications, and additionally, 10.6% of patients suffered from surgical complications. CONCLUSIONS: The electrothermal bipolar vessel sealer is suitable and safe for laparoscopic sigmoid and rectal resections. The use of the device probably reduces the operative time. PMID- 18158800 TI - Stepwise rising CO2 insufflation as an ischemic preconditioning method. AB - BACKGROUND: The pneumoperitoneum (Pp) is associated with ischemia and reperfusion (I/R) injury and oxidative stress. Various ischemic-preconditioning (IP) methods were used to reduce ischemic injury in intra-abdominal organs. In this experimental, randomized, controlled trial with a blind assessment of the outcome, we evaluated the effects of a new IP method, stepwise rising CO(2) insufflation, on oxidative stress and inflammatory cytokine response. METHODS: Twenty-one rats were divided into three groups. Rats in the control group were subjected to general anesthesia for only 60 minutes. The stepwise group was subjected to 5 mm Hg for 10 minutes, 10 mm Hg for 10 minutes, and 15 mm Hg of CO(2) insufflation for 60 minutes without deflation. In the Pp15 group, the pressure of CO(2) insufflation was fixed at 15 mm Hg for 60 minutes without deflation. Liver and blood samples were examined to determine malondialdehyde (MDA), the antioxidant, superoxide dismutase (SOD), and inflammatory cytokine (tumor necrosis factor-alpha [TNF-alpha], interleukin-6 [IL-6]) levels. Histopathologic scores of liver tissue were examined in all groups. RESULTS: The highest plasma and liver MDA, TNF-alpha, and IL-6 values were in the Pp15 group, followed by the stepwise and control groups. However, plasma and liver SOD levels determined in the control group were significantly higher, compared to stepwise and Pp15 groups. The lowest plasma and liver levels of SOD were in the Pp15 group, followed by the stepwise and control groups. Significantly higher histopathologic scores were found in the Pp15 group, followed by the stepwise and control groups, as well as MDA and inflammatory cytokine (TNF-alpha, IL-6) levels. CONCLUSIONS: We concluded that the stepwise rising CO(2) insufflation method may be an alternative IP method that may lead to a reduction in I/R injury. PMID- 18158801 TI - Outcome analysis of laparoscopic cholecystectomy in patients aged 80 years and older with complicated gallstone disease. AB - BACKGROUND: The aim of this study was to evaluate the outcome of laparoscopic cholecystectomy (LC) in octogenarians with complicated gallstone disease. MATERIALS AND METHODS: This study was a retrospective analysis of prospectively collected data of 51 patients aged 80 years or older who underwent an LC for complicated gallstone disease between 2001 and 2006. This group was compared with 41 octogenarian patients with uncomplicated gallstone disease. RESULTS: There were 51 patients (26 men) with a median (range) age of 87 years (range, 80-93) who underwent an LC for complicated biliary disease, including acute cholecystitis in 29 (57%), gallstone pancreatitis in 14 (27%), cholangitis in 4 (7.8%), and obstructive jaundice in 4 (7.8%). Significantly more patients in the complicated disease group underwent preoperative endoscopic retrograde cholangiopancreatography (33.3% vs. 12.1%; P = 0.026) The median operative time was 110 minutes (range, 55-165) and this was significantly longer, compared to the uncomplicated disease group (P = 0.031). Postoperative morbidity was 27.3%, a significantly higher rate, compared to the uncomplicated group (27.4% vs. 7.3%; P = 0.015) with zero mortality. The conversion rate was 7.8% and this was not significantly different from the uncomplicated disease group. The median length of hospital stay was 6 days, which was significantly longer than the uncomplicated disease group (P = 0.021). CONCLUSIONS: The LC can be performed with acceptable morbidity in octogenarians with complicated gallstone disease. Early treatment of gallstone disease in this age group could further improve outcomes. PMID- 18158802 TI - Transduodenal local resection for low-risk group ampulla of vater carcinoma. AB - BACKGROUND: Carcinoma of the ampulla of Vater has a more favorable prognosis, compared to other malignant tumors of the periampullary region, because it usually presents with symptoms in the early stage. However, treatment by local resection only of the ampullary carcinoma remains controversial. The aim of this study was to evaluate the treatment results of the ampulla of Vater carcinoma according to different types of operation in low-risk-group patients. METHODS: We retrospectively reviewed the medical records of 17 low-risk-group patients among a total of 102 patients with ampulla of Vater carcinoma who had underwent curative surgery from 1992 to 2002. All specimens were critically reviewed by a single expert pathologist, and the relationship between surgical outcomes and operation type was assessed. RESULTS: The low-risk group was comprised of 10 men and 7 women with a median age of 57.8 years. Thirteen of 17 patients underwent a pancreaticoduodenectomy (PD) or a pylorus preserving pancreaticoduodenectomy (PPPD), while 4 patients underwent a transduodenal local resection (TDLR). The operation time was significantly shorter in the TDLR group, compared to the PD or PPPD groups. Among the 17 patients, there was only 1 case of recurrence in the inguinal area 33 months after the pancreaticoduodenectomy. CONCLUSIONS: Transduodenal local resection is a comparable mode of operation for low-risk group patients with Ampulla of Vater carcinoma. In particular, it is essential to evaluate the invasion depth in preoperative endoscopic ultrasonography, cell differentiation in preoperative biopsy, and positivity of resection margin accurately by using frozen section during the operation. PMID- 18158803 TI - Laparoscopic management of varicocele using bipolar cautery versus open high ligation technique: a randomized, clinical trial. AB - PURPOSE: The aim of this investigation was to compare laparoscopic management of varicocele using bipolar cautery with open high ligation technique. MATERIALS AND METHODS: A total of 100 patients were operated on for left-sided varicoceles. To evaluate the clinical efficiency of the treatment, the patients were divided randomly into two groups. A laparoscopic varicocelectomy (LV) was performed on 50 patients, and an open varicocelectomy (OV) was performed on 50 patients. Outcome data examined operating time, relapse rate, presence of hydrocele, wound complications, postoperative length of stay, and pain control. In both groups, the operations were performed with the preservation of lymphatics and artery with ligation of the veins in the retroperitoneum above the internal inguinal ring. RESULTS: The operative time was 17.2 +/- 9.8 minutes in LV versus 31.02 +/- 12.8 minutes; in OV (P = .00). In LV versus OV, relapse rates were 8% versus 4% (P > 0.05), hydrocele occurrence was 14% versus 24% (P > 0.05), and wound complication was 0.00% versus 6%. Hospital stay was an average of 12 hours in both groups. Postoperative analgesic use was 3.6 +/- 0.4 mg in LV versus 3.5 +/- 0.5 mg in OV (P > 0.05). CONCLUSIONS: The clinical efficacy of LV using bipolar cautery is superior to traditional OV. PMID- 18158804 TI - Do various mesh placement techniques affect the outcome in totally extraperitoneal hernia repair? What is the role of the surgeon? AB - BACKGROUND: Various modifications of mesh placement are currently used in total extraperitoneal (TEP) groin hernia repair. The aim of this study was to compare three different variants of mesh placement with respect to rate of complications and clinical outcome. METHODS: A series of 397 consecutive patients with a total of 534 preperitoneal groin hernia TEP repairs performed by four surgeons at a single institution between 1999 and 2003 were retrospectively analyzed. The mean follow-up was 19.7 +/- 7.5 months. A single-mesh technique was used in cases of hernial orifice <1.5 cm. Larger hernial defects were closed either in a double mesh or a modified double-mesh placement technique. The three placement techniques were compared with respect to hospital stay, operative time, early and late complications, return-to-work time, and recurrence rate. RESULTS: The modified double-mesh technique was associated with the longest hospital stay, the longest operative time, the slowest return to work, and significantly higher rates of early (5.6% vs. 4.6% vs. 2.9%) and late (19.1% vs. 11.3% vs. 7.9%) postoperative complications, when compared to double-and single-mesh placement. Overall recurrence rate was 1.3% after a mean follow-up of 19.7 months. The larger the experience of a surgeon with his preferred technique, the shorter the operative time and hospital stay were. CONCLUSIONS: Mesh placement techniques appeared to have a direct impact on clinical outcome and hospital stay. The modified double-mesh technique showed the worst postoperative results, independent of the surgeon's experience. Which mesh placement technique is most appropriate for complex hernias remains to be answered by further randomized, controlled trials. PMID- 18158805 TI - Laparoscopic management of acutely incarcerated femoral hernia. AB - Incarcerated femoral hernia is a common surgical emergency condition. Diagnosis is always obvious and straightforward by clinical examination, and open surgical repair is the mainstay of treatment. In the era of minimally invasive surgery, laparoscopic repair of femoral hernia has been shown to be feasible and safe. However, laparoscopic repair of acutely incarcerated femoral hernia has gained little discussion in the past. In this paper, we report the results of 8 consecutive cases of strangulated femoral hernia that was successfully managed by the laparoscopic approach. PMID- 18158806 TI - Laparoscopic splenectomy: ligasure versus EndoGIA: a comparative study. AB - BACKGROUND: Laparoscopic splenectomy (LS) is considered the gold standard for the treatment of hematologic disease of the spleen. Intraoperative bleeding is the main complication and main cause of conversion during LS. In this paper, we present our experience with 112 LS procedures, comparing two different techniques and instruments for spleen dissection and hilar vascular control. METHODS: We have performed a total of 112 LS procedures in 9 years in two affiliated university hospitals. There were 48 males and 64 females (median age, 32.8 years). Indications were as follows: idiopathic thrombocytopenic purpura (ITP) in 42 patients, hereditary spherocitosis in 28, 6 with beta-talassemia, 1 with hemolitic anemia, 27 with lymphoma, 2 with leukemia, 3 with splenic cysts, and 3 PPT cases. Associated procedures were present in 25 cases (17 cholecystectomy and 4 liver/lymphnodal biopsies + lysis of adhesions). Patients were divided in two groups according to the method of dissection and hilar section (group 1, endostapler + monopolar coagulation with 38 patients; group 2, the Ligasure Vessel Sealing System (Valleylab, Boulder, CO) with 74 patients). RESULTS: Groups were well matched according to age, indications, spleen sizes, and procedures associated. Conversion rates (10 cases, 9%; 5 in each group), postoperative hospital stay (median 4 days in each group), and intra- and postoperative complications were similar in the 2 groups without any mortality in each group, while intraoperative blood loss (loss of 100 mL in 55 patients in group 2, 78%; and in 21 patients in group 1, 52%), transfusion rates (8 patients in group 1 versus 4 patients in group 2), and operative mean time (150 minutes in group 1 versus 125 minutes in group 2) were minor in the Ligasure Vessel Sealing System group, with a statistical significance. CONCLUSIONS: The approach to spleen dissection and hilum section was safe and efficacy in each case, otherwise the use of Ligasure results in a gain of time. Furthermore the average intraoperative bleeding and average transfusion rates of this series were lower in group 2. PMID- 18158807 TI - Laparoscopic resection of schwannoma in the anomaly of obturator nerve. AB - Most retroperitoneal schwannomas are benign tumors originating from schwann cells and are relatively rare, especially schwannomas arising in the obturator nerve. In this paper, we describe a 44-year-old woman with a retroperitoneal schwannoma in the anomaly of the obturator nerve, which was treated by laparoscopy without any complications. PMID- 18158808 TI - Total laparoscopic pancreaticoduodenectomy and reconstruction for a cholangiocarcinoma of the bile duct. AB - BACKGROUND: In this paper, we report on our experience with a totally laparoscopic pancreatico-duodenectomy performed for a cholangiocarcinoma of the lower third of the bile duct. METHODS: The patient was placed in the steep reverse Trendelenberg, Lloyd-Davis position. The procedure was performed with six laparoscopic ports, using similar steps to the open approach, with the use of an ultrasonic cutting and coagulating instrument for dissection and endoscopic linear stapling devices for the bile duct, intestinal, and gastroduodenal artery division. Reconstruction was done on a single loop by an intracorporeally sutured pancreaticojejunostomy, hepaticojejunostomy, and a stapled gastroenterostomy. The resection specimen was placed in a bag and retrieved through a 5-cm Pfannenstiel incision. RESULTS: Histology confirmed a T3 N1 R0 cholangiocarcinoma with the involvement of 1 of 17 lymph nodes. Twelve months following surgery, he remains well, having completed a course of adjuvant chemotherapy. CONCLUSIONS: Although the operation was technically demanding, it can be safely performed with a good oncologic result. PMID- 18158809 TI - Abdominal wall hematoma after laparoscopic surgery: early treatment with selective arterial transcatheter embolization. AB - Bleeding from the site of a trocar insertion is a relatively frequent complication after laparoscopic surgery, despite precautionary techniques aimed at reducing such events. It may result in a hemoperitoneum, requiring emergency reintervention or large hematomas of the abdominal wall. In the latter case, although conservative treatment is possible, hospital stay is prolonged and large amounts of blood-bank resources are consumed. Early diagnosis and selective embolization of the bleeding vessel may reduce the severity and consequences of this complication. PMID- 18158810 TI - Laparoscopic pyeloureterostomy in ureteral duplication with lower polar ureteropelvic junction obstruction: easy and effective. AB - Ureteropelvic junction (UPJ) obstruction in ureteral duplication is not uncommon. Ureteropyelostomy is a safe, easy surgical option when the adequate length of the distal ureter is not available for pyeloplasty. In this paper, we present the case of a 26-year-old male who was treated with laparoscopic ureteropyelostomy for a symptomatic lower moiety UPJ obstruction. PMID- 18158811 TI - Laparoscopy-assisted resection of ileoileal intussusception caused by intestinal lipoma. AB - Intussusception is the second most common abdominal emergency in children. In contrast, it is rare in adults. Adult intussusception represents only 1%-3% of patients with bowel obstruction. Although 95% of intussusception in children is idiopathic, merely 7% of adult intussusception is considered idiopathic. Owing to vague symptoms and signs, the preoperative diagnosis of adult intussusception is difficult. Once adult intussusception is diagnosed, surgical intervention is indicated because about half of both colonic and small intestinal intussusceptions are caused by malignant lesions. In this paper, we describe a case of ileoileal intussusception caused by an intestinal lipoma that was diagnosed preoperatively by computed tomography scans and was treated successfully by laparoscopy-assisted surgery. The patient was discharged uneventfully 4 days after the operation. We recommend laparoscopy-assisted surgery as a feasible therapeutic option for adult intussusception. PMID- 18158812 TI - Laparoscopic total gastric vertical plication in morbid obesity. AB - BACKGROUND: The aim of this study was to introduce a new technique, total gastric vertical plication (TGVP), as a restrictive operation. It has the same result of weight loss as others with minimal risk of complication and very low cost, especially in developing countries. METHODS: This technique was used by one surgeon in private hospitals during 3 years in Tehran, Iran. Patients were placed in the supine position with a 30-degree reverse Trendelenburg position. Trocars were inserted based on an ergonomic assessment (three 5 mm and one 10 mm). After the release of the greater curvature, continuous sutures were used with 00 nylon from the fondus to 3 cm of the pylorus. A vertical plication was performed in one or two layers. Distance between the stitch and lesser curvature was 2 cm in the anterior and posterior and between each stitch, all of them getting extra mucosal (far away from acid effect) owing to mild tension on the sutures that cut mucosa and put on a submucosa layer. RESULTS: TGVP was performed in 100 cases (mean age, 32; standard error of the mean = 2.1); mostly female (F/M = 76/24) and with average body mass index of 47 (36-58). The mean weight loss in our patients was 21.4% of excessive weight loss (EWL) 1 month after the operation, 54% after 6 months (72 cases), 61% after 12 months (56 cases), 60% after 24 months (50 cases), and 57% after 36 months (11 cases). The average time of follow-up was 18 months. The mean time of operation was 98 (70-152) minutes and all of the patients were discharged from the hospital after an average of 1.3 days (range, 1 4). The main postoperative complications were permanent vomiting, intracapsular liver hematoma, hypocalcemia at early postoperative period, hepatitis, leakage at the suture line, and acute gastric perforation. The volume of the stomach in this condition was 100 cc, but just one half of it was effective. If more than 50 cc was used, a painful condition would occur. CONCLUSIONS: The percentage of EWL in this technique is comparable to other restrictive methods, but EWL appears more rapidly. Early postoperative complications of this method are minimal, without any important late complications. This technique needs more expertise and is more time consuming. A long-term follow-up is advised. PMID- 18158813 TI - Retrieval of large colonic polyps: simple use of proctoscope. AB - Retrieval of polyps during colonoscopic procedures can be technically difficult and time consuming. This is particularly the case when attempting to retrieve large polyps intact through the anal canal of an anxious patient with a hypertonic sphincter. In this paper, we describe a simple technique that permits the complete removal of a large polyp intact by using a readily available theater instrument. The success rate of this procedure can be improved with the aid of simple instrument like a proctoscope. PMID- 18158814 TI - Comparison of the effectiveness of metoclopramide, ondansetron, and granisetron on the prevention of nausea and vomiting after laparoscopic cholecystectomy. AB - BACKGROUND AND GOALS: A relatively high incidence of postoperative nausea and vomiting (PONV) occurs in patients undergoing a laparoscopic cholecystectomy. Prophylaxis of PONV is usually achieved with a single-dose antiemetic drug administered during the surgical procedure. The aim of the current study was to compare the antiemetic activity of different 5-hydroxytryptamine-3 receptor antagonists with that of metoclopramide. MATERIALS AND METHODS: In a randomised, double-blind study, 75 patients received the following: Group M, 10 mg metoclopramide; Group K, 40 mcg . kg(-1) granisetron; and Group Z, 15 mcg . kg( 1) ondansetron intravenously (IV) diluted in 20 cc 0.9% NaCl (n = 25 of each) i.v. immediately before the induction of anesthesia. The standard general anesthetic technique, which consisted of sevoflurane in air-oxygen and a fentanyl perfusion, was used. Nausea, vomiting, and safety assessments were performed continuously during the first 24 hours after anesthesia. RESULTS: There were no statistically significant differences for demographic data, American Society of Anesthesiology (ASA), operation duration, or anesthesia time among the three groups (P > 0.05). Evaluated nausea and vomiting scores in the first 3-hour period revealed that each of the drugs had a similar antiemetic effect (P > 0.05). Nausea and vomiting scores, evaluated between the 4-24 hours, also revealed that the group M scores were obviously higher than groups K and Z (P < 0.001). A comparison of incidences of dose administrations were statistically not significant among the groups (P > 0.05). CONCLUSIONS: Granisetron, when given prophylactically, resulted in a significantly lower incidence of PONV than metoclopramide and ondansetron, whereas metoclopramide was ineffective. Garnisetron may be an effective treatment in the proflaxy of PONV. PMID- 18158815 TI - Primary pull-through for Hirschsprung's disease: comparison of open and laparoscopic-assisted procedures. AB - Rectosigmoid Hirschsprung's disease is usually amenable to minimally invasive primary neonatal pull-through. This may be performed either entirely transanally or with laparoscopic assistance for biopsies with or without colonic mobilization. In our center, all dissection is performed transanally; laparoscopy is used for obtaining colonic biopsies and orientation of the pulled-through bowel segment. In this paper, we describe our initial experience of a consecutive cohort of 20 one-stage laparoscopic-assisted endorectal pull-through (LAEPT) procedures. A historic consecutive cohort of 22 infants who underwent the same open endorectal pull-through (OPT) with open transabdominal mobilization was used for comparison. Age at operation and mean theater time were not significantly different. The mean postoperative stay was significantly reduced in the laparoscopic group (LAEPT 3.8 days vs. OPT 9.5 days; P = 0.0002). Readmission and enterocolitis rates in the first postoperative year did not differ significantly. LAEPT permits early intraoperative biopsies with a visualization of the pull through to prevent twisting of the bowel. PMID- 18158816 TI - Laparoscopic primary repair of a pediatric transanal impalement injury: a case report. AB - Conventional treatment of anorectal impalement injuries involves an open exploration and either a fecal diversion and/or a primary repair dictated by the degree of soft-tissue disruption and contamination. In this paper, the authors present the case report of a successful primary laparoscopic repair of an accidental transanal intraperitoneal impalement injury in a child. PMID- 18158817 TI - Laparoscopic excision of Mullerian structures in a neonate with mixed gonadal dysgenesis: a case report. AB - Excision of Mullerian structures in children with disorders of sexual differentiation is an operative challenge. We report our experience with laparoscopic excision of Mullerian structures in a neonate with mixed gonadal dysgenesis. The salient features of the procedure were excellent visualisation and ease of dissection. PMID- 18158819 TI - Laparoscopy: an excellent tool in the management of primary omental torsion in children. AB - INTRODUCTION: Primary omental torsion (POT) is an uncommon acute condition, often occurring in obese children. The clinical presentation usually mimics that of acute appendicitis, and preoperative radiologic imaging may not be helpful in the diagnosis. In this paper, we report our experience of using laparoscopy in diagnosing and treating POT in children. MATERIALS AND METHODS: A retrospective review of all cases of POT from 1998 to 2006 was performed. The efficacy and safety of using the laparoscope in the management of omental torsion was assessed. RESULTS: There were 5 boys with a mean age of 8.8 years (range, 5-11) included in the study. The majority of the patients were overweight, and all presented with abdominal pain without other gastrointestinal symptoms. In all patients, there was marked localized tenderness at the right side but without rebound tenderness or guarding. Preoperative investigations did not help in the diagnosis in all cases. A laparoscopic examination was performed and, in all cases, the diagnosis of POT was accurately made. The omentum was either adherent to the anterior abdominal wall or to the ascending colon. In all cases, the twisted omentum was successfully removed by the laparoscopic technique. Rapid recovery was universally observed, with the disappearance of pain and a rapid resumption of diet and discharge from the hospital on the first postoperative day. Histology showed gangrenous omentum in all cases. All the patients were well and had good cosmetic results on the follow-up. CONCLUSIONS: Laparoscopy is an excellent tool for both diagnosing and treating omental torsion in children. PMID- 18158818 TI - Minimal invasive surgery in pediatric solid tumors. AB - BACKGROUND: There is only limited experience of using the minimally invasive surgery (MIS) technique in resecting pediatric solid tumors. In this paper, we report our experience of using the MIS technique in the management of pediatric solid tumors. METHODS: A retrospective review was undertaken on all children who had undergone MIS for their solid tumors between 1995 and 2005. RESULTS: Over a 10-year period, there were 38 patients who had undergone MIS for tumor resection. The mean age at the time of surgery was 7.5 years (range, 1 day to 15 years). There were 22 ovarian tumors, 4 sacrococcygeal tumors, 3 adrenal tumors, 3 retroperitoneal tumors, 1 kidney tumor, 1 liver mass, 1 intra-abdominal testicular tumor, and 3 intrathoracic masses. Thirty of 38 patients had undergone a successful resection using the MIS technique (78.9%). Eight patients required a conversion to the open procedure because of limited intraperitoneal space in 7 and excessive bleeding in 1. Of the 28 successfully MIS-resected intra-abdominal tumors, 18 required enlargement of the umbilical incision and 5 required an additional Pfannenstiel incision for tumor retrieval. Enlargement of the thoracic port site for specimen retrieval was required in the 2 successfully MIS-resected intrathoracic masses. The mean operation time was 171 minutes (range, 45-275). There was no postoperative complication encountered. On an average follow-up of 3.1 years, there was no recurrence observed, even in the 7 patients with malignant tumors, and all patients with successful MIS tumor excision had good cosmetic results. CONCLUSIONS: With the advance of laparoscopic instruments and techniques, a variety of pediatric solid tumors can be resected safely by the MIS technique. This has the potential benefit of a more rapid postoperative recovery and better cosmetic results. The role of the MIS technique in resecting malignant tumors is uncertain, as the number of cases in the current series is too small to draw any conclusion. PMID- 18158820 TI - Thoracoscopy and solid tumors in children: a multicenter study. AB - PURPOSE: The aim of this study was to evaluate the efficacy and safety of thoracoscopy in the treatment of solid thoracic tumors in children. METHODS: A retrospective, multicenter review of 139 thoracoscopies was performed. The procedures were either resection of pulmonary lesions or biopsy. Complications, histologic biopsy results, and outcomes were reviewed. RESULTS: One hundred thirty-nine procedures were performed in 134 children. There were 72 males and 62 females, with a mean age of 9.2 years (3 months to 17 years). The mean follow-up was 4.5 years. Thirty-eight primitive tumors were managed, 20 of which had a complete thorascopic resection, and nine biopsies were performed. In 9 additional cases, an open conversion was necessary. Among the 20 complete resections, 17 of the lesions were neurogenic tumors (i.e., neuroblastoma or ganglioneuroma). Three complications occurred: 2 chylothoraces and 1 with Horner syndrome. All three complications regressed. The mean follow-up was 3 years, and no recurrence has been noted. Fifty-three metastatic lesions were managed: 29 had a complete resection, 11 had a biopsy, and an open conversion was necessary in 13 cases. Among the complete resections, more than half were metastases from bone tumors. The mean follow-up was approximately 4 years and 5 pulmonary recurrences developed. Four deaths occurred, with 2 having had lung metastases. Forty-eight malignant hemopathies were managed; 9 biopsies for primitive tumors were performed. There were 36 secondary lesions, of which 5 were completely resected; 30 were biopsied and 1 required an open conversion. CONCLUSIONS: Thoracoscopy is a safe, effective approach for the evaluation and resection of solid lung tumors and for biopsy. Thoracoscopic resection of metastatic lesions is reasonable for nephroblastoma, but a thoracotomy is suggested for other metastases. PMID- 18158821 TI - Primary omental infarction: laparoscopic approach in two pediatric cases: a case review. AB - Two cases of pediatric acute abdomen owing to omental infarction are described in this paper, which were successfully treated laparoscopically. Owing to the objective rareness and absence of typical symptoms, the diagnosis was often neither made nor considered preoperatively. For other emergency situations, laparoscopy has been shown to be both a diagnostic and a therapeutic tool of a rare pathology found also in the pediatric patients. PMID- 18158822 TI - Minimal access surgical repair of Morgagni hernia: the fate of the unresected hernia sac. AB - Treatment of retrosternal diaphragmatic (Morgagni) hernia is composed of a simple surgical closure of the retrosternal opening either conventionally by open abdominal or thoracic approaches, or more recently, by using minimal access surgery (MAS). Clinical experience using the latter approach is very limited in children. Removal of the hernia sac is a controversial issue, since the sac is said to carry the risk of cyst formation or show spontaneous resolution. This issue represents a 7-year-old boy with Morgagni hernia that was successfully repaired by the MAS approach. The hernia sac was not resected because of adhesion to the adjacent tissues. The patient was readmitted with fluid accumulation within the remaining sac 2 months after the operation. A complete spontaneous resolution was observed within 2 months under conservative follow-up. To our knowledge, the temporary complication described in this paper has not been published in the literature. PMID- 18158823 TI - Non-alcoholic fatty liver disease: the new epidemic and the need for novel nutritional approaches. AB - The epidemic of non-alcoholic fatty liver disease (NAFLD) in the United States is staggering, and there is an enormous void in our understanding of the clinical epidemiology other than the common themes of obesity and insulin resistance. There is also a public health need to better define effective treatments of NAFLD, including dietary interventions and appropriate nutritional supplements. There is, however, a wealth of basic science that helps to set the stage for defining the mechanisms leading to liver pathology. In this article we will attempt to put these concepts in perspective to highlight the need for future research including the use of medicinal food. PMID- 18158824 TI - Clinical potential of Spirulina as a source of phycocyanobilin. AB - Recent research reveals that free bilirubin functions physiologically as a potent inhibitor of NADPH oxidase activity. The chromophore phycocyanobilin (PCB), found in blue-green algae and cyanobacteria such as Spirulina, also has been found to be a potent inhibitor of this enzyme complex, likely because in mammalian cells it is rapidly reduced to phycocyanorubin, a close homolog of bilirubin. In light of the protean roles of NADPH oxidase activation in pathology, it thus appears likely that PCB supplementation may have versatile potential in prevention and therapy -- particularly in light of rodent studies demonstrating that orally administered Spirulina or phycocyanin (the Spirulina holoprotein that contains PCB) can exert a wide range of anti-inflammatory effects. Until PCB-enriched Spirulina extracts or synthetically produced PCB are commercially available, the most feasible and least expensive way to administer PCB is by ingestion of whole Spirulina. A heaping tablespoon (about 15 g) of Spirulina can be expected to provide about 100 mg of PCB. By extrapolating from rodent studies, it can be concluded that an intake of 2 heaping tablespoons daily would be likely to have important antioxidant activity in humans -- assuming that humans and rodents digest and absorb Spirulina-bound PCB in a comparable manner. An intake of this magnitude can be clinically feasible if Spirulina is incorporated into "smoothies" featuring such ingredients as soy milk, fruit juices, and whole fruits. Such a regimen should be evaluated in clinical syndromes characterized and in part mediated by NADPH oxidase overactivity in affected tissues. PMID- 18158825 TI - Soy isoflavones as safe functional ingredients. AB - In recent years, isoflavones have increased in popularity as an alternative to estrogen therapy, particularly after the Women's Health Initiative demonstrated an increased risk of breast cancer, stroke, and heart attacks in response to estrogen and progesterone intervention. Isoflavones are heterocyclic phenols with structural similarity to estradiol-17beta and selective estrogen receptor modulators. Actions at the cellular level depend on the target tissue, receptor status of the tissue, and the level of endogenous estrogen. Clinical studies of soy-based diets evaluating the relation between soy consumption and serum lipid concentrations revealed that soy consumption significantly decreased total cholesterol, low-density lipoprotein cholesterol, and triglyceride levels. Epidemiological studies suggest a protective effect of soy protein on breast tissue as evidenced by the lower rates of breast cancer in East Asian countries where soy is a predominant part of the diet. Soy products also alleviate menopausal symptoms by reducing hot flashes. However, whether these biological effects of soy products originated from isoflavones is not clear. Furthermore, data available from human studies on the effect of isoflavones on osteoporosis are limited, and additional studies are needed to support a role in osteoporosis prevention. To date, no adverse effects of short- or long-term use of soy proteins are known in humans, and the only adverse effects known are those reported in animals. In conclusion, isoflavones are biologically active compounds, and current data are insufficient to draw definitive conclusions regarding the use of isoflavones as an alternative to estrogen for hormone replacement in postmenopausal women. Large, long-term intervention studies examining adverse effects and disease outcomes are needed before definitive conclusion can be drawn. PMID- 18158826 TI - Induction of apoptosis in HT-29 colon cancer cells by phloretin. AB - Phloretin, which is present in apples and pears, has been found to inhibit the growth of several cancer cells and induce apoptosis of B16 melanoma and HL60 human leukemia cells. The present study examined whether and how phloretin induces apoptosis of HT-29 human colon cancer cells. Phloretin (0-100 micromol/L) substantially decreased viable cell number and induced apoptosis of HT-29 cells in a dose-dependent manner. Western blot analysis of total cell lysates revealed that phloretin increased the protein levels of Bax but had no effect on Bcl-2. In addition, phloretin induced cleavage of caspase-8, -9, -7, and -3 and poly(ADP ribose) polymerase. Furthermore, phloretin increased the levels of cytochrome c and Smac/Diablo in the cytosol. The present results indicate that phloretin inhibits HT-29 cell growth by inducing apoptosis, which may be mediated through changes in mitochondrial membrane permeability and activation of the caspase pathways. PMID- 18158827 TI - Methanolic extracts of Plocamium telfairiae induce cytotoxicity and caspase dependent apoptosis in HT-29 human colon carcinoma cells. AB - Natural marine products have recently become the focus of increased research interest, due to their potential pharmacological activities. Therefore, we have screened 50 varieties of marine seaweed and determined that the methanolic extracts from Plocamium telfairiae (PTE) exhibited a cytotoxic effect against HT 29 human colon carcinoma cells. In this study, we report on the cytotoxic activity and mechanism of PTE-induced apoptosis in HT-29 cells. The treatment of HT-29 cells with various PTE concentrations resulted in the inhibition of growth and the induction of apoptosis in a dose-dependent manner, as determined by the results of a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay, a lactate dehydrogenase release assay, a morphological assay, and a colony formation assay. Interestingly, we also detected apoptotic bodies on Hoechst staining and attempted to determine whether the PTE-induced apoptosis involved the caspase pathway, using a caspase colorimetric assay. The activation of caspases-8, -9, -3, and -7 and the specific proteolytic cleavage of poly(ADP ribose) polymerase were detected over the course of apoptosis induction. Our results showed that PTE may function as a chemopreventive and/or chemotherapeutic agent in colon carcinoma cells via the reduction of cell viability and the induction of apoptosis. PMID- 18158828 TI - Effects of soy pinitol on the pro-inflammatory cytokines and scavenger receptors in oxidized low-density lipoprotein-treated THP-1 macrophages. AB - Pinitol, a methylated form of D-chiro-inositol, acts as a insulin mediator. We investigated the effects of soy pinitol on the factors involved in foam cell formation using differentiated THP-1 macrophages. Pinitol slightly inhibited the lipid-laden foam cell formation by oxidized low-density lipoprotein (oxLDL) in a dose-dependent manner. Tumor necrosis factor-alpha and monocyte chemoattractant protein-1 releases were significantly reduced by pinitol treatment (0.05-0.5 mM), whereas interleukin-1beta and interleukin-8 secretions were significantly reduced in low-dose pinitol (0.05 or 0.1 mM) and 0.5 mM pinitol-treated cells, respectively, compared to no pinitol-treated cells. Gene expressions of CD36 and CD68 were significantly down-regulated by 0.05-0.5 mM pinitol compared to the oxLDL-treated control cells. Matrix metalloproteinase-9 gene expression was significantly decreased in 0.05-0.5 mM pinitol-treated cells compared to the no pinitol-treated macrophages. We conclude that pinitol has some inhibitory effects on foam cell formation by reducing lipid accumulation, secretion, and expression of some cytokines and macrophage scavenger receptor expression via its insulin like action. PMID- 18158829 TI - Antidiabetic properties of 2,5-dihydroxy-4,3'-di(beta-D-glucopyranosyloxy)-trans stilbene from mulberry (Morus bombycis koidzumi) root in streptozotocin-induced diabetic rats. AB - We investigated the antidiabetic properties of 2,5-dihydroxy-4,3-di(beta-D glucopyranosyloxy)-trans-stilbene (DGTS) isolated from Morus bombycis Koidzumi in streptozotocin (STZ)-induced diabetic rats. The DGTS prevented the increase in aspartate aminotransferase, alanine aminotransferase, and blood urea nitrogen levels in serum of diabetic rats. At doses of 200-800 mg/kg, DGTS improved hyperglycemia in the rats, and the hypoglycemic effect of DGTS was comparable to that of tolbutamide. The histological observations showed that DGTS prevented atrophy of pancreatic beta-cells and vascular degenerative changes in the islets. DGTS reversed STZ-induced diabetes and had antioxidant activity in assays of FeCl(2)/ascorbic acid-induced lipid peroxidation in the rats. Levels of cytochrome P450 2E1 mRNA, as measured by reverse transcription-polymerase chain reaction, were lower in the livers of the DGTS-treated rats than those of the control group. These results suggest that DGTS might be beneficial in the treatment of type 1 diabetes. PMID- 18158830 TI - Effects of solid-state fermented rice on lipid metabolism and antioxidant status in high-cholesterol-fed rats. AB - We investigated the effect of solid-state fermented rice cultured with Basidiomycota (sangwhang) and Monascus ruber on lipid metabolism and antioxidant activity. Forty 4-week-old male Sprague-Dawley rats were fed high cholesterol diets in which carbohydrate sources in the treatment groups consisted of non fermented rice and sangwhang or M. ruber rice at 80% and 20%, respectively, for 5 weeks. Supplementation with sangwhang and M. ruber rice had no effect on growth and food intakes in high-cholesterol-fed rats. The plasma triglyceride concentration was not significantly different among the groups. Supplementation with M. ruber rice resulted in lower plasma and hepatic cholesterol concentrations and atherogenic index compared to the control group, while the plasma high-density lipoprotein-cholesterol concentration was elevated. In addition, fermented rice cultured with M. ruber-supplemented animals had greater bile acid excretion. The M. ruber groups had significantly lower plasma and hepatic levels of thiobarbituric acid-reactive substances than the control group. Moreover, hepatic antioxidant enzyme activities, including catalase and superoxide dismutase, were significantly higher in the M. ruber group. In conclusion, fermented rice, especially M. ruber rice, was very effective for improving the lipid metabolism and reducing oxidative stress by up-regulating the hepatic antioxidant enzymes in high-cholesterol-fed rats. PMID- 18158831 TI - Food pattern analysis and prevalence of cardiovascular disease risk factors among elderly people from Mediterranean islands. AB - The effect of various foods on the development of cardiovascular disease (CVD) has already been investigated. We performed a food pattern analysis and evaluated the association between the consumption of various patterns and the prevalence of CVD risk factors among elderly people from Mediterranean islands (the MEDIS study). During 2005-2006, 300 men and women from Cyprus, 142 from Mitilini, 100 from Samothraki, and 104 from Kefalonia islands (65-100 years old) were enrolled. CVD risk factors (i.e., hypertension, diabetes, hypercholesterolemia, and obesity) were assessed through standard procedures. All participants were asked about their usual frequency of consumption of various foods through a semiquantitative food frequency questionnaire, and food pattern analysis using the principal components analysis (PCA) method was then performed. PCA extracted five components that explained the 56.53% of the total variation in intake: i.e., a food pattern (component 1) that was loaded mainly on low-fat products, a high glycemic index and high-fat pattern (component 2), a pattern that included consumption of cereals and sweets (component 3), a pattern that was characterized by the intake of dairy products and fruits (component 4), and a pattern that was characterized by the consumption of alcoholic beverages (component 5). Ordinal logistic regression analysis revealed that component 1, component 3, and component 5 were associated with lower likelihood of having increased burden of CVD (P < .01), irrespective of various potential confounders. Food pattern analysis revealed the current nutritional status of our elderly participants, and provided a pathway for reducing the burden of CVD risk factors among these people. PMID- 18158832 TI - Preventive effects of ACTICOA powder, a cocoa polyphenolic extract, on experimentally induced prostate hyperplasia in Wistar-Unilever rats. AB - Plant extracts are useful in the management of benign prostatic hyperplasia (BPH). This study investigates whether ACTICOA (Barry Callebaut France, Louviers, France) powder (AP), a cocoa polyphenolic extract, could prevent prostate hyperplasia induced by testosterone propionate (TP) in rats. Male Wistar-Unilever rats were randomly divided in four groups of 12 rats: one negative control group receiving subcutaneous injections of corn oil and treated with vehicle and three groups injected subcutaneously with TP and treated with the vehicle (positive control) or AP at 24 (AP24) and 48 (AP48) mg/kg/day. Treatments were given orally and started 2 weeks before the induction of prostate hyperplasia. The influence of TP and AP on body weights and food and water consumption of rats was examined. On day 36, rats were sacrificed, and the prostates were removed, cleaned, and weighed. The prostate size ratio (prostate weight/rat body weight) was then calculated. TP significantly influenced the body weight gain of the rats and their food and water consumption, while AP at both doses tested reduced significantly these differences. TP significantly increased prostate size ratio (P < .001), and this induced increase was significantly inhibited in AP-treated rats in comparison with positive controls (P < .001) in a dose-dependent manner. We conclude that AP can prevent TP-induced prostate hyperplasia and therefore may be beneficial in the management of BPH. PMID- 18158833 TI - Therapeutic effect of ACTICOA powder, a cocoa polyphenolic extract, on experimentally induced prostate hyperplasia in Wistar-Unilever rats. AB - Benign prostatic hyperplasia (BPH) is a non-malignant enlargement of the prostate that results in obstructive lower urinary tract symptoms. Plant extracts are frequently used to treat BPH rather than therapeutics that can cause severe side effects. ACTICOA() (Ba0rry Callebaut France, Louviers, France) powder (AP) is a cocoa polyphenolic extract, and we have shown in a previous study that oral treatment with AP prevented prostate hyperplasia. This study investigated whether AP could improve established prostate hyperplasia using the same testosterone propionate (TP)-induced prostate hyperplasia model in rats. Male Wistar-Unilever rats were randomly divided in four groups of 12 rats: one group injected with corn oil and orally treated with the vehicle (negative control) and three groups injected subcutaneously with TP and orally treated with the vehicle (positive control) or AP at 24 (AP24) and 48 (AP48) mg/kg/day. Treatments started 1 week after the start of the induction of prostate hyperplasia and lasted for 2 weeks. The influence of TP and AP on body weights, food and water consumptions, plasma polyphenolic concentration, and serum dihydrotestoterone (DHT) level of rats was examined. At completion of the study, rats were sacrificed, and the prostates were removed, cleaned, and weighed. The prostate size ratio (prostate weight/rat body weight) was then calculated. TP significantly influenced the body weight gain of the rats and their food and water consumptions, while AP reduced significantly these differences in a dose-dependent manner. AP significantly reduced serum DHT level and prostate size ratio in comparison with positive controls also dose-dependently. In conclusion, AP orally administered was effective for reducing established prostate hyperplasia, especially at the dose of 48 mg/kg/day. PMID- 18158834 TI - Topical anti-inflammatory activities of Vitis rotundifolia (muscadine grape) extracts in the tetradecanoylphorbol acetate model of ear inflammation. AB - The ability of muscadine grape skin, seed, or combined skin and seed extracts to inhibit mouse ear inflammation, edema, and polymorphonuclear leukocyte infiltration was tested following topical application of 12-O tetradecanoylphorbol 13-acetate (TPA). Ethanolic extracts of skins, seeds, or a combination of these from purple (Ison) cultivars were applied to both ears of female Swiss mice 30 minutes after TPA (2 microg per ear) administration. Control mice were treated with indomethacin or 50% ethanol vehicle 30 minutes after TPA. Ear thickness was measured before TPA and at 4 and 24 hours post-TPA administration to assess ear edema. Ear punch biopsies were collected at 24 hours and weighed as a second marker of edema. Myeloperoxidase (MPO) (EC 1.11.1.7) activity was measured in each ear punch biopsy as an index of neutrophil infiltration. Extracts of muscadine skin, seed, and combination treatments significantly reduced ear edema, ear biopsy weight, and MPO activity compared to TPA vehicle control. There was no significant difference in anti-inflammatory activity of the skin and seed extracts. However, an additive effect was observed with the combination treatment that was statistically similar to the anti inflammatory activity of indomethacin treatment. It can be concluded that muscadine skin, seed, and combination skin/seed extracts exhibit significant topical anti-inflammatory properties. PMID- 18158835 TI - Pomegranate juice effects on cytochrome P450S expression: in vivo studies. AB - Beneficial health effects have recently been claimed for pomegranate juice. In vitro and in vivo studies have demonstrated its anti-atherosclerotic capacity, chemoprevention and chemotherapy of prostate cancer, and antiproliferative, apoptotic, and antioxidant activity, among others. On the other hand, there is a complex interplay between tumor initiation, promotion, and progression and xenobiotic biotranformation. This led us to investigate the effect of pomegranate juice consumption on cytochrome P450 (CYP) activity and expression. For this purpose, male mice consumed this fruit juice for 4 weeks, and pentobarbital induced sleeping time and total hepatic CYP content, activity, and expression were evaluated. Moreover, the activity of CYP isoform 2E1 and expression of the main CYP isoforms, namely, CYP1A1/2, CYP2E1, and CYP3A, were also assessed. It was found that pomegranate juice consumption decreased total hepatic CYP content as well as the expression of CYP1A2 and CYP3A. Prevention of procarcinogen activation through CYP activity/expression inhibition may be involved in pomegranate juice's effect on tumor initiation, promotion, and progression. PMID- 18158836 TI - Phenolic compounds and antioxidant activity of Italian extra virgin olive oil Monti Iblei. AB - The profile of phenolic compounds, antioxidant capacity, oxidative stability, and chemical characteristics (free acidity, peroxide value, specific extinction K232 and K270 values, and DeltaK) of 22 commercial extra virgin olive oil (EVOO) samples coming from the denomination of protected origin (DPO) Monti Iblei and obtained from olives harvested in the period September-December 2005 in the production area of the province of Siracusa (Sicily, Italy) were evaluated. The content of total phenols, expressed as gallic acid equivalents, ranged from 14.80 to 121.20 mg/100 g, with a mean value of 53.72 mg/100 g, mainly attributable to deacetoxyligstroside aglycone, deacetoxyoleuropein aglycone, oleuropein aglycone, and ligstroside aglycone. The mean values of Trolox equivalent antioxidant capacity (TEAC) and of oxidative stability were 54.76 and 11.99 hours, respectively. Both TEAC and oxidative stability were positively correlated to the phenol content and to the percentage of inclusion of the olive cultivar "Tonda Iblea." The high mean content of phenols, besides conferring prolonged oxidative stability, likely confers to the DPO Monti Iblei EVOO marked potential beneficial effects for human health. PMID- 18158837 TI - Inhibition of tyrosine phosphorylation of vascular endothelial growth factor receptors in human umbilical vein endothelial cells: a potent anti-angiogenic lipid-rich extract from shark. AB - We have previously reported that an ethanolic extract of dried shark muscle mixed with olive oil (shark muscle-olive oil [SMO]) has potent anti-angiogenic activity and that this extract appears to inhibit the binding of vascular endothelial growth factor (VEGF) to its receptor(s). In this study, we investigated the effects of SMO on the phosphorylation of VEGF receptor(s) in human umbilical vein endothelial cells (HUVECs). In vitro cell proliferation assays showed that SMO significantly reversed the VEGF-promoted increase in HUVEC proliferation. Western blot analysis revealed that SMO treatment markedly inhibited the VEGF-promoted tyrosine phosphorylation of VEGF receptor-2 (KDR) and VEGF receptor-1 (Flt-1) in a dose-dependent manner. These results demonstrated that SMO might interfere with or block the binding of VEGF with its receptors, and thereby inhibit the VEGF receptor(s) signal transduction pathway and so inhibit angiogenesis. PMID- 18158838 TI - A 5-month open study with long-chain polyunsaturated fatty acids in dyslexia. AB - This open pilot study investigated effects of a docosahexaenoic acid (DHA)-rich supplement on learning ability in a group of 20 dyslexic children in Sweden. Children formally diagnosed as dyslexic took eight capsules per day of a long chain polyunsaturated fatty acid (LC-PUFA) supplement containing high-DHA fish oil and evening primrose oil. Subjective assessments by the children and their parents were completed at baseline and 6, 12, and 20 weeks after supplementation. Quantitative evaluation by word-chain test was completed before and after 4 months of supplementation to measure word decoding (speed of reading) and letter decoding (motoric-perceptual speed). Subjective parent and child assessments showed increasing numbers of positive responders over time in reading speed, general schoolwork, and overall perceived benefit. Significant improvements were observed in reading speed and motor-perceptual velocity. Thirteen of 17 children had a significant improvement on the word-chain test (P < .04). Reading speed improved by 60% from 1.76 +/- 0.29 before the study to 2.82 +/- 0.36 after supplementation (P < .01 by Wilcoxon sign test). Motoric-perceptual velocity improved by 23% from a stanine value of 3.76 +/- 0.42 to 4.65 +/- 0.66 after supplementation (P < .05 by Wilcoxon sign test). Thus LC-PUFA supplementation for 5 months provides positive and clear beneficial effect on variables usually impaired by dyslexia. PMID- 18158839 TI - In vitro antioxidant and free radical scavenging activity of Cyperus rotundus. AB - Cyperus rotundus (Family Cyperaceae) is used both as a functional food and as a drug. In this study, the antioxidative potential of a hydroalcoholic extract of C. rotundus (CRE) was evaluated by various antioxidant assays, including antioxidant capacity by the phosphomolybdenum method, total antioxidant activity in linoleic acid emulsion systems, 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide, hydroxyl radicals, and nitric oxide (NO) scavenging. We further evaluated the reducing potential of the extract as well as Fe(2+)/ascorbate induced lipid peroxidation in rat liver homogenate. These various antioxidant activities were compared to standard antioxidants such as butylated hydroxytoluene, tocopherol, L-ascorbic acid, and catechin. Total phenolic and flavonoid content of CRE was also determined by a colorimetric method. The extract exhibited high reduction capability and powerful free radical scavenging, especially against DPPH and superoxide anions as well as a moderate effect on NO. CRE also showed inhibited lipid peroxidation in rat liver homogenate induced by Fe(2+)/ascorbate and prevented deoxyribose degradation in both non-site-specific and site-specific assays showing the hydroxyl radical scavenging and metal chelating activity of the hydroalcoholic extract. Moreover, the peroxidation inhibiting activity of CRE was demonstrated in the linoleic acid emulsion system. These results clearly established the antioxidative potency of C. rotundus, which may account for some of the medical claims attributed to this plant. PMID- 18158840 TI - Antioxidant effect of Hemidesmus indicus on ethanol-induced hepatotoxicity in rats. AB - The antioxidant effect of the ethanolic root extract of Hemidesmus indicus, an indigenous Ayurvedic medicinal plant used in soft drinks in India, was studied in rats with ethanol-induced hepatotoxicity. Administering 20% ethanol (5 g/kg of body weight/day) for 60 days to male Wistar rats resulted in significantly decreased body weight and increased liver/body weight ratio. The liver marker enzymes, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatae (ALP), gamma-glutamyl transpeptidase (GGT), and lactate dehydrogenase (LDH), were elevated. In addition, the levels of plasma, erythrocyte, and hepatic thiobarbituric acid-reactive substances (TBARS), hydroperoxides (LOOH), and conjugated dienes (CD) were also elevated in ethanol-fed rats as compared to those of the experimental control rats. Decreased activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), vitamin C, and alpha-tocopherol (vitamin E) were also observed in ethanol-administered as compared to control rats. Ethanolic root extract of H. indicus was administered at a dose of 500 mg/kg of body weight/day for the last 30 days of the experiment to rats with ethanol-induced liver injury, which significantly increased body weight, significantly decreased the liver/body weight ratio, AST, ALT, ALP, GGT, and LDH activities, and also the levels of TBARS, LOOH, and CD, significantly elevated the activities of SOD, CAT, GPx, and GSH in plasma, erythrocytes, and liver, and also increased levels of plasma and liver vitamin C and vitamin E at the end of the experimental period as compared to those of untreated ethanol-administered rats. Thus, our data indicate that treatment with H. indicus extract offers protection against free radical-mediated oxidative stress in plasma, erythrocytes, and liver of animals with ethanol induced liver injury. PMID- 18158841 TI - Evaluation of in vitro antioxidant activity of Sida rhombifolia (L.) ssp. retusa (L.). AB - Sida rhombifolia (L.) ssp. retusa (L.) is widely used in Ayurvedic medicine for the treatment of fever as well as a diuretic. The comparative antioxidant potentials of ethanol extract of roots, stems, leaves, and whole plant were studied. Estimation of total polyphenolic content and high-performance thin-layer chromatography profile were determined. Further inhibition of oxygen-derived free radicals, viz., assays for free radical scavenging, reducing power, superoxide anion scavenging, nitric oxide scavenging, and anti-lipid peroxidation, were performed. All the antioxidant activities were compared with standard antioxidants such as butylated hydroxyanisole and alpha-tocopherol acetate. Extracts were found to be good scavengers of the 1,1-diphenyl-2-picrylhydrazyl radical in the order root > leaves > whole plant > stem with 50% inhibitory concentrations of 546.1, 852.8, 983.8, and 1,222.5 microg/mL, respectively. All extracts of this plant showed effective free radical scavenging activity, reducing power, and superoxide scavenging activity. Only root extract inhibited lipid peroxidation in rat liver and brain homogenate. All these antioxidant properties were concentration dependent. In addition, total polyphenolic contents of all the extracts were determined as gallic acid equivalents. The highest antioxidant activity was observed in root extract. The results obtained from the current study indicate that S. rhombifolia ssp. retusa is a potential source of natural antioxidants. PMID- 18158842 TI - Antifatigue effect of Rubus coreanus Miquel extract in mice. AB - The antifatigue properties of six Korean medicinal herb extracts were studied by evaluating forced swimming capacity and biochemical parameters in ICR mice. The treatment groups were orally administered 30% ethanolic extracts (500 mg/kg/day) of Rubus coreanus Miquel, Cyperus rotundus Linn., Acanthopanax sessiliflorus, Saururus chinensis Baili, Epimedium koreanumNakai, or Houttuynia cordata Thunb. for 4 weeks. Swimming time to exhaustion was found to be longer for the group fed R. coreanus than for the control group (P < .05). No significant differences were found in the plasma levels of either glucose or lactate between the control group and the group fed R. coreanus, which swam longer than the control. The plasma ammonia levels were significantly lower in the groups fed R. coreanus and A. sessiliflorus, when compared to the control group (P < .05). No significant differences were found in gastrocnemius muscle or liver glycogen content between the control group and any treatment group. These results suggest that R. coreanus extract, and none of the other herbs, has antifatigue effects in mice, as demonstrated by the increased forced swimming capacity and decreased plasma ammonia accumulation. PMID- 18158843 TI - The effects of selected dietary bioflavonoid supplementation on dental caries in young rats fed a high-sucrose diet. AB - Recent evidence suggests that certain bioflavonoids reduce dental caries and cariogenic bacteria incidence. The present study evaluates two separate, but related, dietary trials -- trial 1, 0.09%, 0.18%, 0.36%, and 0.72% dietary naringenin (NAR) supplementation; and trial 2, 0.57% dietary rutin (R), quercetin (Q), and naringin (N) supplementation-on dental caries formation in 40 different male albino rats, at the expense of dextrose, for periods of 42 days. All rats were fed 40% sucrose. In dietary trial 1, rats were evaluated for dental caries, dental plaque accumulation, and saliva flow rates using oneway analysis of variance, post hoc Tukey's test, Kruskal-Wallis test, and Spearman's correlations. In dietary trial 2, rats were evaluated for occlusal dental caries only using a Kruskal-Wallis H test and analysis of variance. A 5% level of statistical significance was adopted throughout. In dietary trial 1, NAR showed a statistically significant effect on dental caries, plaque, and saliva flow rate reduction compared with the control group (P < .05-.01). An inverse dose dependent relationship was established among the NAR experimental groups and control group. Dietary NAR supplementation significantly reduced dental caries formation, possibly because of reduced dental plaque accumulation. In dietary trial 2, statistically significant reductions in occlusal caries were observed for R, Q, and N in the maxillary molars and for Q and N in the mandibular molars compared with the control group (P < .05). Significant associations were observed among the experimental groups and maxillary (P < .05) and mandibular (P < .01) occlusal dental caries. Hence, selected bioflavonoids may show promise as an alternative means of reducing dental caries. PMID- 18158844 TI - Anti-ulcerogenic effects of the flavonoid-rich fraction from the extract of Orostachys japonicus in mice. AB - Orostachys japonicus (Crassulaceae) herbal preparations have been used to treat gastric ulcer or gastric cancer disease in Korean folk medicine. To demonstrate the effects of the methanol (MeOH) extract of O. japonicus and its fractions on gastric lesions and pain, the MeOH extract was fractionated into triterpene-rich and flavonoid-rich (FRF) fractions. Second, the fractions were subjected to analgesic assays including hot plate and writhing assays and anti-ulcerogenic assays in HCl/ethanol-induced- and indomethacin/bethanechol-induced ulcer models in mice. In this experiment, it was found that the FRF most significantly reduced ulcerative indices and pain in mice, although the MeOH extract was also effective. Oral administration of the FRF highly reduced the diameter of gastric lesion induced by HCl/ethanol (inhibitory effect, 53%) and by indomethacin/bethanechol (inhibitory effect, 36%) at the 100 mg/kg dose. In addition, oral administration of 200 mg/kg FRF markedly increased the reaction time in the hot plate test by 52% and decreased stretching episodes (45%) in the writhing test. These results suggest that the active component of O. japonicus exhibiting the potent anti-ulcerative and antinociceptive effect is included in the FRF. The anti-ulcerative effects of the MeOH extract and the FRF were also supported by gastric juice and gastric acid volumes and pH in pylorus-ligated mice. Taken together, these results provide evidence-based support for the traditional use of O. japonicus for gastric disease. PMID- 18158845 TI - Antioxidant properties of the methanol extracts from the leaves of Paullinia pinnata. AB - Paullinia pinnata is an African woody vine widely used in traditional medicine for the treatment of malaria. In order to throw more light into its mechanisms of pharmacological actions, the in vitro antioxidant activities of the methanol extract of its leaves were evaluated using different testing systems. Its scavenging activities on 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis-3 ethylbenzothiazoline-6-sulfonic acid radicals as well as its ferric ion reducing power were assessed. The results showed that P. pinnata possessed strong scavenging activity and moderate reducing power. The total phenol, flavonoid, and proanthocyanidin contents of the extracts were very close to those reported for most medicinal plants and showed good correlation with its antioxidant activities. These properties are probably part of the reasons why P. pinnata is effective in folk medicine. PMID- 18158846 TI - Sahara honey shows higher potency against Pseudomonas aeruginosa compared to north Algerian types of honey. AB - Six varieties of honey from different regions in Algeria were used to determine their potency against Pseudomonas aeruginosa. Four varieties originated from northern Algeria, and two from the Sahara. Three types of media were used. On nutrient agar the minimum inhibitory concentration (MIC) of the four northern varieties ranged between 30% (vol/vol) and 31% (vol/vol), while the MIC of the Sahara varieties was 11% (vol/vol) and 14% (vol/vol). On King A agar the MIC of the four northern varieties ranged from 25% (vol/vol) to 31% (vol/vol), whereas the MIC of the two varieties of Sahara honey was 12% (vol/vol) and 15% (vol/vol). On nutrient broth the MIC of the northern varieties ranged from 10% (vol/vol) to 21% (vol/vol), whereas the MIC of the two varieties of Sahara honey was 9% (vol/vol). The botanic flora of Sahara is known in Algeria for its medicinal uses, and thus the higher potency of the Sahara honey is most probably due to antibacterial substances in its plant derivates. These findings suggest that Sahara honey could be used for managing wounds and burns, which are mostly infected by P. aeruginosa. PMID- 18158847 TI - Gas chromatography/mass spectrometry analysis of Laurus nobilis essential oil composition of northern Cyprus. AB - The chemical composition of the essential oil isolated from the leaves of the Laurus nobilis plant (from the Northern Cyprus Mountains) by hydrodistillation was analyzed by gas chromatography-mass spectrometry. Of the 81 compounds representing 98.74% of total oil, monocyclic monoterpenes such as 1,8-cineole (58.59%), alpha-terpinyl acetate (8.82%), and terpinene-4-ol (4.25%) were the main components. Bicyclic monoterpenes such as alpha- and beta- pinene (3.39 3.25%) and sabinene (3.32%) were also identified. The acyclic monoterpenes linalool (0.19%) and myrcenol (0.10%) were present in smaller amounts, and so were the sesquiterpenes. o-Cymene (1.30%) and p-cymene (1.83%) were the main, while cumin aldehyde (0.24%), dimethylstyrene (0.08%), eugenol (0.16%), methyl eugenol (0.05%), and carvacrol (0.05%) were found as minor, aromatic compounds of laurel oil. PMID- 18158848 TI - Fourteen weeks of treatment with Viscofiber increased fasting levels of glucagon like peptide-1 and peptide-YY. AB - Fermentable dietary fiber has been shown to cause fat loss and to increase peptide-YY (PYY) and glucagon-like peptide 1 (GLP-1) levels in rodents. In single meal tests, humans have an increase in PYY and GLP-1 to dietary fiber, but the response of these hormones to longer-term treatment is not known. Viscofiber (Cevena Bioproducts Inc., Edmonton, AB, Canada) is a high-viscosity fermentable dietary fiber made by a proprietary process from oats and barley. Seven healthy overweight and obese subjects were treated with a calorie-restricted diet, a lifestyle change program, and 4 g of Viscofiber/day for 16 weeks. Hunger, satiety, PYY, and GLP-1 were measured before and 1 hour after a standard meal test before and at week 14 of the study. Hunger and satiety were measured by Visual Analog Scales. PYY and GLP-1 were measured by radioimmunoassay and enzyme linked immunosorbent assay, respectively. Weight was reduced 3.07 +/- 3.13 kg (P < .05) over the 16 weeks. Fasting PYY increased 8.67 +/- 6.62 pg/mL (P < .05) and fasting GLP-1 increased 2.67 +/- 0.84 pmol/L (P < .01) at 14 weeks compared to baseline. Satiety increased 1.78 +/- 1.43 cm (P < .01) at the 1-hour post-meal time point on week 14 compared to the study baseline. We conclude that 14 weeks of treatment with Viscofiber at 4 g/day along with a lifestyle change program and diet causes weight loss and increases fasting PYY, fasting GLP-1, and satiety at 1 hour following a standard meal, which extends the single meal test observations in humans. PMID- 18158849 TI - Platelet reactivity in male smokers following the acute consumption of a flavanol rich grapeseed extract. AB - Epidemiological studies suggest that a high dietary intake of flavanols, a subclass of flavonoids, is associated with reduced risk of vascular disease. Clinical studies have also shown that the consumption of certain flavanol-rich foods (e.g., cocoa, tea, red wine), as well as intake of the individual flavanol (-)-epicatechin, can result in improvement in a number of parameters associated with vascular disease, including improved endothelial function, reduced platelet reactivity, and reduced oxidative stress. The present study assessed the effects of a flavanol-rich supplement on platelet reactivity and plasma oxidant defense in a group of smokers, a population at an elevated risk for vascular disease. Male smokers were randomly assigned to a placebo (n = 10) or a flavanol-rich grapeseed extract (FRGSE; n = 13) group, and after an overnight fast, blood samples were collected before and at 1, 2, and 6 hours following consumption of the placebo or supplement. The FRGSE supplement, but not the placebo, significantly decreased ADP-stimulated platelet reactivity at 1, 2, and 6 hours following intake (P < .05) compared to baseline levels. Similarly, the supplement, but not the placebo, decreased epinephrine-stimulated platelet reactivity 2 hours following consumption. Plasma antioxidant capacity (total radical trapping antioxidant potential), lipid oxidation (plasma thiobarbituric acid-reactive substances), and serum uric acid concentrations were not affected in either group. Thus smokers may obtain some health benefits from the consumption of certain flavanol-rich foods, beverages, and supplements. PMID- 18158850 TI - Increase of bioavailability of coenzyme Q(10) and vitamin E. AB - Commercial coenzyme Q(10) (CoQ(10)) and alpha-tocopherol (vitamin E) formulations often show poor intestinal absorption. Delivery of CoQ(10) and vitamin E was enhanced when used with a new formulation, NanoSolve (Lipoid GmbH, Ludwigshafen, Germany), as shown by an open, comparative monocenter, crossover study of 24 volunteers. Plasma CoQ(10) and vitamin E were determined from predose until +14 hours. To compare bioavailability, corrected maximum concentration, time to reach maximum concentration, and area under the curve from 0 to 14 hours were assessed. The NanoSolve test formulation contained 100 mg of CoQ(10) and 120 mg of vitamin E. The pure substances in hard gelatin capsules served as the reference. Although identical amounts of CoQ(10) and vitamin E were administered, absolutely higher serum concentrations of the active ingredients were achieved by the NanoSolve formulation than by the pure materials in gelatin capsules. The bioavailability of CoQ(10) increased fivefold after administration of the NanoSolve formulation, and the bioavailability of vitamin E was enhanced 10-fold both compared to the pure substances. PMID- 18158851 TI - Nutritional and anti-nutritional compositions of Cleome rutidosperma, Lagenaria siceraria, and Cucurbita maxima seeds from Nigeria. AB - Flours produced from the seeds of Cleome rutidosperma (DC), Lagenaria siceraria (Mol.), and Cucurbita maxima (Duch) were evaluated for proximate and elemental compositions. Preliminary phytochemical screening was also carried out on the seed flours, which revealed the presence of alkaloids, steroids, pentose, and reducing sugars in all the seed types. Tannins, flavonoids, and cardiac glycosides were also detected in only C. rutidosperma seed. The results also showed that the seeds of C. rutidosperma, L. siceraria, and C. maxima contained carbohydrates (74.43%, 45.93%, and 24.30%), fats (7.20%, 38.92%, and 51.49%), and proteins (11.73%, 8.93%, and 16.80%), respectively. The major mineral contents of the three defatted seed flours were found to include potassium (39.00 ppm, 19.50 ppm, and 39.00 ppm), sodium (23.00 ppm, 11.50 ppm, and 11.50 ppm), and calcium (18.00 ppm, 12.00 ppm, and 15.00 ppm), respectively. They were also found to contain zinc (0.013 ppm, 0.04 ppm, and 0.084 ppm) and iron (0.014 ppm, 0.028 ppm, and 0.032 ppm), respectively. The proximate and mineral compositions suggest that the seeds are potential sources of carbohydrate, fats, protein, and micro- and macrominerals and may find use as human food or for incorporation into livestock feed. However, the seeds were revealed to also contain slight traces of lead, although at such very low concentrations they may be artifacts. This calls for further research into possible lead toxicity after prolonged consumption of these seeds. PMID- 18158853 TI - Tumorigenesis and embryonic stem cell-derived therapy. PMID- 18158854 TI - A novel monoclonal antibody (STRO-3) identifies an isoform of tissue nonspecific alkaline phosphatase expressed by multipotent bone marrow stromal stem cells. AB - Numerous studies support the concept that the nonhemopoietic cells of the bone marrow (BM), are derived from a population of multipotent bone marrow stromal stem cells (BMSSCs), which reside in perivascular niches within the bone marrow. These BMSSCs are thought to give rise not only to more cells that are phenotypically and functionally identical but also differentiated, lineage committed mesenchymal progeny, including chondrocytes, smooth muscle cells, adipocytes, and osteoblasts. Recently, we have generated a novel monoclonal antibody (mAb) (designated STRO-3) that reacts with a minor subset of STRO-1(+) cells contained within adult BM aspirates and does not react with CD34(+) hemopoietic stem cells. Our results also show that STRO-3 identifies a high proportion of BMSSCs that possess extensive proliferative and multilineage differentiative capacity. Using retroviral expression cloning, we determined that STRO-3 binds to tissue nonspecific alkaline phosphatase (TNSALP), a cell-surface glycoprotein usually associated with cells of the osteoblast lineage. Studies presented here suggest that in addition to being expressed by osteoblasts, TNSALP may also represent a marker of immature BMSSCs in vivo. Finally, these studies suggest that antibodies to TNSALP may be used as an effective single marker of enrichment of BMSSCs from various tissues. PMID- 18158857 TI - Morbidity at elementary school entry differs by sex and level of residence urbanization: a comparative cross-sectional study. AB - BACKGROUND: Health is vital to a child's learning in school and success in life. Therefore, early physical examination, and follow-up if necessary, would bring parents' attention to their child's health and would likely improve outcomes. The purposes of this study are twofold: to assess the health status of first-graders and to examine the health status differences between sexes, levels of residence urbanization, and quantity of available medical resources. METHODS: This is a comparative descriptive study. Data from the 2002 Student Entry Physical Examination (SEPE) and Student Medical History Inventory (SMHI) were obtained from 203 public and private elementary schools in northern Taiwan where a population of 53,053 students was included. Frequencies, independent sample t test, one-way ANOVA along with Scheff's post hoc test, and Pearson's correlation were conducted using SPSS. RESULTS: This study showed that 13.7% of students had at least one diagnosed disease from the SMHI reported by parents. Moreover, the SEPE indicated that 79.5% students had at least one health concern. Dental caries, myopia, and obesity were the most prevalent health problems among the first-graders (69.6%, 27.1%, and 9.5%, respectively). Research results show that there were significant differences in the prevalence of dental caries, myopia, and obesity between different sexes and among levels of urbanization. However, the quantity of available medical resources made no significant difference. CONCLUSION: Elementary school entry physical examination is an important way to detect students' health problems. It is suggested that school health interventions consider students' health profiles along with their sex and level of urbanization in planning. More research is needed to find the risk factors of the health problems. Additionally, the creation of a school health committee is suggested to implement and evaluate the entry health examination program. PMID- 18158859 TI - Differential relations between two types of contact and implicit and explicit racial attitudes. AB - Contact with out-group members has been associated with more favourable explicit attitudes towards the out-group in general, largely via the mediation of reduced intergroup anxiety. In addition, there is now a growing body of evidence suggesting that contact relates to automatically activated evaluations termed implicit attitudes. However, research has not fully illuminated the mechanisms through which contact with outgroup members impacts on implicit attitudes. A study investigating this issue assessed White participants' (N=105) explicit attitudes, implicit attitudes, intergroup anxiety, and contact quantity and quality about Asians. Greater contact quality was related to more positive explicit attitudes, while contact quantity was associated with more positive implicit attitudes. Both effects were mediated by reduced intergroup anxiety. PMID- 18158858 TI - Characterization of the Trichomonas vaginalis surface-associated AP65 and binding domain interacting with trichomonads and host cells. AB - BACKGROUND: AP65 is a prominent adhesin of Trichomonas vaginalis that mediates binding of parasites to host vaginal epithelial cells (VECs). AP65 with no secretion signal sequence, membrane targeting peptide, and anchoring motif was recently found to be secreted. RESULTS: We first wanted to demonstrate surface association of AP65 to the parasite followed by the identification of the binding epitope interacting with both organisms and VECs. AP65 was found to bind to trichomonads, but not to trypsin-treated parasites, in an auto-ligand assay, suggesting the existence of a surface protein associating with AP65. Since rabbit antiserum IgG antibodies reactive with epitopes localized to the N-terminal region of AP65 inhibit the attachment of live parasites to VECs, we hypothesized that the binding domain was localized to this region. We subcloned five overlapping fragments of AP65 called c1 through c5, and expression of recombinant clones was confirmed with antibodies to AP65. Each purified recombinant protein was then tested for binding activity using an established ligand assay, and fragment c1 with the first twenty-five amino acids in the N-terminal domain was required for binding to VECs and, surprisingly, also to parasites. Importantly, c1 competed with the binding of AP65 to both cells types. CONCLUSION: T. vaginalis AP65 is a secreted, surface-associated protein and a model is proposed to explain how this secreted protein functions as an adhesin. PMID- 18158860 TI - It depends what you do in the laboratory. PMID- 18158861 TI - Core beliefs and narcissistic characteristics among eating-disordered and non clinical women. AB - OBJECTIVES: Narcissism is a personality trait that can interfere with the application of evidence-based therapies for the eating disorders, influencing collaboration and the patient's willingness to take responsibility for participating in change. In order to understand and work with this personality characteristic, it is important to understand the cognitions that underpin the traits concerned. DESIGN: This study examined the associations between schema level core beliefs and narcissism in eating disorders. Narcissism was conceptualized in terms of both its core element (entitlement and grandiosity) and the narcissistic defences (bad you and poor me attitudes). METHODS: Validated measures of the different elements of narcissism and of core beliefs were completed by 80 eating-disordered patients and 70 non-clinical comparison women. Multiple regression analyses were used to determine the core beliefs associated with each aspect of narcissism. RESULTS: The pattern of association differed across the two groups. Among the eating-disordered women, different core beliefs were associated with core narcissism and with each of the two defences. CONCLUSIONS: Unconditional schema-level beliefs are associated with narcissistic personality traits in the eating disorders suggesting that these therapy interfering personality characteristics might be addressed by modifying the relevant core beliefs, thus making it possible to work more directly with the eating disorder itself. PMID- 18158862 TI - Molecular markers and targets for colorectal cancer prevention. AB - Colorectal cancer is the third most prevalent cancer in the world. If detected at an early stage, treatment often might lead to cure. As prevention is better than cure, epidemiological studies reveal that having a healthy diet often protects from promoting/ developing cancer. An important consideration in evaluating new drugs and devices is determining whether a product can effectively treat a targeted disease. There are quite a number of biomarkers making their way into clinical trials and few are awaiting the preclinical efficacy and safety results to enter into clinical trials. Researchers are facing challenges in modifying trial design and defining the right control population, validating biomarker assays from the biological and analytical perspective and using biomarker data as a guideline for decision making. In spite of following all guidelines, the results are disappointing from many of the large clinical trials. To avoid these disappointments, selection of biomarkers and its target drug needs to be evaluated in appropriate animal models for its toxicities and efficacies. The focus of this review is on the few of the potential molecular targets and their biomarkers in colorectal cancers. Strengths and limitations of biomarkers/surrogate endpoints are also discussed. Various pathways involved in tumor cells and the specific agents to target the altered molecular biomarker in biomolecular pathway are elucidated. Importance of emerging new platforms siRNAs and miRNAs technology for colorectal cancer therapeutics is reviewed. PMID- 18158863 TI - Advances in the genetics of Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder affecting a significant proportion of the ageing population. The etiology is unknown and it is likely due to a multifactorial interaction of genes and the environment on the background of ageing. Findings in the last decade suggest that the contribution of genetics to familial forms of PD is much greater than previously appreciated. Twelve loci are now associated with highly penetrant autosomal dominant or recessive PD, and causative mutations have been identified in eight genes with mutation carriers often characterized by a phenotype indistinguishable from idiopathic disease. To date, PD pharmacotherapy is symptomatic only and does not slow disease progression. Understanding how genetic mutations cause familial PD is likely to clarify molecular mechanisms underlying PD in general and will provide a guide for the development of novel therapies, both preventative and palliative, applicable to all forms of parkinsonism. This review outlines the advances in the study of the genetic background of PD and their possible clinical implications. PMID- 18158864 TI - The interaction of small molecules with phospholipid membranes studied by 1H NOESY NMR under magic-angle spinning. AB - The interaction of small molecules with lipid membranes and the exact knowledge of their binding site and bilayer distribution is of great pharmacological importance and represents an active field of current biophysical research. Over the last decade, a highly resolved 1H solid-state NMR method has been developed that allows measuring localization and distribution of small molecules in membranes. The classical solution 1H NMR NOESY technique is applied to lipid membrane samples under magic-angle spinning (MAS) and NOESY cross-relaxation rates are determined quantitatively. These rates are proportional to the contact probability between molecular segments and therefore an ideal tool to study intermolecular interactions in membranes. Here, we review recent 1H MAS NOESY applications that were carried out to study lateral lipid organization in mixed membranes and the interaction of membranes with water, ethanol, small aromatic compounds, peptides, fluorescence labels, and lipophilic nucleosides. PMID- 18158865 TI - Adenovirus viral interleukin-10 inhibits adhesion molecule expressions induced by hypoxia/reoxygenation in cerebrovascular endothelial cells. AB - AIM: To investigate the effects of recombinant adenovirus encoding viral interleukin-10 (vIL-10), a potent anti-inflammatory cytokine, on adhesion molecule expressions and the adhesion rates of leukocytes to endothelial cells in cerebrovascular endothelial cells injured by hypoxia/reoxygenation (H/R). METHODS: A recombinant adenovirus expressing vIL-10 (Ad/vIL-10 (or the green fluorescent protein (Ad/GFP) gene was constructed. A cerebrovascular endothelial cell line bEnd.3 was pretreated with a different multiplicity of infection (MOI) of Ad/vIL-10 or Ad/GFP and then exposed to hypoxia for 9 h followed by reoxygenation for 12 h. The culture supernatants were tested for the expression of vIL-10 and endogenous murine IL-10 (mIL-10) by ELISA. The effects of Ad/vIL-10 on monocyte-endothelial cell adhesion were represented as the adhesion rate. Subsequently, the expressions of intercellular adhesion molecule 1(ICAM-1) and vascular cell adhesion molecule 1( VCAM-1) in the endothelial cells after treatment with Ad/vIL-10 and H/R were analyzed by Western blotting and real-time PCR. RESULTS: vIL-10 was expressed in cultured bEnd.3 after Ad/vIL-10 transfection and was significantly increased by H/R. Ad/vIL-10 or Ad/GFP did not affect the mIL-10 level. H/R increased the mIL-10 expression, but insignificantly. Monocyte- endothelial cell adhesion induced by H/R was significantly inhibited by pretreatment with Ad/vIL-10(MOI:80). ICAM-1, and VCAM 1 in bEnd.3 and were significantly increased after H/R, while pretreatment with Ad/vIL-10 (MOI: 80) significantly inhibited their expressions. Ad/GFP did not markedly affect monocyte- endothelial adhesion and the expressions of ICAM-1 and VCAM-1 induced by H/R. CONCLUSION: Ad/vIL-10 significantly inhibits the upregulation of endothelial adhesion molecule expressions and the increase of adhesion of monocytes- endothelial cells induced by H/R, indicating that vIL-10 gene transfer is of farreaching significance in the therapy of cerebrovascular inflammatory diseases, and anti-adhesion treatment may reduce H/R injury. PMID- 18158866 TI - Astragaloside IV inhibits spontaneous synaptic transmission and synchronized Ca2+ oscillations on hippocampal neurons. AB - AIM: To investigate the changes in the spontaneous neuronal excitability induced by astragaloside IV (AGS-IV) in the cultured hippocampal network. METHODS: Hippocampal neurons in culture for 9-11 d were used for this study. The spontaneous synaptic activities of these hippocampal neurons were examined by Ca2+ imaging and whole-cell patch-clamp techniques. In total, 40 mg/L AGS-IV dissolved in DMSO and 2 mL/L DMSO were applied to the neurons under a microscope while the experiments were taking place. RESULTS: AGS-IV inhibited the frequencies of synchronized spontaneous Ca2+ oscillations to 59.39%+/- 3.25%(mean+/-SEM), the spontaneous postsynaptic currents to 43.78%+/- 7.72%(mean+/-SEM), and the spontaneous excitatory postsynaptic currents to 49.25%+/- 7.06%(mean+/-SEM) of those of the control periods, respectively, at 16 min after the AGSIV applications. AGS-IV also decreased the peak values of the voltage-gated K+ and Na+ channel currents at that time point. CONCLUSION: These results indicate that AGS-IV suppresses the spontaneous neuronal excitabilities effectively. Such a modulation of neuronal activity could represent new evidence for AGS-IV as a neuroprotector. PMID- 18158867 TI - Identification of chronic myocardial infarction with extracellular or intravascular contrast agents in magnetic resonance imaging. AB - AIM: To determine whether extracellular or intravascular contrast agents could detect chronic scarred myocardium in magnetic resonance imaging (MRI). METHODS: Eighteen pigs underwent a 4 week ligation of 1 or 2 diagonal coronary arteries to induce chronic myocardial infarction. The hearts were then removed and perfused in a Langendorff apparatus. Eighteen hearts were divided into 2 groups. The hearts in groups I (n=9) and II (n=9) received the bolus injection of Gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA, 0.05 mmol/kg) and gadolinium- based macromolecular agent (P792, 15 micromol/kg), respectively. First pass T2* MRI was acquired using a FLASH sequence. Delayed enhancement T1 MRI was acquired with an inversion recovery prepared TurboFLASH sequence. RESULTS: Wash-in of both agents resulted in a sharp and dramatic T2* signal loss of scarred myocardium similar to that of normal myocardium. The magnitude and velocity of T2* signal recovery caused by wash-out of extracellular agents in normal myocardium was significantly less than that in scarred myocardium. Conversely, the T2* signal of scarred and normal myocardium recovered to plateau rapidly and simultaneously due to wash-out of intravascular agents. At the following equilibrium, extracellular agent enhanced T1 signal intensity was significantly greater in scarred myocardium than in normal myocardium, whereas there was no significantly statistical difference in intravascular agent-enhanced T1 signal intensity between scarred and normal myocardium. CONCLUSION: After administration of extracellular agents, wash-out T2* first-pass and delayed enhanced T1 MRI could identify scarred myocardium as a hyperenhanced region. Conversely, scarred myocardium was indistinguishable from normal myocardium during first-pass and the steady state of intravascular agents. PMID- 18158868 TI - Hypoxic preconditioning attenuates hypoxia/reoxygenation-induced apoptosis in mesenchymal stem cells. AB - AIM: Mesenchymal stem cells (MSC) are a promising candidate for cardiac replacement therapies. However, the majority of transplanted MSC are readily lost after transplantation because of poor blood supply, ischemia-reperfusion, and inflammatory factors. We aimed to study the effects of hypoxia preconditioning (HPC) on hypoxia/reoxygenation-induced apoptosis of MSC. METHODS: Three generations of MSC were divided into 6 groups, including the normal group, hypoxia-reoxygenation (H/R) group, cyclosporine A (CsA), and the HPC 10 min, 20 min, and 30 min groups. The apoptotic index, cell viability, mitochondrial membrane potential, translocation of Bcl-2 and bax, extracellular regulated kinase (ERK), Akt, hypoxia-inducible factor 1-alpha, and the vascular endothelial growth factor (VEGF) were tested after H/R treatment. RESULTS: HPC decreased the apoptotic index and increased the viability induced by H/R. Moreover, HPC markedly stabilized mitochondrial membrane potential, upregulated Bcl-2 and VEGF expressions, and increased the phosphorylation of ERK and Akt. As a positive control, CsA has the same function as HPC, except for promoting ERK and Akt phosphorylation and upregulating VEGF. CONCLUSION: HPC had a protective effect against MSC apoptosis induced by H/R via stabilizing mitochondrial membrane potential, upregulating Bcl-2 and VEGF, and promoting ERK and Akt phosphorylation. HPC has implications for the development of novel stem cell protective strategies. PMID- 18158869 TI - Promotion of cell proliferation by HBXIP via upregulation of human telomerase reverse transcriptase in human mesenchymal stem cells. AB - AIM: We previously found that the hepatitis B X-interacting protein (HBXIP) was able to promote the proliferation of cells. Telomerase activity is known to be critical in cellular senescence and its level is modulated by the regulation of the telomerase catalytic subunit, telomerase reverse transcriptase (TERT), at both the transcriptional and post-transcriptional levels. To investigate the mechanism of promoting proliferation by HBXIP, the effect of HBXIP on human TERT (hTERT) was investigated in human mesenchymal stem cells (hMSC). METHODS: BMMS-03 cells and hMSC from the bone marrow of a 4-month-old elicited fetus, were transiently transfected with the pcDNA3-hbxip plasmid encoding the HBXIP gene and pSilencer-hbxip plasmid encoding RNA interference (RNAi) targeting HBXIP mRNA, followed by the examination of the hTERT promoter reporter gene by luciferase assay, and the detection of telomerase activity by telomeric repeat amplication protocol, respectively, as well as the expression levels of hTERT, c-Myc, and Bcl 2 by Western blot analysis. RESULTS: The overexpression of HBXIP led to a significant upregulation of hTERT promoter activity, telomerase activity, and the expression levels of hTERT, c-Myc, and Bcl-2 in BMMS-03 cells. RNAi targeting HBXIP mRNA produced the opposite results completely. CONCLUSION: Our data demonstrated that HBXIP significantly stimulated the transcription and expression of hTERT and increased the activity of telomerase in BMMS-03 cells, which provides a new insight into the mechanism of promoting cell proliferation by HBXIP. PMID- 18158870 TI - Synergistic actions of diacylglycerol and inositol 1,4,5 trisphosphate for Ca2+ dependent inactivation of TRPC7 channel. AB - AIM: The aim of the present study was to explore the mechanism for the Ca2+- dependent inactivation of the canonical transient receptor potential (TRPC) 7 channel expressed in human embryonic kidney 293 cells. METHOD: The whole-cell patch-clamp technique was used in the study. RESULTS: With Ca2+-free external solution, the perfusion of 100 micromol/L carbachol to, or dialysis of the cell with 100 micromol/L guanosine 5'-3-O-(thio)triphosphate (GTPgammaS), induced large inward currents, respectively. These currents were rapidly inhibited by the addition of 1 mmol/L Ca2+ into the bath, and recovery from this inhibition was only partial after the Ca2+ removal, unless vigorous intracellular Ca2+ buffering with 10 mmol/L 1,2 bis(2- aminophenoxy)ethane-N,N,No,No-tetraacetic acid (BAPTA) (plus 4 mmol/L Ca2+) was employed. In contrast, the current induced by a membrane permeable analog of diacylglycerol (DAG), 1-oleoyl-2-acetyl-sn-glycerol (OAG; 100 micromol/L) did not undergo the inhibition persisting after Ca2+ removal. Interestingly, the inclusion of inositol 1,4,5 trisphosphate (IP3; 100 micromol/L) in the patch pipette rendered the OAG-induced current susceptible to the persistent Ca2+-mediated inhibition independent of the IP3 receptor in the majority of the tested cells, as evidenced by the inability of heparin and thapsigargin in reversing the effect of IP3. CONCLUSION: The present results suggest that Ca2+ entry via the activated TRPC7 channel plays a critical role in inactivating the channel where the cooperative actions of DAG and IP3 are essentially involved. PMID- 18158871 TI - Resistin induces insulin resistance, but does not affect glucose output in rat derived hepatocytes. AB - AIM: The aim of the present study was to observe the effects of resistin on insulin sensitivity and glucose output in rat-derived hepatocytes. METHODS: The rat hepatoma cell line H4IIE was cultured and stimulated with resistin; supernant glucose and glycogen content were detected. The insulin receptor substrate (IRS) 1 and IRS-2, protein kinase B/Akt, glycogen synthase kinase-3beta(GSK-3 beta), the suppressor of cytokine signaling 3 (SOCS-3) protein content, as well as the phosphorylation status were assessed by Western blotting. Specific antisense oligodeoxynucleotides directed against SOCS-3 were used to knockdown SOCS-3. RESULTS: Resistin induced insulin resistance, but did not affect glucose output in rat hepatoma cell line H4IIE. Resistin attenuated multiple effects of insulin, including insulin-stimulated glycogen synthesis and phosphorylation of IRS, protein kinase B/Akt, as well as GSK-3beta. Resistin treatment markedly induced the gene and protein expression of SOCS-3, a known inhibitor of insulin signaling. Furthermore, a specific antisense oligodeoxynucleotide directed against SOCS-3 treatment prevented resistin from antagonizing insulin action. CONCLUSION: The major function of resistin on liver is to induce insulin resistance. SOCS-3 induction may contribute to the resistin-mediated inhibition of insulin signaling in H4IIE hepatocytes. PMID- 18158872 TI - Hypomethylation of interleukin-4 and -6 promoters in T cells from systemic lupus erythematosus patients. AB - AIM: DNA methylation regulates gene expression, and hypomethylation is associated with abnormal T-cell function in systemic lupus erythematosus (SLE). However, little is known about the methylation levels of the interleukin (IL)-4 and -6 promoters in SLE patients. METHODS: T cells were isolated from 20 SLE patients and 10 healthy controls, activated in vitro in the presence or absence of 5- azacytidine (5-azaC), and their IL-4 and -6 transcripts were characterized using semiquantitative RT-PCR. Following bisulfate modification of their genomic DNA, the levels of DNA methylation in the IL-4 or -6 promoter were determined by nested PCR and direct sequencing. RESULTS: The levels of IL-4 and -6 mRNA transcripts were significantly higher in SLE T cells, as compared with that in the controls. Furthermore, the treatment of healthy T cells with 5-azaC demethylated the CpG islands in the IL-4 or -6 promoter and increased IL-4 and -6 mRNA transcriptions. Importantly, the hypomethylation of the CpG islands in the IL-4 and -6 promoters displayed in SLE patients was similar to that of healthy T cells treated with 5-azaC. Finally, the hypomethylation levels of the CpG islands in the IL-4 and -6 promoters in lupus patients were significantly correlated to the IL-4 and -6 expressions. CONCLUSION: The hypomethylation of the CpG islands of the IL-4 and -6 promoters accrued in T cells from SLE patients and was associated with the severity of SLE at the clinic. PMID- 18158873 TI - Anti-inflammatory effect of honokiol is mediated by PI3K/Akt pathway suppression. AB - AIM: In this study, we investigated the regulatory effects of honokiol on various inflammatory events mediated by monocytes/macrophages (U937/RAW264.7 cells) and lymphocytes (splenic lymphocytes and CTLL-2 cells) and their putative action mechanism. METHODS: In order to investigate the regulatory effects, various cell lines and primary cells (U937, RAW264.7, CTLL-2 cells, and splenic lymphocytes) were employed and various inflammatory events, such as the production of inflammatory mediators, cell adhesion, cell proliferation, and the early signaling cascade, were chosen. RESULTS: Honokiol strongly inhibited various inflammatory responses, such as: (i) the upregulation of nitric oxide (NO), prostaglandin E2 and TNF-alpha production and costimulatory molecule CD80 induced by lipopolysaccharide (LPS); (ii) the functional activation of beta1-integrin (CD29) assessed by U937 cell-cell and cell-fibronectin adhesions; (iii) the enhancement of lymphocytes and CD8+CTLL-2 cell proliferation stimulated by LPS, phytohemaglutinin A (PHA), and concanavalin A or interleukin (IL)-2; and (iv) the transcriptional upregulation of inducible NO synthase, TNF-alpha, cyclooxygenase 2, IL-12, and monocyte chemoattractant protein (MCP)-1. These anti-inflammatory effects of honokiol seem to be mediated by interrupting the early activated intracellular signaling molecule phosphoinositide 3-kinase (PI3K)/Akt, but not Src, the extracellular signal-regulated kinase, and p38, according to pharmacological, biochemical, and functional analyses. CONCLUSION: These results suggest that honokiol may act as a potent anti-inflammatory agent with multipotential activities due to an inhibitory effect on the PI3K/Akt pathway. PMID- 18158874 TI - Dynamic effects of autophagy on arsenic trioxide-induced death of human leukemia cell line HL60 cells. AB - AIM: To evaluate the contribution of an autophagic mechanism to the As2O3- induced death of human acute myeloid leukaemia cell line HL60 cells. METHODS: The growth inhibition of HL60 cells induced by As2O3 was assessed with 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assay. The activation of autophagy was determined with monodansylcadaverine labeling and transmission electron microscope. The role of autophagy in the As2O3-induced death of HL60 cells was assessed using autophagic and lysosomal inhibitors. Immunofluorescence, flow cytometry, and Western blot analysis were used to study the apoptotic and autophagic mechanisms. RESULTS: After treatment with As2O3, the proliferation of HL60 cells was significantly inhibited and the formation of autophagosomes increased. The blockade of autophagy maturation with the autophagy specific inhibitor 3-methyladenine (3-MA) or the lysosome-neutralizing agent NH4Cl 1 h before As2O3 potentiated the As2O3-induced death of HL60 cells. In contrast, 3-MA attenuated As2O3-induced death when administered 30 min after As2O3. 3-MA and NH4Cl also inhibited As2O3-induced upregulation of microtubule associated protein 1 light chain 3, the protein required for autophagy in mammalian cells. Following As2O3, lysosomes were activated as indicated by increased levels of cathepsins B and L. The apoptotic response of HL60 cells to As2O3 was suggested by the collapse of mitochondrial membrane potential, release of cytochrome c from mitochondria, and the activation of caspase-3. Pretreatment with 3-MA prior to As2O3 amplified these apoptotic signals, while posttreatment with 3-MA 30 min after As2O3 attenuated the apoptotic pathways. CONCLUSION: Autophagy plays complex roles in the As2O3-induced death of HL60 cells; it inhibits As2O3-induced apoptosis in the initiation stage, but amplifies the As2O3 mediated apoptotic program if it is persistently activated. PMID- 18158875 TI - Phosphorylation of beta-actin by protein kinase C-delta in camptothecin analog induced leukemic cell apoptosis. AB - AIM: This study was conducted to reveal new proteins involved in acute myeloid leukemia (AML) cell apoptosis. METHODS: Using camptothecin analog NSC606985- induced leukemic U937 cell apoptosis as a model, this study performed a differential proteomic analysis during apoptosis induction. The significantly modulated protein was underwent further investigation in the apoptotic process. RESULTS: We found that beta-actin protein presented two different spots on the two-dimensional electrophoresis (2-DE) map, which shared similar molecular weight and different pI. Those two spots demonstrated contrary changes (disappeared on the basic-end and increased on the acid-end spot) during apoptosis induction, although the total level of beta-actin kept constant. This observation was further confirmed by immunoblot analysis on 2-DE gel. When NSC606985-treated cell lysate was incubated with alkaline phosphotase, beta-actin on the basic-end spot was restored, indicating increased phosphorylation of beta-actin during NSC606985 induced apoptosis. Moreover, the polymerization of actin also decreased after NSC606985 treatment. The increased beta-actin phosphorylation and decreased actin polymerization was antagonized by pre-treatment of rottlerin, a specific protein kinase C-delta (PKC delta) inhibitor. CONCLUSION: All these results indicate that beta-actin was phosphorylated during apoptosis induction, which was mediated by activated PKC delta. PMID- 18158876 TI - [Centers of intravenous admixtures: one more advancement in hospital care]. PMID- 18158877 TI - [External periareolar incision for subdermal mastectomy in men with gynecomastia]. AB - BACKGROUND: Gynecomastia describes a benign increase of the mammary gland in men. When medical treatment fails, symptoms and psychological alterations persist. Subdermal mastectomy is the definitive treatment and can be achieved by different incisions, each with potential complications. We undertook this study to present clinical characteristics of 11 patients with gynecomastia and the results obtained with subdermal mastectomy by means of external periareolar incision. METHODS: A descriptive cohort study in male patients with gynecomastia was carried out in a third-level medical care hospital. Patients were treated with subdermal mastectomy by means of external periareolar incision. RESULTS: There were 11 male patients with an average age of 19 years (range: 11-60 years), 3 patients (27.2%) with bilateral gynecomastia and 8 patients (72.7%) with unilateral gynecomastia. Average time of evolution was 22 months (range: 16-48 months), 9 patients (81.8%) reported pain, 11 patients (100%) reported psychological alterations with cutaneous alteration, 11 patients (100%) had normal secondary sexual characteristics, 1 patient (9%) had supernumerary nipple development, and 11 patients (100%) had well-defined lesions. According to Simon's classification: seven patients (63.6%) were classified as grade 1, three patients (27.2%) as grade 2 and one patient (9.09%) as grade 3. Each patient had a subdermal mastectomy with external periareolar incision, 11 patients (100%) had a histopathological report of gynecomastia; 1 patient (9.09%) displayed keloid healing and none displayed complications inherent to the surgical procedure. CONCLUSIONS: Mastectomy by means of external periareolar incision is useful in the treatment of gynecomastia. PMID- 18158878 TI - [Eradication of Helicobacter pylori: triple treatment scheme plus Lactobacillus vs. triple treatment alone]. AB - BACKGROUND: We undertook this study to determine the effectiveness of triple treatment associated with Lactobacillus casei strain Shirota against Helicobacter pylori, in comparison with triple conventional treatment without Lactobacillus. METHODS: We designed a clinical comparative randomized study that included patients of both sexes infected by Helicobacter pylori. Patients were studied with endoscopy, biopsy and serology. Control group received traditional triple treatment with clarithromycin, amoxicillin and omeprazole, and the active group received the same treatment plus Lactobacillus casei strain Shirota. With the breath test at 8 weeks we determined the eradication of Helicobacter pylori. RESULTS: In the active group we included 31 patients. After treatment, in 29 persons (94%) it was possible to eradicate the Helicobacter pylori. On the other hand, the control group was comprised of 33 patients who received only the traditional triple treatment; eradication was achieved in 25 patients (76%). Significant differences (p<0.05) were observed in favor of active group treatment. CONCLUSIONS: Traditional triple treatment plus Lactobacillus casei strain Shirota is associated with a higher frequency of eradication of Helicobacter pylori than triple treatment alone. PMID- 18158879 TI - [Water immersion for adjuvant treatment of refractory ascites in patients with liver cirrhosis]. AB - BACKGROUND: Head-out water immersion has been proposed as an adjuvant treatment in refractory ascites and hepatorenal syndrome. We undertook this study to present the results of management of patients with refractory ascites. METHODS: We included 10 patients with diagnosis of hepatic cirrhosis and refractory ascites. Variables were measured in four stages: stage I (basal); II (at the end of water immersion); III (72 h after water immersion); IV (1 week after water immersion concludes). Clinical and laboratory variables were measured and included general exams and renal function tests. Friedman test was used for statistics to establish differences between variables at the end of stage IV. We considered statistical significance when p<0.05. RESULTS: Median age was 53.8 years, corresponding to seven men and three women with a Child's classification of B or C. Statistically significant variables were weight (p=0.02) and abdominal circumference (p=0.003), as a result of an increased urine output (p=0.03) and glomerular filtration rate (p<0.002). Renal plasma rate increased until stage III, returning to basal level in stage IV. Serum potassium levels decreased but the difference was marginal (p=0.052). During follow-up, two patients died as a consequence of liver insufficiency. CONCLUSIONS: Head-out water immersion showed a decrease in weight and abdominal circumference, which means reduction of ascites. There was a transitory improvement in renal function. No collateral events were reported. Water immersion could be proposed as an adjuvant treatment in patients with refractory ascites and liver cirrhosis. PMID- 18158880 TI - [Incidence, diagnosis and treatment of colovesical fistula]. AB - BACKGROUND: Enterovesical fistula, also known as vesicoenteric fistula, is an abnormal communication of the vesical bladder with a segment of the digestive tract. We undertook this study to describe diagnostic and therapeutic methods to treat colovesical fistula (CVF) in patients who attended the Coloproctology Unit of the Gastroenterology Service of the General Hospital in Mexico City. METHODS: This is a descriptive study in CVF patients carried out from January 2001 to June 2006; descriptive statistics were used for analysis of information. RESULTS: Eleven patients were identified (10 males and 1 female). Average age was 54.72 years (range: 39-73 years). Time from onset of symptoms to diagnosis was on average 11.9 months. The most frequent signs and symptoms were fecaluria, pneumaturia, dysuria, hematuria and chronic abdominal pain in hypogastric and left iliac regions. Nine patients were submitted to sigmoidectomy and primary colorectal anastomosis. Hartmann procedure was carried out in one patient with restoration of intestinal transit 6 weeks later. In one patient, a loop colostomy was built as a first operation, with sigmoidectomy with fistula resection as a second operation, and restoration of intestinal transit as the third. CONCLUSIONS: Surgery is the only treatment that assures cure and avoids relapses. Sigmoidectomy and primary anastomosis must be considered as the treatment of choice. Mortality, although low, continues being a negative factor when surgery is indicated in these patients. PMID- 18158881 TI - [Complex anal fistula treated with cutting seton]. AB - BACKGROUND: Anorectal sepsis is a frequent condition presenting in the office of colorectal specialists. The acute phase presents as an abscess and the chronic phase as a fistula. We undertook this study to report the experience of the Coloproctology Unit of the General Hospital of Mexico in the management of complex anal fistula with cutting seton. METHODS: From May 1999 to April 2004, 11,731 clinical cases were reviewed. Inclusion criteria were patients with complex anal fistula treated using the cutting seton technique and 6 months minimum follow-up after withdrawal of seton. RESULTS: There were 742 fistulous patients among which 50 fulfilled inclusion criteria. There were 44 males and 6 females. Fistulas were classified as high transsphincteric in 41 patients, suprasphincteric in 8 patients, and high intersphincteric in one patient. On average, patients were followed-up during 9.67 postoperative examinations and on average there were 4.55 adjustments. The average permanence of seton was 7.02 months. CONCLUSIONS: Management of complex anal fistula continues to be a challenge for surgeons. Cutting seton is an appropriate surgical option for patients with no alteration of continence and is useful for patients with high transsphincteric, suprasphincteric and, in some cases, extrasphincteric fistula. In women with low anterior transsphincteric fistula, this option must be considered as an alternative. PMID- 18158882 TI - [Development of an animal model of myelomeningocele and options for prenatal treatment in Macaca mulatta]. AB - BACKGROUND: Myelomeningocele is the most common congenital malformation of the central nervous system, with a reported incidence of 4.4-4.6 cases/10,000 live births in the U.S. Experimental studies demonstrated that the neurological deficit is not a cause for a primary defect in neurulation but is probably due to the progressive mechanical and chemical damage of amniotic liquid into the nerve tissue, which remained without protection during gestation. We undertook this study to develop an animal model in myelomeningocele in Macaca mulatta and compare the results of prenatal management between the colocations of an impermeable silicone mesh and closed with skin. METHODS: A total of nine macaques were surgically intervened, randomly assigned between 12 and 14 weeks of gestation. A laminectomy was carried out in a fetus with bone marrow exposure. The defect was closed with skin or mesh. Three fetuses were left completely exposed. Descriptive statistics were done by ANOVA. RESULTS: Of the four surviving fetuses (44%), none presented complications in mobility in the somatosensory evoked potential, hydrocephalus, or in sphincter control. Only one fetus developed bony deformities. The majority of time of the surgical procedure was due to the use of mesh and the least amount of time was involved in skin closing (p<0.005). CONCLUSIONS: Macaca mulatta is an appropriate model for reproducing myelomeningocele and to evaluate the different options for prenatal management. Colocation of an impermeable silicone mesh is feasible, protecting the spine from amniotic liquid, with results similar to skin closure. PMID- 18158883 TI - [Anti-hyperalgesic effect of one combination of morphine and gabapentin in neuropathic pain induced by chronic constriction injury in rat]. AB - BACKGROUND: Neuropathic pain is associated with disease or injury to the peripheral or central nervous system, which is considered particularly difficult to treat due to its diverse etiology and underlying physiopathological mechanisms. Recent experimental and clinical data support the potential of pharmacotherapy using a combination of drugs for neuropathic pain. METHODS: In order to assess a possible synergistic anti-hyperalgesic interaction, the anti hyperalgesic effects of morphine and gabapentin, single-dose administered either separately or in combination, were determined using the von Frey test in a rat model of neuropathic pain (Bennett model). RESULTS: Time course analysis showed that morphine (3.2 mg/kg s.c.) and gabapentin (17.8 mg/kg s.c.) individually reached their maximum effect at 60 min after treatment, producing an anti hyperalgesic effect of 51.7+/-10.5% and 55.0+/-11.7%, respectively, whereas the combination morphine + gabapentin (3.2+17.8 mg/kg s.c.) produced an almost total anti-hyperalgesic effect at 30 min (96.7+/-2.1%) and at 60 min showed 100% anti hyperalgesia. This anti-hyperalgesic effect remained during 180 min of observation. Analysis of global effects as area under the curve of time course showed that the nature of the anti-hyperalgesic interaction of the analyzed dose had an additive effect. There was no significant difference observed in the theoretical sum of anti-hyperalgesic effect produced by each drug alone (225.4+/ 29.1 area units, au) compared with the corresponding effects produced by the combination of drugs (263.33+/-3.3 au). CONCLUSIONS: These findings are useful in determining the type of interaction that these drugs produce using this combination ratio in neuropathic pain. PMID- 18158884 TI - [Antinociceptive effects of the combination metamizol + morphine in rats with intense pain (arthritic gout-type pain produced with AU)]. AB - BACKGROUND: The antinociceptive effects of metamizol and morphine administered either separately or in combination were determined in the "Pain-Induced Functional Impairment Model in the Rat" (PIFIR antinociceptive model). METHODS: Intense nociception (or intense pain) was induced by the intra-articular injection of uric acid (50%) in the right hind limb inducing its dysfunction. Animals then received analgesic agents, and the recovery of functionality over time was assessed as an expression of antinociception. RESULTS: Metamizol (177.8 mg/kg s.c.) or morphine (3.2 mg/kg s.c.) separately resulted in a lower antinociceptive effect (22.1+/-5.4 area units [au] and 31.8+/-9.4 au, respectively). Moreover, the combination of metamizol (177.8 mg/kg) with morphine (3.2 mg/kg) resulted in a potentiation (293.7+/-16.6 au). The antinociceptive effect observed using the combination was significantly greater than expected on the basis of addition of the individual effects. The percent change in antinociceptive effects, using the combination, was 444.9%. CONCLUSIONS: This represents the first study to show that metamizol + morphine can produce potentiation of their antinociceptive effects in intense pain. PMID- 18158885 TI - [Villous adenoma of the rectum with severe hydroelectric alterations. Report of two cases]. AB - BACKGROUND: Approximately 10% of all colorectal adenomas are constituted by villous adenomas. Their relationship with hydroelectrolytic depletion is rare. We report two cases with villous adenoma that presented hydroelectrolytic depletion with clinical and surgical management, exclusively. CLINICAL CASES: Case 1. Patient was a 76-year-old female with hypertension and 3 months evolution of symptoms such as asthenia, adynamia, unexplained weight loss, and abundant mucus with diarrhea. Serum potassium value was 2.2 mEq/l . Upon rectal exam we found a sessile, exophitic soft tumor with irregular surface of approximately 10 cm in diameter. We also performed a transanal resection of tumor reporting villous adenoma. The patient was discharged from the hospital at the 4th postoperative day with potassium values within normal limits. Case report 2. Patient was a 76 year-old female with diabetes and hypertension of long evolution. She reported a 4-month clinical evolution with non-bloody diarrhea and abundant mucus. She reported nausea, vomiting and no unexplained weight loss. Serum potassium value was 2.1 mEq/l . During the rectal exam we identified a sessile, polypoid, 5-cm diameter tumor that did not involve deep planes. In addition, we carried out a transanal resection of the polyp. The patient was discharged from the hospital on the 5th postoperative day. Potassium value was 4.3 mEq/l. CONCLUSIONS: Size and location of the villous adenoma are related to the production of mucus secretory diarrhea. The inhibiting well-known indomethacin of the prostaglandins has been used to decrease the mucus secretion. In every patient with presence of mucus, persistent diarrhea and occasional rectal bleeding of 1 month, it is necessary to carry out lower endoscopy to rule out the presence of villous adenoma. PMID- 18158886 TI - [Lumbar hernia. Case report and literature review]. AB - BACKGROUND: Lumbar hernia is a rare abdominal wall defect that usually presents spontaneously after trauma or lumbar surgery or, less frequently, during infancy (congenital). Few reports have been published in the literature describing congenital lumbar hernia. CASE REPORT: We present the case of a patient with congenital lumbar hernia and a review describing the regional anatomy, laparoscopic techniques as a surgical alternative, and the impact of modern imagenology in diagnosis, confirmation and demarcation of the aponeurotic defect, in order to update information and to provide the surgeon with the tools for optimal perioperative preparation and the best operative technique for this rare disease. We describe here the case of a 5-month-old Mexican female infant with a right lumbar bulging. Ultrasonographic findings showed an aponeurotic defect and an ipsilateral renal agenesia. Open surgical repair was carried out with identification of the defect in the superior or Grynfellt-Lesshaft's triangle and the inferior triangle as well, and a polypropylene mesh was placed below the posterior abdominal sheath. CONCLUSIONS: Congenital lumbar hernia usually originates in the superior triangle or Grynfellt-Lesshaft's triangle and is frequently associated with the lumbocostovertebral syndrome. Surgery is always indicated, with utilization of prosthetic material or muscular flaps as the best repair technique. PMID- 18158887 TI - [Practice guidelines for pain management in Mexico]. AB - BACKGROUND: It has been documented that pain, in its diverse modalities, is the most common cause of medical attention in Mexico. Due to the increased frequency, pain management has been under consideration in health programs. On the other hand, inadequate pain management can cause severe physical, psychoaffective, and socioeconomic repercussions for patients, families, and public health services. Despite this panorama, there has been no agreement to establish better diagnostic and therapeutic methods. METHODS: Three consensus groups were reunited in different times; those were integrated by medical experts from private and public institutions and form diverse states of the Mexican Republic. To assure the development of these practice guidelines, these experts had experience in the assessment and treatment of painful conditions. Following the methodology used for other consensus groups, diverse meetings were held to review medical evidence about the assessment and treatment of acute, perioperative and cancer pain. RESULTS: A series of recommendations were obtained and classified according to their methodological strength. CONCLUSIONS: As a result of these meetings, a series of recommendations based on the medical evidence were obtained. These recommendations are outlined in three practice guidelines that are intended to allow Mexican practitioners to provide optimal management for painful conditions. PMID- 18158888 TI - [Three personages in one old book]. AB - It is always of interest, especially for medical practitioners, to learn about the lives of prominent scientists and their contributions to medicine. Briefly, we present here the highlights of three of those persons: Hermann von Helmholtz, one of the greatest minds in physics, physiology, optics, neurology, and developer of the ophthalmoscope, etc., along with two Mexican physicians, Manuel Carmona y Valle, prominent 19th century physician who was named "father" of ophthalmology in Mexico and Rafael Silva y Zayas, one of the brightest ophthalmologists of the first half of the 20th century in our country, eventually brought together, in communion, in one old book. PMID- 18158890 TI - [Follicular cancer of the thyroid and diffuse toxic goitre. Report of a case]. PMID- 18158891 TI - Ribozyme catalysis revisited: is water involved? AB - Enzymatic catalysis by RNA was discovered 25 years ago, yet mechanistic insights are emerging only slowly. Thought to be metalloenzymes at first, some ribozymes proved more versatile than anticipated when shown to utilize their own functional groups for catalysis. Recent evidence suggests that some may also judiciously place structural water molecules to shuttle protons in acid-base catalyzed reactions. PMID- 18158892 TI - Cutting proteins within lipid bilayers: rhomboid structure and mechanism. AB - Rhomboids were only discovered to be novel proteases in 2001, but progress on understanding this newest family of intramembrane proteases has been rapid. They are now the best characterized of these rather mysterious enzymes that cleave transmembrane domains within the lipid bilayer. In particular, the biochemical analysis of solubilized rhomboids and, most recently, a flurry of high-resolution crystal structures, have led to real insight into their enzymology. Long-standing questions about how it is possible for a water-requiring proteolytic reaction to occur in the lipid bilayer are now answered for the rhomboids. Intramembrane proteases, which control many medically important biological processes, have made the transition from rather heretical outsiders to novel enzymes that are becoming well understood. PMID- 18158893 TI - FOXO3a is activated in response to hypoxic stress and inhibits HIF1-induced apoptosis via regulation of CITED2. AB - FOXO transcription factors are important regulators of cell survival in response to a variety of stress stimuli, among which are oxidative stress, DNA damage, and nutrient deprivation. Here we report a role for FOXO3a under conditions of hypoxic stress. In response to hypoxia, FOXO3a transcript levels accumulate in an HIF1-dependent way, resulting in enhanced FOXO3a activity. We show that transcription of CITED2, a transcriptional cofactor that functions in a negative feedback loop to control HIF1 activity, is induced by FOXO3a during hypoxia. In fibroblasts as well as in breast cancer cells, FOXO3a inhibits HIF1-induced apoptosis by stimulating the transcription of CITED2, which results in reduced expression of the proapoptotic HIF1 target genes NIX and RTP801. Thus, by fine tuning HIF1 activity, FOXO3a plays an important role in the survival response of normal and cancer cells in response to hypoxic stress. PMID- 18158894 TI - Myosin VI walks "wiggly" on actin with large and variable tilting. AB - Myosin VI is an unconventional motor protein with unusual motility properties such as its direction of motion and path on actin and a large stride relative to its short lever arms. To understand these features, the rotational dynamics of the lever arm were studied by single-molecule polarized total internal reflection fluorescence (polTIRF) microscopy during processive motility of myosin VI along actin. The axial angle is distributed in two peaks, consistent with the hand-over hand model. The changes in lever arm angles during discrete steps suggest that it exhibits large and variable tilting in the plane of actin and to the sides. These motions imply that, in addition to the previously suggested flexible tail domain, there is a compliant region between the motor domain and lever arm that allows myosin VI to accommodate the helical position of binding sites while taking variable step sizes along the actin filament. PMID- 18158895 TI - rRNA modifications in an intersubunit bridge of the ribosome strongly affect both ribosome biogenesis and activity. AB - The presence of nucleotide modifications in rRNA has been known for nearly 40 years; however, information about their roles is sparse. Here, we describe the consequences of depleting modifications from an intersubunit bridge (helix 69) of the ribosomal large subunit in yeast. Helix 69 interacts with both A and P site tRNAs and contains five modifications. Blocking one to two modifications has no apparent effect on cell growth, whereas loss of three to five modifications impairs growth and causes the broadest defects observed thus far for modification loss in any ribosome region. Major effects include the following: (1) reduced amino acid incorporation rates in vivo (25%-60%); (2) increased stop codon readthrough activity; (3) increased sensitivity to ribosome-based antibiotics; (4) reduced rRNA levels (20%-50%), due mainly to faster turnover; and (5) altered rRNA structure in the ribosome. Taken together, the results indicate that this subset of rRNA modifications can influence both ribosome synthesis and function and in synergistic ways. PMID- 18158896 TI - Intranuclear distribution and local dynamics of RNA polymerase II during transcription activation. AB - Transcription activation causes dramatic changes in a gene's compaction and macromolecular associations and, in some cases, triggers the translocation of the gene to a nuclear substructure. Here, we evaluate the location, movement, and transcriptional dynamics of Drosophila heat shock (HS) genes both by two-photon microscopy in live polytene nuclei and by FISH in diploid nuclei. The different HS loci occupy separate nuclear positions. Although these loci decondense upon HS, they do not undergo a detectable net translocation nor are they preferentially localized to the nuclear periphery or interior. Additionally, fluorescence recovery after photobleaching reveals that, shortly after HS, newly recruited RNA polymerase II (Pol II) enters elongation via an "efficient entry" mode, which is followed by the progressive establishment of transcription "compartments" at Hsp70 loci where concentrated Pol II is used in a "local recycling" mode. Pol II at highly transcribed developmental loci exhibits dynamics resembling combinations of these Hsp70 transcription modes. PMID- 18158897 TI - An allosteric path to transcription termination. AB - Transcription termination signals in bacteria occur in RNA as a strong hairpin followed by a stretch of U residues at the 3' terminus. To release the transcript, RNA polymerase (RNAP) is thought to translocate forward without RNA synthesis. Here we provide genetic and biochemical evidence supporting an alternative model in which extensive conformational changes across the enzyme lead to termination without forward translocation. In this model, flexible parts of the RNA exit channel (zipper, flap, and zinc finger) assist the initial step of hairpin folding (nucleation). The hairpin then invades the RNAP main channel, causing RNA:DNA hybrid melting, structural changes of the catalytic site, and DNA clamp opening induced by interaction with the G(trigger)-loop. Our results envision the elongation complex as a flexible structure, not a rigid body, and establish basic principles of the termination pathway that are likely to be universal in prokaryotic and eukaryotic systems. PMID- 18158898 TI - Interplay of chromatin modifiers on a short basic patch of histone H4 tail defines the boundary of telomeric heterochromatin. AB - Dot1 (Disruptor of telomeric silencing-1) is a histone H3 lysine 79 methyltransferase that contributes to the establishment of heterochromatin boundary and has been linked to transcription elongation. We found that histone H4 N-terminal domain, unlike other histone tails, interacts with Dot1 and is essential for H3 K79 methylation. Furthermore, we show that the heterochromatin protein Sir3 inhibits Dot1-mediated methylation and that this inhibition is dependent on lysine 16 of H4. Sir3 and Dot1 bind the same short basic patch of histone H4 tail, and Sir3 also associates with the residues surrounding H3 K79 in a methylation-sensitive manner. Thus, Sir3 and Dot1 compete for the same molecular target on chromatin. ChIP analyses support a model in which acetylation of H4 lysine 16 displaces Sir3, allowing Dot1 to bind and methylate H3 lysine 79, which in turn further blocks Sir3 binding/spreading. This draws a detailed picture of the succession of molecular events occurring during the establishment of telomeric heterochromatin boundaries. PMID- 18158899 TI - Role of the conserved Sir3-BAH domain in nucleosome binding and silent chromatin assembly. AB - Silent chromatin domains in Saccharomyces cerevisiae represent examples of epigenetically heritable chromatin. The formation of these domains involves the recruitment of the SIR complex, composed of Sir2, Sir3, and Sir4, followed by iterative cycles of NAD-dependent histone deacetylation and spreading of SIR complexes over adjacent chromatin domains. We show here that the conserved bromo adjacent homology (BAH) domain of Sir3 is a nucleosome- and histone-tail-binding domain and that its binding to nucleosomes is regulated by residues in the N terminus of histone H4 and the globular domain of histone H3 on the exposed surface of the nucleosome. Furthermore, using a partially purified system containing nucleosomes, the three Sir proteins, and NAD, we observe the formation of SIR-nucleosome filaments with a diameter of less than 20 nm. Together, these observations suggest that the SIR complex associates with an extended chromatin fiber through interactions with two different regions in the nucleosome. PMID- 18158900 TI - A NASP (N1/N2)-related protein, Sim3, binds CENP-A and is required for its deposition at fission yeast centromeres. AB - A defining feature of centromeres is the presence of the histone H3 variant CENP A(Cnp1). It is not known how CENP-A(Cnp1) is specifically delivered to, and assembled into, centromeric chromatin. Through a screen for factors involved in kinetochore integrity in fission yeast, we identified Sim3. Sim3 is homologous to known histone binding proteins NASP(Human) and N1/N2(Xenopus) and aligns with Hif1(S. cerevisiae), defining the SHNi-TPR family. Sim3 is distributed throughout the nucleoplasm, yet it associates with CENP-A(Cnp1) and also binds H3. Cells defective in Sim3 function have reduced levels of CENP-A(Cnp1) at centromeres (and increased H3) and display chromosome segregation defects. Sim3 is required to allow newly synthesized CENP-A(Cnp1) to accumulate at centromeres in S and G2 phase-arrested cells in a replication-independent mechanism. We propose that one function of Sim3 is to act as an escort that hands off CENP-A(Cnp1) to chromatin assembly factors, allowing its incorporation into centromeric chromatin. PMID- 18158901 TI - Distinct roles of chromatin-associated proteins MDC1 and 53BP1 in mammalian double-strand break repair. AB - Phosphorylated histone H2AX ("gamma-H2AX") recruits MDC1, 53BP1, and BRCA1 to chromatin near a double-strand break (DSB) and facilitates efficient repair of the break. It is unclear to what extent gamma-H2AX-associated proteins act in concert and to what extent their functions within gamma-H2AX chromatin are distinct. We addressed this question by comparing the mechanisms of action of MDC1 and 53BP1 in DSB repair (DSBR). We find that MDC1 functions primarily in homologous recombination/sister chromatid recombination, in a manner strictly dependent upon its ability to interact with gamma-H2AX but, unexpectedly, not requiring recruitment of 53BP1 or BRCA1 to gamma-H2AX chromatin. In contrast, 53BP1 functions in XRCC4-dependent nonhomologous end-joining, likely mediated by its interaction with dimethylated lysine 20 of histone H4 but, surprisingly, independent of H2AX. These results suggest a specialized adaptation of the "histone code" in which distinct histone tail-protein interactions promote engagement of distinct DSBR pathways. PMID- 18158902 TI - UmuD and RecA directly modulate the mutagenic potential of the Y family DNA polymerase DinB. AB - DinB is the only translesion Y family DNA polymerase conserved among bacteria, archaea, and eukaryotes. DinB and its orthologs possess a specialized lesion bypass function but also display potentially deleterious -1 frameshift mutagenic phenotypes when overproduced. We show that the DNA damage-inducible proteins UmuD(2) and RecA act in concert to modulate this mutagenic activity. Structural modeling suggests that the relatively open active site of DinB is enclosed by interaction with these proteins, thereby preventing the template bulging responsible for -1 frameshift mutagenesis. Intriguingly, residues that define the UmuD(2)-interacting surface on DinB statistically covary throughout evolution, suggesting a driving force for the maintenance of a regulatory protein-protein interaction at this site. Together, these observations indicate that proteins like RecA and UmuD(2) may be responsible for managing the mutagenic potential of DinB orthologs throughout evolution. PMID- 18158903 TI - Tes, a specific Mena interacting partner, breaks the rules for EVH1 binding. AB - The intracellular targeting of Ena/VASP family members is achieved via the interaction of their EVH1 domain with FPPPP sequence motifs found in a variety of cytoskeletal proteins, including lamellipodin, vinculin, and zyxin. Here we show that the LIM3 domain of Tes, which lacks the FPPPP motif, binds to the EVH1 domain of Mena, but not to those of VASP or Evl. The structure of the LIM3:EVH1 complex reveals that Tes occludes the FPPPP-binding site and competes with FPPPP containing proteins for EVH1 binding. Structure-based gain-of-function experiments define the molecular basis for the specificity of the Tes-Mena interaction. Consistent with in vitro observations, the LIM3 domain displaces Mena, but not VASP, from the leading edge and focal adhesions. It also regulates cell migration through a Mena-dependent mechanism. Our observations identify Tes as an atypical EVH1 binding partner and a regulator specific to a single Ena/VASP family member. PMID- 18158904 TI - Ribosome binding of a single copy of the SecY complex: implications for protein translocation. AB - The SecY complex associates with the ribosome to form a protein translocation channel in the bacterial plasma membrane. We have used cryo-electron microscopy and quantitative mass spectrometry to show that a nontranslating E. coli ribosome binds to a single SecY complex. The crystal structure of an archaeal SecY complex was then docked into the electron density maps. In the resulting model, two cytoplasmic loops of SecY extend into the exit tunnel near proteins L23, L29, and L24. The loop between transmembrane helices 8 and 9 interacts with helices H59 and H50 in the large subunit RNA, while the 6/7 loop interacts with H7. We also show that point mutations of basic residues within either loop abolish ribosome binding. We suggest that SecY binds to this primary site on the ribosome and subsequently captures and translocates the nascent chain. PMID- 18158905 TI - Crystal structure of human XLF: a twist in nonhomologous DNA end-joining. AB - DNA double-strand breaks represent one of the most severe forms of DNA damage in mammalian cells. One pathway for repairing these breaks occurs via nonhomologous end-joining (NHEJ) and depends on XRCC4, LigaseIV, and Cernunnos, also called XLF. Although XLF stimulates XRCC4/LigaseIV to ligate mismatched and noncohesive DNA ends, the mechanistic basis for this function remains unclear. Here we report the structure of a partially functional 224 residue N-terminal fragment of human XLF. Despite only weak sequence similarity, XLF(1-170) shares structural homology with XRCC4(1-159). However, unlike the highly extended 130 A helical domain observed in XRCC4, XLF adopts a more compact, folded helical C-terminal region involving two turns and a twist, wrapping back to the structurally conserved N terminus. Mutational analysis of XLF and XRCC4 reveals a potential interaction interface, suggesting a mechanism for how XLF stimulates the ligation of mismatched ends. PMID- 18158906 TI - Identification of a new nicotinamide binding site in a sirtuin: a reassessment. PMID- 18158907 TI - Dual-fluorophore quantitative high-throughput screen for inhibitors of BRCT phosphoprotein interaction. AB - Finding specific small-molecule inhibitors of protein-protein interactions remains a significant challenge. Recently, attention has grown toward "hot spot" interactions where binding is dominated by a limited number of amino acid contacts, theoretically offering an increased opportunity for disruption by small molecules. Inhibitors of the interaction between BRCT (the C-terminal portion of BRCA1, a key tumor suppressor protein with various functions) and phosphorylated proteins (Abraxas/BACH1/CtIP), implicated in DNA damage response and repair pathways, should prove to be useful in studying BRCA1's role in cancer and in potentially sensitizing tumors to chemotherapeutic agents. We developed and miniaturized to a 1536-well format and 3-mul final volume a pair of fluorescence polarization (FP) assays using fluorescein- and rhodamine-labeled pBACH1 fragment. To minimize the effect of fluorescence artifacts and to increase the overall robustness of the screen, the 75,552 compound library members all were assayed against both the fluorescein- and rhodamine-labeled probe-protein complexes in separate but interleaved reactions. In addition, every library compound was tested over a range of concentrations following the quantitative high-throughput screening (qHTS) paradigm. Analyses of the screening results led to the selection and subsequent confirmation of 16 compounds active in both assays. Faced with a traditionally difficult protein-protein interaction assay, by performing two-fluorophore qHTS, we were able to confidently select a number of actives for further studies. PMID- 18158908 TI - One-pot fluorescent labeling of xyloglucan oligosaccharides with sulforhodamine. AB - Xyloglucan oligosaccharides fluorescently labeled with sulforhodamine have proved to be a valuable tool in the assessment of transglycosylating activity of plant xyloglucan endotransglucosylase/hydrolase (XTH; EC 2.4.1.207). Here we describe a simple and fast procedure for their preparation. Accordingly, the starting xyloglucan-derived oligosaccharides are in the first step converted to their corresponding 1-amino-1-deoxyalditols (glycamines) by incubation with ammonium acetate and NaCNBH(3) at 80 degrees C for 2-4 h, and in the second step, the glycamines are reacted with Lissamine rhodamine B sulfonyl chloride to obtain fluorescently labeled derivatives of the oligosaccharide glycamines. All operations are carried out in a single centrifuge tube and the products from the individual reaction steps are isolated on the basis of their differential solubility in organic solvents. Using the described protocol, the whole procedure can be accomplished in less than 24 h. The sulforhodamine-labeled xyloglucan oligosaccharides thus obtained proved suitable as substrates for a sensitive fluorescence assay of the transglycosylating activity of XTH. PMID- 18158909 TI - Analysis of functionalization of methoxy-PEG as maleimide-PEG. AB - The design of the extension arm-facilitated PEGylation (EAFP) of proteins takes advantage of the high selective and quantitative aspects of the thiol-maleimide reaction. However, the efficiency of EAFP with hemoglobin varied with the batches of maleimide-PEG. The low level of functionalization of monomethoxy-PEG (mPEG) as maleimide-PEG has been now investigated as the potential source of this variation. New chemical approaches for the estimation of the functionalization of mPEG using the reaction of the thiol groups of glutathione, dithiothreitol, and hemoglobin with maleimide-PEG have been developed. The single-step modular approach to the synthesis of maleimidophenyl-PEG (MPPEG) that involved the condensation of p-maleimidophenyl isocyanate with mPEG has been optimized to generate a product with an overall purity of 80%. The NMR approach correlates well with the estimates made by the new chemical approaches. Commercial maleimide PEG reagents synthesized using multiple steps exhibited a lower level of functionalization as reflected by these chemical estimations. The better functionalization of MPPEG increases the efficiency of EAFP as reflected by the generation of hexaPEGylated Hb and the masking of the D antigen of RBCs. This new EAFP protocol is expected to improve the cost effectiveness of the generation of hexaPEGylated Hb, PEGylated albumin, and PEGylated RBCs as new PEGylated therapeutics. PMID- 18158910 TI - Up-regulation of hyaluronan synthase genes in cultured human epidermal keratinocytes by UVB irradiation. AB - Hyaluronan controls keratinocyte proliferation and regeneration. We examined effect of UV on the expression of hyaluronan synthases (HASs) and hyaluronidases in cultured normal human newborn foreskin epidermal keratinocytes, NHEK(F). HAS3 mRNA was expressed predominantly and HAS2 mRNA expressed in lesser amounts and both were up-regulated after a single irradiation with moderate UVB but hyaluronidases was unchanged. Increased accumulation of hyaluronan in the culture medium mirrored the UVB-induced increase in the mRNA levels of HAS3 and HAS2. Unexpectedly, hyaluronan derived from UVB-irradiated and non-irradiated cells had identical size distribution. Increased expression of KGF and IL-1beta was detected just prior to the increase of HAS3 and HAS2 mRNAs after UVB irradiation. Antibody-neutralization study revealed that KGF and/or IL-1beta were at least involved in the up-regulation of HAS3 and HAS2 expressions. UVB-irradiated cells may enhance hyaluronan production to maintain homeostasis through up-regulation of HAS3 and HAS2 genes via cytokine response mechanism. PMID- 18158911 TI - Ascaris suum cytochrome b5, an adult-specific secretory protein reducing oxygen avid ferric hemoglobin. AB - The anaerobic parasitic nematode Ascaris suum has an oxygen-avid hemoglobin in the perienteric fluid, the biological function of which remains elusive. Here, we report that Ascaris cytochrome b5 is expressed specifically in the intestinal parasitic stage and is secreted into the perienteric fluid, thus co-localizing with Ascaris hemoglobin. We also found that cytochrome b5 reduces Ascaris non functioning ferric methemoglobin more efficiently than mammalian methemoglobin. Furthermore, a computer graphics model of the electron transfer complex between Ascaris cytochrome b5 and Ascaris hemoglobin strongly suggested that these two proteins are physiological redox partners. Nitric oxide has been reported to react easily with oxygen captured in hemoglobin to form nitrate, but not toxic free radicals, which may result in production of methemoglobin for the cytochrome b5 to regenerate functional ferrous hemoglobin. Therefore, our findings suggest that Ascaris cytochrome b5 is a key redox partner of Ascaris hemoglobin, which acts as an antioxidant. PMID- 18158912 TI - Dysfunction of the ER chaperone BiP accelerates the renal tubular injury. AB - Tubular-interstitial injury plays a key role in the progression of chronic kidney disease. Although endoplasmic reticulum (ER) stress plays significant roles in the development of chronic diseases such as neurodegenerative disease, cardiomyopathy and diabetes mellitus, its pathophysiological role in chronic renal tubular cell injury remains unknown. BiP is an essential chaperone molecule that helps with proper protein folding in the ER. Recently, we have produced a knock-in mouse that expresses a mutant-BiP in which the retrieval sequence to the ER is deleted in order to elucidate physiological processes that are sensitive to ER functions in adulthood. The heterozygous mutant-BiP mice showed significant tubular-interstitial lesions with aging. Furthermore, proteinuria induced by chronic protein overload accelerated the tubular-interstitial lesions in the mutant mice, accompanying caspase-12 activation and tubular cell apoptosis. These results suggest that the ER stress pathway is significantly involved in the pathophysiology of chronic renal tubular-interstitial injury in vivo. PMID- 18158913 TI - Magnetic single-enzyme nanoparticles with high activity and stability. AB - Magnetic single-enzyme nanoparticles (SENs) encapsulated within a composite inorganic/organic polymer network were fabricated via the surface modification and in situ aqueous polymerization of separate enzyme molecule. The resultant nanoparticles were characterized by transmission electron microscope (TEM), Fourier transform infrared (FTIR) spectrometer and X-ray diffraction (XRD). These particles are almost spherical in shape and have a unique size of about 50 nm in diameter. Electrical and magnetic measurements reveal that the magnetic SENs have a conductivity of 2.7 x 10(-3)Scm(-1), and are superparamagnetic with a saturation magnetization of 14.5e microg(-1) and a coercive force of 60Oe. Compared with free enzyme, encapsulated enzyme exhibits a strong tolerance to the variation of solution pH, high temperature, organic solvent and long-term storage, thus showing significantly enhanced enzyme performance and stability. PMID- 18158914 TI - Molecular modelling of miraculin: Structural analyses and functional hypotheses. AB - Miraculin is a plant protein that displays the peculiar property of modifying taste by swiching sour into a sweet taste. Its monomer is flavourless at all pH as well as at high concentration; the dimer form elicits its taste-modifying activity at acidic pH; a tetrameric form is also reported as active. Two histidine residues, located in exposed regions, are the main responsible of miraculin activity, as demonstrated by mutagenesis studies. Since structural data of miraculin are not available, we have predicted its three-dimensional structure and simulated both its dimer and tetramer forms by comparative modelling and molecular docking techniques. Finally, molecular dynamics simulations at different pH conditions have indicated that at acidic pH the dimer assumes a widely open conformation, in agreement with the hypotheses coming from other studies. PMID- 18158915 TI - Split mCherry as a new red bimolecular fluorescence complementation system for visualizing protein-protein interactions in living cells. AB - Bimolecular fluorescence complementation (BiFC) is a recently developed technique for detection of protein-protein interactions in living cells. In this study, a new red BiFC system was developed by splitting mCherry, a mutant monomeric red fluorescent protein, into two fragments between amino acids 159-160 and was verified using a pair of interacting proteins, SV40 large T antigen (LTag), and human p53 protein. By combined use of the mCherry-based red BiFC system with a Venus-based yellow BiFC system, the interaction between LTag and p53 as well as the interaction between sp100 and promyelocytic leukemia protein (PML), were detected simultaneously in Vero cells. The brilliant redness, short maturation time, and the long excitation and emission wavelengths (587/610 nm) of mCherry make the new BiFC system an excellent candidate for analyzing protein-protein interactions in living cells and for studying multiple protein-protein interactions when coupled with other BiFC systems. PMID- 18158916 TI - Hypotonic stimuli enhance proton-gated currents of acid-sensing ion channel-1b. AB - Acid-sensing ion channels (ASICs) are strong candidates for mammalian mechanoreceptors. We investigated whether mouse acid-sensing ion channel-1b (ASIC1b) is sensitive to mechanical stimuli using oocyte electrophysiology, because ASIC1b is located in the mechanosensory stereocilia of cochlear hair cells. Hypotonic stimuli that induced membrane stretch of oocytes evoked no significant current in ASIC1b-expressing oocytes at pH 7.5. However, acid (pH 4.0 or 5.0)-evoked currents in the oocytes were substantially enhanced by the hypotonicity, showing mechanosensitivity of ASIC1b and possible mechanogating of the channel in the presence of other components. Interestingly, the ASIC1b channel was permeable to K(+) (a principal charge carrier for cochlear sensory transduction) and the affinity of the channel for amiloride (IC(50) (inhibition constant)=approximately 48.3 microM) was quite similar to that described for the mouse hair cell mechanotransducer current. Taken together, these data raise the possibility that ASIC1b participates in cochlear mechanoelectrical transduction. PMID- 18158917 TI - Regulation of rnt-1 expression mediated by the opposing effects of BRO-1 and DBL 1 in the nematode Caenorhabditis elegans. AB - During development of Caenorhabditis elegans, expression of the RUNX homolog, rnt 1, is tightly regulated both spatially and temporally. In this study, we investigated the mechanism underlying the temporal regulation of rnt-1. We found that rnt-1 contained evolutionarily conserved consensus RUNX binding sequences within one of its introns, and that RNT-1 bound to these intronic sequences both in vitro and in vivo in the presence of BRO-1, suggesting that RNT-1 together with BRO-1 represses its own transcription. Fine deletion and substitution experiments revealed a binding site within the intron that was critical for rnt-1 regulation. Importantly, we found that the TGFbeta homolog, DBL-1, was required for counteracting the repressive activity of BRO-1 at postembryonic stages. Accordingly, ectopic expression of DBL-1 induced transcription of rnt-1 in the lateral hypodermis and other tissues even at the postembryonic stages. Taken together, our data suggest that rnt-1 expression is regulated by the balance between DBL-1-mediated activation and BRO-1-mediated repression at the postembryonic stages. PMID- 18158918 TI - Nanogel DDS enables sustained release of IL-12 for tumor immunotherapy. AB - For a valid cytokine immunotherapy of malignancies, a suitable delivery system that ensures slow-release of cytokines is required, because short half-life in vivo of the molecules ruins therapeutic efficacy while causing severe systemic toxic effects. We previously showed that the cholesterol-bearing pullulan (CHP) based hydrogel nanoparticles, or nanogel, encapsulates, stabilizes and releases various molecules. Here we applied this nanogel to administration in vivo of interleukin-12 (IL-12). Recombinant murine IL-12 (rmIL-12) was successfully incorporated into CHP nanogel simply by incubated with CHP at room temperature. After subcutaneously injected into mice, the CHP/rmIL-12 complex led to a prolonged elevation in IL-12 concentration in the sera. Repetitive administrations of the CHP/rmIL-12, but not rmIL-12 alone, induced drastic growth retardation of preestablished subcutaneous fibrosarcoma without causing any serious toxic event. The present study proposes a novel therapeutic intervention technology, taking advantage of slow and sustained release of bioactive cytokines from the self-assembling biocompatible nanoparticles. PMID- 18158919 TI - TGF-beta induces the differentiation of bone marrow stem cells into immature cardiomyocytes. AB - Transforming growth factor-beta(1) (TGF-beta(1)) was found to induce the myogenic differentiation of bone marrow stem cells (BMSC). We investigated the morphological and electrophysiological properties of differentiated cells, and the mechanisms related to TGF-beta(1)-induced myogenic differentiation. We found that TGF-beta increased the expression of the cardiac transcription factors, GATA 4 and NKx-2.5, in BMSC after 1-3 days of cultivation. TGF-beta also induced the expressions of cardiac myosin, troponins, and ANP after 3-14 days of cultivation. However, the Ca(2+) transient was relatively weak, the connexin-43 expression was irregular, and spontaneous beating was not detected within 28 days observation. TGF-beta stimulation up-regulated most of the TGF-beta BMP signaling pathway genes, including TGFBI, ACVR2B, and phosphorylated Smad-2 and Smad-3, within 24h. TGF-beta induced the differentiation of BMSC into immature cardiomyocytes by activating the TGF-beta BMP signaling pathway. PMID- 18158920 TI - Canonical Wnt signaling maintains the quiescent stage of hepatic stellate cells. AB - It is well known that hepatic stellate cells (HSC) develop into cells, which are thought to contribute to liver fibrogenesis. Recent data suggest that HSC are progenitor cells with the capacity to differentiate into cells of endothelial and hepatocyte lineages. The present study shows that beta-catenin-dependent canonical Wnt signaling is active in freshly isolated HSC of rats. Mimicking of the canonical Wnt pathway in cultured HSC by TWS119, an inhibitor of the glycogen synthase kinase 3beta, led to reduced beta-catenin phosphorylation, induced nuclear translocation of beta-catenin, elevated glutamine synthetase production, impeded synthesis of alpha-smooth muscle actin and Wnt5a, but promoted the expression of glial fibrillary acidic protein, Wnt10b, and paired-like homeodomain transcription factor 2c. In addition, canonical Wnt signaling lowered DNA synthesis and hindered HSC from entering the cell cycle. The findings demonstrate that beta-catenin-dependent Wnt signaling maintains the quiescent state of HSC and, similar to stem and progenitor cells, influences their developmental fate. PMID- 18158921 TI - Distribution of zinc, copper and iron in biological samples of Pakistani myocardial infarction (1st, 2nd and 3rd heart attack) patients and controls. AB - BACKGROUND: The pathogenesis of some heart diseases has been associated with changes in the balance of certain trace elements. We examined the association of iron, copper and zinc between biological samples (scalp hair, whole blood and urine) and mortality from myocardial infarction (MI) patients of (first, second and third heart attack). METHODS: The biological samples were from 130 MI patients (77 male and 53 female, age range 45-60 years) and 61 healthy age matched controls (33 male and 28 female). The metals in the biological samples were measured by the flame atomic absorption spectrophotometry, prior to microwave assisted acid digestion. The validity of the methodology was checked by the biological certified reference materials. RESULTS: During this study, 78% of the 32 patients aged >50 years, registered after the third MI attack died. In these subjects the concentration of Fe and Cu were increased by 0.83% and 3.12% in the scalp hair while in blood samples 9.7% and 22.5% were enhanced respectively, as compared to those who tolerated 3rd MI attack (p=0.072). The concentrations of Zn in whole blood and scalp hair samples were lower in MI patients as compared to normal subjects. CONCLUSION: Deficiency of zinc and high concentration of copper and iron may play a role in the development of heart disease. PMID- 18158922 TI - Induction of podoplanin by transforming growth factor-beta in human fibrosarcoma. AB - Podoplanin/aggrus is increased in tumors and its expression was associated with tumor malignancy. Podoplanin on cancer cells serves as a platelet-aggregating factor, which is associated with the metastatic potential. However, regulators of podoplanin remain to be determined. Transforming growth factor-beta (TGF-beta) regulates many physiological events, including tumorigenesis. Here, we found that TGF-beta induced podoplanin in human fibrosarcoma HT1080 cells and enhanced the platelet-aggregating-ability of HT1080. TGF-beta type I receptor inhibitor (SB431542) and short hairpin RNAs for Smad4 inhibited the podoplanin induction by TGF-beta. These results suggest that TGF-beta is a physiological regulator of podoplanin in tumor cells. PMID- 18158923 TI - High affinity interactions between red blood cell receptors and synthetic Plasmodium thrombospondin-related apical merozoite protein (PTRAMP) peptides. AB - Plasmodium falciparum thrombospondin-related apical merozoite protein (PTRAMP) has a thrombospondin related (TSR) domain which in many proteins has been reported as a fragment involved in pathogen-host and cell-interactions. Receptor ligand studies using eighteen non-overlapping 20-aminoacid-long synthetic peptides from this protein were carried out to determine regions involved in parasite invasion of red blood cells (RBC). Two high activity binding peptides (HABPs) were determined, 33405 (21YISSNDLTSTNLKVRNNWEH40) and 33413 (180LEGPIQFSLGKSSGAFRINY199), presenting high dissociation constants and positive cooperativity. One of the HABPs displayed a modified Plasmodium export element (PEXEL), suggesting that this protein could be involved in the merozoite cytoplasmic reticulum, parasitophorous vacuole, red blood cell (RBC) cytosol, and probably infected RBC (iRBC) membrane transport of some other molecules and nutrients. Enzymatic treatment of RBCs increased HABP 33405 binding to them whilst it decreased HABP 33413 binding. Merozoite invasion assays revealed that HABPs have around 57% ability to inhibit new RBC invasion. Circular dichroism revealed the presence of possible alpha-helical elements in both HABPs structures. RBC binding interaction specificity and the presence of a PEXEL motif make these 2 HABPs good candidates for being included in further studies to develop a new multi-antigenic, multi-stage, subunit-based, chemically synthesised, anti-malarial vaccine. PMID- 18158924 TI - Allergic contact dermatitis to topical antibiotics: Epidemiology, responsible allergens, and management. AB - Topical antibiotics are widely used to treat cutaneous, ocular, and otic infections. Allergic contact dermatitis to topical antibiotics is a rare but well documented side effect, especially in at-risk populations. The purpose of this article is to review the epidemiology, responsible allergens, and management of allergic contact dermatitis to topical antibiotics. LEARNING OBJECTIVE: After completing this learning activity, participants should be able to describe the epidemiology of allergic contact dermatitis related to topical antibiotics; show knowledge of the most common allergenic topical antibiotics; and understand the allergenic cross-reactivity pattern amongst topical antibiotics. PMID- 18158925 TI - Nephrogenic systemic fibrosis with septal panniculitis mimicking erythema nodosum. AB - Nephrogenic systemic fibrosis/nephrogenic fibrosing dermopathy is an idiopathic fibrosing disorder recently described in patients with renal disease. The typical histology in nephrogenic systemic fibrosis consists of haphazardly arranged dermal collagen bundles with clefting, collagenous change which extends into the subcutaneous septa, mucin deposition, fibroblast proliferation, and increased elastic fibers with minimal inflammation. We present a 65-year-old female with classic clinical features whose biopsy demonstrated unique histologic features of septal pannicilitis with lymphocytic aggregates and Miescher's radial granulomas mimicking erythema nodosum. PMID- 18158926 TI - Benign alveolar ridge keratosis (oral lichen simplex chronicus): A distinct clinicopathologic entity. AB - Benign alveolar ridge keratosis is a common benign white papule or plaque that occurs on the keratinized gingiva of the maxillary or mandibular alveolar ridge that is probably traumatic/frictional in origin, with characteristic histologic features, similar to those of lichen simplex chronicus of the skin. This is a retrospective study of 108 consecutive specimens displaying characteristic histopathologic features of benign alveolar ridge keratosis accessioned during a 36-month period. There was a male:female ratio of 3.7:1. It occurred on the attached gingiva, with the retromolar area and the edentulous alveolar ridge involved in 51% and 49% of cases, respectively; 19% were bilateral and all bilateral cases were on the retromolar pad. Detailed clinical information was available on 27 cases by a mail-in questionnaire. Histologically, the lesions were characterized by moderate to marked hyperorthokeratosis and wedge-shaped hypergranulosis. The epithelium exhibited slight surface papillomatosis and acanthosis in the form of long, tapered rete ridges that frequently anastomosed at the base. There was generally insignificant inflammation. These features are similar if not identical to lichen simplex chronicus of the skin, a benign condition caused by chronic irritation. Ten randomly selected cases were immunostained for p16INK4A(p16), a tumor suppressor protein expressed in dysplastic epithelium. All lesions were negative for p16. Benign alveolar ridge keratosis is a specific clinicopathologic entity that should be removed from the category of leukoplakia as is currently the practice for clinical white lesions with a specific, consistently recognizable histologic appearance. PMID- 18158927 TI - Basal cell (trichoblastic) carcinoma common expression pattern for epithelial cell adhesion molecule links basal cell carcinoma to early follicular embryogenesis, secondary hair germ, and outer root sheath of the vellus hair follicle: A clue to the adnexal nature of basal cell carcinoma? AB - BACKGROUND: Basal cell carcinoma (BCC) is still viewed by many dermatologists as a tumor of the interfollicular epidermis, although references were made early in the dermatopathologic literature to the resemblance of BCC to the hair follicle. OBJECTIVE: Our aim was to characterize the common expression pattern for the epithelial cell adhesion molecule (Ep-CAM) in BCCs, various stages of follicular embryogenesis, and adult hair follicles and, thereby, in analogy point to the similarity between BCC and the hair follicle. METHODS: We studied immunohistochemically 16 superficial BCCs for Ep-CAM and compared the expression pattern with that during hair follicle, nail, and eccrine gland development in human embryos and fetuses. In addition, we examined terminal scalp and vellus hair follicles. RESULTS: All BCCs expressed Ep-CAM similar to the early stages of the embryonic human hair follicle, the secondary hair germ, and the outer root sheath of the vellus hair follicle. The embryonic nail organ and the adult anagen hair follicles were completely negative. LIMITATIONS: The conclusions are based on the similarity in the immunohistochemical expression profile for a single adhesion molecule. CONCLUSION: BCC expresses the cell-cell adhesion molecule Ep CAM similar to the embryonic hair germ, the secondary hair germ of the terminal hair follicle, and the outer root sheath of the vellus hair follicle. We suggest that this may be a clue to the adnexal nature of BCC and propose that BCC is the most primitive follicular tumor. PMID- 18158928 TI - The human genome: Mutating from science fiction to reality. AB - Dialogues in Dermatology, a monthly audio program from the American Academy of Dermatology, contains discussions between dermatologists on timely topics. Commentaries from Dialogues Editor-in-Chief Warren R. Heymann, MD, are provided after each discussion as a topic summary and are provided here as a special service to readers of the Journal of the American Academy of Dermatology. PMID- 18158929 TI - Conflict of interests. PMID- 18158930 TI - Television turning more teens toward tanning? PMID- 18158931 TI - Trauma and erosion in circumscribed hypokeratosis. PMID- 18158932 TI - Epoetin and nephrogenic systemic fibrosis. PMID- 18158933 TI - Comment on acne and glycemic index. PMID- 18158934 TI - Vitamin D: the sun as an essential source. PMID- 18158935 TI - Sparing of previously irradiated skin from erlotinib-induced acneiform rash. PMID- 18158936 TI - Further discussions on choosing the number of animals for an experiment. PMID- 18158937 TI - Number of samples - hypothesis testing. AB - This note reviews "further discussions on choosing the number of animals for an experiment". Particular emphasis will be given to the table therein. PMID- 18158938 TI - Dopamine D2 receptor radiotracers [(11)C](+)-PHNO and [(3)H]raclopride are indistinguishably inhibited by D2 agonists and antagonists ex vivo. AB - INTRODUCTION: In vitro, the dopamine D2 receptor exists in two states, with high and low affinity for agonists. The high-affinity state is the physiologically active state thought to be involved in dopaminergic illnesses such as schizophrenia. The positron emission tomography radiotracer [(11)C](+)-PHNO ([(11)C](+)-4-propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol), being a D2 agonist, should selectively label the high-affinity state at tracer dose and therefore be more susceptible to competition by agonist as compared to the antagonist [(3)H]raclopride, which binds to both affinity states. METHODS: We tested this prediction using ex vivo dual-radiotracer experiments in conscious rats. D2 antagonists (haloperidol or clozapine), a partial agonist (aripiprazole), a full agonist [(-)-NPA] or the dopamine-releasing drug amphetamine (AMPH) were administered to rats prior to an intravenous coinjection of [(11)C](+)-PHNO and [(3)H]raclopride. Rats were sacrificed 60 min after radiotracer injection. Striatum, cerebellum and plasma samples were counted for (11)C and (3)H. The specific binding ratio {SBR, i.e., [%ID/g (striatum)-%ID/g (cerebellum)]/(%ID/g (cerebellum)} was used as the outcome measure. RESULTS: In response to D2 antagonists, partial agonist or full agonist, [(11)C](+)-PHNO and [(3)H]raclopride SBRs responded indistinguishably in terms of both ED(50) and Hill slope (e.g., (-)-NPA ED(50) values are 0.027 and 0.023 mg/kg for [(11)C](+) PHNO and [(3)H]raclopride, respectively). In response to AMPH challenge, [(11)C](+)-PHNO and [(3)H]raclopride SBRs were inhibited to the same degree. CONCLUSIONS: We have shown that the SBRs of [(11)C](+)-PHNO- and [(3)H]raclopride do not differ in their response to agonist challenge. These results do not support predictions of the in vivo binding behavior of a D2 agonist radiotracer and cast some doubt on the in vivo applicability of the D2 two-state model, as described by in vitro binding experiments. PMID- 18158939 TI - In vitro binding of [(11)C]raclopride with ultrahigh specific activity in rat brain determined by homogenate assay and autoradiography. AB - OBJECTIVE: The aim of this study was to characterize the in vitro binding of [(11)C]raclopride with ultrahigh specific activity (SA) in the striatum and cerebral cortex of rat brain. METHODS: [(11)C]Raclopride, a dopamine D(2) receptor ligand, with an ultrahigh SA of 4880+/-2360 GBq/micromol (132+/-64 Ci/micromol, n=25) was synthesized. In vitro binding experiment was performed using brain homogenate assay and autoradiography (ARG). RESULTS: In vitro homogenate assay demonstrated that high SA [(11)C]raclopride (2520-6340 GBq/micromol; 68-171 Ci/micromol) had two-affinity (high and low) binding sites in the striatum and cerebral cortex of rat brain. In the striatum, K(d,high) and B(max,high) values were 0.005+/-0.002 nM and 0.19+/-0.04 fmol/mg tissue, respectively, while K(d,low) and B(max,low) values were 2.2+/-1.0 nM and 35.8+/ 16.4 fmol/mg tissue, respectively. In the cerebral cortex, K(d,high) and B(max,high) values were 0.061+/-0.087 nM and 0.2+/-0.2 fmol/mg tissue, respectively, while K(d,low) and B(max,low) values were 2.5+/-3.2 nM and 5.5+/ 4.8 fmol/mg tissue, respectively. On the other hand, only one binding site was found in the striatum and no binding site was identified in the cerebral cortex using low SA [(11)C]raclopride (44 GBq/micromol; 1.2 Ci/micromol). In vitro ARG for the rat brain using high SA [(11)C]raclopride (6212 GBq/micromol; 168 Ci/micromol) gave a coronal image of the striatum and cerebral cortex with a higher signal/noise ratio than using low SA [(11)C]raclopride (40 GBq/micromol; 1.1 Ci/micromol). CONCLUSION: Using ultrahigh SA [(11)C]raclopride for the in vitro homogenate assay, we succeeded in detecting two-affinity binding sites of [(11)C]raclopride, not only in the striatum but also in the cerebral cortex of rat brain. PMID- 18158940 TI - A change of in vivo characteristics depending on specific activity of radioiodinated (+)-2-[4-(4-iodophenyl)piperidino]cyclohexanol [(+)-pIV] as a ligand for sigma receptor imaging. AB - The radioiodinated (+)-p-iodovesamicol [(+)-pIV], which shows a high binding affinity for sigma-1 (sigma-1) receptors, is prepared by an exchange reaction. The specific activity (SA) is fairly low and therefore is insufficient for clinical use. In this study, we prepared (+)-[(125)I]pIV with a high SA from tributylstannyl precursor and compared the in vivo characteristics between high and low SA by imaging sigma-1 receptors in the central nervous system. In the biodistribution study, a difference in brain accumulation was observed between the two methods. At 30 min postinjection, the brain accumulation (1.58%ID/g) of low SA [0.6-1.1 TBq/mmol (16-30 Ci/mmol)] (+)-[(125)I]pIV was higher than that (1.34%ID/g) of high SA [>88.8 TBq/mmol (>2400 Ci/mmol)] (+)-[(125)I]pIV. In the blocking study, the brain uptake of high SA (+)-[(125)I]pIV was reduced more significantly by the coadministration of sigma ligands such as pentazocine, haloperidol or SA4503 than that of low SA (+)-[(125)I]pIV. These results showed that nonspecific binding of high SA (+)-[(125)I]pIV in the brain was lower than that of low SA (+)-[(125)I]pIV, and high SA (+)-[(125)I]pIV bound more specifically to sigma-1 receptors in the brain than low SA (+)-[(125)I]pIV. In contrast, in the blood-binding study, high SA (+)-[(125)I]pIV (58.4%) bound to blood cells with higher affinity than low SA (+)-[(125)I]pIV (46.0%). In metabolite studies, blood metabolites of high SA (+)-[(125)I]pIV (57.3+/-3.5%) were higher than those of low SA (+)-[(125)I]pIV (45.5+/-4.1%) at 30 min postinjection. Higher SA may be apt to bind to blood cells with higher affinity and to be metabolized faster. PMID- 18158941 TI - Evaluation of [(11)C]rofecoxib as PET tracer for cyclooxygenase 2 overexpression in rat models of inflammation. AB - BACKGROUND: Overexpression of cyclooxygenase type 2 (COX-2) is triggered by inflammatory stimuli, but it also plays a prominent role in the initiation and progression of various diseases. This study aims to investigate [(11)C]rofecoxib as a positron emission tomography (PET) tracer for COX-2 expression. METHODS: [(11)C]Rofecoxib was prepared by methylation of its sulphinate precursor. Regional brain distribution and specific binding of [(11)C]rofecoxib in healthy rats was studied by ex vivo biodistribution and autoradiography. Regional brain distribution and PET imaging studies were also performed on rats with severe encephalitis, caused by nasal infection with herpes simplex virus (HSV). Finally, ex vivo biodistribution and blocking studies were carried in rats with a sterile inflammation, induced by intramuscular turpentine injection. RESULTS: [(11)C]rofecoxib brain uptake in control animals corresponded with the known distribution of COX-2. Pretreatment with NS398 significantly reduced tracer uptake in the cingulate/frontopolar cortex, whereas the reduction in hippocampus approached significance. Ex vivo autoradiography also revealed preferential tracer uptake in hippocampus and cortical areas that could be blocked by NS398. In HSV-infected animals, [(11)C]rofecoxib uptake was moderately increased in all brain regions, but it could not be blocked with indomethacin. Yet, some PET images revealed increased tracer uptake in brain areas with microglia activation. In turpentine-injected animals, [(11)C]rofecoxib uptake in inflamed muscle was not higher than in control muscle and could not be blocked with NS398. Indomethacin caused a slight reduction in muscle uptake. CONCLUSIONS: Despite the apparent correlation between [(11)C]rofecoxib uptake and COX-2 distribution in healthy rats, [(11)C]rofecoxib could not unambiguously detect COX-2 overexpression in two rat models of inflammation. PMID- 18158942 TI - (R)-N-Methyl-3-(3-(125)I-pyridin-2-yloxy)-3-phenylpropan-1-amine: a novel probe for norepinephrine transporters. AB - Alterations in serotonin and norepinephrine neuronal functions have been observed in patients with major depression. Several antidepressants bind to both serotonin transporters and norepinephrine transporters (NET). The ability to image NET in the human brain would be a useful step toward understanding how alterations in NET relate to disease. In this study, we report the synthesis and characterization of a new series of derivatives of iodonisoxetine, a known radioiodinated probe. The most promising, (R)-N-methyl-3-(3-iodopyridin-2-yloxy) 3-phenylpropylamine (PYINXT), displayed a high and saturable binding to NET, with a K(d) value of 0.53+/-0.03 nM. Biodistribution studies of (R)-N-methyl-3-(3 (125)I-pyridin-2-yloxy)-3-phenylpropan-1-amine in rats showed moderate initial brain uptake (0.54% dose/organ at 2 min) with a relatively fast washout from the brain (0.16% dose/organ at 2 h) as compared to [(125)I]INXT. The hypothalamus (a NET-rich region)-to-striatum (a region devoid of NET) ratio was found to be 2.14 at 4 h after intravenous injection. Preliminary results suggest that this improved iodinated ligand, when labeled with (123)I, may be useful for mapping NET-binding sites with single photon emission computed tomography in the living human brain. PMID- 18158943 TI - Synthesis, radiolabeling and baboon SPECT imaging of 2beta-carbomethoxy-3beta-(3' [(123)I]iodophenyl)tropane ([(123)I]YP256) as a serotonin transporter radiotracer. AB - To develop a potential SPECT probe to evaluate the integrity of the serotoninergic system (5-HTT) whose dysfunction is linked to several disease conditions such as Parkinson's disease, Alzheimer's disease and depression, we report the synthesis, radiolabeling and in vivo baboon imaging of 2beta carbomethoxy-3beta-(3'-[(123)I]iodophenyl) tropane (YP256, 6). The radiolabeling was performed by iododestannylation using sodium [(123)I]iodide and peracetic acid. Although the ligand displayed high selectivity for 5-HTT over dopamine transporter in vitro, SPECT imaging in baboons did not reveal selective 5-HTT accumulation in brain in vivo. PMID- 18158944 TI - Preparation and first evaluation of [(18)F]FE@SUPPY: a new PET tracer for the adenosine A(3) receptor. AB - INTRODUCTION: Changes of the adenosine A(3) receptor subtype (A3AR) expression have been shown in a variety of pathologies, especially neurological and affective disorders, cardiac diseases and oncological and inflammation processes. Recently, 5-(2-fluoroethyl) 2,4-diethyl-3-(ethylsulfanylcarbonyl)-6 phenylpyridine-5-carboxylate (FE@SUPPY) was presented as a high-affinity ligand for the A3AR with good selectivity. Our aims were the development of a suitable labeling precursor, the establishment of a reliable radiosynthesis for the fluorine-18-labeled analogue [(18)F]FE@SUPPY and a first evaluation of [(18)F]FE@SUPPY in rats. METHODS: [(18)F]FE@SUPPY was prepared in a feasible and reliable manner by radiofluorination of the corresponding tosylated precursor. Biodistribution was carried out in rats, and organs were removed and counted. Autoradiography was performed on rat brain slices in the presence or absence of 2 Cl-IB-MECA. RESULTS: Overall yields and radiochemical purity were sufficient for further preclinical and clinical applications. The uptake pattern of [(18)F]FE@SUPPY found in rats mainly followed the described mRNA distribution pattern of the A3AR. Specific uptake in brain was demonstrated by blocking with a selective A3AR agonist. CONCLUSION: We conclude that [(18)F]FE@SUPPY has the potential to serve as the first positron emission tomography tracer for the A3AR. PMID- 18158945 TI - Feasibility studies of 4'-[methyl-(11)C]thiothymidine as a tumor proliferation imaging agent in mice. AB - This study reports on the radiosynthesis and feasibility studies of 4'-[methyl (11)C]thiothymidine ([methyl-(11)C]S-dThd) as a tumor proliferation imaging agent. [Methyl-(11)C]S-dThd was synthesized by rapid methylation of corresponding 5-trimethylstannyl- or 5-tributylstannyl-precursor via a palladium-promoted Stille cross-coupling reaction with [(11)C]methyl iodide. The decay-corrected radiochemical yields of [methyl-(11)C]S-dThd synthesized by the corresponding 5 trimethylstannyl-precursor and 5-tributylstannyl-precursor based on [(11)C]CO(2) were 18.9% and 14.5%, respectively. The radiochemical purity of [methyl-(11)C]S dThd was always greater than 99%. The specific activities of [methyl-(11)C]S-dThd synthesized by the corresponding 5-trimethylstannyl-precursor and 5 tributylstannyl-precursor were 47 GBq/mumol and 121 GBq/mumol, respectively, at the end of the synthesis. The total synthesis time was 30 min after the end of bombardment. The comparison between in vivo distribution of [methyl-(14)C]S-dThd and that of [methyl-(3)H]FLT showed that tracer uptake was comparable in nonproliferating tissues. In contrast, [methyl-(14)C]S-dThd showed significantly higher uptake in proliferating tissues than did [methyl-(3)H]FLT. [Methyl-(11)C]S dThd uptake levels in five different tumor tissues were well correlated with the DNA synthesis levels determined by [2-(14)C]thymidine DNA incorporation. At 30 min after injection, plasma analysis found 95% of the activity in unmetabolized form. The microPET imaging of the C6 glioma xenograft showed significantly high uptake in the tumor and urinary bladder, followed by the intestine and marrow. Our results demonstrated that the tumor uptake of [methyl-(11)C]S-dThd was higher than that of [methyl-(3)H]FLT and was well correlated with the DNA synthesis level. Consequently, 4'-[methyl-(11)C]thiothymidine has promise for the imaging of tumor cell proliferation by positron emission tomography. PMID- 18158946 TI - Improved synthesis and metabolic stability analysis of the dopamine transporter ligand [(18)F]FECT. AB - INTRODUCTION: [2'-[(18)F]Fluoroethyl (lR-2-exo-3-exe)-8-methyl-3-(4-chlorophenyl) 8-azabicyclo[3.2.1]-octane-2-carboxylate] ([(18)F]FECT) is a positron emission tomography (PET) tracer for imaging the dopamine transporter (DAT) in vivo. We report an improved radiosynthesis procedure and affinity data and have analyzed both brain tissue and plasma samples for the presence of radiometabolites as a function of time post intravenous injection of [(18)F]FECT to rats. METHODS: The radiosynthesis of [(18)F]FECT was carried out using [(18)F]fluoroethyltriflate ([(18)F]FEtOTf) as a labeling agent. The affinity of FECT for DAT was determined in vitro by binding experiments on rat striatal membranes. Three rats were injected with [(18)F]FECT and blood samples were collected at 1 or 3 h post injection (p.i.). Plasma was separated and analyzed using reversed-phase high performance liquid chromatography (RP-HPLC). Similarly, cerebrum and cerebellum were isolated after sacrifice of the animals at 3 h p.i. of the tracer and homogenized. HPLC analysis was performed on extracts of both samples to examine the presence of metabolites. RESULTS: The radiochemical yield for [(18)F]FECT was 85% relative to the starting activity of [(18)F]FEtOTf. The inhibitory constant (K(i)) of FECT for DAT was found to be 6 nM. The fraction of radioactivity corresponding to intact [(18)F]FECT was 93% in plasma at both 1 and 3 h p.i. and 96% in cerebrum as well as cerebellum samples at 3 h p.i. CONCLUSIONS: FECT has a high affinity for the dopamine transporter. [(18)F]FECT was found to be stable in vivo and the amount of radiolabeled metabolites in plasma and brain at 3 h p.i. is negligible. Hence, [(18)F]FECT can be used for the in vivo quantification of DAT using PET. PMID- 18158947 TI - A novel gallium bisaminothiolate complex as a myocardial perfusion imaging agent. AB - The development of new myocardial perfusion imaging agents for positron emission tomography (PET) may improve the resolution and quantitation of changes in regional myocardial perfusion measurement. It is known that a (68)Ge/(68)Ga generator can provide a convenient source of PET tracers because of the long physical half-life of (68)Ge (271 days). A new ligand, 7,8-dithia-16,24-diaza trispiro[5.2.5.2.5.3]pentacosa-15,24-diene, which consists of a N(2)S(2) chelating core incorporated into three cyclohexyl rings, was prepared. To test feasibility and potential utility, the N(2)S(2) ligand was successfully labeled and tested with (67)Ga (half-life=3.26 day; gamma=93.3, 184.6 and 300.2 keV), which showed >92% radiochemical purity. The corresponding "cold" Ga complex was synthesized, and its structure containing a pyramidal N(2)S(2) chloride core was elucidated with X-ray crystallography. In vivo biodistribution of this novel (67)Ga complex, evaluated in normal rats, exhibited excellent heart uptake and retention, with 2.1% and 0.9% initial dose/organ at 2 and 60 min, respectively, after an intravenous injection. Autoradiography was performed in normal rats and in rats that had the left anterior descending coronary artery permanently ligated surgically. Autoradiography showed an even uptake of activity in the normal heart, and there was a distinctively lower uptake in the damaged side of the surgically modified heart. In conclusion, the new N(2)S(2) ligand was readily prepared and labeled with radioactive (67)Ga. Biodistribution in rats revealed high initial heart uptake and relatively high retention reflecting regional myocardial perfusion. PMID- 18158948 TI - A (99m)Tc(CO)(3)-labeled pyrazolyl-alpha-melanocyte-stimulating hormone analog conjugate for melanoma targeting. AB - Melanoma primary tumors can be, in most cases, removed surgically, whereas there is no satisfactory treatment for metastatic melanoma, being almost always lethal at this stage. Therefore, early detection of primary melanoma tumors is essential. The finding that melanocortin-1 receptor (MC1R) is overexpressed in isolated melanoma cells and melanoma tissues led to the radiolabeling of several alpha-melanocyte-stimulating hormone (alpha-MSH) analogs for early detection and treatment of melanoma. We have coupled the alpha-MSH analog Ac-Nle-Asp-His-d-Phe Arg-Trp-Gly-Lys-NH(2), through the epsilon-amino group of Lys(11), to a pyrazolyl containing chelator (pz). The resulting pz-alpha-MSH analog reacted with the fac [(99m)Tc(CO)(3)](+) moiety, giving [Ac-Nle(4),Asp(5),d-Phe(7),Lys(11)(pz (99m)Tc(CO)(3))]alpha-MSH(4-11) in high yield, high specific activity and high radiochemical purity. This radioconjugate, which presents remarkable stability in vitro, exhibited time- and temperature-dependent internalization (4 h at 37 degrees C; 56.7% maximum internalization) and high cellular retention (only 38% was released from the cell after 5 h) in murine melanoma B16F1 cells. A significant tumor uptake [4.2+/-0.9%ID/g, at 4 h postinjection (p.i.)] was also obtained in melanoma-bearing C57BL6 mice. The in vivo affinity and specificity of the radioconjugate to MC1R were demonstrated by receptor-blocking studies with the potent NDP-MSH agonist (63.5% reduction in tumor uptake at 4 h p.i.). PMID- 18158949 TI - Properties of [(111)In]-labeled HIV-1 tat peptide radioimmunoconjugates in tumor bearing mice following intravenous or intratumoral injection. AB - INTRODUCTION: Our objective was to evaluate the tumor and normal tissue distribution and nuclear importation properties of [(111)In]-mouse IgG (mIgG) conjugated to tat peptides (GRKKRRQRRRPPQGYG) in athymic mice with subcutaneous BT-474 human breast cancer xenografts. METHODS: Tumor and normal tissue uptake was compared after intravenous (iv) or intratumoral injection of [(111)In]-mIgG tat and [(111)In]-mIgG. Area under the curve (AUC) was estimated for blood, liver, spleen, kidneys and tumor. Nuclear localization was measured by subcellular fractionation and estimated by microdosimetry. Imaging studies were performed with a gamma-camera. RESULTS: [(111)In]-mIgG-tat was eliminated from the blood and normal tissues two- to threefold more rapidly after iv injection than [(111)In]-mIgG. Tumor uptake was 4-5% injected dose per gram (%ID/g). Tumor radioactivity after intratumoral injection was initially very high (146-154 %ID/g), but declined 12- to 14-fold by 144 h postinjection. There was greater retention of [(111)In]-mIgG-tat in BT-474 tumors after intratumoral than iv injection, and the AUC (610+/-157 %ID h) was threefold greater than for intratumorally injected [(111)In]-mIgG (200+/-37 %ID h). Tat peptides increased nuclear localization of [(111)In]-mIgG after iv injection in tumor, kidney and liver cells, but only in tumor cells after intratumoral injection. Tumors were not imaged after iv administration but were predominant with intratumorally injected [(111)In]-mIgG and [(111)In]-mIgG-tat. Estimated radiation doses to the nucleus of tumor cells from intratumoral [(111)In]-mIgG-tat were 2.8x10(3) mGy/MBq and were 15-fold higher than for iv injection. CONCLUSION: [(111)In] labeled tat immunoconjugates may have potential for imaging intracellular epitopes or localized Auger electron radiotherapy of tumors. PMID- 18158950 TI - Coligand effects on the solution stability, biodistribution and metabolism of the (99m)Tc-labeled cyclic RGDfK tetramer. AB - In this study, we present the evaluation of two new ternary ligand (99m)Tc complexes [(99m)Tc(HYNIC tetramer)(tricine)(L)] [L=isonicotinic acid (ISONIC) and 2,5-pyridinedicarboxylic acid (PDA)] as potential radiotracers for tumor imaging. Athymic nude mice bearing MDA-MB-435 human breast cancer xenografts were used to evaluate their biodistribution and metabolic properties. Solution stability data showed that [(99m)Tc(HYNIC tetramer)(tricine)(L)] (L=ISONIC and PDA) had significant decomposition (14% and 35%, respectively) at 6 h in the absence of excess ISONIC or PDA coligand. Biodistribution data clearly showed that [(99m)Tc(HYNIC tetramer)(tricine)(PDA)] had a much lower uptake in most organs of interest than [(99m)Tc(HYNIC tetramer)(tricine)(ISONIC)] during the 2-h study period. Results from metabolism studies revealed that approximately 50% of [(99m)Tc(HYNIC tetramer)(tricine)(ISONIC)] remained intact in fecal samples at 120 min postinjection, whereas only 10% of [(99m)Tc(HYNIC tetramer)(tricine)(PDA)] remained intact in fecal samples. The extent of metabolism correlated well with radiotracer solution stability. The results from this and our previous studies clearly demonstrated that coligands [trisodium triphenylphosphine-3,3',3''-trisulfonate (TPPTS), ISONIC and PDA] have a significant impact on the tumor uptake, excretion kinetics and metabolism of the (99m)Tc-labeled cyclic RGDfK tetramer. Among the three radiotracers evaluated in this tumor-bearing animal model, [(99m)Tc(HYNIC tetramer)(tricine)(TPPTS)] remained the best with respect to blood clearance, tumor uptake and target/background ratios. PMID- 18158951 TI - In vitro and in vivo studies with [(18)F]fluorocholine on digestive tumoral cell lines and in an animal model of metastasized endocrine tumor. AB - PURPOSE: The aim of this study was to investigate (a) in vitro the relationship between [(18)F]fluorocholine ([(18)F]FCH) uptake and cell growth in endocrine cell lines and (b) in vivo the uptake of [(18)F]FCH by tumoral sites in an animal model of metastasized endocrine tumor. METHODS: In vitro studies were conducted on three endocrine and two nonendocrine digestive tumoral cell lines. The proliferative ratio was estimated using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay. The uptake of [(18)F]FCH and that of [(18)F]fluorodeoxyglucose ([(18)F]FDG) were measured before and after cytotoxic therapy. [(18)F]FCH biodistribution was studied in nude mice and in an endocrine xenografted mice model. RESULTS: The [(18)F]FCH uptake in tumoral cell lines was related to their proliferative capacities as measured by the MTT assay in basal conditions. After cytotoxic therapy, the IC(50) values calculated with the [(18)F]FCH incorporation test were very close to those determined with the MTT assay. Biodistribution studies showed that [(18)F]FCH was predominantly concentrated in the liver and kidney of nude mice. In the STC-1 xenografted animal model, the uptake of [(18)F]FCH in the primary tumor was only 1.1%. On autoradiography and micro-positron emission tomography, there was no uptake of [(18)F]FCH in liver metastases but there was a significant uptake of [(18)F]FDG. CONCLUSIONS: In vitro studies suggested that the incorporation of [(18)F]FCH in endocrine tumor cell lines was related to their growth capacities; however, in vivo studies conducted in an endocrine xenografted animal model showed an uptake of [(18)F]FCH in hepatic metastases lower than that in normal liver cells. An influence of the microenvironment or a competition phenomenon for [(18)F]FCH uptake between normal liver and endocrine tumor cells cannot be excluded. PMID- 18158952 TI - In vivo measurement of cell proliferation in canine brain tumor using C-11 labeled FMAU and PET. AB - INTRODUCTION: Noncatabolized thymidine analogs are being developed for use in imaging DNA synthesis. We sought to relate a labeling index measured by immunohistochemical staining bromodeoxyuridine (BUdR) technique to the uptake of (11)C 2'-fluoro-5-methyl-1-beta-d-arabinofuranosyluracil (FMAU) measured with positron emission tomography (PET) in a brain tumor model. METHODS: Adult beagles (n=8) with implanted brain tumors received [(11)C]FMAU and dynamic imaging with arterial sampling. Six dogs were then infused with BUdR (200 mg/m(2)) and sacrificed. Tumor time-activity curves (TACs) obtained from computed-tomography defined regions of interest were corrected for partial volume effects and crosstalk from brain tissue. Tissue was analyzed for the percentage of tumor volume occupied by viable cells and by viable cells in S-phase as identified by BUdR staining. PET/[(11)C]FMAU and BUdR were compared by linear regression analysis and analysis of variance, as well as by a nonparametric rank correlation test. RESULTS: Tumor standardized uptake values (SUVs) and tumor-to-contralateral brain uptake ratios at 50 min were 1.6+/-0.4 and 5.5+/-1.2 (n=8; mean+/-S.E.M.), respectively. No (11)C-labeled metabolites were observed in the blood through 60 min. Tumor TACs were well described with a three-compartment/four-parameter model (k(4)=0) and by Patlak analysis. Parametric statistical analysis showed that FMAU clearance from plasma into tumor Compartment 3 (K(FMAU)) was significantly correlated with S-phase percent volume (P=.03), while tumor SUV was significantly correlated with both S-phase percent volume and cell percent volume (P=.02 and .03, respectively). Patlak slope, K(FMAU) and tumor SUV were equivalent with regard to rank correlation analysis, which showed that tumor uptake and trapping of FMAU were correlated with the volume density of dividing cells (P=.0003) rather than nondividing cells (P=.3). CONCLUSIONS: Trapping of [(11)C]FMAU correlated with tumor growth rate, as measured by direct tissue analysis with BUdR in a canine brain tumor model, suggesting that [(11)C]FMAU is useful for the imaging of cell proliferation in cancers. PMID- 18158953 TI - Anesthesia condition for (18)F-FDG imaging of lung metastasis tumors using small animal PET. AB - Small animal positron emission tomography (PET) with (18)F-FDG has been increasingly used for tumor imaging in the murine model. The aim of this study was to establish the anesthesia condition for imaging of lung metastasis tumor using small animal (18)F-FDG PET. METHODS: To determine the impact of anesthesia on (18)F-FDG distribution in normal mice, five groups were studied under the following conditions: no anesthesia, ketamine and xylazine (Ke/Xy), 0.5% isoflurane (Iso 0.5), 1% isoflurane (Iso 1) and 2% isoflurane (Iso 2). The ex vivo counting, standard uptake value (SUV) image and glucose SUV of (18)F-FDG in various tissues were evaluated. The (18)F-FDG images in the lung metastasis tumor model were obtained under no anesthesia, Ke/Xy and Iso 0.5, and registered with CT image to clarify the tumor region. RESULTS: Blood glucose concentration and muscle uptake of (18)F-FDG in the Ke/Xy group markedly increased more than in the other groups. The Iso 2 group increased (18)F-FDG uptake in heart compared with the other groups. The Iso 0.5 anesthesized group showed the lowest (18)F-FDG uptake in heart and chest wall. The small size of lung metastasis tumor (2 mm) was clearly visualized by (18)F-FDG image with the Iso 0.5 anesthesia. CONCLUSION: Small animal (18)F-FDG PET imaging with Iso 0.5 anesthesia was appropriate for the detection of lung metastasis tumor. To acquire (18)F-FDG PET images with small animal PET, the type and level of anesthetic should be carefully considered to be suitable for the visualization of target tissue in the experimental model. PMID- 18158954 TI - State of the art review: HLA matching and outcome of unrelated donor umbilical cord blood transplants. PMID- 18158955 TI - Alemtuzumab-induced resolution of refractory cutaneous chronic graft-versus-host disease. AB - A patient with extensive cutaneous chronic graft-versus-host disease (cGVHD) affecting 100% of the body surface, with painful ulcerations that involved 20% of it, was treated unsuccessfully during 9 months with steroids, cyclosporine-A (CSa), sirolimus, tacrolimus, mychophenolate mofetil (MMF), infliximab, and rituximab. Twenty-one months after the allograft the patient was started on alemtuzumab, 10 mg/day subcutaneously, for 6 consecutive days every 4 weeks. Seven months after starting the treatment, 100% of the ulcers had disappeared, as had the pain. To our knowledge, there are no reports of the use of alemtuzumab in the treatment of extensive, ulcerated, refractory cutaneous cGVHD. The data presented here suggest that this agent may be useful in some patients with refractory forms of cGVHD. PMID- 18158956 TI - Alemtuzumab for the treatment of steroid-refractory acute graft-versus-host disease. AB - The treatment of steroid-refractory acute graft-versus-host disease (aGVHD) remains a clinical challenge, for which no standard therapy exists. Alemtuzumab is a humanized anti-CD52 monoclonal antibody (mAb) that has been successfully used as part of conditioning regimens for hematopoietic stem cell transplantation (HSCT) to prevent GVHD. The purpose of this study was to evaluate the safety and efficacy of alemtuzumab in treating steroid-refractory aGVHD (>or=grade II) following HSCT. Eighteen patients received subcutaneous alemtuzumab 10 mg daily on 5 consecutive days. Response was assessed at day 28 following initiation of alemtuzumab. Eight patients had grade II aGVHD, 8 had grade III, and 2 had grade IV. The main organ involved was the liver in 4 patients, gastrointestinal (GI) tract in 5, skin in 3, skin and liver in 3, and skin and GI tract in 3. Fifteen patients (83%) responded to alemtuzumab, including 6 (33%) with complete response. All 3 unresponsive patients died of GVHD. Ten of 15 responders are alive at median follow-up of 11 months (range: 3-24). Infections occurred in 14 patients, including cytomegalovirus (CMV) reactivation in 11. Grade 3 neutropenia and thrombocytopenia occurred in 6 and 4 patients, respectively. Alemtuzumab was well tolerated, and induces promising response rates in steroid-refractory aGVHD. PMID- 18158957 TI - Importance of interleukin-7 in the development of experimental graft-versus-host disease. AB - Interleukin (IL)-7 promotes both thymopoiesis and mature T lymphocyte survival and proliferation in experimental murine models of hematopoietic stem cell (HSC) transplantation. Because HSC products for transplantation also may contain IL-7 responsive mature T lymphocytes, we examined whether IL-7 is necessary for the induction of GVHD after allogeneic bone marrow transplantation (BMT). Lethally irradiated C57BL6J (B6) and B6.IL-7(-/-) (both H2K(b)) recipient mice were co transplanted with T cell-depleted (TCD) bone marrow cells and lymph nodes (LNs) from either congenic B6.SJL (CD45.1(+)) or allogeneic BALB/c (H2K(d)) donor mice. After transplantation, the recipient mice were subcutaneously injected with either human recombinant IL-7 or phosphate-buffered saline (PBS) for 60 days. No evidence of GVHD was detected in the congenic recipients or in the allogeneic B6/IL-7(-/-) recipients treated with PBS; in contrast, significantly increased rates of GVHD-related mortality and morbidity were found in the allogeneic B6.IL 7(-/-) recipients treated with IL-7. The proliferation and number of donor T cells were significantly lower at day 30 post-BMT in the PBS-treated B6.IL-7(-/-) recipients compared with the IL-7-treated B6.IL-7(-/-) mice. These experiments demonstrate that IL-7 is an important factor in the development of GVHD, presumably by supporting the survival, proliferation, and possibly activation of alloreactive donor-derived T cells in the recipients. PMID- 18158958 TI - A prognostic score for patients with acute leukemia or myelodysplastic syndromes undergoing allogeneic stem cell transplantation. AB - Allogeneic hematopoietic stem cell transplantation (SCT) has the potential to cure patients with acute leukemia or myelodysplastic syndromes (MDS), but a number of prognostic factors can influence the outcome of transplantation. At present, no transplantation-specific risk score exists for this patient population. We propose a simple scoring system for patients with acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), or MDS, based on a retrospective analysis of 445 patients undergoing SCT at our institution (divided into training and validation subsets). The score depends on 5 variables: age, disease, stage at transplantation, cytogenetics, and pretransplantation ferritin. It divides patients into 3 groups of comparable size, with 5-year overall survival (OS) of 56% (low risk), 22% (intermediate risk), and 5% (high risk). This prognostic score could be useful in making treatment decisions for individual patients, in stratifying patients entering clinical trials, and in adjusting transplantation outcomes across centers under the new federal reporting rules. PMID- 18158959 TI - Long-term outcomes of autologous stem cell transplantation for follicular non Hodgkin lymphoma: effect of histological grade and Follicular International Prognostic Index. AB - Although results of autologous stem cell transplantation (SCT) for recurrent follicular non-Hodgkin lymphoma (NHL) have been previously reported, the long term results and evaluation of prognostic factors in a large patient population receiving this therapy are difficult to find in the literature. To address these issues, we evaluated 248 patients with recurrent follicular NHL treated with high dose chemotherapy and autologous SCT between 7/87 and 6/03. According to the World Health Organization (WHO) classification system, 64 patients (26%) had follicular NHL grade 1 (FL 1), 98 (40%) had FL 2, and 86 (35%) had FL 3. At the time of transplantation, 88 of the patients (35%) had a Follicular Lymphoma International Prognostic Index (FLIPI) score of low risk, 87 (35%) had an intermediate-risk FLIPI score, 37 (15%) had a high-risk FLIPI score, and 36 (15%) had at least 1 missing value, preventing calculation of the FLIPI score. The 5 year overall survival (OS) for all patients was 63%, and the 5-year progression free survival (PFS) was 44%. In a multivariate analysis, a histological grade of FL 3, a high-risk FLIPI score at the time of transplantation, and having received 3 or more previous chemotherapy regimens were significant factors for predicting a worse OS. In addition, the use of a transplantation regimen including a monoclonal antibody decreased the relative risk of progressive lymphoma. These data suggest that transplantation earlier in the course of the disease for patients with follicular lymphoma with use of a monoclonal antibody-based regimen may lead to improved outcomes. PMID- 18158960 TI - Long-term outcome after bone marrow transplantation for aplastic anemia using cyclophosphamide and total lymphoid irradiation as conditioning regimen. AB - We retrospectively studied 49 patients in a single institute to evaluate the long term outcome of total lymphoid irradiation (TLI) conditioning for allogeneic stem cell transplantation (allo-SCT) to treat aplastic anemia (AA). Most of the patients had received transfusions and had undergone previous treatment, with 33 receiving related transplants and 16 receiving unrelated transplants. Conditioning consisted of cyclophosphamide (Cy; 200 mg/kg) plus TLI (750 cGy) for related transplantation and Cy plus total body irradiation (TBI; 500 cGy) and TLI (500 cGy) for unrelated transplantation. Antithymocyte globulin (ATG) was added for 6 of the unrelated transplantations. Graft-versus-host-disease (GVHD) prophylaxis consisted mainly of cyclosporine (CSA) and methotrexate (MTX). Graft failure developed in 2 patients (4.1%). With a median follow-up of 7 years, overall survival (OS) was 81% and was not statistically significantly different between the patients receiving related transplants and those receiving unrelated transplants. In multivariate analyses, a history of previous treatment with ATG was the sole factor associated with a worse survival rate, and the interval from diagnosis to treatment was not prognostic. The incidence of acute (grade II to IV) GVHD (aGVHD) was 23%, and that of chronic GVHD (cGVHD) was 29%. Female-to male transplantation was the sole factor associated with chronic GVHD. B cell lymphoproliferative disorder developed only after the ATG-containing conditioning. No other secondary malignancies developed after long-term follow up. Our findings suggest that TLI conditioning is feasible and effective for patients with AA. PMID- 18158961 TI - High response rate to donor lymphocyte infusion after allogeneic stem cell transplantation for indolent non-Hodgkin lymphoma. AB - The role of donor lymphocyte infusion (DLI) in the management of lymphoid malignancies after allogeneic stem cell transplantation (SCT) has not been clearly characterized. There is emerging evidence pointing to the effectiveness of this approach, particularly in patients with low-grade disease, although to date this has been reported only in small numbers of patients, and thus the utility of this treatment remains uncertain. A total of 28 patients with low grade lymphoid malignancies previously treated with allogeneic SCT received a total of 68 infusions of donor lymphocytes. The diagnoses were indolent non Hodgkin lymphoma (NHL; n = 23) and transformed NHL (n = 5), and the indications for DLI were progressive disease with or without mixed chimerism (MC) (n = 17) and persistent MC alone (n = 11). Escalating doses of cells were administered in the absence of graft-versus-host disease (GVHD) or continued disease progression, until stable full donor chimerism or disease response were achieved. The cumulative response rates after DLI to treat progressive disease and persistent MC were 76.5% and 91.6%, respectively. The major toxicity resulting from the use of donor lymphocytes was GVHD. The cumulative incidence of acute grade II-IV disease was 15%, and that of extensive chronic disease was 31%; there were no deaths resulting from GVHD. Seven patients had graft-versus-lymphoma responses without significant GVHD. These data support the existence of a clinically significant graft-versus-tumor effect in indolent NHL and suggest that this is an effective treatment for progressive disease after allogeneic SCT. PMID- 18158962 TI - Dose-reduced busulfan, cyclophosphamide, and autologous stem cell transplantation for human immunodeficiency virus-associated lymphoma: AIDS Malignancy Consortium study 020. AB - Intensive chemotherapy for human immunodeficiency virus (HIV)-associated non Hodgkin lymphoma (NHL) and Hodgkin lymphoma (HL) has resulted in durable remissions in a substantial proportion of patients. High-dose chemotherapy and autologous stem cell transplantation (AuSCT), moreover, has resulted in sustained complete remissions in selected patients with recurrent chemosensitive disease. Based on a favorable experience with dose-reduced high-dose busulfan, cyclophosphamide, and AuSCT for older patients with non-HIV-associated aggressive lymphomas, an AIDS Malignancy Consortium multicenter trial was undertaken using the same dose-reduced busulfan and cyclophosphamide preparative regimen with AuSCT for recurrent HIV-associated NHL and HL. Of the 27 patients in the study, 20 received an AuSCT. The median time to achievement of an absolute neutrophil count (ANC) of >or= 0.5 x 10(9)/L was 11 days (range, 9-16 days). The median time to achievement of an unsupported platelet count of >or= 20 x 10(9)/L was 13 days (range, 6-57 days). One patient died on day +33 posttransplantation from hepatic veno-occlusive disease (VOD) and multiorgan failure. No other fatal regimen related toxicity occurred. Ten of 19 patients (53%) were in complete remission at the time of their day +100 post-AuSCT evaluation. Of the 20 patients, 10 were alive and event-free at a median of 23 weeks post-AuSCT. Median overall survival (OS) was not reached by 13 of the 20 patients alive at the time of last follow up. This multi-institutional trial demonstrates that a regimen of dose-reduced high-dose busulfan, cyclophosphamide, and AuSCT is well tolerated and is associated with favorable disease-free survival (DFS) and OS probabilities for selected patients with HIV-associated NHL and HL. PMID- 18158963 TI - Recipient NOD2/CARD15 variants: a novel independent risk factor for the development of bronchiolitis obliterans after allogeneic stem cell transplantation. AB - Bronchiolitis obliterans (BO) is a serious complication after allogeneic stem cell transplantation. We hypothesized that single nucleotide polymorphisms (SNPs) of the NOD2/CARD15 gene (=NOD2/CARD15 variants) contribute to changes in host defense and subsequent alloreaction, leading to BO. We analyzed 427 donor recipient pairs for the association of NOD2/CARD15 variants (SNP8 [Arg702Trp], SNP12 [Gly908Arg], and SNP13 [Leu1007fsinsC]) with BO occurrence. Overall, 11 patients (2.6%) developed BO. The cumulative incidence of BO rose from 1.3% in donor-recipient pairs without mutation to 18.7% in pairs with donor or recipient NOD2/CARD15 variants (P < .001). Recipient NOD2/CARD15 variants alone led to BO in 22.3% (P < .001), whereas donor variants alone associated with BO in 13.2% (P = .04). Multivariate analysis proved recipient but not donor NOD2/CARD15 variants to be a novel independent risk factor for BO development, and NOD2/CARD15 typing may help identify patients at increased risk for BO. PMID- 18158964 TI - Donor killer immunoglobulin-like receptor (KIR) genotype-patient cognate KIR ligand combination and antithymocyte globulin preadministration are critical factors in outcome of HLA-C-KIR ligand-mismatched T cell-replete unrelated bone marrow transplantation. AB - We previously reported the potent adverse effects of killer immunoglobulin-like receptor (KIR) ligand mismatch (KIR-L-MM) on the outcome of T cell-replete unrelated hematopoietic stem cell transplantation (UR-HSCT) through the Japan Marrow Donor Program. Other UR-HSCT studies have yielded inconsistent results. To address this discrepancy, we evaluated candidate factors contributing to the effects of KIR-L-MM on transplantation outcomes in retrospectively selected hematologic malignancy cases with uniform graft-versus-host disease (GVHD) prophylaxis (n = 1489). KIR-L-MM in the graft-versus-host direction (KIR-L-MM-G) was associated with a higher incidence of acute GVHD (aGVHD; P < .002) and a lower overall survival (OS; P < .0001) only without the preadministration of antithymocyte globulin (ATG). Furthermore, in KIR-L-MM-G, the donor KIR2DS2 gene with the patient cognate C1 ligand was associated with a higher incidence of aGVHD (P = .012). Multivariate analysis by Cox proportional hazard models suggested that donor 2DS2 and ATG preadministration were critical factors in grade III-IV aGVHD (hazard ratio = 1.96; 95% confidence interval = 1.01-3.80; P = .045, and hazard ratio = 0.56; 95% confidence interval = 0.31-0.99; P = .047, respectively). These results indicate that the adverse effects of KIR-L-MM-G depend on combination of donor-activating KIR genotype-patient cognate KIR ligand type and no ATG preadministration, thereby suggesting the importance of these factors in UR-HSCT and in leukemia treatment using natural killer (NK) cell alloreactivity. PMID- 18158965 TI - Once-daily intravenous busulfan with therapeutic drug monitoring compared to conventional oral busulfan improves survival and engraftment in children undergoing allogeneic stem cell transplantation. AB - Because of intra- and interindividual variability, bioavailability, and pharmacokinetics of busulfan (Bu) in children, oral busulfan without therapeutic drug monitoring (TDM) is assumed to be associated with higher graft failure rates as well as higher toxicity (eg, veno-occlusive disease [VOD]). This study compares the outcome of hematopoietic stem cell transplantation (HSCT) of 2 groups: 1) 30 patients who received myeloablation with once-daily intravenous (i.v.) dose-targeted busulfan (BUdtIV) based on TDM and 2) 30 patients who received the current practice of untargeted oral busulfan (BUPO). Patients received a 3-hour infusion of Bu at a first dose of 120 mg/m(2) (age >or=1 year) or 80 mg/m(2) (<1 year), or BUPO 1 mg/kg 4 times daily. Both regimens were continued for 4 days. The target area under the curve (AUC) was defined as 17,500 microg *h/l. BUdtIV resulted in higher event-free survival (EFS) and survival rates compared to BUPO (EFS: 30% versus 83%, P < .001, survival: 53% versus 83%, P = .016). BUdtIV was associated with more cases of VOD. TDM was feasible in routine clinical practice. The results show that i.v. Bu using TDM is preferable over oral Bu in children undergoing allogeneic stem cell transplantation, especially in those at high risk for graft failure/relapse. PMID- 18158966 TI - Depletion of alloreactive donor T lymphocytes by CD95-mediated activation-induced cell death retains antileukemic, antiviral, and immunoregulatory T cell immunity. AB - In allogeneic hematopoietic stem cell transplantation (AHSCT) graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect are closely but not invariably linked. Thus, harnessing donor lymphocyte mediated GVL immunity and separating it from GVHD is of particular interest. Based on results obtained in murine models we have explored the CD95-mediated activation-induced cell death (AICD) strategy to selectively deplete alloreactivity in human donor T lymphocytes in vitro. Following stimulation of CD3(+) T cells isolated from HLA A* 0201-positive donors with HLA or minor histocompatibility antigen mismatched hematopoietic or nonhematopoietic cells in the presence of agonistic anti-CD95 antibody, we achieved efficient and selective allodepletion across major and minor histocompatibility mismatched barriers. Residual alloreactivity was in the range of 10% and 25% using hematopoietic cells and primary keratinocytes as alloantigen-presenting cells, respectively. CD8(+) T cells specific for HLA-A * 0201-associated cytomegalovirus (CMV), Epstein-Barr virus (EBV), and Wilms tumor 1 peptide epitopes were retained at significant numbers within the allodepleted donor lymphocyte subsets. Additionally, CD4(+) FoxP3(+) regulatory T cells persisted after the allodepletion procedure. Our results show that AICD induced by an agonistic anti-CD95 antibody might be useful to generate allodepleted donor lymphocyte products with preserved beneficial immune functions for patients undergoing AHSCT. PMID- 18158967 TI - Clinical evaluation of oral chronic graft-versus-host disease. AB - Oral chronic graft-versus-host disease (cGVHD) is a significant and serious complication following allogeneic hematopoietic stem cell transplantation (HSCT). The purpose of this study was to characterize the distribution, type, and extent of lesions and their correlation with patient-reported symptoms such as pain and discomfort. The effect of time since transplantation on these measures was also assessed. Consecutive patients with oral cGVHD referred to the Center for Oral Disease at Brigham and Women's Hospital, Boston, MA, were evaluated over a 2-year period. Subjective data included the responses to 4 targeted symptom questions (yes/no) and a visual analog scale pain score (0-10). Objective data included the location and extent of reticulation, erythema, and ulcerations using a previously published scoring system as well as time since HSCT. Multiple linear regression analyses were performed using SAS. We evaluated 27 patients, for a total of 79 clinic visits (median 2, range: 1-8). The median time since HSCT was 18 months (range: 5-157 months). The buccal and labial mucosa and tongue were the sites of 93% of all ulcerations, 72% of all erythematous lesions, and 76% of all reticular lesions, and were the most frequently affected sites. The gingiva, floor of mouth, and hard and soft palate were infrequently affected. Although uncommon, ulceration of the soft palate was the objective finding most highly correlated with increased pain (P < .0001), and there was a generalized significant trend for increased pain scores with increased extent of ulceration. Overall, 95% of pain scores were or=1.0 x 10(9)/L compared to patients ALC-30 <1.0 x 10(9)/L. Higher ALC-60, ALC 100, ALC-180, and ALC-365 did not predict superior OS and PFS. Patients with early-stage disease had significantly higher OS (ISS stages I, II, and III: 96, 53, and 29 months, respectively; P = .0023) and PFS (ISS stages I, II, and III: 55.5, 31, and 12 months, respectively; P = .027) compared to patients with advanced-stage disease at diagnosis. On univariate analysis, the type of initial chemotherapy (melphalan, VAD, PCAB), lymphocyte count on day of leukapheresis, and the lymphocyte dose infused (LY-DO) significantly influenced lymphocyte recovery following ASCT. Patients who received higher lymphocyte dose (LY-DO) >or=0.2 x 10(9)/kg had higher median ALC-15 (0.25 versus 0.19 x 10(9)/L; P = .3), ALC-30 (1.20 versus 0.99 x 10(9)/L; P = .08), ALC-60 (1.90 versus 1.01 x 10(9)/L; P = .013), ALC-100 (1.58 versus 1.03 x 10(9)/L; P = .016), and ALC-180 (1.33 versus 1.01 x 10(9)/L; P = .1), compared to patients who received LY-DO <0.2 x 10(9)/kg. In summary, our data suggest that infusing large numbers of lymphocytes improves lymphocyte recovery post-ASCT, and that higher ALC-30 is associated with better PFS and OS. These data suggest that a threshold number of CD34(+) cells should not be the only parameter considered for an adequate PBSC collection- perhaps a certain number of lymphocytes should be aimed for as well. PMID- 18158969 TI - Acute renal failure after nonmyeloablative stem cell transplantation in adults. AB - Acute renal failure (ARF) after myeloablative stem cell transplantation (SCT) is a well-established problem. Little is known about ARF after nonmyeloablative SCT. The aim of the present study was to assess the incidence of ARF and to analyze risk factors for ARF. Moreover, we wanted to study whether ARF influenced survival. We performed a retrospective cohort study of 150 adults who received nonmyeloablative SCT (fludarabine 30 mg/m(2)/day for 3 days and/or total-body irradiation (TBI) 200 cGy). ARF was categorized into grade 0 (no ARF), grade 1 (decrease in glomerular filtration rate >or=25% and doubling in serum creatinine), and grade 2 plus (> tripling in serum creatinine). ARF grade 2-2 plus developed in 49 of 150 patients (33%) after a median of 37 days, 14 patients (9%) had ARF grade 2 plus. No patient required dialysis. Risk factors at baseline for ARF grade 2-2 plus were a history of autologous transplantation (P = .008), the absence of vascular disease (P = .012) lower serum creatinine (P < .001), and higher glomerular filtration rate (P < .001). Acute graft-versus-host disease (aGVHD) grade III-IV was the only complication that was associated with ARF (P = .035). Overall mortality at 1 year was 23%. Patients with ARF grade 2-2 plus had significantly higher mortality compared to ARF grade 0-1 (P = .006). This was largely attributable to a diminished survival in patients with ARF grade 2 plus, who had a mortality rate of 71% caused by, among others, progression of malignancy and GVHD. This makes severe ARF an indicator for decreased survival. PMID- 18158970 TI - Continuing erythropoietin during peripheral blood stem cell collection in myeloma: can it reduce toxicity of autologous transplants? PMID- 18158976 TI - Perioperative and adjuvant treatments for gastric cancer. Definitive new standards of care or are we still on the way? PMID- 18158977 TI - Friendly bacterium. PMID- 18158978 TI - The oestrogen-dependent biology of breast cancer. Sensitivity and resistance to aromatase inhibitors revisited: a molecular perspective. AB - Endocrine treatment of breast cancer was the first molecular targeted anti-cancer therapy to reach clinical practice. Among the several options that share the common denomination of hormonal treatment, aromatase inhibitors (AIs) in the postmenopausal setting show the highest efficacy rates. These drugs have become the standard of care both in the advanced and adjuvant scenarios. Nevertheless resistance to AIs either upfront or after initial clinical response is almost a universal feature whenever tumour excision is not possible. Multiple reports have established the role of alternative pro-growth signalling pathways in the acquisition of resistance to the oestradiol deprivation that AIs produce. However the first clinical trials addressing the double blockade of both the oestrogen and other growing factor pathways raise some concerns on the efficacy of this approach. This review presents the evidence on the molecular events underpinning the response and resistance to AIs and suggests some key issues to consider when designing clinical research projects in this context. PMID- 18158979 TI - Epidermal stem cells in skin homeostasis and cutaneous carcinomas. AB - Skin and squamous epithelia regulate water and heat homeostasis and constitute our first barrier of protection against pathogens. Cells from the outermost layer of the skin, the cornified envelope (stratum corneum), are constantly being shed, imposing a constant demand for replenishment to maintain homeostasis. Hair follicles and sebaceous glands provide protective hair growth and skin sebum, and continuously undergo cycles of growth and regression. The outstanding ability of the epidermis, hair follicles and sebaceous glands to self-renew relies on a population of adult stem cells that are maintained throughout our life span. In this review we will provide an overview of our current knowledge about epidermal stem cells, and some of the molecular mechanisms that identify them and dictate their behaviour. We will also summarise our view on the possible link between adult epidermal stem cells and cancer stem cells within skin and squamous neoplasias. The potential of epidermal stem cells in regenerative medicine and for designing targeted antitumoral therapies will be discussed. PMID- 18158980 TI - Natural products as leads to anticancer drugs. AB - Throughout history, natural products have afforded a rich source of compounds that have found many applications in the fields of medicine, pharmacy and biology. Within the sphere of cancer, a number of important new commercialised drugs have been obtained from natural sources, by structural modification of natural compounds, or by the synthesis of new compounds, designed following a natural compound as model. The search for improved cytotoxic agents continues to be an important line in the discovery of modern anticancer drugs. The huge structural diversity of natural compounds and their bioactivity potential have meant that several products isolated from plants, marine flora and microorganisms can serve as "lead" compounds for improvement of their therapeutic potential by molecular modification. Additionally, semisynthesis processes of new compounds, obtained by molecular modification of the functional groups of lead compounds, are able to generate structural analogues with greater pharmacological activity and with fewer side effects. These processes, complemented with high-throughput screening protocols, combinatorial chemistry, computational chemistry and bioinformatics are able to afford compounds that are far more efficient than those currently used in clinical practice. Combinatorial biosynthesis is also applied for the modification of natural microbial products. Likewise, advances in genomics and the advent of biotechnology have improved both the discovery and production of new natural compounds. PMID- 18158981 TI - Deposition-associated diseases related with a monoclonal compound. AB - Up to 3% of adults over 50 years of age show a monoclonal peak values in blood or urine. Findings and prognosis will be distinct in view of the nature of this factor. In B-cell neoplasias (multiple myeloma, Waldestrom macroglobulinaemia, chronic myeloid leukaemia and non-Hodgkin lymphoma) the clinical pattern is dominated by the systemic effects produced by the expansion of the malign clone; the monoclonal protein may result in hyperviscosity syndrome or renal damage. On the other hand, there are other less frequent processes called diseases associated to monoclonal components, where the main clinical manifestations and prognosis depend of the biological effects of the monoclonal protein. With reference to this last group, which is the objective of this revision, no bone lesions, anaemia or a greater tendency to infections usually occur when compared with the first group. Even so, there are some cases of interposition between both groups: for instance, type IgM immunoglobulin present in Waldestrom macroglobulinaemia may have cold agglutinin activity, and in the case of multiple myeloma, the clone may secrete amyloidogenic light chains. PMID- 18158982 TI - The oncology acute toxicity unit (OATU): an outpatient facility for improving the management of chemotherapy toxicity. AB - OBJECTIVE: To provide an outpatient facility to improve the management of chemotherapy toxicity in cancer patients. PATIENTS AND METHODS: We set up an oncology acute toxicity unit (OATU) to improve toxicity management. A telephone helpline was the initial contact which filters out inappropriate non-toxicity related events. Patients were provided an information booklet describing the possible side effects of the chemotherapy and the helpline telephone number. A specialist nurse received the calls and consulted the doctor if necessary. Depending on requirements, the patient's problem was resolved by telephone, or a consultation visit at the OATU was arranged. RESULTS: Between February 1999 and August 2001, 1126 patients made 2007 contacts with the OATU. The most common tumours were breast (26%), colorectal (20%) and lung (20%). The telephone helpline was used in 87% of contacts and 37% were considered inappropriate. Of the 1263 appropriate contacts, the most frequent chemotherapy schedules that had been administered were 5FU-leucovorin (11.2%) and CMF (10.4%). The most frequent side effects were fever (35.5%), diarrhoea (18.5%), mucositis (16.2%) and emesis (13%). The problem was resolved by telephone in 48% of cases and 52% required attendance in the OATU, of which 40% required hospital admission, i.e., 21.1% of the initial appropriate helpline contacts. The most frequent reason was Grade 3-4 neutropenic fever (56.5%). CONCLUSIONS: The OATU enables prompt and efficient access of patients to medical oncology facilities in the event of toxicity due to chemotherapy. Unnecessary emergency room use is avoided while oncology outpatient and hospitalisation facilities are optimised. PMID- 18158983 TI - Evidence-based estimation and radiotherapy utilisation rate in Andalusia. AB - INTRODUCTION: The objective of this study was to estimate the theoretical needs based on evidence- of radiotherapy treatments (RDT) in Andalusia, compare these needs with actual use of RDT in 2006 and analyse their evolution from 2003. MATERIALS AND METHODS: Correlation between quantitative variables was analysed with Pearson's correlation coefficient. This dealt with differences between administered/estimated treatments and treatments carried out in years with the Student's t-distribution, and the Xi2 test among qualitative variables. RESULTS: In Andalusia, the evidence-based rate of cancer irradiation is 55%. Eighty-five percent of theoretical treatments were administered in 2006. From this group, 107% were in gynaecological tumours, 100% in breast cancer cases, 71% in head and neck cancer and 48% in lung cancers; differences in the last two conditions were significant (p<0.01). As for regional distribution, differences were reported with reference to irradiation rates (p<0.0002) and resource distribution. In the last three years, an increment of 17% was observed in treatments conducted in public hospitals. The rate increased from 61% (with regard to optimal values) to 85% in 2006; in a parallel way, an increment was seen in therapy units (from 22 to 26) and radiation oncologists (from 57 to 69). CONCLUSIONS: Despite the increment of irradiation rates seen in the last years, there is still a serious underutilisation of RDT for some cancer types (lung, head and neck cancer), as well as a great variability in the use of RDT between hospitals. PMID- 18158984 TI - Clinical course of high-grade glioma patients with a "biopsy-only" surgical approach: a need for individualised treatment. AB - INTRODUCTION: 'Biopsy-only' high-grade glioma (HGG) patients get limited benefit from post-operative treatments, and as a group, negatively impact median survival outcomes. MATERIAL AND METHODS: We retrospectively evaluated clinical characteristics, treatment and overall survival of HGG patients with a 'biopsy- only' surgical approach diagnosed between 1997 and 2005 at a University Hospital in Spain. RESULTS: In 31% of 294 suspected gliomas, only a diagnostic biopsy was undertaken. Reasons for 'biopsy-only' for all patients were either location in eloquent areas: (motor area 18.7%, language area 25,3%, basal ganglia 7.7%, visual area 4.4%) or extension of the disease (corpus callosum invasion 14.3% and multicentricity/multifocality 28.6%). Seventy-four patients (80.4%) were HGG: 26% of all grade IV and 49% of all grade III tumours. For these patients, post operative Karnofsky Performance Status of over 70%, median age and median survival were, respectively: 64 and 70%, 60.7 and 57 years old, and 23.1 and 42.7 weeks (p=0.0006). Patients lived longer if post-operative treatment was given, in all grades (p<0.0001). Nineteen patients (25.6%) died within 42 days after surgery. Only 60% of them initiated radiotherapy and 10% of them did not complete it. However, tumour grade, radiotherapy and temozolomide- based chemotherapy were independently associated with longer survival in multivariate analysis (p<0.05). CONCLUSION: Almost one third of HGG patients can undergo only a biopsy and not debulking surgery. Although radiotherapy improves survival, only 50% of them complete the treatment. An individualised approach to these patients is needed to facilitate a correct analysis of therapy results. New therapies must be investigated in these patients. PMID- 18158985 TI - Peritoneal carcinomatosis secondary to carcinoid tumour. AB - Carcinoid tumours are neuroendocrine neoplasms that can appear in every location of the digestive tract. They are low aggressive tumours, although they often produce local invasion and hepatic metastases, whose resection allows long-term survival. We report a case of a 64-year-old man with ileal carcinoid tumour, that underwent ileal resection and metastasectomy of one lesion in liver segment II. Surgical findings indicated peritoneal carcinomatosis. Carcinoid dissemination as peritoneal carcinomatosis has been rarely described in the literature. Cytoreductive surgery, always when complete resection is aimed, achieves asymptomatic long-term survivals. PMID- 18158986 TI - Renal primary angiosarcoma. AB - Angiosarcomas account for 2% of all soft tissue sarcomas and of them, primary renal angiosarcomas represent 1%. Twenty-four cases have been published in the English specialised literature. We report the second case to be described in a middle-aged female, with pulmonary metastases at diagnosis, and fatal outcome despite surgery and chemotherapy. PMID- 18158987 TI - [Neuroprotective effects of vasoactive intestinal peptide against rat cerebral ischemia]. AB - OBJECTIVE: To explore the neuroprotective effect of vasoactive intestinal peptide (VIP) in rat ischemic brain injury. METHODS: VIP was administered via intracerebroventricular injection in SD rats prior to focal cerebral ischemia by intraluminal occlusion of the middle cerebral artery. The infarct volume was assessed with TTC staining, and immunohistochemistry was performed to analyze the S100beta expression in the cerebral tissue, with the serum concentrations of S100beta detected by double-antibody sandwich enzyme-linked immunosorbent assay. RESULTS: After VIP injection, the relative infarct volume in the rats with cerebral ischemia was significantly reduced by 32.3% as compared with the volume in the control group on day 1 (P<0.05), and the number of S100beta-positive cells was significantly decreased in the cerebral tissue (P<0.05). The injection also resulted in significantly decreased serum S100beta concentrations in the rats (P<0.05). CONCLUSION: VIP injection can reduce the infarct volume in rats with focal cerebral ischemia, suggesting the neuroprotective effect of VIP in brain ischemia possibly by reducing S100beta overexpression. PMID- 18158988 TI - [Automatic extraction and segmentation of the intracranial structure from cerebral computed tomography]. AB - OBJECTIVE: This paper describes a new method for extracting and segmenting intracranial structure from serial images of cerebral computerized tomography automatically. A region growing- and morphology-based approach was first developed to extract intracranial structures from the serial images of cerebral computerized tomography, and focusing on the problems of parameter initialization of the expectation maximization (EM) algorithm, an improved EM algorithm based on parameter- limited GMM was presented to segment the intracranial structures successfully. Experimental results of the algorithm showed that this method was effective for all cerebral computerized tomography images from bottom to top of the cerebrum. PMID- 18158989 TI - [Cellular localization of HCBP1 and its interaction with HCV core protein in vivo]. AB - OBJECTIVE: To analyze the interaction of hepatitis C virus (HCV) core protein with HCBP1 and observe the expression and cellular localization of HCBP1. METHODS: The cDNA fragments encoding HCV core protein and HCBP1 were amplified by PCR and subsequently cloned into pGEM T vector, respectively. After sequence verification, the two recombined vectors were respectively subcloned into two hybrid plasmids, pM and pVP16. pM-core, pVP16- HCBP1 and the reporter vector pG5CAT were co-transfected into COS-7 cells, and the interaction between HCV core protein and HCBP1 was assayed by detecting CAT gene expression after 48 h. The expression and subcellular localization of the fusion protein in the transfected COS-7 cells were analyzed by Western blotting and fluorescence microscopy, respectively. RESULTS: CAT-ELISA showed that the absorbance of the co transfection group was significantly higher than that o f the negative control groups but lower than that of the positive control group. Western blotting confirmed the expression of fusion protein in the transfected COS-7 cells. Fluorescence microscopy showed that the fusion protein was distributed mainly in the cytoplasm, and in contrast, diffuse EGFP expression was detected in COS-7 cells transfected with the empty vector. CONCLUSION: Mammalian two-hybrid assay confirms the capacity of HCBP1 to bind HCV core protein, and the expression vector for HCBP1-EGFP fusion gene has been constructed successfully and expressed in COS-7 cells. PMID- 18158990 TI - [MicroRNAs can be expressed in cardiomyocyte-like cells differentiated from human mesenchymal stem cells]. AB - OBJECTIVE: To investigate the expression of representative heart-specific primary microRNAs (pri-miRNAs) in the cardiomyocyte-like cells differentiated from human mesenchymal stem cells (hMSCs). METHODS: The phenotype of hMSCs isolated was identified by flow cytometry using monoclonal antibodies against FITC-conjugated CD29, CD34, and CD11b. The third-passage hMSCs were induced to differentiate into cardiomyocyte-like cells by 5-azacytidine and indirect coculture with neonatal rat myocytes, respectively. Immunocytochemical analysis was performed to detect the expression of the cardiac-specific proteins, namely cardiac troponin I (cTnI) and sarcomeric alpha-actinin, in the cardiomyocyte-like cells differentiated from hMSCs. RT-PCR and DNA sequencing were used to identify the expression of the 5 representative heart-specific pri-miRNAs. RESULTS: High hMSC marker CD29 expression rate (98.87%) and low hematopoietic cell markers CD34 (5%) and CD11b (0.4%) expression rates were identified in the hMSCs isolated. cTnI and sarcomeric alpha-actinin expression occurred in the hMSCs following induction with the 2 differentiation-inducing methods. miRNA-143 and -181 expressions were induced in the hMSCs by 5-azacytidine and miRNA-143, -181, -206, and -208 expressions were induced by indirect coculture with neonatal rat myocytes, but pri-miRNA-1-2 expression failed to be induced by these two induction methods. CONCLUSION: Expressions of the representative heart-specific pri-miRNAs in different patterns can be induced in cardiomyocyte-like cells differentiated from hMSCs by 5-azacytidine and indirect coculture with neonatal rat myocytes. PMID- 18158991 TI - [Main factors affecting the effect of photodynamic therapy against human esophageal cancer cells in vitro]. AB - OBJECTIVE: To evaluate the tumor cell-killing effect of photodynamic therapy against human esophageal cancer cells in vitro and identify the main factors affecting the effect. METHODS: Human esophageal cancer Eca-109 cells were incubated for 24 h in vitro with hematoporphyrin derivative (HpD) and Photofrin at different concentrations prior to exposure to a light energy density of 15 J/cm(2) delivered from a DIOMED 630 PDT system. The cell killing effect was also evaluated for different HpD concentrations combined with 3 light energy densities (10, 30, and 50 J/cm(2)), respectively. The cell survival rate was measured using MTT assay, and fluorescence spectrometry was used to detect the intracellular photosensitizer fluorescence of the tumor cells after incubation with HpD for 4 h. RESULTS: The cell survival rate after incubation with the two photosensitizers at different concentrations were significantly different, and under the 3 different light energy densities, incubation of the cells with different HpD concentrations also resulted in significantly different cell survival rates (P<0.05). At the 4 low photosensitizer concentrations and with different light energy densities, the cell survival rates were similar (P>0.05), but the 4 higher photosensitizer concentrations resulted in significant difference in the cells survival (P<0.05). Correlation analysis showed that the intracellular photosensitizer concentration was positively correlated to the photosensitizer concentrations in cell incubation (r=0.997). CONCLUSION: When the light source remains constant, the light energy density, the kinds of photosensitizers and their concentrations are the main factors affecting the Eca-109 cell-killing effect of PDT. PMID- 18158992 TI - [Value of plasma tissue factor, tissue factor pathway inhibitor and factor VII assessments in patients with acute myocardial and cerebral infarction]. AB - OBJECTIVE: To study the clinical implications of changes in plasma tissue factor (TF), tissue factor pathway inhibitor (TFPI) and factor VII (FVII) after the onset of acute myocardial infarction (AMI) and acute cerebral infarction (ACI). METHODS: Sixty-nine patients with AMI, 71 with ACI and 50 age-matched healthy volunteers were enrolled in this study. Blood samples were obtained from the healthy subjects and from the patients at the early stage of AMI and ACI onset for examination of plasma TF and TFPI activity using chromogenic assay, and the plasma TF and TFPI antigens were measured by enzyme-linked immunosorbent assay (ELISA). The plasma FVII coagulation activity (FVII:C) was also measured, and the plasma FVIIa determined using soluble TF assay. RESULTS: Compared with the healthy control group, AMI patients had significantly enhanced plasma TF and TFPI activities and elevated TF and TFPI antigen levels (P<0.05), with also markedly increased FVIIa (P<0.05) but comparable FVII:C (P>0.05). In ACI patients, the plasma TF activity and antigen were obviously increased in comparison with the control group (P<0.05), but plasma TFPI activity and antigen were lowered (P<0.05), and both the FVII:C and FVIIa were markedly higher (P<0.05). Significant differences were noted in plasma TF and TFPI activities and their antigen levels as well as in FVII:C, but not in FVIIa between AMI and ACI patients. CONCLUSION: V Following the onset of AMI and ACI, TF pathway is initiated and the risk of thrombogenesis increases, and the assessment of TF pathway is therefore of value for understanding the development of the condition. PMID- 18158993 TI - [Effect of Coriolus versicolor polysaccharide B on membrane glycosaminoglycans and cellular glutathione changes in RAW264.7 macrophages exposed to angiotensin II]. AB - OBJECTIVE: To investigate the effect of Coriolus versicolor polysaccharide B (CVP B) on increased membrane glycosaminoglycans (GAG) expression and intracellular glutathione (GSH) of RAW264.7 macrophages exposed to angiotensin II (Ang II). METHODS: The plasma membrane of RAW264.7 macrophages exposed to Ang II treatment was isolated by ultracentrifugation, and the membrane GAG expression was analyzed using 1, 9-dimethylmethylene blue (DMMB) spectrophotometric assay for sulfated GAG. The intracellular reduced GSH was determined using fluorophotometry. RESULTS: The GAG content in the macrophage membranes increased by up to 54% following cell exposure to 1.0 micromol/L Ang II, whereas in presence of 1.0 micromol;/L Ang II, CVP-B at 1, 10, and 50 microg/ml decreased the GAG content by 13%, 43% (P<0.01), and 52% (P<0.01), respectively. The macrophage GSH activity decreased by 69% following incubation with 1.0 micromol;/L Ang II for 24 h, and CVP-B treatment at 1, 10, and 50 microg/ml in presence of 1.0 micromol;/L Ang II resulted in significant increment of GSH activity by 31%(P<0.05), 104% (P<0.01), and 168% (P<0.01), respectively. CONCLUSION: These data provide the first evidence that CVP-B inhibits elevated GAG expression in RAW264.7 macrophage membrane induced by Ang II. PMID- 18158994 TI - [Influence of chronic HBV infection in the husband on the outcome of IVF-ET treatment]. AB - OBJECTIVE: To investigate the outcome of in vitro fertilization and embryo transfer (IVF-ET) in couples with the husband positive for chronic infection of hepatitis B virus (HBV). METHODS: This study involved 102 infertile couples receiving IVF-ET with the husbands(but not the wives) positive for hepatitis B surface antigen (HBsAg), and another 204 couples negative for HBsAg receiving the treatment served as the control group. The cumulative embryo score, fertilization rate, cleavage rate, rate of good quality embryos, implantation rate, clinical pregnancy rate, first trimester and late miscarriage rates, delivery rate, and neonatal malformation rate were recorded and compared between the two groups. RESULTS: Between the HBsAg-positive and the control groups, the cumulative embryo score (52.8-/+18.7 vs 55.4-/+16.9), insemination rate (66.9% vs 66.1%), cleavage rate (97.6% vs 97.2%), rate of good quality embryos (34.0% vs 37.1%), implantation rate (40.9% vs 34.6%), clinical pregnancy rate (56.9% vs 50%), first trimester miscarriage rate (6.9% vs 5.9%) and late pregnancy miscarriage rate (8.6% vs 4.9%), delivery rate (40.2% vs 43.6%) and neonatal malformation rate (0 vs 0) were all similar (P>0.05;). CONCLUSION: Chronic HBV infection in the husband might not affect the outcome of IVF-ET treatment. PMID- 18158995 TI - [Fingerprinting of Fuzhisan by high-performance liquid chromatography with ultraviolet and evaporative light scattering detector]. AB - OBJECTIVE: To establish a method for fingerprinting of Fuzhisan (FZS, a traditional Chinese medicinal preparation) using high-performance liquid chromatography with ultraviolet and evaporative light scattering detector (HPLC UV/ELSD) to allow simultaneous determination of 5 major constituents in the preparation. METHODS: HPLC-UV/ELSD analysis was performed on water AlltechC18 column (5 microm, 4.6 mm x 250 mm) with a mixture of acetonitrile (A) and 0.1% acetice acid water (B) as the mobile phase. The solvent A gradient for elution was 0, 12%; 25, 20%; 30, 20%; 75, 30%; 105, 40%; 120, 80%; 130, 12%, with the flow rate of 1.0 ml/min; and the column temperature at 30 degrees . The detective wavelength was 335 nm, drift tube temperature was 80 degrees , pressure of nebulizer gas was 25 psi. The similarities between the HPLC-UV/ELSD fingerprints of the 12 extracts were calculated using similarity evaluation software. RESULTS: The fingerprint of FZS was established and the 5 major constituents were identified. The complementarity between the fingerprints of UV and ELSD was analyzed, showing good correlation between 12 batches of FZS. CONCLUSION: The method for fingerprinting can simultaneously characterize the main chemical constituents in FZS and allows stable, effective and comprehensive quality control and evaluation of FZS for a single sample. PMID- 18158996 TI - [Construction and identification of a plasmid with EGFP reporter gene for enhancer function analysis]. AB - OBJECTIVE: To construct a plasmid vector with EGFP reporter gene for functional analysis of enhancers. METHODS: EGFP DNA was amplified by PCR from plasmid pEGFP N1 DNA and subcloned into plasmid PGL3-promoter backbone without luc(+) gene to construct the enhancer-identifying vector pEGFP-enhancer. Different copies of hypoxia response element (HRE) sequence were synthetized and subcloned into the multiple cloning site of the plasmid pEGFP-enhancer. Using Lipofectamine 2000, the recombined pEGFP-HRE and pEGFP-5HRE plasmids were transfected into the Hela cells respectively. After hypoxic or normoxic cell culture, EGFP expression in the cells was detected by flow cytometry and fluorescence microscopy. RESULTS: After hypoxic exposure, the fluorescence intensity of EGFP in the Hela cells transfected with the plasmid increased with the enhancer HRE copies, while the fluorescence intensity underwent no significant changes after normoxic cell culture. CONCLUSION: we have successfully constructed the enhancer expression vector plasmid pEGFP-enhancer, which can identify the activity of the enhancers through EGFP expression. PMID- 18158997 TI - [Effects of soluble matrix of nacre on bone morphogenetic protein-2 and Cbfa1 gene expressions in rabbit marrow mesenchymal stem cells]. AB - OBJECTIVE: To provide insights into the mechanisms and pathways of osteogensis by observing the effects of water-soluble matrix of nacre (WSM) on bone morphogenetic protein-2 (BMP-2) and Cbfa1 gene expressions in rabbit marrow mesenchymal stem cells (BMSCs). METHODS: New Zealand rabbit BMSCs cultured in vitro were stimulated with different concentrations of WSM extracted at low temperature, and the activity of AKP in the cells was evaluated with the dose effect curve generated. BMP-2 and Cbfa1 gene expressions in rabbit BMSCs exposed to WSM were assayed with one-step RT-PCR. RESULTS: The activity of AKP in rabbit BMSCs increased after stimulation with different concentrations of WSM, and the effects were the most obvious with the WSM concentration ranging from 150 to 200 microg/ml. BMP-2 gene expression in the BMSCs increased after WSM exposure, but which did not result in obvious changes in Cbfa1 gene expression. CONCLUSION: WSM induces differentiation of rabbit BMSCs towards osteoblasts by increasing BMP-2 gene expression, in which process Cbfa1 gene does not seem to play a significant role. PMID- 18158998 TI - [Small interfering RNA-mediated nuclear factor-kappaB P65 suppression induces apoptosis of hepatic carcinoma SMMC-7721 cells]. AB - OBJECTIVE: To study the mechanism of hepatic carcinoma cell apoptosis induced by small interfering RNA (siRNA)-mediated nuclear factor-kappaB (NF-kappaB) P65 silencing. METHODS: Hepatic carcinoma SMMC-7721 cells were exposed to liposome mediated transfection with NF-kappaB P65 siRNA synthesized by in vitro transcription, and the cells with empty liposome transfection and those without particular treatment served as the control groups. The expression of NF-kappaB P65 in the cells was detected by Western blotting, the cell viability examined by MTT assay, and the cell apoptosis assessed by flow cytometry. Immunohistochemistry was used to examine the expressions of Bcl-2 and Bax. RESULTS: siRNA transfection significantly inhibited the expression of NF-kappaB P65 in SMMC-7721cells, with inhibition rates of 64.74% compared with the untreated cells and of 34.52% compared with the liposome-treated cells. The siRNA treated SMMC-7721 cells also exhibited significant decrease in cell proliferation by 33.39% and 27.23% in comparison with the untreated and liposome-treated cells, respectively. NF-kappaB P65 siRNA induced obvious cell apoptosis with down regulated Bcl-2 and up-regulated Bax expressions. CONCLUSION: NF-kappaB p65 siRNA can induce SMMC-7721 cell apoptosis via the Bcl-2/Bax pathway. PMID- 18158999 TI - [Protective effect of proteasome inhibitor MG-132 in rats with severe acute pancreatitis and lung injury]. AB - OBJECTIVE: To observe the protective effect of the proteasome inhibitor MG-132 in rats with severe acute pancreatitis (SAP) and the associated lung injury. METHODS: In rat models of the SAP established with injection of 5% sodium taurocholate into the biliary-pancreatic duct, the changes of the serum amylase and myeloperoxidase (MPO) activity in the pancreatic and lung tissues were evaluated. The pathological changes of the pancreatic and lung tissues were also observed. RESULTS: MG-132 significantly decreased serum amylase, pancreatic weight/body weight ratio, and pancreatic and pulmonary myeloperoxidase activity (P<0.05). Histopathological examinations revealed milder edema, cellular damage, and inflammation in the pancreatic and lung tissues of rats pretreated with the peptide (P<0.05). CONCLUSION: MG-132 ameliorates SAP and the associated lung injury in rats. PMID- 18159000 TI - [Protective effect of ketamine against acute rat lung injury induced by liopolysaccharide and its mechanism]. AB - OBJECTIVE: To investigate the protective effects of different concentrations of ketamine against acute lung injury induced by lipopolysaccharide (LPS) in rats and its mechanism. METHODS: Forty-eight male Wistar rats were randomized into 4 equal groups, namely the control group, LPS group, ketamine group I (5 mg/kg), and ketamine group II (10 mg/kg). The neutrophil count, protein contents in the bronchoalveolar lavage fluid (BALF) and the wet/dry lung weight ratio were measured 4 h after LPS injection. TNF-alpha, IL-8, NO, iNOS and NF-kappaB were also measured in the lung tissues. RESULTS: In LPS group, the neutrophil count, protein contents in BALF, the wet/dry lung weight ratio and the levels of tumor necrosis factor-alpha(TNF-alpha), interleukin-8 (IL-8), and NO were all significantly increased compared with the control group (P<0.01). The mRNA expression of iNOS and the protein expression of NF-kappaB were also increased in LPS groups. Ketamine treatment attenuated the increase in wet/dry lung weight ratio, neutrophil count, and protein contents in BALF in a dose-dependent manner. Ketamine also dose-dependently inhibited the production of TNF-alpha, IL-8 , and NO and lowered iNOS mRNA and NF-kappaB protein expression. CONCLUSION: Ketamine can offer protection against LPS-induced acute lung injury in rats by inhibiting the expression of NF-kappaB and attenuating the production of the inflammatory cytokines. PMID- 18159001 TI - [Infrared radiation and magnetic field therapy ameliorates cartilage damage in rabbits with knee osteoarthritis]. AB - OBJECTIVE: To evaluate the effect of infrared radiation and magnetic field therapy on cartilage damage in rabbits with knee osteoarthritis. METHODS: Knee osteoarthritis was induced in 24 adult New Zealand rabbits by prolonged fixation of the knee joint in extension for 6 weeks. The rabbits were subsequently randomized into control group (without treatment), infrared therapy group, magnetic field therapy group and the combined infrared and magnetic field therapy group. At the end of the first, second and third weeks of the therapy, respectively, 2 rabbits from each group were sacrificed to observe the general changes and histopathology of the condylar cartilage of the femur, and the findings were assessed using Mankin scores. RESULTS: Compared with other groups, the rabbits in the combined therapy group showed significantly milder cartilage damage (including injury of the cartilage surface and chondrocyte's proliferation and disarrangement) with significantly lower Mankin scores (P<0.05). No significant differences were found in the findings between the two groups with exclusive infrared or magnetic field therapy (P>0.1). CONCLUSION: Combined infrared and magnetic field therapy can effectively alleviate cartilage destruction, shortens the disease course and enhance the therapeutic effects in rabbits with knee osteoarthritis. PMID- 18159002 TI - [Effect of hypoxic preconditioning on the learning and memory ability and expressions of survivin and HSP-70 proteins in rats with focal cerebral ischemia/reperfusion injury]. AB - OBJECTIVE: To study the neuroprotective effect of hypoxic preconditioning on reperfusion injury following ischemia and its molecular mechanism. METHODS: Forty eight rats were randomized into 3 groups, namely the sham operated group, ischemia/reperfusion (I/R) group, and I/R following hypoxic preconditioning group (HP+I/R). In the latter two groups, the rats were subjected to middle cerebral artery occlusion (MACO) for 3 h followed by reperfusion for 24 h to induce cerebral I/R injury. The learning and memory ability of the rats 24 h after reperfusion was assessed using Y-maze test. Immunohistochemistry was performed to quantify the expressions of survivin and HSP-70 proteins in the rat brain tissues. RESULTS: The number of survivin- and HSP-70-positive cells in the brain tissues was significantly different between HP+I/R group and IR and the sham operated groups (P<0.05), and following I/R injury, the rats in HP+I/R group showed much better performance in the Y-maze test than those in I/R group. CONCLUSION: Hypoxic preconditioning can protect the ischemic brain against reperfusion injury, promote recovery of the learning and memory ability and neurological functions following the injury. Up-regulation of the expressions of survivin and HSP-70 proteins might be one of the molecular mechanisms for this neuroprotective effect. PMID- 18159003 TI - [Expression of proliferating cell nuclear antigen in severely damaged small intestinal mucosa due to high-dose 5-FU exposure]. AB - OBJECTIVE: To detect the expression of proliferating cell nuclear antigen (PCNA) in severely damaged intestinal mucosa due to high-dose 5-FU exposure. METHODS: Thirty-two adult C57BL/6J mice were subjected to daily intraperitoneal high-dose 5-FU injection at 150 mg/kg for 5 consecutive days, and on days 1, 3, and 5, the mice were sacrificed to obtain the small intestinal tissue for HE straining and immunohistochemistry for detecting PCNA expression. Another 8 mice with intraperitoneal PBS injection served as the control group. RESULTS: High-dose 5 FU exposure of the mice resulted in severe intestinal mucous damage, with complete destruction of the villi and crypts and significantly increased cells positive for PCNA expression (P<0.01). CONCLUSION: High-dose 5-FU treatment can significantly increase the PCNA index, and the cells expressing PCNA can be closely associated with regeneration of the severely damaged mucosa due to the exposure. PMID- 18159004 TI - [Diagnostic value of 64-slice spiral CT coronary angiography for restenosis after bypass surgery]. AB - OBJECTIVE: To assess the accuracy of 64-slice spiral CT in diagnosis of restenosis of coronary artery bypass grafts (CABG) and native coronary arteries in patients after bypass surgery. METHODS: Fifty-eight patients receiving bypass surgery with totally 140 CABG (43 arterial and 97 venous grafts) were examined using 64-slice spiral CT. CABG and all native coronary arteries with a diameter of >1.5 mm were evaluated for the presence of significant stenoses (>50% diameter reduction) in comparison with the results by coronary angiography as the golden standard. RESULTS: Of the 140 CABG examined, 38 were occluded and 104 remained patent, all of which were accurately identified by 64-slice spiral CT. The sensitivity of CT for restenosis detection in the patent graft was 100% (18/18) with a specificity of 95.2% (80/84). In the segmental evaluation of the native coronary arteries, the sensitivity of the CT in identifying significant stenosis in the evaluable segments (90%) was 84% (87/103) with a specificity of 74% (384/518). The accuracy of CT in detecting the presence of at least 1 stenosis in the CABG, distal runoff vessels or nongrafted arteries was 91% (53/58). CONCLUSION: CT allows noninvasive angiographic evaluation of both the native coronary arteries and bypass grafts after bypass surgery. PMID- 18159005 TI - [Transarterial oily chemoembolization combined with interstitial laser thermotherapy for treatment of hepatocellular carcinoma]. AB - OBJECTIVE: To evaluate the therapeutic effects and adverse effects of transarterial oily chemoembolization combined with interstitial laser thermotherapy (TOCE+ILT) in the treatment of hepatocellular carcinoma. METHODS: Totally 120 patients with hepatocellular carcinoma were randomized into two groups and received interventions with TOCE+ILT or TOCE combined with percutaneous ethanol injection (TOCE+PEI). The treatment was repeated when necessary until the tumor was completely ablated, after which the therapeutic effects were evaluated and the patients were the followed up for observing long term clinical outcome. RESULTS: Of the 120 patients enrolled in this observation, 105 were followed up for two years (54 in TOCE+ILT group and 51 in TOCE+PEI group). The complete tumor necrosis rate of TOCE+ILT group was significantly higher than that of the TOCE+PEI group (84.8% vs 73.9%,Chi(2)=4.405, P=0.036), and TOCE+ILT was associated with a significantly higher negative conversion rate of AFP positivity (77.8% vs 56.1%, Chi(2)=4.592, P=0.032). The 1-year survival rate were similar between two groups, but the 2-year survival rate was significantly higher in patients with TOCE+ILT (79.6% vs 60.8%, Chi(2)=4.477, P=0.034). The hepatic function was comparable between the two groups before treatment, and 1 week after treatment, the ALT level in patients undergoing TOCE+ILT was significantly lower than that in patients with TOCE+PEI (95.90 /+56.06 U/L vs 116.31-/+45.27 U/L, t=2.04, P=0.043). Post-embolization syndrome was observed in the patients in two groups, but no severe adverse events were found. CONCLUSION: TOCE+ILT has good therapeutic effects and mild side effects in the treatment of hepatocellular carcinoma. PMID- 18159006 TI - [Detection of angiotensin II in the maternal-fetal interface in pregnancy-induced hypertension]. AB - OBJECTIVE: To detect angiotensin II (ANG II) in the maternal blood, umbilical cord blood, and at the maternal-fetal interface in women with pregnancy-induced hypertension (PIH) and explore the etiology of PIH and pathophysiologic mechanism of intrauterine growth restriction (IUGR). METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of ANG II in the maternal blood, cord blood, and maternal and fetal placental tissues in 30 women with PIH and 30 with normal pregnancy, and the results were analyzed with independent sample t test and Pearson correlation analysis. RESULTS: ANG II level in the maternal placental tissue homogenate showed no significant difference between women with PIH and normal pregnant women (8.51+/- 4.01 vs 7.76+/-3.47 pg/ml, P>0.05), but women with PIH had significantly higher ANG II in the fetal placental tissue (11.82+/-3.92 vs 9.64+/-2.63 pg/ml, P<0.05). ANG II level was significantly higher in the maternal blood of women with PIH than in normal pregnant women (46.44+/-8.48 vs 32.43+/-5.87 pg/ml, P<0.001), but similar in the cord blood (68.83+/-8.68 vs 72.47+/-8.51 pg/ml; P>0.05). A positive correlation was indicated between the cord blood and maternal peripheral blood ANG II levels in women with PIH (r=0.7379, P<0.05). CONCLUSION: ANG II in the fetal placental tissue is elevated, and the cord blood and maternal peripheral blood ANG II levels are positively correlated in women with PIH. PMID- 18159007 TI - [Impact of fenofibrate on NO and endothelial VCAM-1 expression in hyperlipidemic rats]. AB - OBJECTIVE: To investigate the mechanism of hyperlipidemia- and imflammation induced functional impairment of the endothelium. METHODS: The experiment was conducted using 3 groups of rats fed for 20 weeks with standard chow (control group), high-fat diet and high-fat diet with daily fenofibrate treatment (10 mg/kg, starting since the fifth week), respectively. After 4 and 20 weeks of feeding, respectively, serum lipid level and NO concentration were measured in the rats, and the epithelial vascular cell adhesion molecule-1 (VCAM-1) expression and cell adhesiveness to the aortic endothelium were observed. RESULTS: Compared with the control group, the rats with hyperlipidemia induced by long-term high-fat diet feeding showed lower NO concentration and increased leukocyte accumulation on the endothelial surface, exhibiting also stronger and more extensive endothelial expression of VCAM-1. In contrast, the hyperlipidemic rats with fenofibrate treatment shoed significantly decreased VCAM-1 expression and leukocyte adhesion with recovery of the NO level. CONCLUSION: NO deficiency and activation of inflammation are involved in vascular impairment in rats with high-fat diet-induced hyperlipidemia, and fenofibrate can effectively prevent atherosclerosis by restoring NO concentration and down-regulating VCAM-1 expression in these rats. PMID- 18159008 TI - [Clinical analysis of fine root canal preparation with ProTaper rotary instruments]. AB - OBJECTIVE: To evaluate the clinical effects of fine root canal preparation with rotary ProTaper. METHODS: Ninety-two teeth among 92 patients were treated with root canal therapy which root canals prepared by rotary ProTaper with Glyde gel (contains EDTA) and lateral condensation, including 66 teeth that 141 root canals were fine,17 teeth being root canal therapy and one or more than root canal were omitted,6 maxillary first molar containing second mesiobuccal canal(MB2), 3 maxillary second molar MB2. RESULTS: One hundred and thirty five in 141 fine root canals among 66 teeth were successfully prepared by rotary ProTaper, the prepared successful ratio was 95.74%,X-ray showed that obturator had smooth taper. Two palatal root canals of maxillary first molar were underfilling, 4 root canals appeared ledge at apical one third. Seventeen omitted root canals were successfully prepared and obturated. Five in 9 MB2 were successfully prepared and obturated to root-apex, four in 9 were not to end. No instrument separated in all patients. CONCLUSION: The preparation of fine root canals with rotary ProTaper and EDTA is an effective and safe and limited. Nevertheless, the preventive importance of ledge and perforation should be emphasized. PMID- 18159009 TI - [Effect of short-term intensive treatment with insulin pump on beta cell function and the mechanism of oxidative stress in newly diagnosed type 2 diabetic patients]. AB - OBJECTIVE: To investigate the effect of short-term intensive treatment with insulin pump on beta cell function and the mechanism of oxidative stress in newly diagnosed type 2 diabetic patients. METHODS: Totally 120 newly diagnosed type 2 diabetic patients were treated with insulin pump for 2 weeks. The levels of fasting plasma glucose (FPG), 2-hour postprandial blood glucose (2hPG), homeostatic model assessment of the insulin secretion index and insulin resistance index (HOMA-beta and HOMAIR, respectively), blood malondialdehyde (MDA) and superoxide dismutase (SOD) were measured before and after insulin pump treatment. RESULTS: After insulin pump treatment, FPG, 2hPG, HOMAIR and blood MDA were significantly decreased (P<0.01), while HOMA-beta and blood SOD were significantly increased (P<0.01). CONCLUSION: Short-term intensive treatment with insulin pump can effectively improve beta cell function probably by decreasing oxidative stress in newly diagnosed type 2 diabetic patients. PMID- 18159010 TI - [Optimized isolation and purification of non-typeable Haemophilus influenzae Haps protein]. AB - OBJECTIVE: To optimize the isolation and purification conditions for Hap(s) protein of non-typeable Haemophilus influenzae. METHODS: Hap(s) protein was purified by ammonium sulfate precipitation, dialysis desalting and Hitrap weak cation exchange columns of CM Sepharose Fast Flow. The condition of the elution was optimized for pH and ionic strength, the absorbance at 280 nm of the elution samples were detected, and the targeted protein band in the collected samples was observed by SDS-PAGE electrophoresis. RESULTS: The Hitrap ion exchange column was eluted with buffer 1, which resulted in a baseline distribution of absorbance at 280 nm. Buffer 2 elution of the column resulted in the presence of peak absorbance with trails, which was identified to be constituted by some low molecular weight bands by subsequent SDS-PAGE. In serial column elution with buffer 3 with different ionic strength, a peak absorbance was observed with the ionic strength of 100 mmol/L NaCl, and SDS-PAGE confirmed that the peak was generated by the target protein. No obvious peaks or bands in SDS-PAGE occurred with the other ionic strengths. CONCLUSION: The pH of the buffer only affect the elution of the irrelevant proteins rather than the Hap(s) protein, and elution with the buffer containing 100 mmol/L NaCl can be optimal for eluting the Hap(s) protein. PMID- 18159011 TI - [Landmark-based automatic registration of serial cross-sectional images of Chinese digital human using Photoshop and Matlab software]. AB - OBJECTIVE: This paper describes automatic registration of the serial cross sectional images of Chinese digital human by projective registration method based on the landmarks using the commercially available software Photoshop and Matlab. During cadaver embedment for acquisition of the Chinese digital human images, 4 rods were placed parallel to the vertical axis of the frozen cadaver to allow orientation. Projective distortion of the rod positions on the cross-sectional images was inevitable due to even slight changes of the relative position of the camera. The original cross-sectional images were first processed using Photoshop software firstly to obtain the images of the orientation rods, and the centroid coordinate of every rod image was acquired with Matlab software. With the average coordinate value of the rods as the fiducial point, two-dimensional projective transformation coefficient of each image was determined. Projective transformation was then carried out and projective distortion from each original serial image was eliminated. The rectified cross-sectional images were again processed using Photoshop to obtain the image of the first orientation rod, the coordinate value of first rod image was calculated using Matlab software, and the cross-sectional images were cut into images of the same size according to the first rod spatial coordinate, to achieve automatic registration of the serial cross-sectional images. sing Photoshop and Matlab softwares, projective transformation can accurately accomplish the image registration for the serial images with simpler calculation processes and easier computer processing. PMID- 18159012 TI - [Adenovirus-mediated antisense HSP70 cDNA transfection inhibits the growth of laryngeal carcinoma Hep-2 cells]. AB - OBJECTIVE: To construct a recombinant adenovirus vector carrying antisense heat shock protein 70 (HSP70) cDNA and observe its effect on inhibiting the growth of laryngeal carcinoma Hep-2 cells. METHODS: The HSP70 gene fragment encoding the 5' region was cloned reversely into the shuttle plasmid PAdTrack-CMV, and the resultant plasmid was recombined with the backbone plasmid PadEasy-1 in the E.coli Bj5183 cells to generate the recombinant adenovirus vector. The adenovirus were then packaged and amplified in 293 cells, and the viral titer was determined using GFP. RESULTS: The recombinant adenovirus vector carrying antisense HSP70 cDNA was constructed successfully with a viral titer of 8 x 10(9). HSP70 expression of Hep-2 cells was obviously blocked by antisense HSP70 RNA, and Western blotting and immuohistochemistry demonstrated that cells transfected with antisense HSP70 did not express or express HSP70 at low levels. Flow cytometry presented apoptotic peak in the antisense HSP70-transfected cells, but not in the control cells. CONCLUSION: The recombinant adenovirus vector containing antisense HSP70 cDNA can effectively deliver antisense HSP70 gene into Hep-2 cells, suggesting the great potential of this gene therapy strategy in management of human laryngeal carcinoma. PMID- 18159013 TI - [Early goal-directed therapy lowers the incidence, severity and mortality of multiple organ dysfunction syndrome]. AB - OBJECTIVE: To investigate the effect of early goal-directed therapy (EGDT) on the incidence, severity and mortality of multiple organ dysfunction syndrome (MODS). METHODS: A prospective, randomized controlled trial was performed involving 273 patients in the early stage of shock at risk of potential MODS development. The patients were randomly divided into EGDT group (including 139 patients managed with EGDT) and control group (including 134 patients with conventional empirical therapy). The scores of APACHE II, blood lactate concentration (Lactate(0)) and SOFA scores (SOFA(0)) of the two groups were recorded on admission, and the lactate concentration on the second and fourth day of hospitalization (Lactate(2) and Lactate(4)), and the highest SOFA scores (SOFAT) after admission were also recorded. The discrepancy between the two SOFA scores (SOFA(S)), number of the dysfunctional organ, and the mortality in ICU of the two groups were calculated at the end of the study. RESULTS: The incidence of MODS in the EGDT group was significantly lower than that in control group (P=0.002). The Lactate(2), Lactate(4), SOFA(T), SOFA(S), and the number of dysfunctional organs in EGDT group were also significantly lower (P=0.045, 0.016, 0.009, 0.010, 0.002). EGDT was associated with a significantly lower total mortality rate of MODS than the conventional therapy (P=0.007), and also with a significantly lower mortality rate of MODS after controlling for severe sepsis (P=0.047 and 0.044). CONCLUSION: EGDT can decrease the incidence and severity of MODS, and can effectively decrease the mortality of MODS irrespective of the presence of severe sepsis. PMID- 18159014 TI - [Whole-range encirclement method versus conventional method for microvascular decompression for treatment of hemifacial spasm]. AB - OBJECTIVE: To compare the efficacy and micromanipulation techniques of microvascular decompression with whole-range encirclement method and the conventional method in 65 cases of hemifacial spasm. METHODS: The data of 65 surgically managed cases of hemifacial spasm were retrospectively analyzed for intraoperative findings, surgical techniques, and the patients' outcomes. Microvascular decompression with the conventional approach was performed in 30 patients (group A), and whole-range encircling method was used in the other 35 patients (group B). The patients were also divided into group C (below 40 years, n=27) and group D (above 40 years, n=38) according to the onset age of the symptom. RESULTS: In these patients, 4 patterns of facial nerve compression were identified: simple contact, contact and indentation, adhesion and encasement, and unidentified offending vessels. The offending vessels included the arteries, veins, and vascular loops. In most patients in group C, the arachnoid membrane around the facial nerve thickened and encircled the offending ve;ssel; in group D, the characteristic changes of the vasculature occurred in the offending artery, resulting in its displacement. In groups A and B, the overall efficacy rate was 80% and 97.1%, with recurrence rates of 13.3% and 2.9%, respectively. The major permanent complications in group A included hearing impairment (10.0%) and ataxia (6.7%), whose incidences in group B were 2.9% and 2.9%, respectively. CONCLUSION: Microvascular decompression with whole-range encircling method is a safe approach associated with high cure rate, in which careful avoidance of injuries to the cranial nerves and the penetrating vessels from the root entry/exit zone and the pons ensure good outcome and minimize the likeliness of recurrence and complications. PMID- 18159015 TI - [Rearrangement of immunoglobulin heavy chain variable region genes in human neonates]. AB - OBJECTIVE: To study the rearrangement of immunoglobulin (Ig) heavy chain variable region (V(H)) genes in human neonates with different gestational ages (GA). METHODS: Peripheral blood from the neonates with GA of 27 weeks (4 cases), 28-32 weeks (9 cases), 33-36 weeks (12 cases), and 37-42 weeks (13 cases) was collected. RT-PCR was used to amplify the Ig V(H) gene, and the PCR products were separated by electrophoresis and analyzed using 6% denaturing PAGE gel. RESULTS: All Ig V(H) family genes had several rearranged genes in each GA group, and the neonates with different GA showed no significant difference in the median molecular weight for each rearranged Ig V(H) family gene. CONCLUSION: The neonates with GA of 27-42 weeks exhibit diversity in Ig V(H) gene rearrangement, and for the same Ig V(H) family, the median length of the arranged Ig V(H) genes is independent of the gestational age. PMID- 18159016 TI - [Preliminary study of diffusion tensor imaging in treatment response assessment of major depression]. AB - OBJECTIVE: To assess the value of diffusion tensor imaging (DTI) in therapeutic effect evaluation of major depression. METHODS: Eighteen patients who met the CCMD-3-R criteria for major depression or bipolar disorder (with depressed episode and total score no less than 18 for 17 items of Hamilton Depression Rating Scale) and 13 aged-matched controls were examined by routine magnetic resonance imaging (MRI) and DTI. DTI were used to determine fractional anisotropy (FA) in the preselected white matter regions. All the patients with major depression received treatment with selective serotonine reuptake inhibitor (SSRI) for 6-8 weeks, and the efficacy were assessed by Hamilton Depression Scale, Hamilton Anxiety Scale (HAMA), and Clinical Global Impression (CGI) scale. RESULTS: The total response rate to fluoxetine was 67%, and significant improvement was observed in 56% of the patients while 33% failed to respond after 8 weeks of treatment. The depressed subjects failing to respond to the treatment had a significant lower FA of the frontal white matter than those responding favorably to the treatment and the healthy control subjects. CONCLUSION: DTI may identify the microstructural abnormality in the white matter, which is associated with a low remission rate of major depression. PMID- 18159017 TI - [Biological significance of expression of calcyclin in human pancreatic carcinoma: a tissue microarray-based study]. AB - OBJECTIVE: To investigate the expression of calcyclin in pancreatic carcinoma and its relation to the patients' prognosis. METHODS: Human pancreatic carcinoma tissue microarray was constructed, which contained 63 cores of 3 normal adult pancreas tissues, 6 chronic pancreatitis tissues, 51 pancreatic carcinoma tissues and 3 islet cell carcinoma tissues. Immunohistochemistry was performed to detect the expression of calcyclin in these tissues, and the relationship between calcyclin and the clinicopatholoical features of pancreatic carcinoma was analyzed. RESULTS: The positivity rate of calcyclin in the pancreatic carcinoma tissue was 76.5% (39/51), and calcyclin staining was more intense in the malignant cells than in the benign cells (P=0.007), suggesting a correlation between calcyclin expression and pancreatic carcinoma. No evidence was found for an association of calcyclin expression with the variables including the patients' gender, age at surgery, and tumor grade. Weak staining for calcyclin was noted in chronic pancreatitis tissues. CONCLUSION: Calcyclin expression is related to the pancreatic carcinomas and up-regulation of calcyclin expression is possibly an early event in pancreatic and pragression of development cancer. PMID- 18159018 TI - [Construction and screening of cDNA library of osteosarcoma cells]. AB - OBJECTIVE: To construct a cDNA library of osteosarcoma SAOS-2 cells and screen the proteins interacting with the bone morphogenetic protein-2 (BMP-2) to explore the role of BMP-2 in the development of prostate cancer. METHODS: The total RNA was extracted from SAOS-2 cells, from which poly(A)(+)RNA was purified to generate the cDNA using RT-PCR with primer SMARTIII and primer CDSIIIoligo(dT). The library was constructed and screened by cotransformation of the double stranded cDNA (dscDNA) and linearized pGADT7-Rec with the bait plasmid to the yeast AH109. The proteins interacting with BMP-2 were screened by AH109 mating with the bait strain Y187, and the interaction was confirmed using far-Western blot analysis. RESULTS: The cDNA library of human osteosarcoma SAOS-2 cells was constructed, which was characterized by high diversity and sufficient capacity. The dscDNA size range was between 250-5000 bp, with a cotransformation efficiency of 4.3 x 10(5) and recombination efficiency of 1.9 x 10(6), and 4.3 x 10(5) clones were screened. The mating efficiency was 32% and 1.0 x 10(6) clones were screened by the bait of BMP-2, and 4 positive clones was obtained and sequenced. CONCLUSION: The diversity and capacity of the cDNA library meet the needs for screening genes related to prostate cancer. PMID- 18159019 TI - [Immunodetection of ginsenoside Rb1 in rat serum]. AB - OBJECTIVE: To establish a novel immunoassay for qualitative detection of ginsenoside Rb1 in rat serum. METHODS: Anti-G-Rb1 monoclonal antibody (mAb) was through a hybridoma approach. Rat serum containing G-Rb1 was deproteinized with methanol to prepare the sample for testing, which was loaded onto polyethersulfone (PES) membrane and developed in the mixture of acetonitrile, water and acetic acid (25:75:1). After treatment with NaIO(4), the membrane was transferred to 1% BSA solution for immobilization of G-Rb1. The membrane was subsequently treated with anti-G-Rb1 mAb solution, followed by addition of peroxidase-labeled goat anti-mouse IgG and color development using 4-chloro-1 naphthol-0.03% H(2)O(2). RESULTS: On the PES membrane, a clear blue spot representing G-Rb1 occurred where the rat serum for testing and the standard G Rb1 samples were blotted. The limit of this immunodetection was 0.25 microg. CONCLUSION: This immunoassay has greater specificity and reliability than thin layer chromatography with a sensitivity similar to that of high-performance liquid chromatography, and does not require sophisticated equipment for convenient G-Rb1 detection in rat serum. PMID- 18159020 TI - [Effect of aldosterone on aquaporin and ionophorous protein expressions in guinea pig cochlea]. AB - OBJECTIVE: To study the effect of aldosterone on aquaporin and ionophorous protein expressions in the cochlea of guinea pigs. METHODS: RT-PCR was used to detect the expression of Na-K ATPase and epithelial sodium channel (ENaC) mRNA in the cochlea of guinea pigs 6 h after intraperitoneal aldosterone injection. Immunohistochemistry was used to detect the expression of aquaporin 1(AQP1) in the cochlea 1 month after the injection. RESULTS: Early after aldosterone injection, the expression level of Na-K ATPase beta(1) and beta(3) subunit mRNAs remained unchanged and level of ENaC alpha subunit mRNA was up-regulated in the cochlear lateral wall (P<0.05), and 1 month after the injection, the expression of AQP1 protein was down-regulated (P<0.05). CONCLUSION: The effect of aldosterone on the cochlea is mediated possibly through its action on the genome, and aldosterone may cause ion concentration alterations to induce endolymphatic hydrops. PMID- 18159021 TI - [X-ray irradiation enhances tumor necrosis factor receptor p55 expression in human colorectal cancer cells and inhibits the release of its soluble form in vitro]. AB - OBJECTIVE: To investigate the effects of X ray on human colorectal cancer cells for their tumor necrosis factor receptor-p55 (TNFR-p55) expression and release of soluble soluble TNFR-p55 (sTNFR-p55) in vitro. METHODS: The protein expression of TNFR-p55 in Lovo cells exposed to X-ray was detected using immunohistochemistry, and enzyme-linked immunosorbent assay was used to examine the levels of sTNFR-p55 in the supernatants of the cell culture. The cell apoptosis of the exposed cells was analyzed with flow cytometry, and the changes in cell morphology were observed microscopically. RESULTS: X-ray exposure of cells resulted in a strong increase in TNFR-p55 expression of (P<0.01) and LoVo cell apoptosis (P<0.05). The levels of sTNFR-p55 in the supernatant of cells with X-ray exposure was significantly lowered in comparison with the levels before exposure (P<0.01). Optical microscopy showed that the exposed LoVo cells shrank and became spherical with cytoplasmic condensation and nuclear pyknosis. CONCLUSION: X-ray exposure can induce LoVo cell apoptosis by increasing TNFR-p55 expression on the cell membrane and inhibiting the release of sTNFR-p55 in the supernatants. PMID- 18159022 TI - [Therapeutic effect of sirolimus against chronic allograft nephropathy in kidney transplant recipients]. AB - OBJECTIVE: To investigate the efficacy and safety of sirolimus in management of chronic allograft nephropathy (CAN). METHODS: A retrospective study was conducted involving 31 CAN patients followed up since March 2002, who experienced a change from a calcineurin inhibitor (CNI)-based regimen to a SRL-based regimen. Serum creatinine (Cr) in these patients was compared before and after the regimen change, and the adverse events associated with SRL were analyzed. RESULTS: Till March 2007 when the study closed, 15 patients reached the primary endpoint for resuming dialysis, 8 had improved and 8 had stable renal function. In patients with high Cr(0)(> or =3 mg/L, n=12), 9 resumed dialysis and 2 had improved renal function, but one of the patients with renal improvement eventually died due to infection; in the patients with low Cr(0)(<3 mg/L, n=19), 5 resumed dialysis, 8 had stable renal function and 6 had improved renal function, showing significant difference between the 2 groups (P=0.003). Altogether 14 patients reached the secondary endpoint for ceasing SRL for severe infection (5 patients, of whom 4 resumed dialysis and 1 died of infection) or adverse events associated with SRL (9 patients, of whom 4 resumed dialysis, 2 had stable and 3 had improved renal function). Hyperlipidemia (51.6%), leukocytopenia (41.9%), mouth ulcer (29.0%) and liver function lesion (16.1%) were the commonest adverse events in these patients, and totalling 13 severe adverse events were recorded, including 2 fatal cerebral hemorrhage, 3 fatal infection episodes, and 8 pulmonary and urinary infections that require hospitalization. CONCLUSION: Conversion from a CNI-based to SRL-based regimen can be effective for some CAN cases, especially for those with Cr(0) below 3 mg/L. Attention must be given to adverse events like hyperlipidemia and leukocytopenia, as well as the related cerebral vascular accidents and infections. PMID- 18159023 TI - [Therapeutical efficacy of routine intra-aortic balloon pump support in patients with high-risk acute myocardial infarction undergoing percutaneous coronary intervention]. AB - OBJECTIVE: To assess the value of routine intra-aortic balloon pump (IABP) support in patients with high-risk acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI). METHODS: The clinical data of 41 patients with high-risk AMI undergoing emergency PCI with routine IABP support were retrospectively reviewed, and 38 patients paired with the former group receiving emergency PCI for high-risk AMI without IABP support at the same time were included as the control group. Thirty days after the operation, the two groups were compared for myocardial ischemic events, left ventricular function and major adverse cardiac events (MACE). RESULTS: Patients receiving IABP support had a significantly lower incidence of myocardial ischemic events than those without IABP (4.9% vs 15.8%, P<0.05), and showed greater improvement in the left ventricular function. Significant differences were also observed in the mortality rate, incidence of reinfarction and revascularization rate between the two groups, but not in the rate of MACE. CONCLUSION: Patients undergoing PCI for high risk acute AMI can benefit from routine IABP support in terms of improvement of left ventricular function and reduce myocardial ischemic events and the rate of MACE. These results, however, still await further confirmation by large-scale clinical trials. PMID- 18159024 TI - [Third ventriculostomy for obstructive hydrocephalus: findings in radioisotope cisternography]. AB - OBJECTIVE: To study the findings in radionuclide cisternography in patients with obstructive hydrocephalus following third ventriculostomy. METHODS: Lumbar radionuclide cisternography was performed in 15 patients with obstructive hydro- cephalus before and after third ventriculostomy. RESULTS: The postoperative cisternography identified radiopharmaceutical reflux into the lateral cerebral ventricles with delayed clearance for 24 h, similar to the findings by cisternography of communicating hydrocephalus. CONCLUSION: The cause of ventricular radiopharmaceutical reflux is not yet understood, possibly in association with the reversal of normal cerebrospinal fluid flow though the fenestration on the third ventrical floor. PMID- 18159025 TI - [Diagnosis of aortic intramural hematoma by electron beam CT]. AB - OBJECTIVE: To assess the value of electron beam CT (EBCT) in the diagnosis of aortic intramural hematoma (AIH). METHODS: Twenty-five patients who complained of acute chest and back pain were scanned with an EBCT scanner (Imatron C-150) using contrast-enhanced continuous volume scanning (CVS) for establish the diagnosis of AIH. RESULTS: Seven patients were diagnosed as having Stanford type A, and the others as type B AIH. The direct features of AIH in EBCT included crescent or circular thickening (>5 mm) of the aortic wall without signs of lumen formation resulting from intimal rupture. The indirect features included calcification ingression (7 cases), penetrating ulcer (12 cases), atherosclerosis (18 cases) and leakages (5 cases). The complicating features included pericardial effusion (5 cases), pleural effusion (14 cases), involvement of the large branches (5 cases), aortic dissection (3 cases) and aneurysms (4 cases). CONCLUSION: EBCT can provide important information for the diagnosis and treatment of AIH, and can be useful for follow-up observation of the patients. PMID- 18159026 TI - [Peripheral blood p53 single nucleotide polymorphism analysis for early diagnosis of colorectal carcinoma]. AB - OBJECTIVE: To evaluate the role of p53 gene mutation in colorectal carcinoma and assess the value of peripheral blood p53 single nucleotide polymorphism (SNP) in early diagnosis of colorectal carcinoma. METHODS: NSP in axons 5-8 of p53 gene was detected using ligase detection reaction-polymerase chain reaction (LDR-PCR) in the peripheral blood of 100 patients with colorectal cancer and 100 healthy subjects. RESULTS: The mutation rate of p53 gene was 24% (24/120) in colorectal carcinoma patients and 0% (0/120) in the healthy subjects (P<0.05). CONCLUSION: p53 plays an important role in the carcinogenesis of colorectal carcinoma, and SNP analysis for p53 gene can be helpful in early diagnosis of colorectal carcinoma. PMID- 18159027 TI - [Effect of extracts from rabbit skin inflamed by vaccinia virus in the management of postherpetic neuralgia and on serum interleukin-6 level in aged patients]. AB - OBJECTIVE: To observe the effect of the extracts from rabbit skin inflamed by vaccinia virus for injection, a commercially available agent (Analgecine), in the treatment of postherpetic neuralgia (PHN) in aged patients and its influence on serum interleukin-6 (IL-6) level. METHODS: A total of 44 elderly patients with PHN were randomly divided into two groups. In addition to the conventional antiviral therapy, patients in group A were given analgecine, and those in group B given indomethacin tablet. After the completion of the 14-day treatment course, the therapeutic effects were evaluated using visual analogue scale (VAS), and the changes in serum IL-6 levels after treatment were measured. RESULTS: The score of VAS decreased significantly after treatment in group A (P<0.01), but not in group B (P>0.05). In both groups, the serum IL-6 level before treatment was significantly higher than that in normal control subjects (P<0.001), and lowered significantly after treatment (P<0.01) in group A, but not in group B (P>0.05). CONCLUSION: The extracts from rabbit skin inflamed by vaccinia virus have definite effects for rapid pain relief, and can effectively decrease the serum IL 6 level in elderly patients with PHN. PMID- 18159028 TI - [Endometrial carcinoma in patients below 45 years of age: clinical analysis of 53 cases]. AB - OBJECTIVE: To investigate the clinical characteristics of endometrial carcinoma in patients below 45 years of age. METHODS: The clinical data were collected from 53 patients with endometrial carcinoma below 45 years of age, who were divided into less than 40 year group (group A, 28 cases) and 40 to 45 year group (group B, 25 cases) and their clinical data were compared. RESULTS: A rate of 57.1% (16/28) of the patients in group A were infertile, and 78.5% (22/28) reported irregular menstruation. In group B, the infertility rate was 28.0% (7/25), and 48% (12/25) of the patients were obese, 56% (14/25) had abnormal vaginal bleeding, and 32% (8/25) had diabetes and hypertension. The two groups differed significantly in the infertility rate, number of pregnancies and deliveries and the incidences of obesity, hypertension and diabetes. The pathological classification, grade of differentiation, pathological staging, depth of myometrial invasion, and cervical infiltration status were comparable between the two groups (P>0.05). All the 53 patients received surgical interventions and follow. up for two years, during which 1 patients in group B had relapse. CONCLUSION: Patients with endometrial carcinoma at 40 years of age or below often have infertility, and those over 40 years are exposed to such high-risk factors as obesity, diabetes, and hypertension. Its major clinical symptom of endometrial carcinoma is menstrual disorders. The differentiation, pathological staging, pathological classification of the malignancy are not associated with age in patients below 45 years of age. Early diagnosis often warrants more favorable prognosis. PMID- 18159029 TI - Pharmacists want FDA to create new intermediate drug category. PMID- 18159030 TI - FDA approves new epoetin product: Roche unsure when it can market in United States. PMID- 18159031 TI - Robotic workbench to prepare hazardous drugs. PMID- 18159033 TI - Washout period for immune-modifying drugs before natalizumab therapy. PMID- 18159034 TI - Development of a work-force retention program. PMID- 18159035 TI - Gene-expression assays: new tools to individualize treatment of early-stage breast cancer. AB - PURPOSE: The clinical and economic data for the two currently available gene expression assays are reviewed. SUMMARY: Two gene-expression assays, used to determine the risk of breast cancer recurrence in patients with stage I or II node-negative breast cancer, are currently available. Oncotype DX is an assay performed on RNA extracted from paraffin-embedded tumor tissue. It analyzes the expression of 21 genes: 16 cancer-related genes and 5 reference genes. The results are used to calculate a recurrence score to identify the likelihood of cancer recurrence in patients treated with tamoxifen. The results of two studies evaluating the ability of Oncotype DX to predict the risk of breast cancer recurrence suggest that patients with ER-positive, node-negative breast cancer and a low recurrence score may need only adjuvant treatment with tamoxifen, while intermediate- and high-risk patients may require additional treatment with adjuvant chemotherapy. MammaPrint, an oligonucleotide microassay performed on fresh-frozen tumor samples, analyzes the expression of 70 genes. Studies have found that MammaPrint allows young patients (<61 years) with early-stage breast cancer to be categorized as having a high or low risk of distant metastasis. High risk patients may then be managed with more aggressive therapy. CONCLUSION: Two gene-expression assays, Oncotype DX and MammaPrint, have been developed to determine the risk of breast cancer recurrence in patients with stage I or II node-negative breast cancer. In the future, these tests may be useful in determining the need for systemic adjuvant therapy in such patients. PMID- 18159036 TI - Recent developments in the management of acute respiratory distress syndrome in adults. AB - PURPOSE: Recent developments in the management of acute respiratory distress syndrome (ARDS) in adults are reviewed. SUMMARY: Corticosteroids have been extensively studied in ARDS; however, they have not demonstrated clear benefit in patients with ARDS. Some trials have found increased complications and mortality related to corticosteroid use. The use of conservative fluid management has been associated with significant reductions in morbidity, highlighting the need to avoid fluid over-administration in patients with ARDS. A number of ventilatory strategies have also been studied. Studies have found that higher positive end expiratory pressure settings do not appear to be harmful in patients with ARDS. In an effort to prevent alveolar overdistention, low tidal volume and plateau pressure ventilation is increasingly being used in patients with acute lung injury (ALI). Given the increasing evidence supporting the use of lower tidal volume ventilation, this strategy has become the new standard of care in patients with suspected ALI and ARDS. No clear benefit has been shown in the treatment of ARDS with nitric oxide and surfactant. Prostaglandins and acetylcysteine are not considered useful in the treatment of ARDS, while no conclusions can be drawn regarding the benefits of albuterol on mortality in patients with ARDS. The use of prone positioning should be discouraged in the treatment of ARDS based on its associated risks. CONCLUSION: Early administration of moderate-dosage corticosteroids likely helps decrease the time of ventilator dependence and duration of intensive care unit stay. Conservative fluid management and low tidal volume ventilation are becoming increasingly widespread in the management of patients with ARDS. Nitric oxide, surfactant, prostaglandins, albuterol, acetylcysteine, and prone positioning have not been shown to be beneficial in the treatment of ARDS. PMID- 18159037 TI - Elevated hepatic transaminases associated with telithromycin therapy: a case report and literature review. AB - PURPOSE: A case of mild hepatocellular injury associated with the administration of telithromycin in a patient with no risk factors for hepatotoxicity is presented. SUMMARY: A 44-year-old man with no significant past medical history arrived at the emergency room after six days of high fever, chills, headache, neck stiffness, and back pain. Five days earlier, he visited a family medicine clinic for his symptoms and oral telithromycin 800 mg daily was prescribed for a suspected upper-respiratory-tract infection. The patient stopped taking the drug after three days due to persistent symptoms. On admission, the patient's laboratory tests revealed an aspartate transaminase (AST) concentration of 68 units/L, an alanine transaminase (ALT) value of 155 units/L, and an erythrocyte sedimentation rate of 40 mm/hr. The patient was not taking any long-term medications, had taken only aspirin for his fever, and denied the use of alcohol and illegal drugs. The patient was admitted to the general medical unit with a diagnosis of possible viral hepatitis. His urine culture was negative, and serology tests later revealed no evidence of hepatitis A, B, or C. Ibuprofen, pantoprazole, and enoxaparin were prescribed. On hospital day 2, the patient's AST and ALT concentrations had decreased to 50 and 110 units/L, respectively. By day 3, the patient's symptoms had improved and he remained afebrile. His AST and ALT values had further decreased to 41 and 105 units/L, respectively. He was then diagnosed with acute viral upper-respiratory-tract infection with mild hepatotoxicity associated with telithromycin and was discharged home with orders for follow-up at the family medicine clinic. CONCLUSION: A patient without risk factors for hepatotoxicity developed mild elevations in hepatic transaminases after receiving telithromycin for the treatment of a suspected upper-respiratory tract infection. PMID- 18159038 TI - Influence of medication treatment patterns on resource use for patients hospitalized for schizophrenia. AB - PURPOSE: The influence of medication treatment patterns on resource use for patients hospitalized for schizophrenia was studied. METHODS: Patients hospitalized with a primary diagnosis of schizophrenia or schizo-affective disorder discharged between October 1, 2003, and September 30, 2004, were identified from a large, multihospital database. The duration of dosage adjustment was defined as the period between first and last days of dosage change. Ordinary least-squares regression was used to examine the relationships between labeled dosage-escalation durations, actual durations of dosage adjustment, and length of stay. RESULTS: The study sample consisted of 21,950 hospitalized schizophrenia patients, representing 30,873 hospitalizations from 157 hospitals. The majority of patients were 18-44 years old (56.3%), covered by Medicare (46.6%), and admitted through the emergency room (55.5%). The average length of hospital stay was 10 days, and 30% of patients were readmitted for a related reason within 90 days. The labeled dosage-escalation duration was not correlated with the actual duration of dosage adjustment (r = 0.05, p < 0.0001) or with longer hospital stays (r = -0.03, p < 0.0001). Drug-related factors affecting length of stay included medication changes (p < 0.0001), dose changes (p < 0.0001), use of short-acting intramuscular antipsychotics ( p < 0.0001), and medications to manage extrapyramidal symptoms (p < 0.0001). CONCLUSION: Analysis of information about patients hospitalized for schizophrenia revealed that antipsychotic medication changes, prolonged dosage escalation, and initial use of short-acting intramuscular antipsychotics were significantly associated with a greater than average length of stay. Longer labeled dosage-escalation durations were not associated with longer hospital stays. PMID- 18159039 TI - Enhanced photoemission spectroscopy for verification of high-risk i.v. medications. AB - PURPOSE: The sensitivity and specificity of enhanced photoemission spectroscopy (EPS) for performing an automated final check of compounded i.v. admixtures at a pediatric hospital pharmacy were studied. METHODS: A tabletop EPS device was used to test samples of seven high-risk drug-diluent combinations compounded in the pharmacy; the drugs were vancomycin, lorazepam, morphine, insulin, hydromorphone, gentamicin, and epinephrine. Ten sets of samples were prepared for each drug. Typically, a sample set consisted of dilutions ranging from 10-fold above to 10 fold below the targeted concentration. Testing was performed twice weekly between November 2005 and March 2006. RESULTS: The EPS device detected errors departing from the targeted concentration by 20% or more with a sensitivity of at least 95%. Specificity in distinguishing among test medications at targeted concentrations was 100%. The percentage of passing samples with intermediate concentrations varied among the drugs. CONCLUSION: A tabletop EPS device demonstrated acceptable sensitivity and specificity for validating the identity and concentrations of selected high-risk i.v. medications compounded for pediatric patients. The device may help prevent clinically important medication errors caused by inaccurate compounding. PMID- 18159040 TI - Drug-related problems in hospitalized patients with HIV infection. AB - PURPOSE: The type and number of drug-related problems that commonly occur in hospitalized patients with HIV were studied. METHODS: The medical records of HIV infected patients who were receiving antiretroviral therapy at the time of hospital admission between January 1, 2005, and August 31, 2006, were reviewed. Patients age 18 years or older who had received at least one dose of an antiretroviral for an HIV-related indication during their hospitalization were included in the study. Patients' medical records were evaluated to identify drug related problems and adverse drug events secondary to antiretroviral therapy. RESULTS: Eighty-three patients were eligible for study inclusion. A total of 176 drug-related problems were identified. The most common drug-related problem identified among medication orders reviewed was inappropriate dosing. Of the 251 orders for antiretroviral agents, 57 drugs were inappropriately dosed. The most common drug-related problems among patients were drug-drug interactions and incomplete antiretroviral regimens. There was no significant difference in the mean length of stay between patients with or without drug-related problems. Admission by physicians who were not infectious diseases specialists was an independent risk factor for having at least one drug-related problem during hospitalization (odds ratio, 3.83; 95% confidence interval, 1.08-13.54). CONCLUSION: A majority of HIV-infected patients at one institution had at least one drug-related problem at hospital admission. The most common problem observed among the medication orders reviewed was inappropriate dosing. The most common drug-related problems observed among patients were drug-drug interactions and incomplete antiretroviral regimens. PMID- 18159041 TI - Variation in medication information for elderly patients during initial interventions by emergency department physicians. AB - PURPOSE: Discrepancies between medication information received from elderly patients in the emergency department (ED) at the time of initial diagnosis or therapeutic intervention and information collected by pharmacists from outside caregivers (pharmacies and physicians) were studied. The time from patient arrival at the ED to initial intervention was also studied. METHODS: Written and oral medication histories of ED patients 65 years and older who were seen at an urban academic hospital between June 2003 and May 2004 were collected and represented information available at the time of initial diagnosis and treatment. A clinical pharmacist contacted all relevant physicians and pharmacies identified in the ED and compiled for each patient a medication profile. The times of ED arrival and initial intervention were recorded. RESULTS: For the 98 patients included, the mean discordance in drug names between the two resultant types of drug list exceeded 30%. The classes of medications that varied most frequently included antihypertensives, supplements, analgesics, nitrates, and antiplatelets and anticoagulants. The median time from patient entry to initial diagnostic intervention was 37 minutes, with a range from 1 to 599 minutes. The median time from patient entry to initial therapeutic intervention was 81.5 minutes, with a range from 8 to 598 minutes. CONCLUSION: A pilot study demonstrated that approximately one third of elderly patients' medication information available to the ED physician at the time of the initial diagnostic and therapeutic intervention differed from that obtainable from outside caregivers. PMID- 18159042 TI - Readability of consumer medication information for intranasal corticosteroid inhalers. AB - PURPOSE: The readability of consumer medication information (CMI) inserts accompanying intranasal corticosteroid (INCS) inhalers currently prescribed in the United States was studied. METHODS: INCS inhalers were identified (n = 7) using Epocrates R(x) Pro and English- language CMI was obtained from each inhaler's manufacturer. The CMI was evaluated for reading grade level (using Fry's readability formula) and font size, dimensions (length and width), illustrations (diagrams and figures), and directions for use. RESULTS: The mean +/- S.D. reading grade level of the CMI was 6.9 +/- 0.7 (range, 6-8). The mean +/ S.D. font size was 9.0 +/- 2.2 (range, 6-12). The mean +/- S.D. CMI page length and width were 31.3 +/- 22.5 cm and 14.0 +/- 12.9 cm, respectively. A device overview figure was included in three of the seven educational samples. The mean size of illustrations was 7.9 cm(2) (range, 2.9-25.9 cm(2)), with a mean +/- S.D. of 7.6 +/- 3.2 (range, 3-11) figures per CMI insert. Directions followed a logical, step-by-step chronological sequence in every CMI insert. CONCLUSION: Most CMI for INCS inhalers is written at a reading level slightly higher than recommended, printed in a font size smaller than recommended, and illustrated inadequately for successful patient education. PMID- 18159043 TI - Job-sharing. Drs. Finks and Rogers interviewed by Kate Traynor, M.S., AJHP News writer. PMID- 18159044 TI - Calcium and phosphate compatibility: revisited again. PMID- 18159046 TI - The cover. Madonna II. PMID- 18159047 TI - A piece of my mind. In search of redemption. PMID- 18159048 TI - Studies probe timely cardiac reperfusion: swift treatment needed to boost MI survival. PMID- 18159049 TI - Vaccine takes aim at hypertension. PMID- 18159050 TI - New tool helps primary care clinicians with diagnosis and treatment of COPD. PMID- 18159051 TI - Investigational drug slows bone erosion. PMID- 18159052 TI - Resident duty hour reform and mortality in hospitalized patients. PMID- 18159053 TI - Resident duty hour reform and mortality in hospitalized patients. PMID- 18159054 TI - Resident duty hour reform and mortality in hospitalized patients. PMID- 18159055 TI - Postrecall surveillance following a multistate fusarium keratitis outbreak, 2004 through 2006. PMID- 18159056 TI - Prevalence of pathogenic BRCA1 mutation carriers in 5 US racial/ethnic groups. AB - CONTEXT: Information on the prevalence of pathogenic BRCA1 mutation carriers in racial/ethnic minority populations is limited. OBJECTIVE: To estimate BRCA1 carrier prevalence in Hispanic, African American, and Asian American female breast cancer patients compared with non-Hispanic white patients with and without Ashkenazi Jewish ancestry. DESIGN, SETTING, AND PARTICIPANTS: We estimated race/ethnicity-specific prevalence of BRCA1 in a population-based, multiethnic series of female breast cancer patients younger than 65 years at diagnosis who were enrolled at the Northern California site of the Breast Cancer Family Registry during the period 1996-2005. Race/ethnicity and religious ancestry were based on self-report. Weighted estimates of prevalence and 95% confidence intervals (CIs) were based on Horvitz-Thompson estimating equations. MAIN OUTCOME MEASURE: Estimates of BRCA1 prevalence. RESULTS: Estimates of BRCA1 prevalence were 3.5% (95% CI, 2.1%-5.8%) in Hispanic patients (n = 393), 1.3% (95% CI, 0.6% 2.6%) in African American patients (n = 341), and 0.5% (95% CI, 0.1%-2.0%) in Asian American patients (n = 444), compared with 8.3% (95% CI, 3.1%-20.1%) in Ashkenazi Jewish patients (n = 41) and 2.2% (95% CI, 0.7%-6.9%) in other non Hispanic white patients (n = 508). Prevalence was particularly high in young (<35 years) African American patients (5/30 patients [16.7%]; 95% CI, 7.1%-34.3%). 185delAG was the most common mutation in Hispanics, found in 5 of 21 carriers (24%). CONCLUSIONS: Among African American, Asian American, and Hispanic patients in the Northern California Breast Cancer Family Registry, the prevalence of BRCA1 mutation carriers was highest in Hispanics and lowest in Asian Americans. The higher carrier prevalence in Hispanics may reflect the presence of unrecognized Jewish ancestry in this population. PMID- 18159057 TI - Incidence and prognosis of transient neurological attacks. AB - CONTEXT: Transient neurological attacks (TNAs) are attacks with temporary (<24 hours) neurological symptoms. These symptoms can be focal, nonfocal, or a mixture of both. The prognostic significance of TNAs with focal symptoms (better known as transient ischemic attacks [TIAs]) is well understood. Conversely, hardly anything is known about the prognostic significance of TNAs with nonfocal or mixed symptoms. OBJECTIVE: To study the incidence and prognosis of focal TNAs (or TIAs), nonfocal TNAs, and mixed TNAs. DESIGN, SETTING, AND PARTICIPANTS: The study population comprised 6062 community-dwelling Rotterdam Study participants who were aged 55 years or older and free from stroke, myocardial infarction, and dementia at baseline (1990-1993). They were followed up for events until January 1, 2005. We analyzed the associations between incident TNAs and subsequent adverse events with age- and sex-adjusted Cox regression models. MAIN OUTCOME MEASURES: Stroke, ischemic heart disease, or dementia. RESULTS: During 60 535 person-years, 548 participants developed TNA (282 focal, 228 nonfocal, and 38 mixed). The incidence rate per 1000 person-years was 4.7 (95% confidence interval [CI], 4.1-5.2) for focal TNA, 3.8 (95% CI, 3.3-4.3) for nonfocal TNA, and 0.6 (95% CI, 0.4-0.9) for mixed TNA. Participants with focal TNA were at higher risk of subsequent stroke than participants without TNA (n = 46 vs 540; hazard ratio [HR], 2.14; 95% confidence interval [CI]; 1.57-2.91) but had an equal risk of ischemic heart disease and dementia. Nonfocal TNA patients were at higher risk of stroke (27 vs 540; HR, 1.56; 95% CI, 1.08-2.28) and dementia (30 vs 552; HR, 1.59; 95% CI, 1.11-2.26) than participants without TNA. Mixed TNA patients were at higher risk of stroke (6 vs 540; HR, 2.48; 95% CI, 1.11-5.56), ischemic heart disease (8 vs 779; HR, 2.26; 95% CI, 1.07-4.78), vascular death (8 vs 594; HR, 2.54; 95% CI, 1.31-4.91), and dementia (7 vs 552; HR, 3.46; 95% CI, 1.72-6.98) than participants without TNA. CONCLUSION: Patients who experience nonfocal TNAs, and especially those with mixed TNAs, have a higher risk of major vascular diseases and dementia than persons without TNA. PMID- 18159058 TI - Health of previously uninsured adults after acquiring Medicare coverage. AB - CONTEXT: Uninsured near-elderly adults, particularly those with cardiovascular disease or diabetes, experience worse health outcomes than insured adults. However, the health benefits of providing insurance coverage for uninsured adults have not been clearly demonstrated. OBJECTIVE: To assess the effect of acquiring Medicare coverage on the health of previously uninsured adults. DESIGN AND SETTING: We conducted quasi-experimental analyses of longitudinal survey data from 1992 through 2004 from the nationally representative Health and Retirement Study. We compared changes in health trends reported by previously uninsured and insured adults after they acquired Medicare coverage at age 65 years. PARTICIPANTS: Five thousand six adults who were continuously insured and 2227 adults who were persistently or intermittently uninsured from ages 55 to 64 years. MAIN OUTCOME MEASURES: Differential changes in self-reported trends after age 65 years in general health, change in general health, mobility, agility, pain, depressive symptoms, and a summary measure of these 6 domains; and adverse cardiovascular outcomes (all trend changes reported in health scores per year). RESULTS: Compared with previously insured adults, previously uninsured adults reported significantly improved health trends after age 65 years for the summary measure (differential change in annual trend, +0.20; P = .002) and several component measures. Relative to previously insured adults with cardiovascular disease or diabetes, previously uninsured adults with these conditions reported significantly improved trends in summary health (differential change in annual trend, +0.26; P = .006), change in general health (+0.02; P = .03), mobility (+0.04; P = .05), agility (+0.08; P = .003), and adverse cardiovascular outcomes (-0.015; P = .02) but not in depressive symptoms (+0.04; P = .32). Previously uninsured adults without these conditions reported differential improvement in depressive symptoms (+0.08; P = .002) but not in summary health (+0.10; P = .17) or any other measure. By age 70 years, the expected difference in summary health between previously uninsured and insured adults with cardiovascular disease or diabetes was reduced by 50%. CONCLUSION: In this study, acquisition of Medicare coverage was associated with improved trends in self-reported health for previously uninsured adults, particularly those with cardiovascular disease or diabetes. PMID- 18159059 TI - Does this child have a urinary tract infection? AB - CONTEXT: Urinary tract infection (UTI) is a frequently occurring pediatric illness that, if left untreated, can lead to permanent renal injury. Accordingly, accurate diagnosis of UTI is important. OBJECTIVE: To review the diagnostic accuracy of symptoms and signs for the diagnosis of UTI in infants and children. DATA SOURCES: A search of MEDLINE and EMBASE databases was conducted for articles published between 1966 and October 2007, as well as a manual review of bibliographies of all articles meeting inclusion criteria, 1 previously published systematic review, 3 clinical skills textbooks, and 2 experts in the field, yielding 6988 potentially relevant articles. STUDY SELECTION: Studies were included if they contained data on signs or symptoms of UTI in children through age 18 years. Of 337 articles examined, 12 met all inclusion criteria. DATA EXTRACTION: Two evaluators independently reviewed, rated, and abstracted data from each article. DATA SYNTHESIS: In infants with fever, history of a previous UTI (likelihood ratio [LR] range, 2.3-2.9), temperature higher than 40 degrees C (LR range, 3.2-3.3), and suprapubic tenderness (LR, 4.4; 95% confidence interval [CI], 1.6-12.4) were the findings most useful for identifying those with a UTI. Among male infants, lack of circumcision increased the likelihood of a UTI (summary LR, 2.8; 95% CI, 1.9-4.3); and the presence of circumcision was the only finding with an LR of less than 0.5 (summary LR, 0.33; 95% CI, 0.18-0.63). Combinations of findings were more useful than individual findings in identifying infants with a UTI (for temperature >39 degrees C for >48 hours without another potential source for fever on examination, the LR for all findings present was 4.0; 95% CI, 1.2-13.0; and for temperature <39 degrees C with another source for fever, the LR was 0.37; 95% CI, 0.16-0.85). In verbal children, abdominal pain (LR, 6.3; 95% CI, 2.5-16.0), back pain (LR, 3.6; 95% CI, 2.1-6.1), dysuria, frequency, or both (LR range, 2.2-2.8), and new-onset urinary incontinence (LR, 4.6; 95% CI, 2.8-7.6) increased the likelihood of a UTI. CONCLUSIONS: Although individual signs and symptoms were helpful in the diagnosis of a UTI, they were not sufficiently accurate to definitively diagnose UTIs. Combination of findings can identify infants with a low likelihood of a UTI. PMID- 18159060 TI - Role of professional organizations in regulating physician expert witness testimony. PMID- 18159061 TI - Genetic testing in diverse populations: are researchers doing enough to get out the correct message? PMID- 18159062 TI - Transient neurological attack: a useful concept? PMID- 18159063 TI - JAMA patient page. Transient neurological attacks. PMID- 18159064 TI - Stat bite: Types of research misconduct. PMID- 18159065 TI - Older survivors and cancer care. PMID- 18159066 TI - Anthracyclines in the treatment of HER2-negative breast cancer. PMID- 18159067 TI - Testicular cancer risk in first- and second-generation immigrants to Denmark. AB - BACKGROUND: Immigrant studies offer insights into the relative importance of environment and genes in disease etiology. There is considerable variation in testicular cancer incidence worldwide. We investigated testicular cancer risk in first- and second-generation immigrants to Denmark, a high-incidence country, to evaluate the relative influence of genes and environment and the potential timing of action of environmental factor(s). METHODS: A cohort of 2.1 million men who were born since 1930 and lived in Denmark between 1968 and 2003 was established based on information in the Danish Civil Registration System, which included their immigration histories. Cancer histories were obtained from the Danish Cancer Registry. Testicular cancer risk was estimated as rate ratios (RRs) with 95% confidence intervals (CIs) based on log-linear Poisson regression. RESULTS: Overall, 4216 testicular cancer cases occurred during 43 million person-years of follow-up in 2.1 million men. These included 166 cases among 344,444 direct immigrants to Denmark and 13 cases among 56,189 men born in Denmark to immigrant parents. These first- and second-generation immigrants had RRs of testicular cancer of 0.37 (95% CI = 0.31 to 0.43) and 0.88 (95% CI = 0.51 to 1.53), respectively, compared with men born in Denmark of parents born in Denmark. The rate in first-generation immigrants was not modified by age at immigration or duration of stay and reflected that in the country of origin. CONCLUSION: The testicular cancer risk in first-generation immigrants was lower than that in native-born Danes and reflected that in the countries of origin, whereas the risk in second-generation immigrants was similar to that in natives of Denmark. Together these findings argue for a substantial influence of environmental factors limited to the period early in life, most probably to the period in utero. PMID- 18159068 TI - Absence of cancer diagnosis and treatment in elderly Medicaid-insured nursing home residents. AB - BACKGROUND: Little is known about the effect cancer has on the lives of nursing home patients and the quality of care, including palliative care, delivered to them. METHODS: Using a statewide population-based dataset assembled from the Michigan Tumor Registry and Medicare records, we identified 1907 elderly Medicaid insured nursing home residents who were diagnosed with cancer between 1997 and 2000. Logistic regression models were used to estimate odds ratios (ORs) and relative risks (RRs) according to age, race, sex, income, comorbidity, and cancer site for late or unstaged cancer at diagnosis, death within 3 months of diagnosis, receipt of hospice care, and--for patients diagnosed with early-stage breast, colorectal, lung, or prostate cancer--the likelihood of cancer-directed surgery. All statistical tests were two-sided. RESULTS: Nursing home residents diagnosed with cancer had a preponderance of late or unstaged disease (62%), high mortality within 3 months of diagnosis (48%), and low hospice use if they had distant-stage cancer (28%). Only 22% received cancer-directed surgery, 61% of which was confined to breast cancer patients, and only 6% of patients received chemotherapy and/or radiation. Older age was positively associated with late or unstaged cancer and with death within 3 months of diagnosis. Patients aged 71-75 years were more likely to have cancer-directed surgery than patients aged 86 years and older (OR = 2.83, 95% confidence interval [CI] = 1.26 to 6.32; RR = 1.37, 95% CI = 1.08 to 1.75). African American patients were less likely to receive surgery (OR = 0.51, 95% CI = 0.26 to 0.99; RR = 0.80, 95% CI = 0.62 to 1.03) than white patients. Other demographic characteristics and comorbid conditions had little predictive value with regard to cancer treatment or hospice use in nursing home patients. CONCLUSIONS: Very few cancer services are provided to Medicaid-insured nursing home patients, despite the fact that many of these patients likely experienced cancer-related symptoms and marked physical decline before diagnosis and death. A middle ground between what would be considered guideline treatment practices and the apparent absence of diagnosis and treatment is needed. PMID- 18159069 TI - Inhibition of cancer cell invasion by cannabinoids via increased expression of tissue inhibitor of matrix metalloproteinases-1. AB - BACKGROUND: Cannabinoids, in addition to having palliative benefits in cancer therapy, have been associated with anticarcinogenic effects. Although the antiproliferative activities of cannabinoids have been intensively investigated, little is known about their effects on tumor invasion. METHODS: Matrigel-coated and uncoated Boyden chambers were used to quantify invasiveness and migration, respectively, of human cervical cancer (HeLa) cells that had been treated with cannabinoids (the stable anandamide analog R(+)-methanandamide [MA] and the phytocannabinoid delta9-tetrahydrocannabinol [THC]) in the presence or absence of antagonists of the CB1 or CB2 cannabinoid receptors or of transient receptor potential vanilloid 1 (TRPV1) or inhibitors of p38 or p42/44 mitogen-activated protein kinase (MAPK) pathways. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting were used to assess the influence of cannabinoids on the expression of matrix metalloproteinases (MMPs) and endogenous tissue inhibitors of MMPs (TIMPs). The role of TIMP-1 in the anti-invasive action of cannabinoids was analyzed by transfecting HeLa, human cervical carcinoma (C33A), or human lung carcinoma cells (A549) cells with siRNA targeting TIMP-1. All statistical tests were two-sided. RESULTS: Without modifying migration, MA and THC caused a time- and concentration-dependent suppression of HeLa cell invasion through Matrigel that was accompanied by increased expression of TIMP-1. At the lowest concentrations tested, MA (0.1 microM) and THC (0.01 microM) led to a decrease in invasion (normalized to that observed with vehicle-treated cells) of 61.5% (95% CI = 38.7% to 84.3%, P < .001) and 68.1% (95% CI = 31.5% to 104.8%, P = .0039), respectively. The stimulation of TIMP-1 expression and suppression of cell invasion were reversed by pretreatment of cells with antagonists to CB1 or CB2 receptors, with inhibitors of MAPKs, or, in the case of MA, with an antagonist to TRPV1. Knockdown of cannabinoid-induced TIMP-1 expression by siRNA led to a reversal of the cannabinoid-elicited decrease in tumor cell invasiveness in HeLa, A549, and C33A cells. CONCLUSION: Increased expression of TIMP-1 mediates an anti-invasive effect of cannabinoids. Cannabinoids may therefore offer a therapeutic option in the treatment of highly invasive cancers. PMID- 18159070 TI - Effect of systemic adjuvant treatment on risk for contralateral breast cancer in the Women's Environment, Cancer and Radiation Epidemiology Study. AB - BACKGROUND: Results from randomized trials indicate that treatment with tamoxifen or chemotherapy for primary breast cancer reduces the risk for contralateral breast cancer. However, less is known about how long the risk is reduced and the impact of factors such as age and menopausal status. METHODS: The study included 634 women with contralateral breast cancer (case patients) and 1158 women with unilateral breast cancer (control subjects) from the Women's Environment, Cancer and Radiation Epidemiology Study. The women were younger than age 55 when they were first diagnosed with breast cancer during 1985-1999. Rate ratios (RRs) and 95% confidence intervals (CIs) for contralateral breast cancer after treatment with chemotherapy or tamoxifen were assessed by multivariable adjusted conditional logistic regression analyses. RESULTS: Chemotherapy was associated with a lower risk for contralateral breast cancer (RR = 0.57, 95% CI = 0.42 to 0.75) than no chemotherapy. A statistically significant association between chemotherapy and reduced risk for contralateral breast cancer persisted up to 10 years after the first breast cancer diagnosis and was stronger among women who became postmenopausal within 1 year of the first breast cancer diagnosis (RR = 0.28, 95% CI = 0.11 to 0.76). Tamoxifen use was also associated with reduced risk for contralateral breast cancer (RR = 0.66, 95% CI = 0.50 to 0.88) compared with no use, and the association was statistically significant for 5 years after the first diagnosis. CONCLUSION: The associations between chemotherapy and tamoxifen treatment and reduced risk for contralateral breast cancer appear to continue for 10 and 5 years, respectively, after the initial breast cancer is diagnosed. Ovarian suppression may have a role in the association between chemotherapy and reduced risk for contralateral breast cancer. PMID- 18159071 TI - High-resolution mapping of DNA breakpoints to define true recurrences among ipsilateral breast cancers. AB - BACKGROUND: To distinguish new primary breast cancers from true recurrences, pangenomic analyses of DNA copy number alterations (CNAs) using single-nucleotide polymorphism arrays have proven useful. METHODS: The pangenomic profiles of 22 pairs of primary breast carcinoma (ductal or lobular) and ipsilateral breast cancers from the same patients were analyzed. Hierarchical clustering was performed using CNAs and DNA breakpoint information. A partial identity score developed using DNA breakpoint information was used to quantify partial identities between two tumors. The nature of ipsilateral breast cancers (true recurrence vs new primary tumor) as defined using the clustering methods and the partial identity score was compared with that based on clinical characteristics. Metastasis-free survival was compared among patients with primary tumors and true recurrences as defined using the partial identity score and by clinical characteristics. All statistical tests were two-sided. RESULTS: All methods agreed on the nature of ipsilateral breast cancers for 14 pairs of samples. For five pairs, the clinical definition disagreed with both clustering methods. For three pairs, the two clustering methods were discordant and the one using DNA breakpoints agreed with the clinical definition. The partial identity score confirmed the nature of ipsilateral breast cancers as defined by clustering of DNA breakpoints in 21 of 22 pairs. The difference in metastasis-free survival of patients with new primary tumors and those with true recurrences was not statistically significant when tumors were defined based on clinical and histologic characteristics (5-year metastasis-free survival: 76%, 95% confidence interval [CI] = 52% to 100% for new primary tumors and 38%, 95% CI = 17% to 83% for true recurrences; P = .18; new primary tumor vs true recurrence, hazard ratio = 2.8, 95% CI = 0.6 to 13.7), but the difference was statistically significant when tumors were defined using the partial identity score (5-year metastasis-free survival: 100% for new primary tumors and 29%, 95% CI = 11% to 78% for true recurrences; P = .01). CONCLUSIONS: DNA breakpoint information more often agreed with the clinical determination than CNAs in this population. The partial identity score, which was calculated based on DNA breakpoints, allows statistical discrimination between new primary tumors and true recurrences that could outperform the clinical determination in terms of prognosis. PMID- 18159072 TI - HER2 status and efficacy of adjuvant anthracyclines in early breast cancer: a pooled analysis of randomized trials. AB - BACKGROUND: Adjuvant chemotherapy with anthracyclines improves disease-free and overall survival compared with non-anthracycline-based adjuvant chemotherapy regimens in the treatment of early breast cancer. The role of HER2 status as a marker of anthracycline responsiveness has been explored by subset analyses within randomized clinical trials, with inconsistent results. We performed a pooled analysis of the interaction between HER2 status and the efficacy of adjuvant anthracyclines based on the published subset data. METHODS: We searched literature databases to identify randomized trials that compared anthracycline based with non-anthracycline-based adjuvant chemotherapy regimens in the treatment of early breast cancer and reported efficacy data according to HER2 status. Log hazard ratios (HRs) for disease-free and overall survival were pooled across the studies according to HER2 status by inverse variance weighting. A pooled test for treatment by HER2 status interaction was performed by weighted linear meta-regression. All statistical tests were two-sided. RESULTS: Eight studies (with 6564 randomly assigned patients, of whom 5354 had HER2 status information available) were eligible for this analysis. In HER2-positive disease (n = 1536 patients), anthracyclines were superior to non-anthracycline-based regimens in terms of disease-free (pooled HR of relapse = 0.71; 95% confidence interval [CI] = 0.61 to 0.83; P < .001) and overall (pooled HR of death from any cause = 0.73; 95% CI = 0.62 to 0.85; P < .001) survival. In HER2-negative disease (n = 3818 patients), anthracyclines did not improve disease-free (HR = 1.00; 95% CI = 0.90 to 1.11; P = .75) or overall (HR = 1.03; 95% CI = 0.92 to 1.16; P = .60) survival. The test for treatment by HER2 status interaction yielded statistically significant results: for disease-free survival, the chi-square statistic for interaction was 13.7 (P < .001), and for overall survival, it was 12.6 (P < .001). CONCLUSIONS: The added benefits of adjuvant chemotherapy with anthracyclines appear to be confined to women who have HER2 overexpressed or amplified breast tumors. PMID- 18159073 TI - Knockout mouse creation wins Nobel Prize. PMID- 18159074 TI - Recent conference addresses research integrity on global scale. PMID- 18159075 TI - Reactive oxygen species may have antitumor activity in metastatic melanoma. PMID- 18159076 TI - What happened to the cancer-detecting bra? PMID- 18159078 TI - p63 null mutation protects mouse oocytes from radio-induced apoptosis. AB - Female fertility in mammals is determined by the pool of primordial follicles and low doses of radiation induce a major loss of primordial follicles in the ovary. We investigated the expression of p53 and its homologues, p63 and p73, in the normal and irradiated neonatal ovary. p63 was the only member of the p53 family detected in oocyte nucleus. No p63 transcripts or protein were detected in the early foetal ovary. p63 production began in late pachytene-stage oocytes and peaked in diplotene oocytes in mice and humans. The production of p63 was correlated with meiotic DNA double-strand break repair. Only transactivation (TA) isoforms were present in the ovary, with TAp63 alpha by far the most abundant in terms of mRNA and protein levels. Complete p63 null mutation did not affect normal ovary development. Irradiation rapidly triggered p63 phosphorylation. p63 null mutation prevented the cleavage of caspases-9 and -3 and the follicle loss induced by ionising radiation. Thus, our results evidence that irradiation induced depletion of the primordial follicle pool results from the activation of p63 in quiescent oocytes. PMID- 18159079 TI - High hydrostatic pressure: a new way to improve in vitro developmental competence of porcine matured oocytes after vitrification. AB - The purpose of the present study was to improve cryotolerance using high hydrostatic pressure (HHP) pretreatment of porcine in vitro matured (IVM) oocytes, to facilitate their further developmental competence after parthenogenetic activation. A total of 1668 porcine IVM oocytes were used in our present study. The pressure tolerance and optimal duration of recovery after HHP treatment were determined. Oocytes were treated with either 20 or 40 MPa (200 and 400 times greater than atmospheric pressure) for 60 min, with an interval of 10, 70, and 130 min between pressure treatment and subsequent vitrification under each pressure parameter. Oocytes from all vitrification groups had much lower developmental competence than fresh oocytes (P<0.01) measured as cleavage and blastocyst rates. However, significantly higher blastocyst rates (P<0.01) were obtained in the groups of 20 MPa pressure, with either 70 (11.4+/-2.4%) or 130 (13.1+/-3.2%) min recovery, when compared with the vitrification control group without HHP treatment where no blastocysts were obtained. The influence of temperature at HHP treatment on further embryo development was also investigated. Treatments of 20 MPa with 70 min recovery were performed at 37 degrees C or 25 degrees C. Oocytes pressurized at 37 degrees C had a significantly higher blastocyst (14.1+/-1.4%) rate than those treated at 25 degrees C (5.3+/-1.1%; P<0.01). Our results demonstrate that HHP pretreatment could considerably improve the developmental competence of vitrified pig in vitro matured (IVM) oocytes. The HHP pretreatment will be tested as a means to improve survival and developmental competence at different developmental stages in different species including humans. PMID- 18159080 TI - Hardening of the zona pellucida of unfertilized eggs can reduce polyspermic fertilization in the pig and cow. AB - One of the proposed mechanisms of polyspermy block is an increased resistance of the zona pellucida (ZP) to proteolytic digestion (ZP hardening) as a consequence of cortical granule exocytosis that occurs soon after fertilization. However, evidence is available that the zonae pellucidae of freshly ovulated pig and cow oocytes harden considerably before fertilization. It was thought that such pre fertilization ZP hardening could be involved in the control of polyspermy, and its lack in the oocytes matured in vitro could be one of the reasons for the extremely high incidence of polyspermy in pig in vitro fertilization (IVF). To test this hypothesis, two different types of cross-linking reagents were employed and their effects on ZP hardening and IVF efficiency were examined. The sulfhydryl-reactive cross-linkers produced a slight hardening of ZP (P<0.001) of treated oocytes compared with control oocytes, and totally inhibited sperm penetration into pig oocytes after IVF. In the cow, sperm penetration into eggs was reduced to 10%. It is proposed that formation of disulfide bonds in ZP or blocking of SH groups in the oocyte plasma membrane proteins prevents sperm penetration. An amine-reactive cross-linker was then assayed and produced strong ZP hardening, increasing the incidence of monospermy in both pig and cow oocytes after fertilization. When the cross-linker concentration was optimized, a 45% improvement for pig IVF efficiency was reached. It is proposed that the observed physiological ZP hardening is a mechanism to control polyspermy, differentially affecting various mammalian species and can be imitated by the use of amine reactive cross-linkers during IVF. PMID- 18159081 TI - Monoallelic expression of nine imprinted genes in the sheep embryo occurs after the blastocyst stage. AB - The preimplantation embryos of a range of mammals can be susceptible to disruptions in genomic imprinting mechanisms, resulting in loss of imprinting. Such disruptions can have developmental consequences involving foetal and placental growth such as Beckwith-Wiedemann syndrome in humans and large offspring syndrome in sheep. Our objective was to investigate the dynamics of establishing monoallelic expression of individual sheep imprinted genes post fertilisation. Semi-quantitative RT-PCR was used to amplify cDNA from the sheep blastocyst, day 21 foetus and day 21 chorioallantois, to compare expression levels between biparental and parthenogenetic embryos in order to indicate allelic expression status. In common with other mammals, IGF2, PEG1 and PEG3 were paternally expressed in the day 21 conceptus, while H19, IGF2R, GRB10 and p57KIP were maternally expressed. Interestingly, GNAS was maternally expressed in the foetus, but paternally expressed in the chorioallantois at day 21. Overall, the imprinting of ovine GRB10 and IGF2R was comparable with mouse but not with human. Contrary to the trophoblast-restricted maternal expression in both mouse and human, SASH2 (sheep homologue of Mash2/HASH2) was expressed in the ovine foetus and was biallelically expressed in the chorioallantois. Differential methylation of the H19 CTCF III upstream region and IGF2R DMR2 in the chorioallantois revealed predominantly paternal and maternal methylation respectively, indicating conservation of these imprinting regulatory regions. In blastocysts, IGF2R, GRB10 and SASH2 were expressed biallelically, while the other genes were not detected. Thus, for the majority of ovine imprinted genes examined, monoallelic expression does not occur until after the blastocyst stage. PMID- 18159082 TI - Leukemia inhibitory factor ligand-receptor signaling is important for uterine receptivity and implantation in golden hamsters (Mesocricetus auratus). AB - Blastocyst implantation occurs in the progesterone-primed uterus of hamsters, but not in mice where the progesterone-primed uterus requires estrogen influence. Leukemia inhibitory factor (Lif), an estrogen-regulated gene in mice, is an absolutely needed cytokine for uterine receptivity and implantation in this species. This study aimed to evaluate the importance of Lif ligand-receptor signaling during uterine receptivity and implantation in hamsters. We investigated whether or not the uterine expression patterns of Lif and its receptors, Lif-r and gp130, during the periimplantation period of pregnancy and its hormonal regulation in the ovariectomized hamster correlate with some of the vital phases of uterine changes during early pregnancy. Uterine Lif, Lif-r, and gp130 mRNA expressions were examined by Northern and in situ hybridization. During the uterine preparatory phase for implantation, Lif, Lif-r, and gp130 were expressed either in the gland, luminal epithelium or both. As the implantation process began, Lif expression was minimal, but Lif-r and gp130 extended to the decidual areas. This decidual expression of Lif-r and gp130 was not dependent on the presence of the embryo since these genes were expressed in the suture-induced deciduomata. We also observed that, while the uterine Lif was induced by estrogen, Lif-r and gp130 were induced by progesterone in ovariectomized hamsters. Additionally, we show that a Lif antibody when instilled intraluminally on day 3 of pregnancy reduced the number of implantation sites. Taken together, these data suggest that Lif signaling is important for uterine receptivity and implantation in hamsters. PMID- 18159083 TI - Increased potency of alpha 1-adrenergic receptors to induce inositol phosphates production correlates with the up-regulation of alpha 1d/Gh alpha/phospholipase C delta 1 signaling pathway in term rat myometrium. AB - In the present study, we studied the potential regulation by rat myometrial alpha1-adrenergic receptors (alpha1-AR) of the newly identified Gh alpha protein/phospholipase C delta 1 (PLC delta 1) signaling pathway and compared myometrial inositol phosphates (InsP) production and activity of the uterine circular muscle in response to alpha1-AR activation between mid-pregnancy and term. For this, we quantified the level of rat myometrial alpha1-AR coupling to Gh alpha protein by photoaffinity-labeling, the cytosolic amount of PLC delta 1 enzyme by immunoblotting, and the expression level of alpha1-AR subtypes by RT PCR. The results showed an increased level of alpha1-AR/Gh alpha protein coupling and the amount of PLC delta 1 at term (+147 and +65% respectively, versus mid pregnancy). This was correlated with an up-regulation of alpha 1d-AR subtype (+70% versus mid-pregnancy). Incubation of myometrial strips with phenylephrine (Phe), a global alpha1-agonist, increased InsP production in a dose-dependent manner at both mid-pregnancy and term, but with an enhanced potency (tenfold decrease in EC(50) value) at term. Phe also dose-dependently induced contraction of the circular muscle at both mid-pregnancy and term. However, unlike InsP response, no amelioration of potency was observed at term. Similar results were obtained with the endogenous agonist norepinephrine. Our results show, for the first time, that rat myometrial alpha 1d-AR/Gh alpha/PLC delta 1 signaling pathway is up-regulated at term. This is associated with an increased potency of alpha1-AR to elicit InsP production but not uterine contraction at this period. It is thus hypothesized that alpha1-AR, through activation of Gh alpha/PLC delta 1 system, are not primarily involved in the initiation of labor but may rather regulate responses such as myometrial cell proliferation or hypertrophy. PMID- 18159085 TI - Gas6 and the Tyro 3 receptor tyrosine kinase subfamily regulate the phagocytic function of Sertoli cells. AB - The apoptotic spermatogenic cells and residual bodies are phagocytosed and degraded by Sertoli cells during spermatogenesis. The mechanisms of this process are largely unknown. Here, we demonstrate that Gas6 and its receptors, the Tyro 3 subfamily of receptor tyrosine kinases (RTKs; Tyro 3, Axl, and Mer), regulate the phagocytic function of Sertoli cells. The phagocytic ability of Sertoli cells increased by five times in the presence of Gas6 in serum-free medium when compared with controls. The Sertoli cells lacking Mer showed a 35% reduction in phagocytosis of apoptotic spermatogenic cells when compared with wild-type (WT) controls, whereas the Sertoli cells lacking Tyro 3 or Axl exhibited phagocytic activity comparable with the controls. Notably, the Sertoli cells lacking all three members of the Tyro 3 RTK subfamily showed a dramatic decrease in phagocytic ability of 7.6-fold when compared with WT Sertoli cells. The deficiency in phagocytosis by the triple-mutant Sertoli cells was due to the deficit in binding of the Sertoli cells to apoptotic germ cells. These findings suggest that Mer is responsible for triggering phagocytosis of apoptotic spermatogenic cells by Sertoli cells and that Tyro 3, Axl, and Mer participate in recognizing and binding apoptotic germ cells by Sertoli cells in a redundant manner. Gas6 is a functional ligand of the Tyro 3 RTK subfamily in mediating phagocytic ability of Sertoli cells. PMID- 18159084 TI - Negative energy balance in dairy cows is associated with specific changes in IGF binding protein expression in the oviduct. AB - Negative energy balance (NEB) during early lactation in dairy cows leads to an altered metabolic state that has major effects on the production of IGF family members. Low IGF-I concentrations are associated with poor fertility and therefore we aimed to determine whether NEB exerts a direct effect on IGF expression in the postpartum oviduct. Multiparous Holstein cows were allocated to two treatments (each n=6) designed using differential feeding and milking regimes to produce either mild NEB (MNEB) or severe NEB (SNEB). Animals were slaughtered in week 2 of lactation when divergent metabolic profiles were evident. Oviducts were collected for RNA analysis by real-time RT-PCR and in situ hybridisation. Quantitative measures in oviduct gene expression were obtained for all members of the IGF family (IGF-I/II, IGF-binding proteins (IGFBP) 1-6 and receptors for IGF types 1 and 2), insulin A/B, GH, glucocorticoid and oestrogen alpha/beta. Expression of IGFBP-2 and IGFBP-6 (both of which have a high affinity for IGF-II) was decreased in SNEB relative to MNEB (P<0.05). No other gene was altered by NEB, but IGF-II, IGFBP-3, IGFBP-5 and IGFBP-6 all showed differential expression in different regions of the oviduct. These results indicate that, in addition to low circulating IGF-I after calving, NEB may also influence IGF availability in the oviduct indirectly through changes in specific IGFBP expression. It is possible that the predicted increased signalling by IGF-II may perturb embryo development, contributing to the high rates of embryonic mortality in dairy cows. PMID- 18159086 TI - Impact of social environment on variation in menstrual cycle length in captive female olive baboons (Papio anubis). AB - Factors affecting menstrual cycles and conception were explored for captive female olive baboons. We evaluated the relationship between the social environment and adequacy of the menstrual cycle in 55 non-conceptive and 21 conception cycles from 23 females. More abnormal cycles were expected for low status females, and social stress levels were associated with variation in menstrual cycle length. Mean cycle length was 39.9 days (median=38) with a mean follicular phase duration of 23.7 (median=22) days. The duration of the follicular phase was more variable than that of the luteal phase (mean=15.8 days). The first cycle after postpartum resumption of cycling was not markedly different from subsequent cycles in terms of duration or probability of conception. Dominance rank was one significant factor affecting female fertility. Low-ranking females experienced more cycles prior to conception, longer cycles once cycling was well established and had smaller sexual swellings (anogenital area) than did high-ranking females. Both acute and chronic stresses may play important roles in fertility outcomes for these baboons and further research is needed to understand the role of stress and subtle menstrual cycle abnormalities in female mammal fertility. PMID- 18159087 TI - The onset of puberty in female mice as reflected in urinary steroids and uterine/ovarian mass: interactions of exposure to males, phyto-oestrogen content of diet, and ano-genital distance. AB - Development of puberty in female mice was examined in relationship with the ano genital distance index (AGDI), phyto-oestrogen content of diet and exposure to males post weaning. Throughout gestation and post-natal development, females were exposed to a regular diet or a nutritionally similar diet deficient in phyto oestrogens. After segregation at weaning on the basis of short or long AGDI, an indirect measure of in utero androgen exposure, females were housed alone or underneath two outbred adult males for 2 weeks. Female urinary samples were collected non-invasively throughout this exposure, then assayed for oestradiol, progesterone and creatinine. Females were then killed and uterine and ovarian mass was determined. Urinary oestradiol was substantially reduced in females raised on the phyto-oestrogen-free diet. Oestradiol levels were more dynamic over days in urine of male-exposed females, especially among those on the regular diet. Urinary progesterone was not strongly influenced by diet. Progesterone was more dynamic in urine of male-exposed females, and was generally elevated compared with levels in isolated females, the size of this effect dependent on AGDI, diet and whether the measure was adjusted for creatinine. Urinary creatinine was elevated by the phyto-oestrogen-free diet and reduced by male exposure, tending to decline over days in females exposed to males. Male exposure increased uterine and ovarian mass and was influenced by AGDI in interaction with diet and male exposure. PMID- 18159088 TI - Association of FSH receptor and CYP19A1 gene variations with sterility and ovarian hyperstimulation syndrome. AB - Severe ovarian hyperstimulation syndrome (OHSS) is a potentially life-threatening complication during assisted reproduction technology (ART). The aetiology of this condition is still not fully understood. Several gene variations in the FSH receptor (FSHR) gene have been identified for the very rare cases of spontaneous OHSS. There are only few published data on gene variations in sterility and iatrogenic OHSS and no data regarding aromatase (cytochrome P450 19A1; CYP19A1). Ninety-one ART patients with OHSS, eighty-eight ART patients without OHSS and ninety-seven women with assumed normal fecundity were analysed for the FSHR single nucleotide polymorphism (SNP) gene variations Asn680Ser (rs6166), Ala189Val, Ile160Thr, Thr449Ile (rs28928870) and the CYP19A1 rs10046 locus using real-time PCR. In addition, exon 10 of FSHR of two patients with spontaneous hyperreactio luteinalis (HL) was sequenced. Significantly lower frequencies of homozygous Ser680/Ser680 (P=0.035) and heterozygous Thr160/Ile160 (P=0.039) were found in patients with normal fecundity than those undergoing ART. The Ile160Thr SNP with a frequency of 6.7 and 6.1% in ART patients with and without OHSS respectively does not represent a rare mutation as previously published. There were no differences in the frequencies of all other gene variations. Of two patients with HL, both had homozygous point mutations for Ser680/Ser680 and one was heterozygous for Ile160Thr and CYP19A1 rs10046. The FSHR gene variations Asn680Ser as well as Ile160Thr may be contributing factors in unexplained sterility. The other FSHR coding gene variations and CYP19A1 rs10046 investigated are most likely not involved in the aetiology of iatrogenic OHSS or sterility. PMID- 18159090 TI - Resolving the mechanism of bile acid negative-feedback regulation, a Journal of Lipid Research tradition. PMID- 18159091 TI - Exogenous cytosine deaminase gene expression in Bifidobacterium breve I-53-8w for tumor-targeting enzyme/prodrug therapy. AB - Bifidobacteria are nonpathogenic, anaerobic domestic bacteria with health promoting properties for the host. In our previous study, Bifidobacterium longum (B. longum) were found to be localized selectively and to proliferate within solid tumors after systemic application. Additionally, B. longum transformed by shuttle-plasmid including the cytosine deaminase (CD) gene expressed active CD, converted the prodrug 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU). We also demonstrated antitumor efficacy with a transformant of B. longum in rats. In this study, we found that Bifidobacterium breve (B. breve), the smallest species of human-derived bifidobacterium, expressed the exogenous transgene (CD), that CD enzymatic activity in the transformant of B. breve was much higher, and that the segregational stability of the plasmid was greater than that of B. longum. Thus, numerous transformants of B. breve were detected solely in the tumors after systemic administration. We consider the transformant of B. breve to be more beneficial in our enzyme/prodrug therapy. PMID- 18159092 TI - Redox regulation in the extracellular environment. AB - The oxidizing nature of the extracellular environment is vastly different from the highly reducing nature of the intracellular compartment. The redox potential of the cytosolic compartment of the intracellular environment limits disulfide bond formation, whereas the oxidizing extracellular environment contains proteins rich in disulfide bonds. If not for an extracellular antioxidant system to eliminate reactive oxygen and nitrogen species, lipid peroxidation and protein oxidation would become excessive, resulting in cellular damage. Many reviews have focused on the role of intracellular antioxidants in the elimination of oxidative stress, but this one will focus on the coordinated action of both intracellular and extracellular antioxidants in limiting cellular oxidant stress. PMID- 18159093 TI - Effect of early use of low-dose pravastatin on major adverse cardiac events in patients with acute myocardial infarction: the OACIS-LIPID Study. AB - BACKGROUND: It is unclear whether early initiation of low-dose pravastatin therapy can reduce the occurrence of major adverse cardiac events after acute myocardial infarction (AMI). METHODS AND RESULTS: The study group comprised 353 patients with AMI who had plasma total cholesterol levels of 200-250 mg/dl and triglyceride levels <300 mg/dl. The patients were randomly assigned to either receive pravastatin (10 mg/daily, n=176) or not (n=177). The primary endpoint was a composite of death, nonfatal myocardial infarction (MI), unstable angina (UA), stroke, revascularization, and rehospitalization because of other cardiovascular disease. The follow-up period was 9 months. The primary endpoint occurred in 31 patients (17.9%) in the pravastatin group and 55 patients (31.4%) in the non pravastatin group (relative risk, 0.56; 95% confidence interval, 0.36-0.87). There were no significant differences in the risk of death, nonfatal MI, UA, and stroke between the 2 groups, although the pravastatin group had a lower risk of need for revascularization. CONCLUSION: For patients with AMI, early and low-dose pravastatin therapy (10 mg/daily) reduces recurrent major adverse cardiac events, mostly the requirement for revascularization. PMID- 18159094 TI - Clinical significance of high-molecular weight form of adiponectin in male patients with coronary artery disease. AB - BACKGROUND: It has been reported previously that the measurement of plasma total adiponectin level is clinically useful to estimate the risk of coronary artery disease (CAD). Here, the relevance of high molecular weight (HMW) adiponectin with risk factors for atherosclerosis is investigated METHODS AND RESULTS: A total of 186 consecutive male CAD patients participated in the study and were categorized into quartiles based on their total adiponectin level. The interquartile cut-off points were 4.0, 5.5 and 7.0 microg/ml. The HMW adiponectin levels were significantly lower in the quartile of lower total adiponectin levels both in non-diabetic and diabetic patients. In contrast, low molecular weight adiponectin levels (which were calculated as the Total - HMW) were constant. In univariate analysis, total adiponectin correlated negatively with body mass index and hemoglobin (Hb) A1c, and HMW adiponectin correlated negatively with HbA1c in non-diabetic patients. On the other hand, total and HMW adiponectin correlated positively with high-density lipoprotein-cholesterol (HDL-C) in diabetic patients. Multiple regression analysis revealed that HMW adiponectin correlated negatively with HbA1c in non-diabetic patients, and total and HMW adiponectin correlated positively with HDL-C in diabetic patients. CONCLUSIONS: Change in the HMW isoform reflects a change in total adiponectin level. Measurement of total and HMW adiponectin were equally useful in assessing metabolic risk in CAD patients. PMID- 18159095 TI - Relation of apolipoprotein E polymorphism to markers of early atherosclerotic changes in young adults--the Cardiovascular Risk in Young Finns Study. AB - BACKGROUND: Carotid artery intima-media thickness (IMT), which is measured by ultrasound, is used in the assessment of early atherosclerotic changes, and has been associated with apolipoprotein E (APOE) polymorphism in many studies comprising elderly subjects. However, results are still inconclusive and data relating to young adults are missing. Whether common APOE polymorphism is related to carotid IMT was studied in a population of young adults. Also brachial flow mediated dilatation (FMD) and carotid artery compliance (CAC) were determined to clarify their relation to this genetic factor. METHODS AND RESULTS: A total of 1,188 young adults (aged 24-39 years) participating in the Cardiovascular Risk in Young Finns Study with complete data of common APOE polymorphism underwent a carotid and brachial ultrasound. Patients' lipid levels and blood pressure were also examined. There was no significant association between the APOE phenotypes and carotid IMT, brachial FMD or CAC either in young men or in young women. The results were similar for systolic and diastolic blood pressure. In the same population, the well-known association between APOE phenotypes and lipids was seen. CONCLUSIONS: Common APOE polymorphism does not seem to be an independent genetic determinant of carotid IMT, brachial FMD or CAC. PMID- 18159096 TI - Randomized comparison of cilostazol vs clopidogrel after drug-eluting stenting in diabetic patients--clilostazol for diabetic patients in drug-eluting stent (CIDES) trial. AB - BACKGROUND: Previous studies have shown that cilostazol may not only prevent stent thrombosis, but may also have positive effect in the prevention of restenosis. However, the effect of cilostazol on restenosis after successful deployment of drug-eluting stent (DES) in patients with diabetes mellitus has not been evaluated. METHODS AND RESULTS: A total of 280 patients at 8 clinical sites were randomized. The patients (61.7+/-9.9 years old, 163 males) who underwent successful stenting were randomized to aspirin and cilostazol (group I, n=141, 61.2+/-9.6 years old) vs aspirin and clopidogrel (group II, n=139, 62.0+/-10.0 years old) after 1 month of aspirin, cilostazol, and clopidogrel combination treatment. There were no significant differences in baseline characteristics of the groups. The type of DES implanted did not differ between the groups. There were no differences in angiographic and procedural characteristics of the groups. Major adverse cardiac events, including acute and subacute stent thrombosis within 1 month, did not occur in either group. Cases of angiographic late stent thrombosis were 1 (0.9%) in group I and 1 (0.8%) in group II. Follow-up coronary angiography was performed in 237 patients (84.6%). Mean follow-up duration was 7.1 months. The rate of angiographic restenosis (stent plus 5-mm borders) was 9 (8.0%) in group I and 20 (16.1%) in group II, p=0.041). The minimal luminal diameter at follow-up period in group I was 2.55+/-0.63 mm compared with 2.41+/ 0.83 mm in group II (p=NS). CONCLUSIONS: Combination therapy with aspirin and cilostazol for the prevention of stent restenosis is comparable or superior to that of aspirin and clopidogrel in diabetic patients who undergo DES implantation. PMID- 18159097 TI - Apolipoprotein A5 IVS3+476A allelic variant associates with increased trigliceride levels and confers risk for development of metabolic syndrome in Hungarians. AB - BACKGROUND: Metabolic syndrome consists of multiple risk factors that are increasing the cardiovascular mortality. The T-1131C variant of the apolipoprotein A5 gene, associated with increased triglycerides, has been found to confer risk for cardiovascular diseases and metabolic syndrome. Because other naturally occurring variants of the gene also correlate with elevated triglycerides, the possible role of 2 common variants, the IVS3+G476A and T1259C, with metabolic syndrome was investigated. METHODS AND RESULTS: A total of 213 metabolic syndrome patients and 142 healthy controls were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Serum triglycerides were increased in carriers compared with non-carriers in both groups (p<0.001); serum cholesterol levels were similar in all genotypes. The IVS3+476A allele frequency was increased in metabolic syndrome patients compared with controls (8.05 vs 2.47%; p<0.05), whereas the 1259C allele frequency did not differ between the groups. Multiple logistic regression analyses adjusted for age, gender, serum total cholesterol, acute myocardial infarction and stroke revealed that the IVS3+476A variant confers risk for development of metabolic syndrome (odds ratio =3.529, 95% confidence interval 1.308-9.029, p=0.009), but the 1259C allele had no such an effect. CONCLUSIONS: Carrying the IVS3+473A allele is associated with elevated triglycerides and confers risk for development of metabolic syndrome, a combination that represents increased risk for development of atherogenic vascular diseases. PMID- 18159098 TI - Fat and fiber consumption are associated with peripheral arterial disease in a cross-sectional study of a Japanese-Brazilian population. AB - BACKGROUND: The Western diet plays a role for the epidemics of obesity and related diseases. This study examined a possible association between peripheral arterial disease (PAD) and the dietary components of Japanese immigrants living in Brazil. METHODS AND RESULTS: In this cross-sectional study, 1,267 subjects (aged > or =30 years) with complete dietary, clinical and laboratory data were studied according to a standardized protocol. Ankle-to-brachial index was used to identify subjects with PAD. The overall prevalence of PAD was 14.6%. Subjects with PAD were older, had lower education and higher mean values of blood pressure, triglycerides, and fasting and 2-h plasma glucose levels compared with those without the disease. Among the subjects with PAD, the consumption of fiber from whole grains (3.0 vs 3.4 g, p=0.001) and linoleic acids (11.0 vs 11.7 g, p=0.017) were lower and intake of total (72.8 vs 69.1 g, p=0.016) and saturated fatty acids (17.4 vs 16.3 g, p=0.012) were higher than those without PAD. Results of multiple logistic regression analysis showed a significant association between PAD with high total fat intake, low intake of fiber from fruit and oleic acid, independently of other variables. CONCLUSIONS: Despite limitations in examining the cause - effect relationship, the data support the notion that diet could be important in reducing the occurrence of PAD. PMID- 18159099 TI - Asymptomatic acute ischemic stroke after primary percutaneous coronary intervention in patients with acute coronary syndrome might be caused mainly by manipulating catheters or devices in the ascending aorta, regardless of the approach to the coronary artery. AB - BACKGROUND: Asymptomatic acute ischemic stroke (aAIS) following primary percutaneous coronary intervention (p-PCI) in patients with acute coronary syndrome (ACS) has not been studied in detail. METHODS AND RESULTS: Of 75 patients who underwent p-PCI, 26 (34.7%) developed aAIS as determined by diffusion-weighted magnetic resonance imaging (MRI). Including the approach to the coronary artery (via lower limb or right upper limb), 23 factors were compared between patients with (n=26) and without (n=49) aAIS. Age, hypertension, smoking, plasma glucose levels, Killip grade, right coronary artery (RCA) as culprit vessel, percutaneous coronary intervention (PCI) time, and the frequency of device insertion into the coronary artery differed in a statistically significant manner. However, multivariate analysis showed that the RCA (odds ratio 3.477) and the frequency of device insertion (1.375) were independent factors linked to the incidence of aAIS. Moreover, anterior or posterior location and left or right cerebral circulation of aAIS were equivalent in both approaches. CONCLUSIONS: Cranial MRI images following emergency PCI revealed that 34.7% of the patients with ACS had aAIS that might be caused by manipulating the catheter or devices in the ascending aorta, micro-air bubble embolism during injection, or micro-thrombus embolism derived from the ACS lesions during the PCI procedure. PMID- 18159100 TI - Sirolimus-eluting stents vs bare metal stents for coronary intervention in Japanese patients with renal failure on hemodialysis. AB - BACKGROUND: Accelerated atherosclerosis is a major risk for long-term survivors receiving hemodialysis (HD), with coronary events being the leading cause of mortality. METHODS AND RESULTS: A total of 88 consecutive patients on HD (121 lesions) who underwent percutaneous coronary intervention (PCI) with sirolimus eluting stents (SES) were compared with 78 patients on HD (95 lesions) who received bare metal stents (BMS) in the preceding 1 year. The primary endpoint was angiographic restenosis defined as > or =50% diameter stenosis at 6-8 months follow-up after PCI. The angiographic restenosis rate at follow-up was 22.2% in the SES group and 24.4% in the BMS group. No difference was detected in the restenosis rate between the 2 groups (p=0.73). When including both HD and non-HD patients, the independent predictors for restenosis after SES implantation were treatment with HD (hazard ratio (HR) 3.12; 95% confidence interval (CI) 1.23 7.95; p=0.016), incidence of hyperlipidemia (HR 3.93; 95%CI 1.12-13.7; p=0.032), coronary calcification (HR 2.78; 95%CI 1.12-6.91; p=0.027), and implantation of multi-stents (HR 4.14; 95%CI 1.70-10.1; p=0.0017). CONCLUSIONS: Even if treated with SES, patients with end-stage renal failure on HD are at high risk of restenosis after PCI. PMID- 18159101 TI - Relationship between the serum sCD40L level and aspirin-resistant platelet aggregation in patients with stable coronary artery disease. AB - BACKGROUND: Current evidence supports the central role of inflammation in all phases of the atherosclerotic process, including its thrombotic complications. Increased serum sCD40L may trigger platelet activation, so the aim of the present study was to determine the relation between sCD40L levels and aspirin-resistant platelet aggregation in patients with coronary atherosclerosis. METHODS AND RESULTS: A total of 167 consecutive patients (39-85 years old, 35.9% women) with stable coronary artery disease was enrolled in the study. Platelet function was evaluated by a Platelet Function Analyzer 100 device (PFA-100) with collagen and epinephrine (Col/Epi) and collagen and adenosine diphosphate (ADP) (Col/ADP) cartridges. Aspirin resistance was defined as a closure time (CT) <186 s with Col/Epi cartridges, despite regular aspirin therapy. Serum sCD40L level was determined quantitatively with an ELISA method. Fifty-seven (34.1%) patients had aspirin resistance according to the PFA-100. Mean CT measured with the Col/ADP cartridges was 83+/-18 s (65-101 s). Mean serum sCD40L was 157 pg/ml (6-700 pg/ml) in the entire cohort. Patients with aspirin resistance had a mean serum sCD40L level of 166 pg/ml and patients with aspirin-sensitive platelet aggregation had an sCD40L level of 152 pg/ml (p=0.582). CONCLUSION: The sCD40L level is similar in patients with aspirin-resistant and aspirin-sensitive platelet aggregation according to the PFA-100. There is still need for further studies to elucidate the relationship between aspirin-resistant platelet aggregation and sCD40L, which is now known to be prothrombotic, proinflammatory and to be a risk factor for cardiovascular events. PMID- 18159102 TI - Effectiveness and safety of routine primary angioplasty in patients aged > or =85 years with acute myocardial infarction. AB - BACKGROUND: Although octogenarians constitute a fast growing portion of cardiovascular patients, few data are available on the outcome of very old patients (age >80 years) with ST-segment elevation myocardial infarction (STEMI) undergoing primary angioplasty. METHODS AND RESULTS: Short- and long-term outcomes of 88 consecutive very old (age > or =85 years) patients with STEMI undergoing primary angioplasty were evaluated. In-hospital mortality was 17%, significantly higher in patients with cardiogenic shock (90%; p<0.001), with failure of percutaneous coronary intervention (PCI; p=0.016), with Killip class > or =III on admission (p=0.018), or with chronic renal failure (p=0.033). Major bleeding complications occurred in 11 patients (12%). Multivariable logistic regression analysis identified 3 independent predictors of in-hospital death: age > or =90 years (p=0.018), Killip > or =III on admission (p=0.018), and PCI failure (p=0.025). Multivariable logistic regression analysis identified age > or =90 years (p=0.008), Killip > or =III on admission (p=0.015), and time from symptoms to PCI >12 h (p=0.04) as independent predictors of mortality at long term follow-up. CONCLUSIONS: The low incidence of procedural complications, together with good long term survival, suggest that primary PCI in STEMI patients > or =85 years is safe and efficacious, with a low rate of PCI failure in the presence of a low Killip class on admission, whereas primary PCI is unable to affect the poor prognosis for very old patients with cardiogenic shock. PMID- 18159103 TI - Serum chitotriosidase activity in patients with coronary artery disease. AB - BACKGROUND: Atherosclerosis is considered to be an inflammatory disease in which the initial process is augmented infiltration of monocytes into the vessel wall and their subsequent differentiation from macrophages into lipid-laden foam cells. Chitotriosidase is one of the most quantitative proteins secreted by activated macrophages, so the aim of this study was to investigate the association of the level of serum chitotriosidase activity with atherosclerotic coronary artery disease (CAD). METHODS AND RESULTS: A total of 200 subjects undergoing coronary angiography were divided into 4 subgroups according to the number of diseased vessels and their serum chitotriosidase activity levels were measured. Serum chitotriosidase activity in patients with CAD was significantly higher than in normal control subjects (p<0.001). Serum chitotriosidase activity was also significantly associated with the extent of CAD as defined by the number of stenosed vessels (p<0.001). CONCLUSION: Serum chitotriosidase activity can be considered a strong inflammatory marker of CAD. Moreover, plasma chitotriosidase activity may be also regarded as a quantitative indicator of disease extent, as well as being a marker of disease presence. PMID- 18159104 TI - Clinical study of the acute effects of intravenous nifekalant on the defibrillation threshold in patients with persistent and paroxysmal atrial fibrillation. AB - BACKGROUND: Antiarrhythmic agents are considered to have significant effects on the defibrillation energy requirement, so this study investigated the effects of nifekalant on defibrillation. METHODS AND RESULTS: Forty-two patients with persistent atrial fibrillation (AF) underwent electrical cardioversion via intracardiac electrode catheters prior to and after the intravenous administration of nifekalant. The success rate of the defibrillation and change in the defibrillation threshold using sequential incremental defibrillation energy deliveries was investigated. In addition, the parameters that could predict the beneficial effects of nifekalant were also assessed. Nifekalant significantly decreased the defibrillation energy requirement in 13 of the 42 cases, and nifekalant also converted AF to sinus rhythm with an identical energy to that of the last unsuccessful defibrillation in 21 of 42 cases. The success of defibrillation seemed to be dependent on significant prolongation of the intracardiac atrial electrogram intervals during AF by the nifekalant. CONCLUSIONS: Intravenous nifekalant significantly improved the electrical defibrillation efficacy in patients with persistent AF that was resistant to defibrillation, without any serious adverse effects. PMID- 18159105 TI - Long-term outcome of isolated congenital complete atrioventricular block pacing since neonatal period: experience at a single Japanese institution. AB - BACKGROUND: Few investigators have examined the outcome of patients with isolated congenital complete atrioventricular block (CCAVB) paced from the neonatal period. The present study follows the clinical course and describes the outcome of patients who have been paced with CCAVB since they were neonates. METHODS AND RESULTS: The medical records of 20 patients with CCAVB paced as neonates between 1981 and 2006 were retrospectively studied and reviewed; 18 were diagnosed in utero and 2 at birth. SS-A antibodies were detected in 9 of 14 (64%) mothers tested. The median age at follow-up was 5.6 years (range, 2 days to 21.9 years). Three (15%) of the early neonates died and 3 (15%) died later: 2 from dilated cardiomyopathy (DCM) and 1 from pneumonia. The total mortality rate was 30%. The cumulative probability of survival at 10 years was 73%. Of the 14 survivors, 4 developed DCM, representing a total morbidity of 30%. The cumulative probability of freedom from DCM at 10 years was 59%. CONCLUSIONS: Patients with CCAVB who undergo pacing as neonates have high mortality and high morbidity because of DCM. Left ventricular pacing or resynchronization can benefit patients who develop DCM and left ventricular epicardial pacing is recommended for neonates with CCAVB. PMID- 18159106 TI - Electrophysiologic study-guided amiodarone for sustained ventricular tachyarrhythmias associated with structural heart diseases. AB - BACKGROUND: Although an electrophysiologic study (EPS) and Holter-monitoring are often helpful in evaluating the efficacy of antiarrhythmic drugs in patients with ventricular tachyarrhythmias (ventricular tachycardia/fibrillation (VT/VF)), the efficacy of EPS- or Holter-guided oral amiodarone therapy in Japanese patients is still unclear. METHODS AND RESULTS: EPS was performed 1 month after starting amiodarone, and Holter-monitoring was recorded before and 1 month after amiodarone in 188 patients with sustained VT/VF because of structural heart diseases. In spite of the judgment of EPS (n=89) or Holter (n=75), all patients continued amiodarone. Patients were followed up to 3 years and the primary endpoint was VT/VF recurrence and secondary endpoint was death by all cause. Kaplan-Meier estimated the risk of VT/VF recurrence was significantly smaller with EPS-guided amiodarone (p<0.01) but not with Holter-guided amiodarone. Multivariate Cox hazard analysis revealed that EPS-guided amiodarone was an independent factor suppressing the recurrence of VT/VF (p<0.05, 95% confidence interval =0.15 to 0.96). In the subgroup analysis, EPS-guided amiodarone was effective in patients with relatively well-preserved left ventricular ejection fraction (LVEF > or =0.30) but not in patients with lower LVEF (LVEF <0.30). CONCLUSION: EPS-guided amiodarone was useful for preventing recurrence of VT/VF in patients with a relatively well-preserved LVEF, but not always beneficial in patients with a lower LVEF. PMID- 18159107 TI - Magnetic dispersion of the late repolarization in Brugada syndrome. AB - BACKGROUND: Magnetocardiography (MCG) is a new noninvasive modality for recording cardiac depolarization and repolarization and was used in the present study to evaluate abnormalities in patients with Brugada syndrome (BS). METHODS AND RESULTS: The MCG findings of 10 BS, 21 right bundle branch block (RBBB), and 34 normal patients were compared. On the horizontal spatiotemporal activation graph (STAG), the r' waves were more frequently located on the right side in the RBBB than in the normal (p=0.001) or BS groups (p=0.001). The maximum current angles of the r' wave fell into the northwest axis in all BS patients as compared to having a right axis deviation in 19 of 21 RBBB patients (90.4%, p=0.001). In the magnetic field and current density vector maps during late repolarization, the BS group had a non-dipole pattern more frequently and a higher number of poles compared with the normal (p=0.001) and RBBB groups (p=0.001). CONCLUSIONS: During depolarization, the horizontal STAG location and maximum current angle of the r' wave were beneficial in differentiating BS from RBBB and normal. The magnetic dispersion was a more frequently observed finding in BS patients than in RBBB and normal patients during late repolarization. PMID- 18159108 TI - Effects of antiarrhythmic drugs on inappropriate shocks in patients with implantable cardioverter defibrillators. AB - BACKGROUND: Patients with atrial fibrillation (AF) or congestive heart failure (CHF) are more vulnerable to inappropriate shocks from implantable cardioverter defibrillators (ICDs), but the effect of antiarrhythmic drugs in these patients remains unknown. METHODS AND RESULTS: A total of 55 patients with AF and/or CHF (New York Heart Association functional class > or =III) who had ICDs were divided into 3 groups [amiodarone (n=24), sotalol (n=12), beta-blocker (n=19)] and the cumulative rates of inappropriate shocks were compared. The baseline characteristics of the 3 groups were not significantly different. The 4-year event rate of inappropriate shocks was 27.3% in the amiodarone group, 54.3% in the sotalol group, and 70.6% in the beta-blocker group (amiodarone vs beta blocker: log-rank p=0.003; sotalol vs beta-blocker: log-rank p=0.16; amiodarone vs sotalol: log-rank p=0.29). Amiodarone reduced the risk of inappropriate shocks significantly as compared with beta-blockers (hazard ratio (HR) 0.17; 95% confidence interval (CI) 0.05-0.64; p=0.008), whereas sotalol did not (HR 0.57; 95%CI 0.19-1.68; p=0.3). Amiodarone was discontinued in 4 patients (16.7%) because of pulmonary toxicity and the dose was reduced in 4 patients (16.7%) because of a thyroid function abnormality. CONCLUSIONS: Amiodarone is more effective than sotalol or beta-blockers in preventing inappropriate ICD shocks in patients with AF or CHF, but it has a significant risk of drug-related adverse effects. PMID- 18159109 TI - Local release of catecholamines from the hearts of patients with tako-tsubo-like left ventricular dysfunction. AB - BACKGROUND: The precise mechanism of tako-tsubo-like left ventricular (LV) dysfunction remains unclear, although recent studies have shown that activation of sympathetic tone might be involved. However, local release of catecholamines from cardiac sympathetic efferent neurons in patients with tako-tsubo-like LV dysfunction remains poorly understood. The purpose of this study was to investigate evidence of local release of catecholamines from the hearts of patients with tako-tsubo-like LV dysfunction. METHODS AND RESULTS: Five consecutive patients with tako-tsubo-like LV dysfunction were studied. After confirming LV apical ballooning and a normal coronary angiogram, sampling of blood for the measurement of plasma catecholamine levels was performed at the aortic root (Ao) and coronary sinus (CS). In all 5 cases, increased local release of norepinephrine from the heart was documented (597, 4,238, 2,121, 486, 371 pg/ml at the Ao; 836, 5,719, 3,386, 658, 472 pg/ml at the CS, respectively). CONCLUSIONS: Increased cardiac catecholamines might cause the transient LV apical ballooning in patients with tako-tsubo-like LV dysfunction. PMID- 18159110 TI - Persistently increased serum concentration of heart-type fatty acid-binding protein predicts adverse clinical outcomes in patients with chronic heart failure. AB - BACKGROUND: Heart-type fatty acid-binding protein (H-FABP) is a small cytosolic protein that is released into the circulation when the myocardium is injured. This study examined whether serial measurement of the H-FABP level provides additional prognostic information. METHODS AND RESULTS: Serum H-FABP levels were measured in 113 consecutive chronic heart failure (CHF) patients at both admission and discharge. The following 3 patterns of changes were identified. In 41 patients, H-FABP levels (<4.3 ng/ml) at both admission and discharge were normal (Group 1). The remaining 72 patients had high initial H-FABP levels (> or =4.3 ng/ml) at admission, and in 21 of them (29%), H-FABP decreased to the normal range at discharge (Group 2), whereas 51 had persistently high H-FABP levels despite improvement in symptoms and signs of CHF (Group 3). There were 33 cardiac events (29%) during the follow-up period, and Group 3 had significantly higher cardiac event rates than Groups 1 and 2 (p=0.0002). Group 3 had the highest cardiac risk among the groups (hazard ratio 5.68, p=0.012). CONCLUSION: Serial measurement of the H-FABP level is a new monitoring tool that provides information to guide optimal therapy and management of CHF patients. PMID- 18159111 TI - Mechanisms and clinical implications of asymptomatic mitral regurgitation in patients with left ventricular dysfunction. AB - BACKGROUND: Functional mitral regurgitation (MR) is common in patients with chronic heart failure (CHF). During exercise, hemodynamic changes such as elevation of blood pressure and an increase in the left ventricular end-diastolic dimension may increase MR. Severity of CHF is reflected by exercise tolerance and, therefore, MR during exercise is supposed to indicate the severity of heart failure. The degree of MR increase and left ventricle (LV) shape was quantified during exercise in CHF patients and were compared with exercise tolerance. METHODS AND RESULTS: Twenty patients with CHF (mean age: 63 years) underwent dynamic cycle exercise at steady-state levels of 80% and 150% of the anaerobic threshold (AT). The MR jet area and left atrial (LA) area were measured during exercise and the ratio of MR/LA was calculated. The LV shape was assessed by calculating the ratio of the major to the minor axis. The MR/LA ratio increased during exercise (rest: 16.9+/-6.5%, 80% AT: 21.9+/-8.9%, 150% AT: 30.9+/-11.2%; p<0.01) and the LV shape became more spherical (rest: 1.34+/-0.10, 80% AT: 1.31+/ 0.10, 150% AT: 1.23+/-0.11; p<0.05). There was a negative correlation between MR/LA ratio and the ratio of the major to the minor axis (-0.722, r<0.01). CONCLUSIONS: MR during exercise increases as the severity of CHF increases. Functional MR is correlated with the shape of the LV. PMID- 18159112 TI - Right bundle branch block and impaired left ventricular function as evidence of a left ventricular conduction delay. AB - BACKGROUND: The indications and efficacy of cardiac resynchronization therapy (CRT) have not been sufficiently clarified in patients with right bundle branch block (RBBB). METHODS AND RESULTS: This study included 55 patients with normal QRS morphology and duration (Control-Gr) and 49 patients with complete RBBB (CRBBB-Gr). Using tissue Doppler imaging, the time difference (TD) between the electromechanical delay of the septal wall, left ventricular (LV) lateral wall, and right ventricular free wall were measured. Using tissue tracking imaging, the coefficient of the time variation from the beginning of the QRS to the peak displacement time of 6 regions of the LV (CV-PMDLV) was calculated. The TD between the septal wall and that of the LV lateral wall (TDSEPT-LAT) did not differ between the Control-Gr and RBBB-Gr. However, a significant difference was found in the TDSEPT-LAT between the CRBBB patients with LV systolic dysfunction (ejection fraction (EF) < or =50%) and those with normal LV function (EF >50%; p<0.001). The CV-PMDLV was greater in the CRBBB patients with LV systolic dysfunction than in those with a normal LV function (p<0.05). The RBBB-Gr patients with LV dysfunction and a great TDSEPT-LAT, improved clinically after the CRT. CONCLUSIONS: The presence of RBBB and LV dysfunction may indicate LV dyssynchrony and a heterogeneous mechanical dysfunction. PMID- 18159113 TI - Implications of mutations of activin receptor-like kinase 1 gene (ALK1) in addition to bone morphogenetic protein receptor II gene (BMPR2) in children with pulmonary arterial hypertension. AB - BACKGROUND: Mutations of the bone morphogenetic protein receptor II gene (BMPR2), and 1 mutation of the activin receptor-like kinase 1 gene (ALK1) have been reported in patients with pulmonary arterial hypertension (PAH). METHODS AND RESULTS: A genomic study of ALK1 and BMPR2 was conducted in 21 PAH probands under 16 years of age to study the relationship between the clinical features of the patients and these genes. In all 4 familial aggregates of PAH, 3 ALK1 or 1 BMPR2 mutations were identified. Among 17 probands aged between 4 and 14 years with idiopathic PAH, 2 ALK1 mutations (2/17: 11.8%) and 3 BMPR2 mutations (3/17: 17.6%; 5 mutations in total: 5/17: 29.4%) were found. CONCLUSION: Each proband with the ALK1 mutation developed PAH, as did the probands with the BMPR2 mutation. Hence, it is proposed that ALK1 plays as notable a role as BMPR2 in the etiology of PAH. Furthermore, asymptomatic carriers with the ALK1 mutation within the serine - threonine kinase domain are at risk of developing PAH and hereditary hemorrhagic telangiectasia, so close follow-up is recommended for those individuals. PMID- 18159114 TI - Mortality among persons with a history of kawasaki disease in Japan: mortality among males with cardiac sequelae is significantly higher than that of the general population. AB - BACKGROUND: The long-term prognosis of those having a history of Kawasaki disease (KD) is still unknown. METHODS AND RESULTS: Between July 1982 and December 1992, 52 collaborating hospitals collected data on all patients having a new definite diagnosis of KD. Patients were followed-up until December 31, 2004 or their death. Standardized mortality ratios (SMRs) were calculated based on the Japanese vital statistics data. Of 6,576 patients enrolled, 36 (27 males, 9 females) died and the SMR was 1.14. Despite the high SMRs during the acute phase, the mortality rate was not high after the acute phase for the entire group of patients. Although the SMR after the acute phase was 0.71 for those without cardiac sequelae, 10 males (but none of the females) with cardiac sequelae died during the observation period; and the SMR for the male group with cardiac sequelae was 2.55 (95% confidence interval: 1.23-4.70). CONCLUSIONS: The mortality rate among males with cardiac sequelae because of KD was significantly higher in this cohort than in the general population. On the other hand, those for females with the sequelae and for both males and females without sequelae were not elevated. PMID- 18159115 TI - Whole-body periodic acceleration enhances brachial endothelial function. AB - BACKGROUND: Periodic acceleration in the direction of the spinal axis through repetitive movement increases the shear stress on the vascular endothelium. In the present study it was assessed whether whole-body periodic acceleration with a new device would enhance endothelial function in sedentary adult volunteers. METHODS AND RESULTS: Twenty-six sedentary subjects (44+/-3 years) were randomly assigned to remain sedentary or perform exercise training for 4 weeks, followed by crossover. Periodic acceleration was applied with a horizontal motion platform at 2-3 Hz and approximately +/-2.2 m/s2 for 45 min. Increases in the brachial artery diameter were examined at rest, during reactive hyperemia (flow-mediated dilatation: %FMD) and after sublingual administration of 0.3 mg nitroglycerin (%NTG) using high-resolution ultrasound. All subjects completed the study with no adverse side-effects. There were no significant changes in the resting heart rate or arterial pressure, body weight, or lipid profiles during the study. Although %FMD did not change during the non-training period with periodic acceleration, it significantly increased from 7.3+/-0.4% at baseline to 8.4+/-0.4% after the training period (p<0.05), while %NTG remained unchanged. CONCLUSIONS: Whole-body periodic acceleration with a horizontal motion platform improved vascular endothelial function in sedentary adults. This device might offer an alternative to active exercise for patients whose medical condition limits physical activity. PMID- 18159116 TI - Inhalation exposure to carbon black induces inflammatory response in rats. AB - BACKGROUND: A link between exposure to fine particulate matter and cardiovascular events has been established. Inhaled nanoparticles are thought to pass through the lungs to reach other tissues via systemic circulation and to induce cell or tissue injuries. It was recently shown that long-term exposure to intra-tracheal dispersion of nano-sized carbon black (CB) exacerbates atherosclerotic lesions in low-density lipoprotein receptor-deficient mice. Because intra-tracheal dispersion of CB may be associated with aggregate formation and may not be an ideal method for CB exposure, whole-body inhalation exposure was used in the present study, the aim of which was to examine whether exposure of rats to nano sized CB particles by inhalation leads to translocation of these particles into the circulation, exerting direct adverse effects on extrapulmonary tissues. METHODS AND RESULTS: Sprague-Dawley rats were exposed to a high dose of CB or filtered air for 6 h/day, 5 days a week for a total of 4 weeks. Although the presence of CB was confirmed in pulmonary macrophages, electron microscopic survey did not detect CB in other tissues including liver, spleen and aorta. CB exposure raised blood pressure levels in an exposure-time dependent manner. Levels of circulating inflammatory marker proteins, including monocyte chemoattractant protein-1, interleukin-6, and C-reactive protein, were higher in the CB-treated group than in the controls. CONCLUSION: Evidence of translocation of inhaled CB was not obtained. It is likely that inhaled nano-sized CB particles form aggregations in the lung and do not exert direct adverse effects on extrapulmonary tissues. Air-pollution-mediated cardiovascular events appear to be induced by the low-grade inflammatory response to the accumulation of aggregated nano-sized particles in the lung. PMID- 18159117 TI - Shock wave therapy applied to rat bone marrow-derived mononuclear cells enhances formation of cells stained positive for CD31 and vascular endothelial growth factor. AB - BACKGROUND: This study tested the hypothesis that shock wave (SW) therapy applied to bone marrow-derived mononuclear cells (BMDMNCs) enhances the formation of vascular endothelial growth factor (VEGF) and positively stained CD31 (CD31+) cells, an endothelial phenotype. METHODS AND RESULTS: The BMDMNCs (approximately 1.2 x 10(6) cells/2 femoral bones) were obtained from adult male Sprague-Dawley rats and SW therapy was applied once to BMDMNCs in group I (140 SW: defined as 140 shots in total, given at 0.09 mJ/mm2), group II (280 SW), and group III (560 SW). Group IV was not treated by SW and served as the control group. Six experiments were done in each group. The BMDMNCs were cultured following SW therapy and the supernatants were collected on days 1, 3, 7 and 14 for assessment of VEGF levels. Immunocytochemical staining and flow cytometric measurement were performed on days 0 and 14. Experimental results demonstrated that VEGF levels were significantly higher in groups I-III than in group IV, and in group II than in group I at all intervals, and in group II than in group III on day 14 (all p values <0.005). Additionally, the number of positively stained VEGF cells on days 1, 3 and 14 and the number of newly formed CD31+ cells on day 14 were significantly higher in group II than in group IV (all p values <0.001). CONCLUSIONS: These data suggest that application of SW to BMDMNCs significantly enhanced VEGF production and promoted differentiation of BMDMNCs into endothelial phenotype cells. PMID- 18159118 TI - Heavily calcified coronary lesions preclude strut apposition despite high pressure balloon dilatation and rotational atherectomy: in-vivo demonstration with optical coherence tomography. AB - Heavily calcified lesions (HCL) continue to present challenges that are not always solved by modern low-profile, non-compliant high-pressure balloons, or bladed balloons. Uncrossable or unexpandable lesions need lesion modification, using ablating devices such as rotational atherectomy. Three cases of HCL treated with drug-eluting stents, using a new intravascular imaging device, Optical Coherence Tomography with 10-fold superior resolution and fewer artifacts compared with conventional intravascular ultrasound, are presented. Insights from using this highly sensitive imaging technique outline the high prevalence of persistent stent strut malapposition in this group, despite the use of high pressure dilatation or rotational atherectomy. PMID- 18159119 TI - Intraoperative endotoxin adsorption as a new therapeutic option for mycotic aortic aneurysm. AB - An 82-year-old man with a mycotic aortic aneurysm of the distal arch underwent urgent surgery because of sepsis. The infected aortic arch was excised, replaced with a rifampicin-bonded prosthetic graft, and covered with omentum. Direct hemoperfusion using polymyxin B-immobilized fiber (PMX-DHP) was intraoperatively carried out in parallel with the cardiopulmonary circuit. Intraoperative PMX-DHP dramatically reduced the level of plasma endotoxin, and ameliorated hemodynamic instability and oxygenation, resulting in smooth weaning from cardiopulmonary bypass. Intraoperative endotoxin adsorption is technically simple and easy, effective in hemodynamic stabilization, and so could be a new therapeutic option for mycotic aortic aneurysm. PMID- 18159120 TI - Young woman affected by a rare form of familial connective tissue disorder associated with multiple arterial pulmonary stenosis and severe pulmonary hypertension. AB - A woman with skin findings of a connective tissue disorder, typical of Ehlers Danlos syndrome, was admitted to the Cardiology Division because of signs of congestive heart failure. Electrocardiogram showed sinus tachycardia, signs of right ventricular enlargement and hypertrophy. Echocardiogram showed right ventricular dilatation, and severe tricuspid regurgitation with indirect signs of severe pulmonary systolic hypertension. Chest computed tomography revealed bilateral and diffuse involvement of the peripheral pulmonary arteries, with kinking and elongation of the pulmonary vessels associated with multiple stenoses and post-stenotic dilatation. On artery angiography an elongation of the aortic root with kinking and coiling of the carotid and vertebral vessels was also detected. This young patient exhibited features of arterial tortuosity syndrome, an uncommon connective tissue disorder, with peculiar dysmorphism and clinical signs overlapping Ehlers-Danlos syndrome. PMID- 18159121 TI - Late stent thrombosis in patients receiving ticlopidine and aspirin after sirolimus-eluting stent implantation. AB - BACKGROUND: There is little information about the efficacy of ticlopidine plus aspirin after sirolimus-eluting stent (SES) implantation. METHODS AND RESULTS: The incidence of stent thrombosis was evaluated in 1,029 patients receiving ticlopidine and aspirin after SES deployment. Clinical follow-up was obtained in 98.9% (mean follow-up 17.0+/-7.9 months). Early stent thrombosis was observed in 5 patients (0.49%). There was 1 case each of late (0.1%) and very late stent thrombosis (0.1%). CONCLUSION: Late and very late stent thrombosis in Japanese patients receiving ticlopidine and aspirin after SES deployment occurs infrequently. PMID- 18159122 TI - Letter by Sata et Al regarding article,"Plasma brain natriuretic peptide levels indicating thromboembolism in very elderly patients with non-valvular atrial fibrillation". PMID- 18159124 TI - Identification of novel nuclear localization signals of Drosophila myeloid leukemia factor. AB - Myeloid leukemia factor 1 (MLF1) was first identified as part of a leukemic fusion protein produced by a chromosomal translocation, and MLF family proteins are present in many animals. In mammalian cells, MLF1 has been described as mainly cytoplasmic, but in Drosophila, one of the dMLF isoforms (dMLFA) localized mainly in the nucleus while the other isoform (dMLFB), that appears to be produced by the alternative splicing, displays both nuclear and cytoplasmic localization. To investigate the difference in subcellular localization between MLF family members, we examined the subcellular localization of deletion mutants of dMLFA isoform. The analyses showed that the C-terminal 40 amino acid region of dMLFA is necessary and sufficient for nuclear localization. Based on amino acid sequences, we hypothesized that two nuclear localization signals (NLSs) are present within the region. Site-directed mutagenesis of critical residues within the two putative NLSs leads to loss of nuclear localization, suggesting that both NLS motifs are necessary for nuclear localization. PMID- 18159125 TI - Involvement of p120 carboxy-terminal domain in cadherin trafficking. AB - P120 plays an essential role in cadherin turnover. The molecular mechanism involved, however, remains only partially understood. Here, using a gene trap targeting technique, we replaced the genomic sequence of p120 with HA-tagged p120 cDNA in mouse teratocarcinoma F9 cells. In the p120 knock-in (p120KI) cells, we found that the expression level of p120 was severely reduced and that the expression level of other components of the cadherin-catenin complex was also reduced. The stable expression of various p120 mutants in p120KI cells revealed that the armadillo repeat domain of p120 is sufficient to restore the expression level of E-cadherin. In p120KI cells, internalized E-cadherin was frequently detected as large aggregates. Transient expression of wild-type p120 and mutant p120 lacking the N-terminal region induced both relocalization of E-cadherin at the cell-cell boundaries and the disappearance of cytoplasmic E-cadherin aggregates. Transient expression of mutant p120 lacking the C-terminal region, however, only induced a small increase in E-cadherin signals at the cell-cell boundary. In these cells, the cytoplasmic E-cadherin signals became brighter and the expressed mutant p120 was incorporated in the E-cadherin aggregates. These results suggested the novel function of the p120 C-terminal region in regulating the trafficking of cytoplasmic E-cadherin. PMID- 18159126 TI - Call for papers to theme issue on membrane transporters: an opportunity to boost transporter studies. PMID- 18159127 TI - Xenobiotic transporter-adaptor network. AB - Many types of xenobiotic transporters have been identified. They generally exhibit multispecific recognition of various types of substrates, and mediate membrane permeation of therapeutic agents, thereby playing important roles in drug absorption and disposition. It has recently been proposed that protein protein interactions involving the xenobiotic transporters may affect their function, localization and expression on plasma membranes. So-called adaptor proteins that directly interact with the transporters include PDZ domain containing proteins (PSD95, Dlg and ZO1). These PDZ adaptors have multiple PDZ domains in their structure, and each PDZ domain can interact with the cytosolic region of the transporters, and so it has been hypothesized that transporters are localized within networks consisting of several transporters and adaptors. Interaction with a PDZ adaptor is essential for the cell-surface localization of at least some xenobiotic transporters, and therefore, such interaction could be required for efficiency and fidelity in the vectorial transport of xenobiotics and therapeutic agents in epithelial cells. This review article summarizes recent evidence on the interactions of xenobiotic transporters with adaptor proteins, and presents a working hypothesis concerning their pharmacological significance. PMID- 18159128 TI - Prediction of the metabolic interaction of nateglinide with other drugs based on in vitro studies. AB - Nateglinide is an antidiabetic agent metabolized by CYP2C9 and CYP3A4; hence inhibitors of these CYP isozymes may interact with nateglinide. There are, however, only limited in vitro data on how to predict drug-drug interactions in vivo. We examined the effects of 18 drugs that may be prescribed together with nateglinide (metformin, buformin, aspirin, gemfibrozil, simvastatin, pioglitazone, rosiglitazone, carbamazepine, clarithromycin, gliclazide, clofibrate, fluconazole, bezafibrate, phenylbutazone, nifedipine, famotidine, ibuprofen and miconazole) on the conversion of nateglinide to its major metabolite (N-[trans-4-(1-hydroxy-1-methylethyl)-cyclohexanecarbonyl]-D phenylalanine) using human liver microsomes. Eight compounds showed a<50% inhibitory effect and we estimated the K(i) values for the remaining 10 compounds. Except for fluconazole and miconazole, 1+I(in, max, u)/K(i) calculated from the K(i) values, was approximately 1 and thus the possibility of a drug-drug interaction was considered low. The value for fluconazole suggested the risk of interaction and agreed with the results of clinical studies in which the AUC of nateglinide increased by 48% when it was co-administered with fluconazole. The present study showed that nateglinide metabolism would hardly be affected by the drugs used in this study, except for miconazole and fluconazole that are potent inhibitors of multiple isoforms of CYPs. PMID- 18159129 TI - Decreased renal accumulation and toxicity of a new VCM formulation in rats with chronic renal failure. AB - We previously reported that MEEK, a generic product of vancomycin hydrochloride (VCM), was less nephrotoxic than a conventional preparation (S-VCM) in normal rats at a nephrotoxic dose (400 mg/kg) of VCM.(1)) To infer the clinical significance of this finding, we compared the risk of nephrotoxicity of these two formulations in rats with chronic renal failure in this study. MEEK or S-VCM was given intravenously to two weeks post-5/6 nephrectomy rats, and the pharmacokinetic profile of VCM and pathological evaluation were compared. There were no differences at the daily clinical dose (40 mg/kg), but at the twice the daily clinical dose (80 mg/kg), the mean plasma concentration of VCM was higher after S-VCM administration than after MEEK and the CL(tot) and CL(r) decreased to approximately 60% of those after MEEK. The renal tissue concentration of VCM was 1.5-fold higher at 24hr after S-VCM administration than after MEEK. Pathologically, no marked differences between the findings were observed at 24hr after administration of each formulation. These findings suggest that MEEK reduces renal damage caused by VCM and prevents the iatrogenic aggravation of nephrotoxicity. These results hold out hope that MEEK will permit high-dose administration of VCM, while revealing clinical significance of the nephrotoxicity-reduction by MEEK. PMID- 18159130 TI - In vitro evaluation of photosensitivity risk related to genetic polymorphisms of human ABC transporter ABCG2 and inhibition by drugs. AB - Since porphyrins are regarded as endogenous substrates for the ATP-binding cassette (ABC) transporter ABCG2, it is hypothesized that functional impairment owing to genetic polymorphisms or inhibition of ABCG2 by drugs may result in a disruption of cellular porphyrin homeostasis. In the present study, we expressed ABCG2 genetic variants, i.e., V12M, Q141K, S441N, and F489L, as well as the wild type (WT) in Flp-In-293 cells to examine the hypothesis. Cells expressing S441N and F489L variants exhibited high levels of both cellularly accumulated pheophorbide a and photosensitivity, when those cells were incubated with pheophorbide a and irradiated with visible light. To further elucidate the significance of ABCG2 in cellular porphyrin homeostasis, we observed cellular accumulation and compartmentation of porphyrin and pheophorbide a by means of a new fluorescence microscopy technology, and found that accumulation of porphyrin and pheophorbide a in the cytoplasm compartment was maintained at low levels in Flp-In-293 cells expressing ABCG2 WT, V12M, or Q141K. When ABCG2 was inhibited by imatinib or novobiocin, however, those cells became sensitive to light. Based on these results, it is strongly suggested that certain genetic polymorphisms and/or inhibition of ABCG2 by drugs can enhance the potential risk of photosensitivity. PMID- 18159131 TI - Distinct effects of omeprazole and rabeprazole on the tacrolimus blood concentration in a kidney transplant recipient. AB - Proton-pump inhibitors (PPIs, e.g. omeprazole and rabeprazole) are often administered to transplant patients as a treatment or prophylaxis for ulcers after surgery. Since tacrolimus and PPIs share the CYP3A4 system for metabolism, pharmacokinetic interactions are anticipated when they are administered simultaneously. We present a Japanese male patient who underwent a living-donor kidney transplantation having received tacrolimus, mycophenolate mofetil, and prednisolone for immunosuppression. The concentration/dose (C/D) ratio for tacrolimus was markedly higher during the period of treatment with omeprazole than ranitidine or rabeprazole. The results of liver functional tests were within the normal range during the use of these three antacid drugs. Since the higher C/D ratio for tacrolimus when omeprazole was being administered did not result from a decrease in the elimination of tacrolimus due to hepatic dysfunction, drug interaction between omeprazole and tacrolimus was strongly suspected. The present case indicates that rabeprazole can be used safely in place of omeprazole in kidney transplant recipients receiving tacrolimus. PMID- 18159132 TI - Capsaicin-induced increase of intestinal cefazolin absorption in rats. AB - The effect of capsaicin on intestinal cefazolin absorption was examined by means of an in situ closed loop method in rats to clarify whether the vanilloid receptor (TRPV1) is involved in drug absorption driven by passive diffusion. In control experiments with 1 mg/mL cefazolin, the amount of cefazolin absorbed from the closed loop was 15.3+/-1.5 microg/cm in the rat jejunum. The absorption amount was increased to 22.8+/-0.9 and 23.4+/-2.4 microg/cm when capsaicin was applied with cefazolin at concentrations of 10 and 400 microM, respectively. The enhancing effect of capsaicin on cefazolin absorption was suppressed when ruthenium red, a non-selective inhibitor of transient receptor potential (TRP) cation channels, was intravenously infused into the rat during the experiment. Cefazolin accumulation in the intestinal tissue was not altered in the presence of capsaicin. Collectively, the mechanism accounting for the capsaicin-induced increase in the intestinal cefazolin absorption is probably that capsaicin associating with TRPV1 increases the intrinsic permeability of cefazolin in intestine. PMID- 18159133 TI - Down-regulation of intestinal multidrug resistance-associated protein 2 in long evans cinnamon rats. AB - Wilson's disease is an inherited, autosomal recessive disorder of copper accumulation and toxicity. Lifelong chelation therapy is essential in all Wilson's disease patients. Intestinal absorption of some compounds is limited partly because they are preferentially transported in the secretory direction. Several ATP-binding cassette (ABC) transporters are expressed in the apical membrane of the small intestine and secrete various drugs into the lumen. In this study, we investigated the characteristics of the intestinal efflux ABC transporters in LEC rats. We found that the expression of multidrug resistance associated protein 2 (Mrp2) in the jejunum of Long-Evans Cinnamon (LEC) rats, an animal model for Wilson's disease, is decreased. PMID- 18159134 TI - Genetic variations and frequencies of major haplotypes in SLCO1B1 encoding the transporter OATP1B1 in Japanese subjects: SLCO1B1*17 is more prevalent than *15. AB - A liver-specific transporter organic anion transporting polypeptide 1B1 (OATP1B1, also known as OATP-C) is encoded by SLCO1B1 and mediates uptake of various endogenous and exogenous compounds from blood into hepatocytes. In this study, 15 SLCO1B1 exons (including non-coding exon 1) and their flanking introns were comprehensively screened for genetic variations in 177 Japanese subjects. Sixty two genetic variations, including 28 novel ones, were found: 7 in the 5'-flanking region, 1 in the 5'-untranslated region (UTR), 13 in the coding exons (9 nonsynonymous and 4 synonymous variations), 5 in the 3'-UTR, and 36 in the introns. Five novel nonsynonymous variations, 311T>A (Met104Lys), 509T>C (Met170Thr), 601A>G (Lys201Glu), 1553C>T (Ser518Leu), and 1738C>T (Arg580Stop), were found as heterozygotes. The allele frequencies were 0.008 for 1738C>T (Arg580Stop) and 0.003 for the four other variations. Arg580Stop having a stop codon at codon 580 results in loss of half of transmembrane domain (TMD) 11, TMD12, and a cytoplasmic tail, which might affect transport activity. In addition, novel variations, IVS12-1G>T at the splice acceptor site and -3A>C in the Kozak motif, were detected at 0.003 and 0.014 frequencies, respectively. Haplotype analysis using -11187G>A, -3A>C, IVS12-1G>T and 9 nonsynonymous variations revealed that the haplotype frequencies for (*)1b, (*)5, (*)15, and (*)17 were 0.469, 0.000 (not detected), 0.037, and 0.133, respectively. These data would provide fundamental and useful information for pharmacogenetic studies on OATP1B1-transported drugs in Japanese. PMID- 18159135 TI - Genetic variations of VDR/NR1I1 encoding vitamin D receptor in a Japanese population. AB - The vitamin D receptor (VDR) is a transcriptional factor responsive to 1alpha,25 dihydroxyvitamin D(3) and lithocholic acid, and induces expression of drug metabolizing enzymes CYP3A4, CYP2B6 and CYP2C9. In this study, the promoter regions, 14 exons (including 6 exon 1's) and their flanking introns of VDR were comprehensively screened for genetic variations in 107 Japanese subjects. Sixty one genetic variations including 25 novel ones were found: 9 in the 5'-flanking region, 2 in the 5'-untranslated region (UTR), 7 in the coding exons (5 synonymous and 2 nonsynonymous variations), 12 in the 3'-UTR, 19 in the introns between the exon 1's, and 12 in introns 2 to 8. Of these, one novel nonsynonymous variation, 154A>G (Met52Val), was detected with an allele frequency of 0.005. The single nucleotide polymorphisms (SNPs) that increase VDR expression or activity, 29649G>A, 2T>C and 1592((*)308)C>A tagging linked variations in the 3'-UTR, were detected at 0.430, 0.636, and 0.318 allele frequencies, respectively. Another SNP, -26930A>G, with reduced VDR transcription was found at a 0.028 frequency. These findings would be useful for association studies on VDR variations in Japanese. PMID- 18159136 TI - Examination of the effectiveness of DVD decision support tools for patients with unruptured cerebral aneurysms. AB - Preventative treatments for unruptured cerebral aneurysms include craniotomy, endovascular treatment, and follow up. Since there is no agreement as to the best procedure, it is important to provide adequate information so that the patient and physician can share in the decision-making process. A multi-media DVD was created to inform patients of the facts. This study examined how effectively this DVD changes patients' recognition including knowledge of unruptured cerebral aneurysms. Forty-seven patients with unruptured cerebral aneurysms who sought neurosurgery consultation between December 2005 and February 2006 completed a questionnaire before and after watching the DVD, as well as at 3 months follow up. Before watching the DVD, the average knowledge score was 8.72 out of 15 total points. The average score increased to 12.4 after watching the DVD (p < 0.001). At 3 months follow up, the average score was 10.34, which was still higher than before watching the DVD (p < 0.01). Participants' knowledge about treatment methods also increased after watching the DVD (p < 0.001). Compared to 63.2% who were satisfied with their treatment decision before watching the DVD, 69.6% were satisfied with their decision after watching the DVD. All participants responded that the use of multi-media images was helpful in better understanding treatment options and in making informed decisions. The DVD was favorably accepted as a decision support tool by patients with unruptured cerebral aneurysm and effectively increased patients' knowledge. PMID- 18159137 TI - Effect of raloxifene on cerebral vasospasm following experimental subarachnoid hemorrhage in rats. AB - The effect of raloxifene on cerebral vasospasm following experimental subarachnoid hemorrhage (SAH) was investigated in a rat model. Seven groups of seven rats underwent no SAH, no treatment; SAH only; SAH plus vehicle; SAH plus 3 days intraperitoneal raloxifene treatment; SAH plus 4 days intraperitoneal raloxifene treatment; SAH plus 3 days intrathecal raloxifene treatment; and SAH plus 4 days intrathecal raloxifene treatment. The basilar artery cross-sectional areas were measured at 72 or 96 hours following SAH. The results showed raloxifene decreased SAH-induced cerebral vasospasm in all treatment groups, and suggested no difference between intraperitoneal and intrathecal application, or between 3 days and 4 days of raloxifene treatment. The present study demonstrates that raloxifene is a potential therapeutic agent against cerebral vasospasm after SAH. PMID- 18159138 TI - Prognostic factors in elderly patients with supratentorial malignant gliomas. AB - The prognostic factors were retrospectively analyzed in 30 patients aged 70 years or over with supratentorial malignant gliomas treated by surgery in our hospital. The histological diagnosis was confirmed as grade 3 in 13 patients and grade 4 in 17. Seventeen patients received adjuvant radiation therapy. Only 10 patients underwent chemotherapy. Survival time was analyzed with the Kaplan-Meier method. Prognostic factors were obtained from the Cox proportional hazards model. Univariate analysis showed preoperative Karnofsky performance status (KPS) score of 70 or greater and radiation therapy were significantly associated with longer survival. However, multivariate analysis revealed that preoperative KPS score of 70 or greater was the only independent prognostic factor and radiation therapy lost its significance due to selection bias. Neurological deterioration and medical complications occurred in six and seven patients, respectively. Performance status rather than histological grade is the key prognostic factor in elderly patients with supratentorial malignant gliomas. Patients with good preoperative KPS score should be aggressively treated with extensive resection and radiotherapy. PMID- 18159139 TI - Intracranial pial arteriovenous fistula. AB - A 33-year-old woman presented with a rare intracranial pial arteriovenous fistula manifesting as monoparesis and hypesthesia of the right lower extremity. Computed tomography demonstrated an approximately 10-mm diameter subcortical hematoma in the left postcentral gyrus. Two months after suffering the ictus, angiography demonstrated a pial arteriovenous fistula in the late arterial phase fed by the left paracentral artery and drained into the left precentral vein. No nidus or dural arteriovenous fistula was detected. Left parietal craniotomy was performed and the pial arteriovenous fistula was extirpated by electrocoagulation. Intraoperative angiography demonstrated disappearance of the fistula. She experienced no postoperative neurological deterioration, but hypesthesia of the right leg persisted. Obliteration of the pial arteriovenous fistula was reconfirmed by postoperative angiography. She suffered no rebleeding episodes during the 36-month follow-up period. Pial arteriovenous fistula causing mild symptoms should be treated by flow disconnection because the direct arteriovenous shunt and attendant high blood flow usually results in huge venous varices. To determine whether direct surgery or endovascular treatment is appropriate, the position and shape of the lesion must be known. PMID- 18159140 TI - Spontaneous intracranial hypotension associated with dural sinus thrombosis. AB - A 32-year-old man presented with a rare case of spontaneous intracranial hypotension (SIH) associated with dural sinus thrombosis (DST) manifesting as severe postural headache which was relieved by lying down. Initial cerebrospinal fluid pressure was low. He was treated with hydration and rest under a diagnosis of SIH. Magnetic resonance (MR) imaging after 1 month showed signs of both DST and SIH. However, the patient did not demonstrate any of the classical symptoms associated with DST. The patient underwent an epidural blood patch procedure. His headache was relieved and MR imaging showed improvement of both SIH and DST. This case suggests that epidural blood patch may be effective in some cases of SIH associated with DST. PMID- 18159141 TI - Rapid revascularization after therapeutic parent artery occlusion for a large intracavernous carotid artery aneurysm. AB - A 21-year-old woman presented with an unruptured large intracavernous aneurysm, which was spontaneously revascularized via unusual collateral pathways a short time after extracranial-intracranial bypass and surgical ligation of the proximal internal carotid artery. The patient had been treated for a large basilar trunk aneurysm with intraaneurysmal embolization using Guglielmi detachable coils, and an intracavernous carotid artery aneurysm treated conservatively. Two years later, the patient presented with right abducens nerve palsy, and was referred to our hospital. She had small nevi in the right forehead and eyelid. Cerebral angiography revealed enlargement of the intracavernous aneurysm. Superficial temporal artery-middle cerebral artery bypass followed by surgical carotid artery ligation were performed, and good patency of bypass and disappearance of the aneurysm were confirmed by intraoperative angiography. However, follow-up magnetic resonance angiography and cerebral angiography on the 20th postoperative day revealed revascularization of the internal carotid artery and the intracavernous carotid artery aneurysm via unusual collateral pathways. Subsequently, the recurrent aneurysm and the recanalized internal carotid artery were occluded by endovascular procedures. Histological examination of the nevus showed lack of properly organized vascular structures, and the diagnosis was angiodysplasia. The early development of unusual collateral pathway, and aneurysm formation at a young age might be related to the angiodysplasia. Revascularization is possible within a short time even in cases of intracavernous carotid artery aneurysm successfully treated with surgical ligation of the parent artery. PMID- 18159142 TI - Histological analysis of microbleed after surgical resection in a patient with moyamoya disease. AB - A 65-year-old male with moyamoya disease underwent surgical resection of a microbleed simultaneously with revascularization surgery. Histological examination identified several fragile arteries surrounding the microbleed. Microbleeds detected by T2*-weighted magnetic resonance imaging are considered to be a general marker of vascular vulnerability in cerebral angiopathy with a tendency to bleeding. The microbleeds observed in patients with moyamoya disease probably originate in bleeding from the fragile arteries. PMID- 18159143 TI - Papillary tumor of the pineal region. AB - A 48-year-old female presented with an extremely rare primary tumor of the pineal region with papillary features manifesting as morning headaches persisting for 1 month. Magnetic resonance imaging showed a well-defined mass, with some cystic components, in the region of the pineal gland. The tumor was completely removed through an occipital transtentorial approach in the prone position. Histological examination found a distinctive papillary growth pattern in which the vessels were covered by multiple layers of tumor cells. The histological diagnosis was papillary tumor of the pineal region (PTPR), which has recently been described as a distinct clinicopathological entity requiring careful follow up because the prognosis is not well understood. Postoperatively, the patient has continued to do well, with no recurrence at the 8-month follow-up examination. PTPR should be considered in the differential diagnosis of pineal tumors. PTPR may have been frequently misinterpreted in the past as either ependymoma or choroid plexus papilloma due to the similar morphology. PMID- 18159144 TI - Basal encephalocele associated with hypoplasia of the internal carotid artery. AB - A 22-year-old woman presented with a basal encephalocele associated with hypoplasia of the internal carotid artery (ICA) manifesting as a 6-year history of decreased vision in the right eye. She underwent encephalo-arterio-synangiosis under a diagnosis of hypoplasia of the ICA at age 6 years. Magnetic resonance imaging showed the encephalocele medial to the right temporal lobe. Frontotemporal craniotomy was performed for resection of the encephalocele and repair of the defect of the skull base. The pathogeneses of these developmental anomalies were probably related to developmental failure of the embryonic primordium during the 4th and 10th weeks. PMID- 18159145 TI - Symptomatic Rathke's cleft cyst with cavernous sinus syndrome. AB - An 88-year-old man presented with Rathke's cleft cyst (RCC) manifesting as complete cavernous sinus syndrome. He had no headache, endocrinological symptom, or blood abnormality. Neuroimaging revealed a cystic intrasellar lesion with lateral extension. The patient underwent surgery by a transsphenoidal approach. Histological examination revealed squamous and cuboidal epithelium. The diagnosis was RCC. RCC is rarely symptomatic, but enlargement and compression of the surrounding structures usually causes headache, visual field defects, or symptoms of pituitary dysfunction. The present case shows that RCC may manifest as complete cavernous sinus syndrome. PMID- 18159146 TI - [Review: clinical outcome after living donor liver transplantation in patients with hepatitis C virus-associated cirrhosis]. PMID- 18159147 TI - [Recent etiology and clinical features of acute viral hepatitis in a single center of Korea]. AB - BACKGROUND AND AIMS: The etiology of acute viral hepatitis in Korea has been dynamically changing during the recent years. The aim of this study was to investigate the recent etiology and the clinical features of acute viral hepatitis in a single center of Korea. METHODS: We performed a retrospective analysis of a prospective cohort of 55 patients who were diagnosed with acute viral hepatitis A to E during the period from May 2005 to August 2006. In addition to the clinically acute manifestations, the confirmatory serological tests were performed for the diagnosis of acute hepatitis A, B, C and E. RESULTS: The proportion of patients with acute viral hepatitis A, B, C, E and others were 56.4% (n=31), 12.7% (n=7), 18.2% (n=10), 9.1% (n=5) and 3.6% (n=2), respectively. The mean age of the patients with acute hepatitis A, B, C and E were 29.1+/-4.38, 38.7+/-11.72, 45.3+/-17.62 and 32.4+/-6.58 years, respectively. There was no fatal case. All cases of acute hepatitis B and six out of ten cases of acute hepatitis C recovered spontaneously. Four out of the five patients with acute hepatitis E had no history of travel to endemic area. CONCLUSIONS: The most common etiology of acute viral hepatitis in Korea is hepatitis A virus, and hepatitis C and B virus were the next most common causes. The sporadic cases of acute hepatitis E were not rare, and coinfection of HAV and HEV was observed. A multicenter, prospective study is warranted in the future. PMID- 18159148 TI - [The efficacy and safety of telbivudine in korean patients with chronic hepatitis B]. AB - BACKGROUND AND AIMS: Telbivudine is an L-nucleoside analogue with potent antiviral activity against hepatitis B virus (HBV). Clinical trials have shown that telbivudine is more potent than lamivudine for suppressing virus. METHODS: A total 101 Korean patients among 1,367 patients who participated in the phase III GLOBE trial were randomized in this study. All 101 HBeAg positive or HBeAg negative patients were assigned to treatment with 600 mg of telbivudine or 100 mg of lamivudine once daily. The primary efficacy endpoint (the "therapeutic response") was defined as suppression of the serum HBV DNA to less than 5 log10 copies/mL coupled with either normalization of the serum alanine aminotransferase level or loss of HBeAg. The secondary endpoints included the histologic response, serum HBV DNA reduction, serum alanine aminotransferase normalization and HBeAg loss for the HBeAg positive patients. This analysis includes the data collected at 52 weeks of treatment. RESULTS: Fifty four of 101 patients were assigned to telbivudine treatment and 47 patients were assigned to lamivudine treatment. At week 52, significantly more patients who were treated with telbivudine than those treated with lamivudine had a therapeutic response (83% vs 62%, respectively, P=0.017), their mean serum HBV DNA levels were more reduced (6.6 vs 5.6 log10 copies/mL, respectively, P=0.027), and they more often achieved PCR-undetectable levels of serum HBV DNA (74% vs 34%, P<0.0001). No virologic resistance to telbivudine was detected (0% vs 18%, respectively, P=0.001). Telbivudine was well tolerated and it had a safety profile comparable to lamivudine. CONCLUSIONS: Patients treated with telbivudine achieved earlier and more profound viral suppression than those treated with lamivudine. PMID- 18159149 TI - [Change of HBV DNA level as a predictor of HBeAg loss after lamivudine treatment]. AB - BACKGROUND AND AIMS: Lamivudine is an effective, safe therapeutic agent for the treatment of chronic hepatitis B. The aim of this study was to investigate whether early suppression of the viral load predicts HBeAg loss within 1 year during lamivudine therapy. METHODS: This prospective study encompassed 74 patients (mean age: 37.1 years, male/female: 51/23) who were positive HBeAg, their AST or ALT levels were > or =2 times the upper limit of normal and their HBV DNA was > or =10(5) copies/mL. The patients received lamivudine 100 mg for 12 months with monitoring their HBV DNA, AST, ALT, HBeAg and anti-HBe, and all these tests were performed at pretreatment and 1, 3, 6, 9 and 12 months after treatment. The serum HBV DNA was measured by HBV branched DNA assay. RESULTS: HBeAg loss was observed in 12 patients (16.2%), and 9 patients achieved anti-HBe seroconversion during up to 1 year of lamivudine therapy. The mean time to HBeAg loss was 5.6 months (range: 1-12 months). The posttreatment HBV DNA (<2,000 copies/mL) after 3 month (P=0.008) and 6 month (P=0.012)) were significant predictors of HBeAg loss after 1 year of lamivudine treatment on univariate analysis. Pretreatment HBV DNA, AST/ALT, gender, age and liver cirrhosis had no impact on HBeAg loss. The six-month posttreatment HBV DNA level <2,000 copies/mL was a significant predictor of HBeAg loss on multivariate analysis (P=0.008, odds ratio=0.108). CONCLUSION: We suggest that an HBV DNA level <2,000 copies/mL at 6 month after lamivudine therapy is the most important predictor of HBeAg loss during up to 1 year of lamivudine therapy. PMID- 18159150 TI - [Liver stiffness measurement for the diagnosis of hepatic fibrosis in patients with chronic viral hepatitis]. AB - BACKGROUND AND AIMS: FibroScan is a new medical device that noninvasively measures liver stiffness. The aim of this study was to assess the accuracy of the liver stiffness measurement by FibroScan for making the diagnosis of liver fibrosis in patients with chronic viral hepatitis. METHODS: We studied 103 patients with chronic viral hepatitis B or C and they underwent FibroScan and liver biopsy between October 2005 and August 2006. Liver fibrosis was staged on a 0-4 scale according to the Korean Society of Pathologists Scoring System. The diagnostic accuracy was assessed by analysis of the receiver operator characteristics (ROC). RESULTS: The liver stiffness was 3.5-57.1 kPa (mean: 11.8, SD: 8.9). The mean value of liver stiffness in each fibrosis stage group (F1, F2, F3 and F4) was 5.8+/-1.8 kPa, 11.3+/-6.8 kPa, 11.8+/-6.0 kPa and 23.4+/-16.5 kPa, respectively. Liver stiffness measured by FibroScan showed reliable correlation with the liver fibrosis stage as confirmed by liver biopsy (r=0.56, p<0.001). The AUROC (95% CI) of > or = F2, > or = F3 and F4 was 0.93 (0.86-0.99), 0.72 (0.62 0.82) and 0.80 (0.67-0.92), respectively. The sensitivity and specificity of 7.5 kPa, which was the cutoff value for > or = F2, was 84% and 90%, respectively. CONCLUSIONS: FibroScan is a reliable method for the diagnosis of significant fibrosis (> or =F2) and cirrhosis in patients with chronic liver disease. The liver stiffness measurement by FibroScan showed good diagnostic performance for significant fibrosis. PMID- 18159151 TI - [Five-year survival analysis of a cohort of hepatocellular carcinoma patients who treated at the National Cancer Center, Korea]. AB - BACKGROUND AND AIMS: We investigated the five-year survival outcomes of a large cohort of hepatocellular carcinoma (HCC) patients who were treated at a single institute, and this is a follow-up study of a previous report. METHODS: Nine hundred four HCC patients who were treated at the National Cancer Center Korea were enrolled and they were followed till February 2007. RESULTS: The mean age of the patients was 56.0 years and 731 patients were male. Six hundred seventy-seven (74.9%) patients died and the overall 5-year survival rate (5-YSR) was 23.9%. The 5-YSRs of the patients with modified UICC stage I, II and III were 61.2%, 54.4% and 18.4%, respectively, and the median survival time was 4.3 and 3.7 months for the stage IVa and IVb patients, respectively. For the analysis of the treatment modality, surgical resection showed significantly better outcomes for the five year survival as compared with transcatheter arterial chemoembolization (TACE) for Child-Pugh A patients with modified UICC stage I or II disease (80.1% vs 52.8%, respectively, P<.001), or stage III disease (60.7% vs 17.0%, respectively, P<.001). For patients with advanced stage IVb disease, TACE, systemic chemotherapy and radiotherapy increased the median survival period more than conservative management for the Child-Pugh class A patients. The serum alpha fetoprotein level, portal vein tumor thrombosis, the Child-Pugh class, the tumor stage, the tumor type and symptoms were related to the prognosis. CONCLUSIONS: This study presented, for the first time, the 5-YSRs of a cohort of HCC patients. PMID- 18159152 TI - [Clinical outcome after living donor liver transplantation in patients with hepatitis C virus-associated cirrhosis]. AB - BACKGROUND AND AIMS: Hepatitis C virus (HCV)-associated cirrhosis is an increasingly frequent indication for liver transplantation (LT). However, HCV recurrence is universal and this immediately occurs following LT, which endangers both the graft and patient survival. We investigated the frequency of posttransplant recurrence of HCV infection and the patient-graft survival, and we analyzed the responses to ribavirin and interferon therapy in the patients with recurrent HCV infection after living donor liver transplantation (LDLT). METHODS: We retrospectively reviewed the clinical outcomes of 39 HCV-associated cirrhosis patients who underwent LDLT at Asan Medical Center between August 1992 and June 2006. In this study, the diagnosis of recurrent HCV was made on the basis of increased transaminases and serum HCV RNA levels greater than 10 million IU/mL because protocol liver biopsy was not performed. RESULTS: HCV recurrence was seen in 26 of the 39 LDLT patients (66.7%). 86.7% of recurrence occurred within the first postoperative year. Antiviral treatment was used for all patients with recurrence of HCV. None of the 10 patients receiving ribavirin alone and 9 of 16 patients who received combination therapy with pegylated interferon alpha-2a plus ribavirin became HCV RNA negative and they remained persistently negative during the median follow-up of 24.9 months. Our data indicates that there is no significant factor influencing HCV recurrence except for the recipient's age. The 2-year patient survival for the HCV patients with HCC and those patients without HCC were 81.2% and 81.3%, respectively (P=0.85) and the 2-year graft survival rates were 81.2% and 68.2%, respectively (P=0.29). No patient died from HCV recurrence during the follow-up period. CONCLUSIONS: Combination therapy with ribavirin and interferon appears to improve the outcome of recurrent HCV infected patients after LDLT. PMID- 18159153 TI - [A case of imported Dengue fever with acute hepatitis]. AB - Dengue fever is an acute febrile disease caused by the dengue virus, which belongs to the flaviviridae family, and this virus is transmitted by the bite of the mosquito Aedes aegypti. It occurs in the tropical climates of the South Pacific, Southeast Asia, India, Africa and the subtropical zone of America. Imported cases of Dengue fever and Dengue hemorrhagic fever are rapidly increasing as many Koreans are now traveling abroad. Liver injury is usually detected by laboratory investigation according to a surveillance protocol. Although liver injury by dengue virus has been described in Asia and the Pacific islands, the pathogenic mechanisms are not yet fully clarified. It is usually expressed in a self-limiting pattern and the patient has a complete recovery. We report here on a case of a young woman who presented with general weakness, nausea and significant elevation of the aminotransferase levels, and she was diagnosed with dengue fever. PMID- 18159154 TI - [A case of solitary fibrous tumor of the liver]. AB - A 46-year-old woman was found to have a huge liver mass that was detected by abdominal ultrasonography. Abdominal CT and MRI showed a 10 cm-sized, encapsulated mass occupying the anterior segment of the right hepatic lobe. Extended right hemihepatectomy was performed and pathological examination revealed fibroblast-like spindle cells within dense deposits of collagen. On immunohistochemical staining, these spindle tumor cells showed an intense CD34 immunoreactivity. The patient is alive without evidence of tumor recurrence 7 months after the resection. Solitary fibrous tumor is a very rare neoplasm found in the liver parenchyma, and it has been reported in less than 30 patients in the English literature. We present here the first such case in Korea. PMID- 18159155 TI - [A case of fulminant hepatic failure secondary to hepatic metastasis of small cell lung carcinoma]. AB - Although liver metastasis is commonly found in cancer patients, fulminant hepatic failure secondary to diffuse cancer infiltration into the liver is rare. Liver metastasis-induced fulminant hepatic failure has been reported in patients with primary cancer of the gastrointestinal tract, breast and uroepithelium, and in patients with melanoma and hematologic malignancy. Small cell lung cancer is so highly invasive that hepatic metastasis is common, but rapid progression to fulminant hepatic failure is extremely rare. We report here on a case of a patient who died because of rapid progression to fulminant hepatic failure as a result of hepatic metastasis of small cell lung carcinoma. PMID- 18159156 TI - [Combined hepatocellular-cholangiocarcinoma]. PMID- 18159158 TI - [Esophageal cancer with esophageal perforation]. PMID- 18159159 TI - [Eosinophilic esophagitis]. AB - Eosinophilic esophagitis (EE) is characterized by eosinophilic infiltration of the esophageal wall including mucosa, submucosa, and muscle proper. EE is a condition involving both pediatrics and adults. Patients with EE are predominantly young males, commonly related to atopy. The typical clinical presentation includes dysphasia, food impaction, and symptoms mimicking gastroesophageal reflux disease. Endoscopic examination reveals mucosal fragility, ring or corrugated mucosa, whitish plaques, or small caliber esophagus. Histologic finding of >20 eosinophils/HPF (high power field) is the diagnostic hallmark of EE. Elemental formula, systemic or topical corticosteroid, anti-inflammatory drugs such as leukotriene receptor antagonists, anti interleukin (IL)-5, and anti-IL-13 monoclonal antibodies have been used to manage EE. Esophageal dilation is considered in adult patients with severe obstructive symptoms due to stricture. PMID- 18159160 TI - [p53 Codon 72 and 16-bp duplication polymorphisms of gastric cancer in Koreans]. AB - BACKGROUND/AIMS: p53 gene plays an important role in cell cycle control in response to DNA damage which may increase the probability of mutations leading to carcinogenesis. The role of p53 gene polymorphisms [codon 72 (exon 4) and 16-bp duplication (intron 3)] as potential markers indicating cancer risk remains inconclusive, and the data on gastric cancer are very limited. The aim of this study was to assess the role of p53 gene polymorphisms in the risk of gastric cancer and in the determination of genetic susceptibility to gastric cancer in Koreans. METHODS: We analysed p53 genotypes using a polymerase chain reaction based restriction fragment length polymorphism assay in a population-based case control study in 120 gastric cancer patients and 145 cancer-free controls in Koreans. RESULTS: There was no specific genotype of p53 gene polymorphism in the gastric cancer patients compared to cancer-free controls. In p53 codon 72 and 16 bp duplication polymorphisms, the frequency and distribution of genotypes showed no statistical significance (p=0.7125 and p=0.1659). There was no difference in genotype by histologic subtypes, location of lesion, and age. However, the genotypic distribution in the patient subgroups with a history of heavy cigarette smoking of p53 16-bp duplication polymorphism were significantly different from those of cancer-free controls (p=0.0079). CONCLUSIONS: The p53 codon 72 and 16-bp duplication polymorphisms were not associated with the increased risk of gastric cancer and did not seem to contribute to gastric cancer susceptibility among Koreans. It is possible that p53 16-bp duplication polymorphism modulates the risk of smoking-induced gastric cancer development in Koreans. In order to clarify the associations between specific genotypes and gastric cancer risk, the evaluations of these polymorphisms in other ethnic backgrounds with larger number of patients would be needed. PMID- 18159161 TI - [Histological changes of gastric atrophy and intestinal metaplasia after Helicobacter pylori eradication]. AB - BACKGROUND/AIMS: Long-term Helicobater pylori infection results in atrophic gastritis and intestinal metaplasia, and increases the risk of gastric cancer. However, it is still controversial that eradication of H. pylori improves atrophy or metaplasia. Therefore, we investigated histological changes after the H. pylori eradication in patients with atrophy or metaplasia. METHODS: One hundred seven patients who received successful eradication of H. pylori infection in Hanyang University, Guri Hospital from March 2001 to April 2006, were enrolled. Antral biopsy was taken before the eradication to confirm the H. pylori infection and grade of atrophy or metaplasia by updated Sydney System. After a certain period of time, antral biopsy was repeatedly taken to confirm the eradication and investigate histological changes of atrophy or metaplasia. RESULTS: Mean age of the patients was 55.3+/-11.3, and average follow-up period was 28.7+/-13.9 months. Endoscopic diagnosis included gastric ulcer, duodenal ulcer, non-ulcer antral gastritis. Atrophy was observed in 41 of 91 and their average score was 0.73+/-0.92. After the eradication of H. pylori, atrophy was improved (0.38+/ 0.70, p=0.025). However, metaplasia which was observed in 49 of 107, did not significantly improve during the follow-up period. Newly developed atrophy (7 of 38) or metaplasia (18 of 49) was observed in patients who without atrophy or metaplasia initially. Their average scores were slightly lower than those of cases with pre-existing atrophy or metaplasia without statistical significance. CONCLUSIONS: After the eradication of H. pylori infection, atrophic gastritis may be improved, but change of intestinal metaplasia is milder and may take longer duration for improvement. PMID- 18159162 TI - [The prevalence of esophageal disorders in the subjects examined for health screening]. AB - BACKGROUND/AIMS: Various disorders can be developed in the esophagus. However, esophagus has been less well focused than other gastrointestinal (GI) tracts since the esophageal disorders are relatively uncommon. There has been no report on the prevalence of overall esophageal disorders in Korea. The aim of this study was to evaluate the prevalence of esophageal disorders in health examinee. METHODS: We reviewed retrospectively the endoscopic reports of 6,683 subjects who underwent upper GI endoscopy for screening purpose at Gyeongsang National University Hospital from March, 2005 to May, 2006. RESULTS: Among 6,683 health examinee, 1,154 (17.26%) had esophageal diseases. Gastro-esophageal reflux diseases (GERD) were the most common diseases (14.66%). The prevalence of erosive reflux esophagitis, minor change esophagitis, and Barrett's esophagus was 8.45%, 5.01% and 1.12%, respectively. Hiatal hernia and esophageal submucosal tumor were 2nd and 3rd common diseases (2.0% and 0.6%, respectively). A variety of other esophageal disorders were seen as well. The esophageal disorders with over 0.1% of prevalence were esophageal varices (0.37%), esophageal squamous papilloma (0.31%), esophageal candidiasis (0.25%), nonspecific esophagitis (0.16%), heterotopic gastric mucosa (0.16%), and esophageal vascular ectasia (0.12%) in order. CONCLUSIONS: The overall prevalence of esophageal disorders among health examinee was 17.3%. The GERD was the main esophageal disease and a variety of esophageal diseases were identified. The endoscopists need to take an interest in the esophageal disorders and a community-oriented well organized study should be warranted. PMID- 18159163 TI - [Association of interleukin-12 gene polymorphism with persistence of hepatitis B virus infection and hepatocellular carcinoma]. AB - BACKGROUND/AIMS: Infection with hepatitis B virus (HBV) may result in various conditions. Natural course of HBV infection is influenced by various host immune factors and cytokines play a crucial role in host immune defense. This study was undertaken to investigate the association between HBV persistence and development of hepatocelluar carcinoma (HCC) and single nucleotide polymorphisms (SNPs) of interleukin (IL)-12A. METHODS: Between March 2002 and December 2004, seven hundred thirty Korean patients with HBV infection and 320 healthy individuals who recovered from HBV infection were enrolled. We assessed polymorphisms and haplotype in IL-12A, and the genotype distributions of the HBV clearance and persistence groups were compared in order to investigate the association between HBV persistence and SNPs of IL-12A. Moreover, the genotypic distributions between patients with HCC and without HCC were compared to investigate the association between the development of HCC and SNPs of IL-12A. RESULTS: We asssesed the SNPs of IL-12A at position +6400, +6624 and +7003. On the basis of logistic regression analysis, no statistically significant association with HBV persistence was observed with IL-12A exon 7 +6400, +6624, 3' UTR +7003 SNP and haplotype of IL 12A +6400/+6624/+7003. Furthermore, no statistically significant association of HCC development with IL-12A exon 7 +6400, +6624, 3' UTR +7003 SNP and haplotype of IL-12A +6400/+6624/+7003 was observed. CONCLUSIONS: These results suggest that SNPs and haplotype of IL-12A are not associated with HBV persistence and development of HCC. Further studies are needed to identify the host genetic factors in immune defense including cytokine gene polymorphisms of both IL-12A and IL-12B. PMID- 18159164 TI - [A case of Crohn's disease presenting with free perforation and portal venous gas]. AB - Crohn's disease is characterized by its chronic course and transmural inflammation of gastrointestinal tract. The accompanying fibrous reaction and adhesion to adjacent viscera appears to limit the complication of free perforation. The true incidence of free bowel perforation is difficult to assess, however, the anticipated occurrence rate is 1-2% during the course of illness. Moreover, portal venous gas is also an uncommon event in the natural history of Crohn's disease. Portal venous gas occurs when intraluminal gas from the gastrointestinal tract or gas-forming bacteria enters the portal venous circulation. The finding of portal venous gas associated with Crohn's disease does not always mandate surgical intervention. We experienced a case of Crohn's disease presenting with free perforation and portal venous gas. The literatures on the cases with perforation and portal venous gas associated with Crohn's disease were reviewed. PMID- 18159165 TI - [A case of terminal ileal ulcer of Henoch-Schonlein purpura treated with high dose steroid]. AB - Henoch-Schonlein purpura (H-S purpura) is a systemic small-vessel vasculitis involving skin, joint, gastrointestinal tract, and kidney. It is characterized by the classic tetrad of abdominal pain, arthralgia, typical rash, and renal involvement. All of these clinical findings can occur in any order and at any time over several days to weeks. Gastrointestinal manifestations such as abdominal pain, melena, or hematochezia occur in 45-85% and preceed skin lesions up to 40% in H-S purpura. However, endoscopically proven gastrointestinal lesion is rare because majority of involved sites are small intestine. We report a case of Henoch-Schonlein purpura with terminal ileal ulcer, healed after treatment with high dose steroid, proven by colonoscopy. PMID- 18159166 TI - [Ten cases of Fitz-Hugh-Curtis syndrome]. AB - Fitz-Hugh-Curtis syndrome, a kind of perihepatitis, occurs approximately in 3 to 10 percent of patients with pelvic inflammatory disease. It is not easy to detect in clinical settings due to requirement of invasive methods for diagnosis, for example, like a laparoscopic examination. Now, it has become possible to recognize it easily with the aid of non-invasive methods including an abdominal dynamic CT scan and laboratory tests. Moreover, it can be improved after the oral administration of antibiotics. Therefore, noninvasive diagnosis is desirable. Herein, clinical characteristics of ten cases of Fitz-Hugh-Curtis syndrome are reported, with a review of the literature. PMID- 18159167 TI - [A case of pneumatosis cystoides intestinalis: diagnosed by CT colonoscopy]. AB - Pneumatosis cystoides intestinalis (PCI) is a rare condition characterized by multiple gas filled cysts in the intestinal wall. The diagnosis of PCI is usually made by colonoscopy, histology, or radiologic findings. We report a case of PCI in a 35-year-old man. The patient initially complained of watery diarrhea and abdominal bloating for 2 weeks. Simple abdominal X-ray demonstrated numerous, small, round, air densities on the right upper abdomen along the ascending and proximal transverse colon. Colonoscopy revealed numerous, 5-20 mm sized, sessile polypoid, balloon-like distended, protruding subepithelial masses covered with normal colonic mucosa from cecum to proximal transverse colon. We performed a CT colonoscopy and confirmed PCI with multiple air-filled cystic masses along the colonic wall from cecum to proximal transverse colon. The patient was treated with antibiotics and oxygen inhalation for 2 weeks. Follow-up CT colonoscopy revealed marked regression in the number and size of the air-filled cystic masses. Herein, we report the first case of the PCI in Korea diagnosed by CT colonoscopy. Follow-up evaluation with CT colonoscopy was performed after the treatment of the PCI. CT colonoscopy is a useful non-invasive diagnostic tool for the diagnosis of pneumatosis cystoides intestinalis. PMID- 18159168 TI - [A case of intestinal obstruction caused by strangulated femoral hernia accompanying soft tissue necrosis]. AB - Intestinal obstruction involves a partial or complete blockage of the bowel which results in the failure of intestinal contents to pass through. The mechanical causes of obstruction may include the followings: hernias, postoperative adhesions or scar tissue, impacted feces, gallstones, tumors, granulomatous processes, intussusception, volvulus, foreign bodies, and etc. Hernias are the third leading cause of intestinal obstruction by 10% approximately. However, most hernias are the cases with abdominal wall, inguinal or internal hernia. Femoral, obturator, lumbar, or sciatic hernia as the cause of obsturction is rare. Furthermore, the cases accompanying soft tissue necrosis are seldomly reported. Herein, we report a case of intestinal obstruction caused by strangulated femoral hernia accompanying soft tissue necrosis in a 78-years-old female patient. PMID- 18159170 TI - [Endoscopic resection for inflammatory fibroid polyp of the cecum]. PMID- 18159171 TI - [COX-2 inhibitors in inflammatory bowel disease: friends or foes?]. AB - The cyclooxygenase (COX) is a key enzyme in the conversion of arachidonic acid to prostaglandins. COX-1 is constitutively expressed and is a critical housekeeping gene, whereas COX-2 is rapidly upregulated by growth factors and cytokines and thus responsible for inflammation. COX-2 is frequently overexpressed in colonic adenoma and carcinoma. Specific inhibitors of COX-2 have been shown to induce apoptosis in tumor cells and to inhibit tumor growth in animal models and in humans. Long-standing IBD patients have increased risk of developing colorectal cancer compared to general population. IBD-associated colorectal carcinogenesis is probably promoted by chronic inflammation and closely related to COX-2. In a recent study, powerful chemopreventive ability of selective COX-2 inhibitor was observed in colitis-related colon carcinogenesis in mouse model. But it was reported that even selective COX inhibitors aggravated the DSS-induced colonic inflammation. It is assumed that endogenous PGs are involved in the mucosal defense against DSS-induced colonic ulcerations which are produced by COX-1 at early phase and by COX-2 at late phase. Long-term use of COX-2 inhibitors for the chemoprevention of colitic cancer is needed to define their mechanism of action, that reduce side effects and have specific tumor target. PMID- 18159172 TI - [Changing patterns of antibiotic resistance of Helicobacter pylori in patients with peptic ulcer disease]. AB - BACKGROUND/AIMS: Antibiotic resistance of Helicobacter pylori (H. pylori) is a significant clinical problem because it reduces the efficacy of eradication therapy. The aims of this study were to assess the changing patterns of antibiotic resistance of H. pylori in patients with peptic ulcer diseases and to evaluate the eradication rate in antibiotic resistant H. pylori strains. METHODS: One hundred forty four H. pylori isolates obtained from 466 patients with peptic ulcer disease between June 2001 and December 2005 were examined for antimicrobial resistance. The minimum inhibitory concentration (MIC) of metronidazole was determined by modified broth microdilution method (mBMD) and E test. MICs of clarithromycin and amoxicillin were determined by mBMD, E test, and disc diffusion test. The breakpoints for metronidazole, clarithromycin, and amoxicillin resistance were defined as >8 microg/mL, >1 microg/mL, and > or =1 microg/mL, respectively. RESULTS: Resistance to metronidazole and clarithromycin was detected in 34.7% and 16.7% of H. pylori isolates, respectively. During the recent 5-year study period, amoxicillin-resistant rate of H. pylori was 11.8%, and multi-drug resistance rate of H. pylori was 16.7%. The eradication rate of clarithromycin containing triple therapies was low (7.8%) in clarithromycin resistant H. pylori strains. CONCLUSIONS: The proportions of clarithromycin resistant H. pylori strains have increased significantly over the last 5-years. There is an increasing tendency for the emergence of strains with multi-drug resistance. The increase in clarithromycin-resistant strains results in a decrease in eradication rate for H. pylori. In areas with high clarithromycin resistance, new alternative first-line treatment combination should be considered. PMID- 18159173 TI - [The role of gastric acid in the H. pylori-induced gastritis in mouse]. AB - BACKGROUND/AIMS: This study was designed to investigate the role of gastric acid in the extent of H. pylori-induced gastritis. METHODS: Twenty eight mice were inoculated with live H. pylori. They were allocated into four groups. Mice in group I received no treatment, group II mice were treated with sham injection, group III received 125 microg/kg body weight of pentagastrin, while group IV received 250 microg/kg body weight of pentagastrin subcutaneously three times a week. After 7 months, the mucosal pH, H. pylori density, neutrophils and monocytes infiltration, and the degree of atrophy were assessed in the stomach. RESULTS: In the gastric body, the densities of H. pylori were not different among groups. The degree of neutrophil infiltration was significantly lower in group IV compared to other groups (p<0.05). The degree of monocyte infiltration was also significantly lower in group IV than group III (p<0.05). In the gastric antrum, there was no significant difference of the H. pylori density, neutrophil and monocyte infiltration, and degree of atrophy among the groups. The mice with the gastric mucosal pH lower than mean of 3.2 had significant lower level of H. pylori density (1.4 vs. 2.4, p=0.04), and infiltration of neutrophils (0.9 vs. 2.3, p=0.018), and monocytes (1.2 vs. 1.8; p=0.011) than the those with mucosal pH above 3.2 in the body of stomach. CONCLUSIONS: Gastric acid plays a role in suppressing the proximal propagation of H. pylori-induced gastritis to the body of stomach. PMID- 18159174 TI - [Expressions and clinical significances of c-met, c-erbB-2, COX-2, and IL-6 in the biliary tract cancers]. AB - BACKGROUND/AIMS: c-met, c-erbB-2, interleukin (IL)-6, and cyclooxygenase (COX)-2 expressions are considered to be implicated in the carcinogenesis and progression of cholangiocarcinoma, but the molecular pathogenesis of cholangiocarcinoma is still poorly understood. We aimed to analyze the expressions of each marker and their relationships with clinicopathologic factors. METHODS: One hundred and fourteen tissue samples were obtained from surgically resected specimens from patients with biliary tract cancer. The expressions of c-met, c-erbB-2, COX-2, and IL-6 were examined by immunohistochemically. The expression of each marker and correlations between these markers and clinicopathologic factors were analyzed. RESULTS: The expression rates of each maker were as follows: c-met 34/112 (30.4%), c-erbB-2 5/112 (4.5%), COX-2 53/113 (46.9%), and IL-6 68/113 (60.2%), respectively. c-met expression was more frequently observed in cases with invasion through the adjacent connective tissues (p=0.0263). IL-6 overexpression was more frequently observed in cases with absent lymph node metastasis (p=0.0325). Either c-erbB-2 expression or COX-2 expression was significantly associated with lymph node metastasis (p=0.0442). CONCLUSIONS: The expression of c-met was closely related to the invasiveness of cholangiocarcinoma. Co-expression of c-met, COX-2 and, IL-6 showed a significant correlation with invasiveness and lymph node metastasis and these could be useful marker to guide clinical outcome in patients with cholangiocarcinoma. PMID- 18159175 TI - [A case of acute pancreatitis caused by 5-aminosalicylic acid suppositories in a patient with ulcerative colitis]. AB - Oral 5-aminosalicylic acid (5-ASA) has been known as a first-choice drug for ulcerative colitis. However, hypersensitivity reactions, including pancreatitis, hepatitis, and skin rash, have been reported with 5-ASA. Topical formulations of 5-ASA like suppositories have been rarely reported to induce adverse reactions because of their limited absorption rate. We recently experienced a case of acute pancreatitis caused by 5-ASA suppositories in a patient with ulcerative colitis. A 26-year-old male was admitted with abdominal pain and diagnosed as ulcerative colitis. Acute pancreatitis occurred soon after 24 hours of treatment with oral mesalazine. Drug-induced pancreatitis was suspected and administration of mesalazine was discontinued. Then 5-ASA suppositories were started instead of oral mesalazine. Twenty-four hours after taking 5-ASA suppositories, he experienced severe abdominal pain, fever, and elevation of amylase levels. The suppositories were immediately stopped and symptoms resolved over next 48 hours. Herein, we suggest that, in patients treated with 5-ASA suppositories who complain of severe abdominal pain, drug-induced pancreatitis should be suspected. PMID- 18159176 TI - [A case of Diphyllobothrium latum infection in a patient with abdominal pain]. AB - The growing popularity of eating raw fish has resulted in increase of certain human parasitic infection, such as diphyllobothriasis. Even though, upper and lower gastrointestinal endoscopy reveal no specific abnormality, if a patient complains of persistent abdominal pain, we should consider the possibility of parasitic infection. Careful history taking and stool examination can avoid further invasive study. We report a case of Diphyllobothrium latum infection in a patient with vague abdominal pain who showed normal finding on endoscopy. PMID- 18159177 TI - [A case of appendicular tuberculosis presenting as acute appendicitis]. AB - Tuberculosis may affect primarily all organs and tissues of the body, although some of these show high immunity against the infection. The most common forms of non-pulmonary tuberculosis are tuberculosis of bones and joints (30%), urinary system (24%), lymph nodes (13%), sexual organs (8%), cerebrospinal meninges (4%), and alimentary system (3%). Especially, the commonest presentation of abdominal tuberculosis is ileocecal disease, but isolated appendicular involvement is also rarely seen, occurring in only 1.5% to 3% of cases in the absence of pulmonary or other abdominal involvement. The appendix may either be involved secondary to ileocecal tuberculosis, or to tuberculosis at another site within the abdomen, or may occur in the even, rarer "isolated" form, without the evidence of disease elsewhere. We report a case of acute appendicitis underwent appendectomy and histopathologic examination of appendix revealed appendicular tuberculosis. PMID- 18159178 TI - [A case of malignant gastrointestinal stromal tumor of ileum with liver abscess]. AB - Gastrointestinal stromal tumor (GIST) is an uncommon mesenchymal tumor of the gastrointestinal tract and is generally located in the stomach and small intestine. They usually present with abdominal pain, gastrointestinal bleeding, and palpable mass. Some patients present with rare symptoms that are more common in malignant GIST. Malignant GIST combined with a liver abscess has not been reported yet in the literatures. We report a case of 67-year-old woman who suffered from liver abscess combined by malignant GIST with central necrosis and fistula in the ileum. She complained of fever, chills, and abdominal pain. Abdominal CT scan showed huge liver abscess and ileal mass with air pocket. Small bowel series showed contrast material filling into the ileal GIST mass. An operation was performed and the final diagnosis was malignant GIST of the ileum with invasion into the sigmoid colon and urinary bladder. PMID- 18159179 TI - [A case of benign schwannoma in the ascending colon]. AB - Schwannomas are rare tumors derived from the cells of Schwann which form the neural sheath. Some patients with gastrointestinal schwannoma have been previously reported in the literature. However, schwannomas of the colon are extremely rare. We herein describe a case of schwannoma of the colon. A 49-year old woman was admitted with complaint of abdominal pain and investigations revealed the presence of a 4 cm sized mass in the ascending colon. Following right hemicolectomy, histopathology and immunohistochemistry confirmed the colonic lesion to be a benign schwannoma. There was no evidence of specific complication or recurrence until now. PMID- 18159180 TI - [A case of sclerosing cholangitis showing response to prednisolone]. AB - Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease characterized by progressive fibrosis and destruction of intra- and extrahepatic bile ducts resulting in hepatic failure and death. Only the liver transplantation is the possible treatment for patients to survive. There has been a few reports that steroid is an effective treatment in autoimmune variant sclerosing cholangitis, which is thought to be a familial diseases with different etiology, and steroid responsive biliary strictures be named as immunoglobulin G4 (IgG4) associated cholangitis (IAC). There is no reliable data regarding effective steroid treatment in autoimmune variant sclerosing cholangitis in Korea. We report a case of 32-year-old male with sclerosing cholangitis, who was diagnosed by endoscopic retrograde cholangiopancreatography (ERCP) and liver biopsy, showing favorable response to prednisolone therapy. PMID- 18159181 TI - [Changing patterns of antibiotic resistance of Helicobacter pylori in patients with peptic ulcer disease]. PMID- 18159183 TI - [A case of rectal carcinoid with invasion to proper muscle layer]. PMID- 18159184 TI - [Current application of minimally invasive surgery for gastrointestinal diseases]. AB - The advent of minimally invasive surgery (MIS) brought a major deviation in trend from conventional surgery. Since the introduction of first laparoscopic cholecystectomy in 1985, many operations for gastrointestinal diseases adopted MIS technique in a relatively short period of time. These MIS operations yielded better outcomes when compared to their open counterparts: less pain, shorter hospital stay, faster recovery, and better cosmetics. More complex surgical procedures for benign and malignant diseases of gastrointestinal tract are currently being performed by MIS technique with the improvement in equipment, instrumentation, and surgical skills. At the forefront of MIS, lies robotics. This paper briefly reviews the current status of MIS in the field of gastrointestinal diseases. PMID- 18159185 TI - [Minimally invasive surgery for gastroesophageal reflux disease]. AB - Gastroesophageal reflux disease (GERD) is a chronic disease deteriorating patient's quality of life. With the advent of proton pump inhibitors, treatment failures have decreased considerably. However, surgical therapy offers the potential for cure in more than 90% of patients with GERD. Specific indications for antireflux surgery are: incomplete response to medical therapy, frequent recurrences despite the medical treatment, laryngopharyngeal, and/or respiratory symptoms, and complications of GERD, such as esophageal stricture, erosive esophagitis, esophageal ulcer, and/or Barrett's esophagus. The introduction of laparoscopic surgery in early ninties had a profound impact on many surgical fields, including the treatment of GERD. In this review, laparoscopic Nissen fundoplication is described and controversial topics, such as total vs. partial fundoplication, and the natural history of Barrett's esophagus after antireflux surgery are addressed. PMID- 18159186 TI - [Minimally invasive surgery for esophageal cancer]. AB - Minimally invasive surgery is now rapidly developing and becoming a standard surgical option in some fields. In the past, many thoracic surgeons were reluctant to adopt minimally invasive techniques in esophageal cancer surgery due to concern over the oncologic perspectives and technical difficulties. However, over the last few years, thoracic surgeons have progressively embraced the technical advancements and now many experienced centers have adopt minimally invasive surgery as a primary option for non-advanced esophageal cancer operations. In esophageal cancer surgery, the volume of operation performed in some hospital is closely related to the outcome of patients, and the experiences of surgical team play an important role in minimally invasive surgery. Minimally invasive esophageal surgery (MIES) has steep learning curves, also. The merits of MIES are as follows. The conventional esophageal cancer operation has two or triple incisions, resulting in high postoperative morbidity and mortality. However, postoperative complication in MIES became less frequent than conventional surgery. The patient's satisfaction is high. Mid-term outcomes of MIES have been reported that it is safe and feasible in esophageal cancer and survival curves are similar to those of conventional surgery. Therefore, MIES is a valuable therapeutic modality for both esophageal cancer patients and thoracic surgeons. PMID- 18159187 TI - [The current status and future perspectives of laparoscopic surgery for gastric cancer]. AB - The incidence of early gastric cancer (EGC) has increased to over 50% in Korea and Japan due to rapid advances in diagnostic instrumentation and increased use of mass screening. Considering the excellent prognosies of EGC patients, the quality of life of these patients after treatment has recently been emphasized. For the better quality of life, laparoscopic surgery has emerged as an alternative therapy for EGC patients. Since Kitano et al. first performed laparoscopy-assisted distal gastrectomy (LADG) for early gastric cancer in 1991, it has been performed worldwide, especially in Japan and Korea. A number of reports have presented the excellent short term outcomes; less pain, better cosmetics, faster recovery, and shorter hospital stay. However, there is little evidence of the oncological outcome of laparoscopic gastrectomy as a treatment modality for gastric cancer. Multi-center randomized controlled trials of laparoscopic versus open gastrectomy are needed to establish the future role of laparoscopic surgery in the treatment of patients with gastric cancer. PMID- 18159188 TI - [Minimally invasive surgery based on sentinel node biopsy for gastrointestinal cancer]. AB - Sentinel lymph node biopsy has been found to be highly effective in correctly predicting the nodal status of melanoma and breast cancer patients. Recently, individualized surgical treatments based on the actual metastatic status have been introduced in patients with melanoma and breast cancer. Although several promising results of extensive lymph node dissection have been reported in the field of gastrointestinal (GI) cancer, prognostic benefits of extensive surgery are still not validated by prospective randomized trials. Many feasibility studies favoring sentinel node biopsy in various organs have been reported. Although more evidence from large-scale multicenter clinical trials is required, sentinel lymph node mapping might be also widely acceptable for GI cancer. PMID- 18159189 TI - [Current status of laparoscopic surgery for colorectal cancer]. AB - Laparoscopic surgery for colorectal cancer was first adopted 16 years ago. There are various limitations in performing laparoscopic surgery including the technical complexity and question of positive impact on the long-term oncologic outcome. The purpose of this review is to outline the important issues surrounding the laparoscopic surgery for colorectal cancer based on the most recently published articles. The laparoscopic approach provides the advantages of an illuminated and magnified view, which may be superior to open surgery. There was no significant difference on the oncologic clearance, especially its proportion of positive radial margins to the number of harvested lymph nodes. In addition, laparoscopic surgery for colorectal cancer was associated with earlier recovery of bowel function, need for fewer analgesics, and with a shorter hospital stay compared to open surgery. Long-term oncologic outcome does not appear to be impaired by laparoscopic resection and local recurrence and disease specific survival has been reported to be similar for both laparoscopic and open surgery for colorectal cancer. Laparoscopic surgery for colorectal cancer is feasible and safe when performed by experienced surgeons. The oncologic results of many ongoing prospective randomized controlled trials are eagerly awaited. PMID- 18159190 TI - [Robotic surgery in gastrointestinal surgery]. AB - Robotic surgery is an emerging technology. After adoption of robotic surgery for cholecystectomy in 1997, various general surgical procedures have been performed using surgical robot. In general surgery, robotic surgery is applied to wide range of procedures, however, it is still in its early years. Cholecystectomy, Nissen fundoplication, Heller myotomy, and Roux-en-Y gastric bypass are the most frequently performed robotic operations. Most reports proved that application of robotic technology for general surgery is technically feasible and safe with the help of improved dexterity, better visualization, and high level of precision. However, still the absence of tactile sense and extremely high costs are the problems to be solved. Although robotic surgery has demonstrated some clear benefits compared to conventional surgeries including laparoscopy, it remains to be seen whether these benefits will outweigh the associated disadvantages or problems of robot surgery. Therefore, more prospective randomized study comparing the shot-term and long-term surgical outcomes between robotic and conventional laparoscopic surgery is needed to further define the impact of robotic surgical technology in general surgery. PMID- 18159191 TI - [Jejunal diverticular bleeding diagnosed by double-balloon enteroscopy: a case report and review of literatures]. AB - Jejunal diverticula is a rare disease and an unusual cause of obscure gastrointestinal hemorrhage. Obscure gasterointestinal bleeding is difficult to treat because the bleeding site cannot be identified by routine endoscopy and contrast studies. A wireless capsule endoscopy is not invasive and can visualize the entire small bowel. However, this method has limitations of incapability of taking biopsies and performing endoscopic interventions such as polypectomy or stent insertion. The double-balloon enteroscopy is being used frequently for the diagnosis and management of various small bowel diseases. We report a case of proximal jejunal diverticular bleeding diagnosed by double-balloon enteroscopy and treated with angiographic embolization. PMID- 18159192 TI - [Hemobilia from pancreatic arteriovenous malformation: successful treatment with transportal coil embolization of draining veins]. AB - Pancreatic arteriovenous malformations (AVM) are extremely rare diseases frequently complicated by gastrointestinal hemorrhage. While surgical resection of affected lesion is preferred for the treatment of pancreatic AVM, angiographic intervention can be used as an alternative treatment, especially in surgically high-risk patients. We experienced a patient with pancreatic AVM manifested by hemobilia and biliary sepsis. Superior mesenteric and common hepatic arteriography showed pancreaticoduodenal AVM composed of nidus supplied by numerous fine feeding arteries and of draining veins encircling the common bile duct (CBD). Hemobilia was controlled by transportal coil embolization of draining veins of AVM around the CBD. Herein, we report this case with the review of literatures. PMID- 18159193 TI - [A case of simple type Caroli's disease confined to right lobe of the liver]. AB - Caroli's disease is a rare congenital hepatobiliary disease characterized by multifocal segmental dilatation of intrahepatic bile ducts affecting all or parts of the liver. Two forms of Caroli's disease are described, the pure form and that associated with periportal fibrosis. The disease may diffusely affect the liver or be localized to one lobe or segment. Less than 20% of all reported cases of Caroli's disease are monolobar type. We report a case of simple type Caroli's disease confined to right lobe of the liver in a 22 year old man. He was admitted due to right upper abdominal pain and diagnosed by magnetic resonance cholangiopancreatography. He was treated with right hepatic lobectomy and recovered completely. PMID- 18159194 TI - [A case of acute appendicitis due to intestinal stricture after intestinal tuberculosis treatment]. AB - Intestinal hemorrhage, perforation, obstruction, and fistula formation are the common complications associated with intestinal tuberculosis. However, these complications usually occur in active stage of intestinal tuberculosis. A 45-year old man was diagnosed as intestinal tuberculosis and received anti-tuberculosis medications for 9 months. After the end of treatment, intestinal lesion was cured. However a deformed appendiceal orifice due to hypertrophic sear resulting in symptomatic appendicitis was noted. We report a case of acute appendicitis due to intestinal stricture after the successful treatment of intestinal tuberculosis. PMID- 18159196 TI - The potentials and challenges of patient registries. PMID- 18159197 TI - C-Reactive protein and its implications in systemic lupus erythematosus. AB - C-reactive protein CRP is an acute-phase protein known as a biomarker for inflammation. As such CRP levels have been traditionally used to detect and predict the outcome of infections inflammatory and necrotic processes and to monitor the efficacy of treatment in these conditions. With the development of high sensitivity assays CRP has resurfaced as a very strong predictor in cardiovascular disease and as a mediator of atherosclerosis. The Centers for Disease Control and American Heart Association have elaborated guidelines for the use of CRP in the primary prevention setting and in patients with stable coronary disease or acute coronary syndromes. CRP has been used for differentiation between Systemic Lupus Erythematosus activity and infection in individuals without serositis. More recently CRP has also elicited interest as a therapeutic option in lupus. Murine lupus models treated with CRP have been reported to present delayed Lupus onset decreased antibody levels enhanced survival and reversal of ongoing proteinuria. In this paper we reviewed the multiple roles of CRP particularly in lupus. PMID- 18159198 TI - [Fetal safety profile of drugs used in the treatment of inflammatory rheumatic diseases]. AB - The high prevalence of inflammatory rheumatic diseases in women of childbearing age increases the risk of exposure to antirheumatic agents during conception pregnancy and breast feeding. The decision for pharmacological treatment initiation maintenance should be the result between the severity of maternal disease and the risk benefits with treatment. The aim of this paper was to review recent literature about drug fetal safety profile strength the importance of monitoring the pregnancy in patients with inflammatory rheumatic diseases and stress the need for further research in this area. PMID- 18159199 TI - [Surgical procedures for treatment of the rheumatoid knee]. AB - In the last decade considerable modifications in the surgical procedures recommended for the treatment of rheumatoid knee have been observed. This was due to all the medical developments achieved in pharmacology and therapeutic as well as a significant quality improvement of the rheumatologist s intervention. The synovectomy and namely the total knee arthroplasty represent the most commonly procedures used in the surgical treatment of the rheumatoid knee. An arthroscopic followed by a radionuclide synovectomy can be an appropriate treatment in a knee with an inflammatory arthritis Larsen radiograph grade I II . The ideal patient for synovectomy must present an early disease absence of deformity or instability good range of motion and preserved articular cartilage. On the other hand a total knee arthroplasty represents the only possible operation to treat a rheumatoid knee with a severe bone and cartilage damage Larsen radiograph grade IV V including younger patients. Total knee arthroplasty is actually a successful operation providing pain relief and the restoration of the function. Nevertheless the excellent good short and medium-term results achieved do not resist over time. Similarly to what happens with every other arthoplasty joint replacements the particules that come from the wear of the biomaterials included in its composition are the cause of biological intolerance reactions which can lead to the need of a new implant. The replacement prosthesis raises technical issues related to the reconstruction of bone mass losses where the cryopreserved bone allografts can be recommended. PMID- 18159200 TI - Systemic lupus erythematosus symptom checklist cross-cultural adaptation to Brazilian Portuguese language and reliability evaluation. AB - INTRODUCTION: Reliable and sensitive measurements are needed to evaluate quality of life QOL in patients with systemic lupus erythematosus SLE . No lupus-specific questionnaires are available in the Portuguese language. OBJECTIVES: This study describes the translation cultural adaptation and reliability testing of the Systemic Lupus Erythematosus Symptom Checklist questionnaire in Portuguese. PATIENTS AND METHODS: We evaluated 50 patients with SLE. A protocol was developed for translating and validating the SLE Symptom Checklist in accordance with the proposals in publications report on the methodology for translating questionnaires into foreign languages. RESULTS: With regard to cross-cultural adaptation some adaptations to people s culture had to be made without endangering the main objective of the questions in the original language. The correlation coefficients for internal consistency intraobserver reproducibility and interobserver reproducibility were 0.874 0.925 and 0.917 respectively. CONCLUSIONS: This questionnaire was shown to be reliable when applied to patients with SLE. Studies are currently being carried out to demonstrate its validity and sensitivity to changes. PMID- 18159201 TI - [Diagnostic performance of anti-cyclic citrullinated antibodies and IgM rheumatoid factor in rheumatoid arthritis]. AB - The presence of IgM class rheumatoid factor RF-M in the serum is one of the ACR criteria for the classification of rheumatoid arthritis RA . OBJECTIVES: To compare the sensitivity and specificity of two diagnostic markers for RA in Portuguese patients the 2nd generation anti-cyclic citrullinated peptide antibodies anti-CCP2 and the RF-M. METHODS: Serum samples from 56 patients with established RA 44 women and 12 men mean disease duration 9 5 -8 5 years were evaluated for the presence of anti-CCP2 and RF-M. Results were compared with 93 samples from a control group with identical demographic characteristics 43 from patients with spondylarthropaties and 50 from healthy blood donors . The determination of anti-CCP2 antibodies was performed by enzymatic fluroimunoassay and RF-M was measured by microElisa. RESULTS: Anti-CCP2 antibodies were positive in 40 patients with RA 71 4 at a mean antibody concentration of 350 5 UA ml range 11-1670 UA ml and negative in all controls. The RF-M was positive in 34 patients with RA 60 7 in 4 patients with spondylarthropaties 9 3 and in one blood donor 2 . The sensitivity specificity positive predictive value and negative predictive value for RF-M was 60 7 90 7 89 5 and 63 9 and for anti-CCP2 was 71 4 100 100 and 72 9 respectively. CONCLUSIONS: The anti-CCP2 assay showed an excellent specificity and a global performance superior to that of the RF-M test in this population with established RA. PMID- 18159202 TI - [Perceived pain and weather changes in rheumatic patients]. AB - INTRODUCTION: Rheumatic patients with chronic pain describe in a vivid way the influence of climate on pain and disease activity. Several studies seem to confirm this association. OBJECTIVES: To evaluate and compare in a population of rheumatic patients the perceived influence of weather changes on pain and disease activity METHODS: This is a retrospective cross-sectional study. For three weeks an assisted self-reported questionnaire with nine dimensions and a VAS pain scale was performed on consecutive out-patients in our clinic. RESULTS: 955 patients 787 female 168 male mean age 57.9 years with several rheumatologic diagnosis were evaluated. Overall 70 of the patients believed that the weather influenced their disease and 40 believed that the influence was high. Morning stiffness was influenced in 54 high influenced in 34 . Autumn and Winter were the most influential periods as well as humidity 67 and low temperatures 59 . CONCLUSION: In our study as well as in literature we found that a high percentage of patients 70 perceived that weather conditions influenced their pain and disease. Fibromyalgia patients seemed to be strongly influenced by weather changes. Our study confirms that patients perception on the influence of climate on pain and therefore their disease is an important clinical factor and it should be considered when evaluating rheumatic patients. PMID- 18159203 TI - Portuguese guidelines for the use of biological agents in rheumatoid arthritis- December 2007 update. AB - The authors present the revised version of the Portuguese Society of Rheumatology SPR guidelines for the treatment of rheumatoid arthritis RA with biological therapies. In these guidelines the criteria for introduction and maintenance of biological agents are discussed as well as the contraindications and procedures in case of non-responders. Biological treatment should be considered in RA patients with a disease activity score 28 DAS 28 superior to 3.2 despite treatment with 20mg week of methotrexate MTX for at least 3 months or if such treatment is not possible after 6 months of other conventional disease modifying drug or combination therapy. A DAS 28 score between 2.6 and 3.2 with a significant functional or radiological deterioration under treatment with conventional regimens could also constitute an indication for biological treatment. The follow-up should be performed each 3 months. The response criteria at the end of the first 3 months of treatment are a decrease of 0.6 in the DAS28 score. After 6 months of treatment response criteria is defined as follows for those with an initial DAS28 score superior to 5.1 a reduction of the DAS28 score below 4 is required for those with an initial DAS28 score inferior to 5.1 a decrease of the DAS28 score below 3.2 without a significant functional or radiological worsening is required. Non-responders in accordance to the Rheumatologist s clinical opinion should try a switch to other biological agent tumour necrosis factor alpha antagonist rituximab or abatacept . PMID- 18159204 TI - [Protocol for clinical monitoring of rheumatoid arthritis [PMAR]--December 2007 update]. AB - The authors present the update of a protocol for the clinical follow-up of Rheumatoid Arthritis Patients PMAR which aims to contribute to a standardized clinical observation of these patients particularly when they are being treated with biologic therapies. PMID- 18159205 TI - [Acute rheumatic fever associated with Henoch-Schonlein purpura. Case report and review of the literature]. AB - INTRODUCTION: Henoch-Schonlein purpura is one of the most common vasculitis in children. Some microorganisms have being suggested as possible etiological agents as group A streptococcus. CASE REPORT: R.L.B 7 years old presented with purpuric lesions in lower extremities and buttocks following fever and polyarthritis. After 7 days he arrived in our hospital showing pharyngitis new systolic murmur migratory polyarthritis and palpable purpura. His urinalysis had raised proteins and white cell count hemogram was normal sedimentation rate and streptococcal antibody titer were elevated. Electrocardiography showed a prolonged PR interval and echocardiogram confirmed moderate to severe mitral valve regurgitation. The patient was treated with prednisone 2mg kg day and penicillin G benzathine with clinical and laboratorial improvement. DISCUSSION: Literature reports HSP associated with rheumatic fever and carditis. Our case adds further evidence to the possibility of streptococcus being a causal agent of HSP. PMID- 18159206 TI - [Cystic Rheumatoid Arthritis--case report]. AB - Among the many radiological findings seen in Rheumatoid Arthritis RA small subchondral geodes and erosions are typical. Large geodes are far less common abnormalities and their presence may indicate diagnostic and therapeutic difficulties. We present a case report of a 55-year old woman with seronegative RA that developed a large geode in the knee with extensive joint destruction. PMID- 18159207 TI - [Fibrous dysplasia of bone--a case of polyostotic fibrous dysplasia]. AB - Fibrous Dysplasia FD of bone is a rare non-inheritable congenital disease characterized by a focal proliferation of fibrous tissue in the bone marrow leading to osteolytic lesions deformities and fractures. FD can be presented in a monostotic or polyostotic form this last one is usually more severe. FD primarily affects adolescents and young adults and in some cases in an asymptomatic way. We describe the case of a male patient with long evolution form of polyostotic fibrous dysplasia of bone with delayed clinical presentation. It is also highlighted the importance of an activity and involvement form evaluation as well as possible endocrine abnormalities associated in order to establish the correct treatment and prognosis. PMID- 18159208 TI - [Adverse cutaneous reaction to sulfasalazine]. PMID- 18159209 TI - [Camurati-Engelmann disease]. PMID- 18159210 TI - Increased IL-18 serum levels in patients with juvenile systemic lupus erythematosus. PMID- 18159211 TI - Intraocular rituximab. PMID- 18159213 TI - Identification of non-recurrent submicroscopic genome imbalances: the advantage of genome-wide microarrays over targeted approaches. AB - Genome-wide analysis of DNA copy-number changes using microarray-based technologies has enabled the detection of de novo cryptic chromosome imbalances in approximately 10% of individuals with mental retardation. So far, the majority of these submicroscopic microdeletions/duplications appear to be unique, hampering clinical interpretation and genetic counselling. We hypothesised that the genomic regions involved in these de novo submicroscopic aberrations would be candidates for recurrent copy-number changes in individuals with mental retardation. To test this hypothesis, we used multiplex ligation-dependent probe amplification (MLPA) to screen for copy number changes at eight genomic candidate regions in a European cohort of 710 individuals with idiopathic mental retardation. By doing so, we failed to detect additional submicroscopic rearrangements, indicating that the anomalies tested are non-recurrent in this cohort of patients. The break points flanking the candidate regions did not contain low copy repeats and/or sequence similarities, thus providing an explanation for its non-recurrent nature. On the basis of these data, we propose that the use of genome-wide microarrays is indicated when testing for copy-number changes in individuals with idiopathic mental retardation. PMID- 18159214 TI - IGF2/H19 hypomethylation in Silver-Russell syndrome and isolated hemihypoplasia. AB - Silver-Russell syndrome (SRS) is a clinically and genetically heterogeneous syndrome characterized by severe pre and postnatal growth retardation, body asymmetry and a typical facial phenotype with a triangular face and relative macrocephaly. In 30% of patients, the differentially methylated IGF2/H19 imprinting center region (ICR1) on chromosome 11p15 was found to be hypomethylated, as determined by Southern blot analysis of an HpaII restriction site close to the third CTCF-binding site (CTS3) within ICR1. Using bisulfite treatment and a real-time PCR-based methylation assay (QAMA), we analyzed the third and sixth CTCF-binding sites (CTS3, CTS6) in 5 patients with CTS3 hypomethylation, in 14 patients who were suspected to have SRS but were normal by Southern blot analysis, and in 1 patient with body asymmetry without any other features of SRS or Beckwith-Wiedemann syndrome (BWS). In all 5 patients with CTS3 hypomethylation, in 5 of 14 patients who were judged to be normal at CTS3 by Southern blot analysis and in the patient with isolated body asymmetry, we found CTS3 and CTS6 hypomethylation by QAMA. Using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA), we obtained similar results at four additional ICR1 sites in the CTS6 region. These results show that ICR1 hypomethylation occurs more often in SRS patients than as previously thought as well as in isolated hemihypoplasia. Furthermore, we show that methylation analysis by QAMA and MLPA is more sensitive in detecting ICR1 hypomethylation than Southern blot analysis of CTS3. PMID- 18159215 TI - The I,105V polymorphism in glutathione S-transferase P1, parental smoking and the risk for nonsyndromic cleft lip with or without cleft palate. AB - Genetic variations in the detoxification enzyme glutathione S-transferase P1 (GSTP1) may modify the teratogenicity of lifestyles, such as smoking. We investigated the role of the I105V polymorphism in GSTP1, parental periconception smoking, and their interaction with nonsyndromic cleft lip with or without cleft palate (CL/P) risk in the offspring. The GSTP1 I105V polymorphisms were determined in Dutch non-consanguineous Caucasians comprising of 155 CL/P triads (mother, father, child) and 195 control triads. The analyses were also carried out on complete triads only (n=69 CL/P and n=95 controls). Transmission disequilibrium testing and logistic regression analyses were performed. Neither maternal nor paternal smoking increased CL/P risk; odds ratios (OR): 1.2, 95 confidence intervals (CI)=0.7-2.0 and OR: 1.0, 95% CI=0.6-1.6, respectively. Carriership of the polymorphic Val105 allele in mothers may increase CL/P risk, OR: 1.5, 95% CI=0.96-2.5. Children homozygous for the Val105 allele may show an increased risk of CL/P, OR: 2.2, 95% CI=0.8-6.4. Maternal smoking tended to increase CL/P risk in mothers and children carrying Val105 alleles, OR=1.9, 95% CI=0.9-4.0 and OR=2.2, 95% CI=0.98-4.9, respectively. The highest risk for CL/P in children carrying Val105 alleles with a smoking father was 1.7, 95% CI=0.8 3.5. The GSTP1 I105V polymorphism in mothers and/or children either alone or in combination with maternal smoking may contribute to CL/P risk. Although of borderline significance, these results may underline the importance of smoking cessation in the periconception period for the prevention of CL/P in future generations. PMID- 18159216 TI - Nutrition in Pediatric Cardiomyopathy. AB - Pediatric cardiomyopathies are heterogeneous groups of serious disorders of the heart muscle and are responsible for significant morbidity and mortality among children who have the disease. While enormous improvements have been made in the treatment and survival of children with congenital heart disease, parallel strides have not been made in the outcomes for cardiomyopathies. Thus, ancillary therapies, such as nutrition and nutritional interventions, that may not cure but may potentially improve cardiac function and quality of life, are imperative to consider in children with all types of cardiomyopathy. Growth failure is one of the most significant clinical problems of children with cardiomyopathy with nearly one-third of children with this disorder manifesting some degree of growth failure during the course of their illness. Optimal intake of macronutrients can help improve cardiac function. In addition, several specific nutrients have been shown to correct myocardial abnormalities that often occur with cardiomyopathy and heart failure. In particular, antioxidants that can protect against free radical damage that often occurs in heart failure and nutrients that augment myocardial energy production are important therapies that have been explored more in adults with cardiomyopathy than in the pediatric population. Future research directions should pay particular attention to the effect of overall nutrition and specific nutritional therapies on clinical outcomes and quality of life in children with pediatric cardiomyopathy. PMID- 18159217 TI - Cartography of complex networks: modules and universal roles. AB - Integrative approaches to the study of complex systems demand that one knows the manner in which the parts comprising the system are connected. The structure of the complex network defining the interactions provides insight into the function and evolution of the components of the system. Unfortunately, the large size and intricacy of these networks implies that such insight is usually difficult to extract. Here, we propose a method that allows one to systematically extract and display information contained in complex networks. Specifically, we demonstrate that one can (i) find modules in complex networks and (ii) classify nodes into universal roles according to their pattern of within- and between-module connections. The method thus yields a 'cartographic representation' of complex networks. PMID- 18159218 TI - Fabrication of high aspect-ratio polymer microstructures for large-area electronic portal x-ray imagers. AB - Megavoltage x-ray imaging performed during radiotherapy is the method of choice for geometric verification of patient localization and dose delivery. Presently, such imaging is increasingly performed using electronic portal imaging devices (EPIDs) based on indirect detection active matrix flat panel imagers (AMFPIs). These devices use a scintillating phosphor screen in order to convert incident x rays into optical photons, which are then detected by the underlying active matrix photodiode array. The use of a continuous phosphor introduces a trade-off between x-ray quantum efficiency and spatial resolution, which limits current devices to use only ~2% of the incident x-rays. This trade-off can be circumvented by "segmented phosphor screens", comprising a two-dimensional matrix of optically-isolated cell structures filled with scintillating phosphor. In this work we describe the fabrication of millimeter-thick segmented phosphor screens using the MEMS (micro-electro-mechanical-system) polymer SU-8. This method is capable of being extended to large-area substrates. PMID- 18159219 TI - Oct4 expression is not required for mouse somatic stem cell self-renewal. AB - The Pou domain containing transcription factor Oct4 is a well-established regulator of pluripotency in the inner cell mass of the mammalian blastocyst as well as in embryonic stem cells. While it has been shown that the Oct4 gene is inactivated through a series of epigenetic modifications following implantation, recent studies have detected Oct4 activity in a variety of somatic stem cells and tumor cells. Based on these observations it has been suggested that Oct4 may also function in maintaining self-renewal of somatic stem cells and, in addition, may promote tumor formation. We employed a genetic approach to determine whether Oct4 is important for maintaining pluripotency in the stem cell compartments of several somatic tissues including the intestinal epithelium, bone marrow (hematopoietic and mesenchymal lineages), hair follicle, brain, and liver. Oct4 gene ablation in these tissues revealed no abnormalities in homeostasis or regenerative capacity. We conclude that Oct4 is dispensable for both self-renewal and maintenance of somatic stem cells in the adult mammal. PMID- 18159220 TI - Large Maf Transcription Factors: Cousins of AP-1 Proteins and Important Regulators of Cellular Differentiation. AB - A large number of mammalian transcription factors possess the evolutionary conserved basic and leucine zipper domain (bZIP). The basic domain interacts with DNA while the leucine zipper facilitates homo- and hetero-dimerization. These factors can be grouped into at least seven families: AP-1, ATF/CREB, CNC, C/EBP, Maf, PAR, and virus-encoded bZIPs. Here, we focus on a group of four large Maf proteins: MafA, MafB, c-Maf, and NRL. They act as key regulators of terminal differentiation in many tissues such as bone, brain, kidney, lens, pancreas, and retina, as well as in blood. The DNA-binding mechanism of large Mafs involves cooperation between the basic domain and an adjacent ancillary DNA-binding domain. Many genes regulated by Mafs during cellular differentiation use functional interactions between the Pax/Maf, Sox/Maf, and Ets/Maf promoter and enhancer modules. The prime examples are crystallin genes in lens and glucagon and insulin in pancreas. Novel roles for large Mafs emerged from studying generations of MafA and MafB knockouts and analysis of combined phenotypes in double or triple null mice. In addition, studies of this group of factors in invertebrates revealed the evolutionarily conserved function of these genes in the development of multicellular organisms. PMID- 18159222 TI - Population-Level Right Handedness for a Coordinated Bimanual Task in Chimpanzees: Replication and Extension in a Second Colony of Apes. AB - The purpose of this study was to evaluate the reliability of previously published findings on hand preferences in chimpanzees by evaluating hand use in a second colony of captive chimpanzees. We assessed hand preferences for a coordinated bimanual task in 116 chimpanzees (Pan troglodytes) at the University of Texas M. D. Anderson Cancer Center and compared them to previously published findings in captive chimpanzees at the Yerkes National Primate Research Center. The new sample showed significant population-level right handedness, which is consistent with previously published findings in the Yerkes chimpanzees. Combined data on the 2 chimpanzee colonies, revealed a significant effect of rearing history on hand preference, with wild-caught chimpanzees showing less right-handedness than captive-born mother-reared chimpanzees. We discuss the results in terms of the role of early environment on the development of laterality. PMID- 18159223 TI - Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein. AB - Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9 27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection. PMID- 18159224 TI - A Randomized Evaluation of Multidimensional Treatment Foster Care: Effects on School Attendance and Homework Completion in Juvenile Justice Girls. AB - Despite growing evidence that child welfare youth are at increased risk for juvenile delinquency, little is known about gender-specific processes and effective treatment programs for girls. Multidimensional Treatment Foster Care (MTFC), an empirically validated intervention for child welfare and juvenile justice populations, has demonstrated efficacy in reducing arrest rates in delinquent boys and girls. In this study, the efficacy of MTFC on school attendance and homework completion was examined in juvenile justice girls who were referred to out-of-home care (N = 81). Results from this randomized intervention trial suggest that MTFC was more effective than group care in increasing girls' school attendance and homework completion while in treatment and at 12 months postbaseline. In addition, the previously reported effect of MTFC on reducing girls' days in locked settings was mediated by homework completion while girls were enrolled in the intervention setting. Implications for policy and practice are described. PMID- 18159225 TI - Understanding the Point of Chimpanzee Pointing: Epigenesis and Ecological Validity. AB - Pointing has long been considered to be a uniquely human, universal, and biologically based gesture. However, pointing emerges spontaneously, without explicit training, in captive chimpanzees. Because pointing is commonplace in captive chimpanzees and virtually absent in wild chimpanzees, and because both captive and wild chimpanzees are sampled from the same gene pool, pointing by captive apes is attributable to environmental influences on communicative development. If pointing by captive chimpanzees is so variably expressed in different rearing environments, this suggests that pointing by humans may also be attributable to situational factors that make pointing effective in certain developmental contexts. PMID- 18159226 TI - Item Response Analysis of the Inventory of Depressive Symptomatology. AB - BACKGROUND: Both the clinician (IDS-C(30)) and self-report (IDS-SR(30)) versions of the 30-item Inventory of Depressive Symptomatology have acceptable psychiatric properties and have been used in various clinical studies. These two scales, however, have not been compared using item response theory (IRT) methods to determine whether the standard scoring methods are optimal. METHODS: Data were derived from 428 adult public sector outpatients with nonpsychotic major depressive disorder. The IDS-C(30) and IDS-SR(30) were compared using Samejima's graded response model. RESULTS: A model was constructed jointly fitting the IDS C(30) and IDS-SR(30). An improvement in scale performance was obtained by grouping selected items into domains (specifically sleep, psychomotor, and appetite/weight domains) analogous to the standard scoring of the 16-item Quick Inventory of Depressive Symptomatology. CONCLUSIONS: For the IDS-C(30) and IDS SR(30), standard scoring (ie, computing total score using all individual items) provides simplicity, comparability to published data, and a basis for clinical decision making. The revised scoring method, however, improves the utility of both scales when comparing groups as it provides explicit tests of item parameters. PMID- 18159227 TI - Genome evolution and the emergence of fruiting body development in Myxococcus xanthus. AB - BACKGROUND: Lateral gene transfer (LGT) is thought to promote speciation in bacteria, though well-defined examples have not been put forward. METHODOLOGY/PRINCIPLE FINDINGS: We examined the evolutionary history of the genes essential for a trait that defines a phylogenetic order, namely fruiting body development of the Myxococcales. Seventy-eight genes that are essential for Myxococcus xanthus development were examined for LGT. About 73% of the genes exhibit a phylogeny similar to that of the 16S rDNA gene and a codon bias consistent with other M. xanthus genes suggesting vertical transmission. About 22% have an altered codon bias and/or phylogeny suggestive of LGT. The remaining 5% are unique. Genes encoding signal production and sensory transduction were more likely to be transmitted vertically with clear examples of duplication and divergence into multigene families. Genes encoding metabolic enzymes were frequently acquired by LGT. Myxobacteria exhibit aerobic respiration unlike most of the delta Proteobacteria. M. xanthus contains a unique electron transport pathway shaped by LGT of genes for succinate dehydrogenase and three cytochrome oxidase complexes. CONCLUSIONS/SIGNIFICANCE: Fruiting body development depends on genes acquired by LGT, particularly those involved in polysaccharide production. We suggest that aerobic growth fostered innovation necessary for development by allowing myxobacteria access to a different gene pool from anaerobic members of the delta Proteobacteria. Habitat destruction and loss of species diversity could restrict the evolution of new bacterial groups by limiting the size of the prospective gene pool. PMID- 18159228 TI - Risk assessment of transmission of sporadic Creutzfeldt-Jakob disease in endodontic practice in absence of adequate prion inactivation. AB - BACKGROUND: Experimental results evidenced the infectious potential of the dental pulp of animals infected with transmissible spongiform encephalopathies (TSE). This route of iatrogenic transmission of sporadic Creutzfeldt-Jakob disease (sCJD) may exist in humans via reused endodontic instruments if inadequate prion decontamination procedures are used. METHODOLOGY/PRINCIPAL FINDINGS: To assess this risk, 10 critical parameters in the transmission process were identified, starting with contamination of an endodontic file during treatment of an infectious sCJD patient and ending with possible infection of a subsequent susceptible patient. It was assumed that a dose-risk response existed, with no risk below threshold values. Plausible ranges of those parameters were obtained through literature search and expert opinions, and a sensitivity analysis was conducted. Without effective prion-deactivation procedures, the risk of being infected during endodontic treatment ranged between 3.4 and 13 per million procedures. The probability that more than one case was infected secondary to endodontic treatment of an infected sCJD patient ranged from 47% to 77% depending on the assumed quantity of infective material necessary for disease transmission. If current official recommendations on endodontic instrument decontamination were strictly followed, the risk of secondary infection would become quasi-null. CONCLUSION: The risk of sCJD transmission through endodontic procedure compares with other health care risks of current concern such as death after liver biopsy or during general anaesthesia. These results show that single instrument use or adequate prion-decontamination procedures like those recently implemented in dental practice must be rigorously enforced. PMID- 18159229 TI - Axon myelin transfer of a non-enveloped virus. AB - We showed previously that Theiler's virus, a neurotropic non-enveloped picornavirus of mouse, traffics from the axon of infected neurons into the surrounding myelin. When this traffic is interrupted, as in the shiverer mouse which bears a mutation in the myelin basic protein gene, the virus is unable to persist in the central nervous system. In the present work, we used the Wld(s) mutant mouse, a strain in which axonal degeneration is considerably slowed down, to show that axon to myelin traffic takes place in the absence of axon degeneration. Our results suggest the existence of a mechanism of transfer of axonal cytoplasm into the myelin which Theiler's virus might exploit to ensure its persistence. PMID- 18159230 TI - A tale of switched functions: from cyclooxygenase inhibition to M-channel modulation in new diphenylamine derivatives. AB - Cyclooxygenase (COX) enzymes are molecular targets of nonsteroidal anti inflammatory drugs (NSAIDs), the most used medication worldwide. However, the COX enzymes are not the sole molecular targets of NSAIDs. Recently, we showed that two NSAIDs, diclofenac and meclofenamate, also act as openers of Kv7.2/3 K(+) channels underlying the neuronal M-current. Here we designed new derivatives of diphenylamine carboxylate to dissociate the M-channel opener property from COX inhibition. The carboxylate moiety was derivatized into amides or esters and linked to various alkyl and ether chains. Powerful M-channel openers were generated, provided that the diphenylamine moiety and a terminal hydroxyl group are preserved. In transfected CHO cells, they activated recombinant Kv7.2/3 K(+) channels, causing a hyperpolarizing shift of current activation as measured by whole-cell patch-clamp recording. In sensory dorsal root ganglion and hippocampal neurons, the openers hyperpolarized the membrane potential and robustly depressed evoked spike discharges. They also decreased hippocampal glutamate and GABA release by reducing the frequency of spontaneous excitatory and inhibitory post synaptic currents. In vivo, the openers exhibited anti-convulsant activity, as measured in mice by the maximal electroshock seizure model. Conversion of the carboxylate function into amide abolished COX inhibition but preserved M-channel modulation. Remarkably, the very same template let us generating potent M-channel blockers. Our results reveal a new and crucial determinant of NSAID-mediated COX inhibition. They also provide a structural framework for designing novel M channel modulators, including openers and blockers. PMID- 18159231 TI - Inhibition of Chk1 kills tetraploid tumor cells through a p53-dependent pathway. AB - Tetraploidy constitutes an adaptation to stress and an intermediate step between euploidy and aneuploidy in oncogenesis. Tetraploid cells are particularly resistant against genotoxic stress including radiotherapy and chemotherapy. Here, we designed a strategy to preferentially kill tetraploid tumor cells. Depletion of checkpoint kinase-1 (Chk1) by siRNAs, transfection with dominant-negative Chk1 mutants or pharmacological Chk1 inhibition killed tetraploid colon cancer cells yet had minor effects on their diploid counterparts. Chk1 inhibition abolished the spindle assembly checkpoint and caused premature and abnormal mitoses that led to p53 activation and cell death at a higher frequency in tetraploid than in diploid cells. Similarly, abolition of the spindle checkpoint by knockdown of Bub1, BubR1 or Mad2 induced p53-dependent apoptosis of tetraploid cells. Chk1 inhibition reversed the cisplatin resistance of tetraploid cells in vitro and in vivo, in xenografted human cancers. Chk1 inhibition activated p53-regulated transcripts including Puma/BBC3 in tetraploid but not in diploid tumor cells. Altogether, our results demonstrate that, in tetraploid tumor cells, the inhibition of Chk1 sequentially triggers aberrant mitosis, p53 activation and Puma/BBC3-dependent mitochondrial apoptosis. PMID- 18159232 TI - Molecular analysis of thymopentin binding to HLA-DR molecules. AB - Thymopentin (TP5) triggers an immune response by contacting with T cells; however the molecular basis of how TP5 achieves this process remains incompletely understood. According to the main idea of immunomodulation, we suppose that it would be necessary for TP5 to form complex with human class II major histocompatibility complex DR molecules (HLA-DR) before TP5 interacts with T cells. The uptake of TP5 by EBV-transformed B cells expressing HLA-DR molecules and the histogram of fluorescence intensities were observed by using fluorescent- labeled TP5, testifying the direct binding of TP5 to HLA-DR. The binding specificity was confirmed by the inhibition with unlabeled TP5, suggesting the recognition of TP5 by HLA-DR. To confirm the interaction between TP5 and HLA-DR, the complex formation was predicted by using various modeling strategies including six groups of trials with different parameters, alanine substitutions of TP5, and the mutants of HLA-DR. The results demonstrated that TP5 and its alanine substitutions assumed distinct conformations when they bound to HLA-DR. The observation further showed that there was flexibility in how the peptide bound within the binding cleft. Also, the molecular analysis supplemented a newly important discovery to the effect of Val anchor on TP5 binding HLA-DR, and revealed the important effects of Glu11 and Asn62 on the recognition of TP5. These results demonstrated the capability of TP5 to associate with HLA-DR in living antigen presenting cells (APC), thereby providing a new and promising strategy to understand the immunomodulation mechanism induced by TP5 and to design potential immunoregulatory polypeptides. PMID- 18159233 TI - External validation of a measurement tool to assess systematic reviews (AMSTAR). AB - BACKGROUND: Thousands of systematic reviews have been conducted in all areas of health care. However, the methodological quality of these reviews is variable and should routinely be appraised. AMSTAR is a measurement tool to assess systematic reviews. METHODOLOGY: AMSTAR was used to appraise 42 reviews focusing on therapies to treat gastro-esophageal reflux disease, peptic ulcer disease, and other acid-related diseases. Two assessors applied the AMSTAR to each review. Two other assessors, plus a clinician and/or methodologist applied a global assessment to each review independently. CONCLUSIONS: The sample of 42 reviews covered a wide range of methodological quality. The overall scores on AMSTAR ranged from 0 to 10 (out of a maximum of 11) with a mean of 4.6 (95% CI: 3.7 to 5.6) and median 4.0 (range 2.0 to 6.0). The inter-observer agreement of the individual items ranged from moderate to almost perfect agreement. Nine items scored a kappa of >0.75 (95% CI: 0.55 to 0.96). The reliability of the total AMSTAR score was excellent: kappa 0.84 (95% CI: 0.67 to 1.00) and Pearson's R 0.96 (95% CI: 0.92 to 0.98). The overall scores for the global assessment ranged from 2 to 7 (out of a maximum score of 7) with a mean of 4.43 (95% CI: 3.6 to 5.3) and median 4.0 (range 2.25 to 5.75). The agreement was lower with a kappa of 0.63 (95% CI: 0.40 to 0.88). Construct validity was shown by AMSTAR convergence with the results of the global assessment: Pearson's R 0.72 (95% CI: 0.53 to 0.84). For the AMSTAR total score, the limits of agreement were -0.19+/-1.38. This translates to a minimum detectable difference between reviews of 0.64 'AMSTAR points'. Further validation of AMSTAR is needed to assess its validity, reliability and perceived utility by appraisers and end users of reviews across a broader range of systematic reviews. PMID- 18159234 TI - The oral cavity and age: a site of chronic inflammation? AB - BACKGROUND: Aging may be accompanied by a low grade chronic up-regulation of inflammatory mediators. A variety of endogenous locally released mediators as well as inflammatory cells have been reported in the human oral cavity. The aim of this investigation was to determine the presence of different classes of inflammatory mediators in human saliva and correlate the levels with age. METHODOLOGY AND PRINCIPAL FINDINGS: Unstimulated whole buccal salivary samples were obtained in the morning from 94 healthy volunteers within 30 minutes after waking. None of the participants had taken aspirin in the week prior to the saliva collection. Lysozyme activity, eicosanoid levels (prostaglandin E(2) and leukotriene B(4)) and MMP-9 activity were measured. The antimicrobial activity (lysozyme activity) was not correlated with age whereas PGE(2) levels were markedly correlated with age (r = 0.29; P<0.05; n = 56). Saliva from healthy subjects (< or =40 years) compared with data derived from older volunteers (>40 years) demonstrated a significant increase in the mean values for PGE(2) and MMP 9 activity with age. In addition, significant correlations were observed between LTB(4) and PGE(2) (r = 0.28; P<0.05; n = 56) and between LTB(4) levels and MMP-9 activity in smokers (r = 0.78; P<0.001; n = 15). CONCLUSIONS/SIGNIFICANCE: The presence of significant levels and activity of inflammatory mediators in saliva suggests that the oral cavity of healthy subjects may be in a constant low state of inflammation associated with age. PMID- 18159235 TI - Arrest of cytoplasmic streaming induces algal proliferation in green paramecia. AB - A green ciliate Paramecium bursaria, bearing several hundreds of endosymbiotic algae, demonstrates rotational microtubule-based cytoplasmic streaming, in which cytoplasmic granules and endosymbiotic algae flow in a constant direction. However, its physiological significance is still unknown. We investigated physiological roles of cytoplasmic streaming in P. bursaria through host cell cycle using video-microscopy. Here, we found that cytoplasmic streaming was arrested in dividing green paramecia and the endosymbiotic algae proliferated only during the arrest of cytoplasmic streaming. Interestingly, arrest of cytoplasmic streaming with pressure or a microtubule drug also induced proliferation of endosymbiotic algae independently of host cell cycle. Thus, cytoplasmic streaming may control the algal proliferation in P. bursaria. Furthermore, confocal microscopic observation revealed that a division septum was formed in the constricted area of a dividing paramecium, producing arrest of cytoplasmic streaming. This is a first report to suggest that cytoplasmic streaming controls proliferation of eukaryotic cells. PMID- 18159236 TI - Colorectal hyperplasia and dysplasia due to human carcinoembryonic antigen (CEA) family member expression in transgenic mice. AB - CEA and CEACAM6 are immunoglobulin family intercellular adhesion molecules that are up-regulated without structural mutations in approximately 70% of human cancers. Results in in vitro systems showing tumorigenic effects for these molecules suggest that this correlation could indicate an instrumental role in tumorigenesis. To test whether this applies in vivo, transgenic mice harboring 187 kb of the human genome containing four CEA family member genes including the CEA and CEACAM6 genes were created and their copy numbers increased by mating until colonocyte expression levels reached levels seen in human colorectal carcinomas. The colonocyte surface level of integrin alpha5 and the activation of AKT increased progressively with the expression levels of CEA/CEACAM6. Colonic crypts showed a progressive increase in colonocyte proliferation, an increase in crypt fission, and a strong inhibition of both differentiation and anoikis/apoptosis. All transgenic mice showed massively enlarged colons comprising a continuous mosaic of severe hyperplasia, dysplasia and serrated adenomatous morphology. These results suggest that up-regulated non-mutated adhesion molecules could have a significant instrumental role in human cancer. PMID- 18159237 TI - High levels of MeCP2 depress MHC class I expression in neuronal cells. AB - BACKGROUND: The expression of MHC class I genes is repressed in mature neurons. The molecular basis of this regulation is poorly understood, but the genes are particularly rich in CpG islands. MeCP2 is a transcriptional repressor that binds to methylated CpG dinucleotides; mutations in this protein also cause the neurodevelopmental disease called Rett syndrome. Because MHC class I molecules play a role in neuronal connectivity, we hypothesised that MeCP2 might repress MHC class I expression in the CNS and that this might play a role in the pathology of Rett syndrome. METHODOLOGY: We show here that transiently transfected cells expressing high levels of MeCP2 specifically downregulate cell surface expression of MHC class I molecules in the neuronal cell line N2A and they prevent the induction of MHC class I expression in response to interferon in these cells, supporting our first hypothesis. Surprisingly, however, overexpression of the mutated forms of MeCP2 that cause Rett syndrome had a similar effect on MHC class I expression as the wild-type protein. Immunohistological analyses of brain slices from MECP2 knockout mice (the MeCP2(tm1.1Bird) strain) demonstrated a small but reproducible increase in MHC class I when compared to their wild type littermates, but we found no difference in MHC class I expression in primary cultures of mixed glial cells (mainly neurons and astrocytes) from the knockout and wild-type mice. CONCLUSION: These data suggest that high levels of MeCP2, such as those found in mature neurons, may contribute to the repression of MHC expression, but we find no evidence that MeCP2 regulation of MHC class I is important for the pathogenesis of Rett syndrome. PMID- 18159238 TI - Conditional transgenesis using Dimerizable Cre (DiCre). AB - Cre recombinase is extensively used to engineer the genome of experimental animals. However, its usefulness is still limited by the lack of an efficient temporal control over its activity. We have recently developed a conceptually new approach to regulate Cre recombinase, that we have called Dimerizable Cre or DiCre. It is based on splitting Cre into two inactive moieties and fusing them to FKBP12 (FK506-binding protein) and FRB (binding domain of the FKBP12-rapamycin associated protein), respectively. These latter can be efficiently hetero dimerized by rapamycin, leading to the reinstatement of Cre activity. We have been able to show, using in vitro approaches, that this ligand-induced dimerization is an efficient way to regulate Cre activity, and presents a low background activity together with a high efficiency of recombination following dimerization. To test the in vivo performance of this system, we have, in the present work, knocked-in DiCre into the Rosa26 locus of mice. To evaluate the performance of the DiCre system, mice have been mated with indicator mice (Z/EG or R26R) and Cre-induced recombination was examined following activation of DiCre by rapamycin during embryonic development or after birth of progenies. No recombination could be observed in the absence of treatment of the animals, indicating a lack of background activity of DiCre in the absence of rapamycin. Postnatal rapamycin treatment (one to five daily injection, 10 mg/kg i.p) induced recombination in a number of different tissues of progenies such as liver, heart, kidney, muscle, etc. On the other hand, recombination was at a very low level following in utero treatment of DiCrexR26R mice. In conclusion, DiCre has indeed the potentiality to be used to establish conditional Cre-deleter mice. An added advantage of this system is that, contrary to other modulatable Cre systems, it offers the possibility of obtaining regulated recombination in a combinatorial manner, i.e. induce recombination at any desired time-point specifically in cells characterized by the simultaneous expression of two different promoters. PMID- 18159239 TI - Involvement of skeletal muscle gene regulatory network in susceptibility to wound infection following trauma. AB - Despite recent advances in our understanding the pathophysiology of trauma, the basis of the predisposition of trauma patients to infection remains unclear. A Drosophila melanogaster/Pseudomonas aeruginosa injury and infection model was used to identify host genetic components that contribute to the hyper susceptibility to infection that follows severe trauma. We show that P. aeruginosa compromises skeletal muscle gene (SMG) expression at the injury site to promote infection. We demonstrate that activation of SMG structural components is under the control of cJun-N-terminal Kinase (JNK) Kinase, Hemipterous (Hep), and activation of this pathway promotes local resistance to P. aeruginosa in flies and mice. Our study links SMG expression and function to increased susceptibility to infection, and suggests that P. aeruginosa affects SMG homeostasis locally by restricting SMG expression in injured skeletal muscle tissue. Local potentiation of these host responses, and/or inhibition of their suppression by virulent P. aeruginosa cells, could lead to novel therapies that prevent or treat deleterious and potentially fatal infections in severely injured individuals. PMID- 18159240 TI - Sensitive commercial NASBA assay for the detection of respiratory syncytial virus in clinical specimen. AB - The aim of the study was to evaluate the usability of three diagnostic procedures for the detection of respiratory syncytial virus in clinical samples. Therefore, the FDA cleared CE marked NOW(R) RSV ELISA, the NucliSENS EasyQ RSV A+B NASBA, and a literature based inhouse RT-PCR protocol were compared for their relative sensitivities. Thereby, NASBA turned out to be the most sensitive method with a total number of 80 RSV positive samples out of a cohort of 251 nasopharyngeal washings from patients suffering from clinical symptoms, followed by the inhouse RT-PCR (62/251) and ELISA (52/251). Thus, NASBA may serve as a rapid and highly sensitive alternative for RSV diagnostics. PMID- 18159241 TI - The complete genome sequence and analysis of the epsilonproteobacterium Arcobacter butzleri. AB - BACKGROUND: Arcobacter butzleri is a member of the epsilon subdivision of the Proteobacteria and a close taxonomic relative of established pathogens, such as Campylobacter jejuni and Helicobacter pylori. Here we present the complete genome sequence of the human clinical isolate, A. butzleri strain RM4018. METHODOLOGY/PRINCIPAL FINDINGS: Arcobacter butzleri is a member of the Campylobacteraceae, but the majority of its proteome is most similar to those of Sulfuromonas denitrificans and Wolinella succinogenes, both members of the Helicobacteraceae, and those of the deep-sea vent Epsilonproteobacteria Sulfurovum and Nitratiruptor. In addition, many of the genes and pathways described here, e.g. those involved in signal transduction and sulfur metabolism, have been identified previously within the epsilon subdivision only in S. denitrificans, W. succinogenes, Sulfurovum, and/or Nitratiruptor, or are unique to the subdivision. In addition, the analyses indicated also that a substantial proportion of the A. butzleri genome is devoted to growth and survival under diverse environmental conditions, with a large number of respiration-associated proteins, signal transduction and chemotaxis proteins and proteins involved in DNA repair and adaptation. To investigate the genomic diversity of A. butzleri strains, we constructed an A. butzleri DNA microarray comprising 2238 genes from strain RM4018. Comparative genomic indexing analysis of 12 additional A. butzleri strains identified both the core genes of A. butzleri and intraspecies hypervariable regions, where <70% of the genes were present in at least two strains. CONCLUSION/SIGNIFICANCE: The presence of pathways and loci associated often with non-host-associated organisms, as well as genes associated with virulence, suggests that A. butzleri is a free-living, water-borne organism that might be classified rightfully as an emerging pathogen. The genome sequence and analyses presented in this study are an important first step in understanding the physiology and genetics of this organism, which constitutes a bridge between the environment and mammalian hosts. PMID- 18159242 TI - Up-regulation of intestinal vascular endothelial growth factor by Afa/Dr diffusely adhering Escherichia coli. AB - BACKGROUND: Angiogenesis has been recently described as a novel component of inflammatory bowel disease pathogenesis. The level of vascular endothelial growth factor (VEGF) has been found increased in Crohn's disease and ulcerative colitis mucosa. To question whether a pro-inflammatory Escherichia coli could regulate the expression of VEGF in human intestinal epithelial cells, we examine the response of cultured human colonic T84 cells to infection by E. coli strain C1845 that belongs to the typical Afa/Dr diffusely adhering E. coli family (Afa/Dr DAEC). METHODOLOGY: VEGF mRNA expression was examined by Northern blotting and q PCR. VEGF protein levels were assayed by ELISA and its bioactivity was analysed in endothelial cells. The bacterial factor involved in VEGF induction was identified using recombinant E. coli expressing Dr adhesin, purified Dr adhesin and lipopolysaccharide. The signaling pathway activated for the up-regulation of VEGF was identified using a blocking monoclonal anti-DAF antibody, Western blot analysis and specific pharmacological inhibitors. PRINCIPAL FINDINGS: C1845 bacteria induce the production of VEGF protein which is bioactive. VEGF is induced by adhering C1845 in both a time- and bacteria concentration-dependent manner. This phenomenon is not cell line dependent since we reproduced this observation in intestinal LS174, Caco2/TC7 and INT407 cells. Up-regulation of VEGF production requires: (1) the interaction of the bacterial F1845 adhesin with the brush border-associated decay accelerating factor (DAF, CD55) acting as a bacterial receptor, and (2) the activation of a Src protein kinase upstream of the activation of the Erk and Akt signaling pathways. CONCLUSIONS: Results demonstrate that a Afa/Dr DAEC strain induces an adhesin-dependent activation of DAF signaling that leads to the up-regulation of bioactive VEGF in cultured human intestinal cells. Thus, these results suggest a link between an entero-adherent, pro-inflammatory E. coli strain and angiogenesis which appeared recently as a novel component of IBD pathogenesis. PMID- 18159243 TI - An animal model of emotional blunting in schizophrenia. AB - Schizophrenia is often associated with emotional blunting--the diminished ability to respond to emotionally salient stimuli--particularly those stimuli representative of negative emotional states, such as fear. This disturbance may stem from dysfunction of the amygdala, a brain region involved in fear processing. The present article describes a novel animal model of emotional blunting in schizophrenia. This model involves interfering with normal fear processing (classical conditioning) in rats by means of acute ketamine administration. We confirm, in a series of experiments comprised of cFos staining, behavioral analysis and neurochemical determinations, that ketamine interferes with the behavioral expression of fear and with normal fear processing in the amygdala and related brain regions. We further show that the atypical antipsychotic drug clozapine, but not the typical antipsychotic haloperidol nor an experimental glutamate receptor 2/3 agonist, inhibits ketamine's effects and retains normal fear processing in the amygdala at a neurochemical level, despite the observation that fear-related behavior is still inhibited due to ketamine administration. Our results suggest that the relative resistance of emotional blunting to drug treatment may be partially due to an inability of conventional therapies to target the multiple anatomical and functional brain systems involved in emotional processing. A conceptual model reconciling our findings in terms of neurochemistry and behavior is postulated and discussed. PMID- 18159244 TI - Genome wide association (GWA) study for early onset extreme obesity supports the role of fat mass and obesity associated gene (FTO) variants. AB - BACKGROUND: Obesity is a major health problem. Although heritability is substantial, genetic mechanisms predisposing to obesity are not very well understood. We have performed a genome wide association study (GWA) for early onset (extreme) obesity. METHODOLOGY/PRINCIPAL FINDINGS: a) GWA (Genome-Wide Human SNP Array 5.0 comprising 440,794 single nucleotide polymorphisms) for early onset extreme obesity based on 487 extremely obese young German individuals and 442 healthy lean German controls; b) confirmatory analyses on 644 independent families with at least one obese offspring and both parents. We aimed to identify and subsequently confirm the 15 SNPs (minor allele frequency > or =10%) with the lowest p-values of the GWA by four genetic models: additive, recessive, dominant and allelic. Six single nucleotide polymorphisms (SNPs) in FTO (fat mass and obesity associated gene) within one linkage disequilibrium (LD) block including the GWA SNP rendering the lowest p-value (rs1121980; log-additive model: nominal p = 1.13 x 10(-7), corrected p = 0.0494; odds ratio (OR)(CT) 1.67, 95% confidence interval (CI) 1.22-2.27; OR(TT) 2.76, 95% CI 1.88-4.03) belonged to the 15 SNPs showing the strongest evidence for association with obesity. For confirmation we genotyped 11 of these in the 644 independent families (of the six FTO SNPs we chose only two representing the LD bock). For both FTO SNPs the initial association was confirmed (both Bonferroni corrected p<0.01). However, none of the nine non-FTO SNPs revealed significant transmission disequilibrium. CONCLUSIONS/SIGNIFICANCE: Our GWA for extreme early onset obesity substantiates that variation in FTO strongly contributes to early onset obesity. This is a further proof of concept for GWA to detect genes relevant for highly complex phenotypes. We concurrently show that nine additional SNPs with initially low p values in the GWA were not confirmed in our family study, thus suggesting that of the best 15 SNPs in the GWA only the FTO SNPs represent true positive findings. PMID- 18159245 TI - Noncompaction of the ventricular myocardium is associated with a de novo mutation in the beta-myosin heavy chain gene. AB - Noncompaction of the ventricular myocardium (NVM) is the morphological hallmark of a rare familial or sporadic unclassified heart disease of heterogeneous origin. NVM results presumably from a congenital developmental error and has been traced back to single point mutations in various genes. The objective of this study was to determine the underlying genetic defect in a large German family suffering from NVM. Twenty four family members were clinically assessed using advanced imaging techniques. For molecular characterization, a genome-wide linkage analysis was undertaken and the disease locus was mapped to chromosome 14ptel-14q12. Subsequently, two genes of the disease interval, MYH6 and MYH7 (encoding the alpha- and beta-myosin heavy chain, respectively) were sequenced, leading to the identification of a previously unknown de novo missense mutation, c.842G>C, in the gene MYH7. The mutation affects a highly conserved amino acid in the myosin subfragment-1 (R281T). In silico simulations suggest that the mutation R281T prevents the formation of a salt bridge between residues R281 and D325, thereby destabilizing the myosin head. The mutation was exclusively present in morphologically affected family members. A few members of the family displayed NVM in combination with other heart defects, such as dislocation of the tricuspid valve (Ebstein's anomaly, EA) and atrial septal defect (ASD). A high degree of clinical variability was observed, ranging from the absence of symptoms in childhood to cardiac death in the third decade of life. The data presented in this report provide first evidence that a mutation in a sarcomeric protein can cause noncompaction of the ventricular myocardium. PMID- 18159246 TI - High uptake of exclusive breastfeeding and reduced early post-natal HIV transmission. AB - BACKGROUND: Empirical data showing the clear benefits of exclusive breastfeeding (EBF) for HIV prevention are needed to encourage implementation of lactation support programs for HIV-infected women in low resource settings among whom replacement feeding is unsafe. We conducted a prospective, observational study in Lusaka, Zambia, to test the hypothesis that EBF is associated with a lower risk of postnatal HIV transmission than non-EBF. METHODS AND RESULTS: As part of a randomized trial of early weaning, 958 HIV-infected women and their infants were recruited and all were encouraged to breastfeed exclusively to 4 months. Single dose nevirapine was provided to prevent transmission. Regular samples were collected from infants to 24 months of age and tested by PCR. Detailed measurements of actual feeding behaviors were collected to examine, in an observational analysis, associations between feeding practices and postnatal HIV transmission. Uptake of EBF was high with 84% of women reporting only EBF cumulatively to 4 months. Post-natal HIV transmission before 4 months was significantly lower (p = 0.004) among EBF (0.040 95% CI: 0.024-0.055) than non EBF infants (0.102 95% CI: 0.047-0.157); time-dependent Relative Hazard (RH) of transmission due to non-EBF = 3.48 (95% CI: 1.71-7.08). There were no significant differences in the severity of disease between EBF and non-EBF mothers and the association remained significant (RH = 2.68 95% CI: 1.28-5.62) after adjusting for maternal CD4 count, plasma viral load, syphilis screening results and low birth weight. CONCLUSIONS: Non-EBF more than doubles the risk of early postnatal HIV transmission. Programs to support EBF should be expanded universally in low resource settings. EBF is an affordable, feasible, acceptable, safe and sustainable practice that also reduces HIV transmission providing HIV-infected women with a means to protect their children's lives. TRIAL REGISTRATION: ClinicalTrials.gov NCT00310726. PMID- 18159247 TI - Hypoxia inducible factor (HIF)-1 coordinates induction of Toll-like receptors TLR2 and TLR6 during hypoxia. AB - BACKGROUND: During acute infection and inflammation, dramatic shifts in tissue metabolism are typical, thereby resulting in profound tissue hypoxia. Therefore, we pursued the hypothesis, that tissue hypoxia may influence innate immune responses by transcriptional modulation of Toll-like receptor (TLRs) expression and function. METHODOLOGY/PRINCIPAL FINDINGS: We gained first insight from transcriptional profiling of murine dendritic cells exposed to hypoxia (2% oxygen for 24 h). While transcript levels of other TLRs remained unchanged, we found a robust induction of TLR2 (2.36+/-0.7-fold; P<0.05) and TLR6 (3.46+/-1.56-fold; P<0.05). Additional studies in different cells types and cell-lines including human dendritic cells, monocytic cells (MM6), endothelia (HMEC-1) or intestinal epithelia (Caco-2) confirmed TLR2 and TLR6 induction of transcript, protein and function during hypoxia. Furthermore, analysis of the putative TLR2 and TLR6 promoters revealed previously unrecognized binding sites for HIF-1, which were shown by chromatin immunoprecipitation to bind the pivotal hypoxia-regulating transcription factor HIF-1alpha. Studies using loss and gain of function of HIF-1 confirmed a critical role of HIF-1alpha in coordinating TLR2 and TLR6 induction. Moreover, studies of murine hypoxia (8% oxygen over 6 h) showed TLR2 and TLR 6 induction in mucosal organs in vivo. In contrast, hypoxia induction of TLR2 and TLR6 was abolished in conditional HIF-1alpha mutant mice. CONCLUSIONS/SIGNIFICANCE: Taking together, these studies reveal coordinated induction of TLR2 and TLR6 during hypoxia and suggest tissue hypoxia in transcriptional adaptation of innate immune responses during acute infection or inflammation. PMID- 18159249 TI - Stress preconditioning of spreading depression in the locust CNS. AB - Cortical spreading depression (CSD) is closely associated with important pathologies including stroke, seizures and migraine. The mechanisms underlying SD in its various forms are still incompletely understood. Here we describe SD-like events in an invertebrate model, the ventilatory central pattern generator (CPG) of locusts. Using K(+) -sensitive microelectrodes, we measured extracellular K(+) concentration ([K(+)](o)) in the metathoracic neuropile of the CPG while monitoring CPG output electromyographically from muscle 161 in the second abdominal segment to investigate the role K(+) in failure of neural circuit operation induced by various stressors. Failure of ventilation in response to different stressors (hyperthermia, anoxia, ATP depletion, Na(+)/K(+) ATPase impairment, K(+) injection) was associated with a disturbance of CNS ion homeostasis that shares the characteristics of CSD and SD-like events in vertebrates. Hyperthermic failure was preconditioned by prior heat shock (3 h, 45 degrees C) and induced-thermotolerance was associated with an increase in the rate of clearance of extracellular K(+) that was not linked to changes in ATP levels or total Na(+)/K(+) ATPase activity. Our findings suggest that SD-like events in locusts are adaptive to terminate neural network operation and conserve energy during stress and that they can be preconditioned by experience. We propose that they share mechanisms with CSD in mammals suggesting a common evolutionary origin. PMID- 18159248 TI - Ultrasonic communication in rats: can playback of 50-kHz calls induce approach behavior? AB - Rats emit distinct types of ultrasonic vocalizations, which differ depending on age, the subject's current state and environmental factors. Since it was shown that 50-kHz calls can serve as indices of the animal's positive subjective state, they have received increasing experimental attention, and have successfully been used to study neurobiological mechanisms of positive affect. However, it is likely that such calls do not only reflect a positive affective state, but that they also serve a communicative purpose. Actually, rats emit the highest rates of 50-kHz calls typically during social interactions, like reproductive behavior, juvenile play and tickling. Furthermore, it was recently shown that rats emit 50 kHz calls after separation from conspecifics. The aim of the present study was to test the communicative value of such 50-kHz calls. In a first experiment, conducted in juvenile rats situated singly on a radial maze apparatus, we showed that 50-kHz calls can induce behavioral activation and approach responses, which were selective to 50-kHz signals, since presentation of 22-kHz calls, considered to be aversive or threat signals, led to behavioral inhibition. In two other experiments, we used either natural 50-kHz calls, which had been previously recorded from other rats, or artificial sine wave stimuli, which were identical to these calls with respect to peak frequency, call length and temporal appearance. These signals were presented to either juvenile (Exp. 2) or adult (Exp. 3) male rats. Our data clearly show that 50-kHz signals can induce approach behavior, an effect, which was more pronounced in juvenile rats and which was not selective to natural calls, especially in adult rats. The recipient rats also emitted some 50-kHz calls in response to call presentation, but this effect was observed only in adult subjects. Together, our data show that 50-kHz calls can serve communicative purposes, namely as a social signal, which increases the likelihood of approach in the recipient conspecific. PMID- 18159250 TI - Empirical comparison of Simple Sequence Repeats and single nucleotide polymorphisms in assessment of maize diversity and relatedness. AB - While Simple Sequence Repeats (SSRs) are extremely useful genetic markers, recent advances in technology have produced a shift toward use of single nucleotide polymorphisms (SNPs). The different mutational properties of these two classes of markers result in differences in heterozygosities and allele frequencies that may have implications for their use in assessing relatedness and evaluation of genetic diversity. We compared analyses based on 89 SSRs (primarily dinucleotide repeats) to analyses based on 847 SNPs in individuals from the same 259 inbred maize lines, which had been chosen to represent the diversity available among current and historic lines used in breeding. The SSRs performed better at clustering germplasm into populations than did a set of 847 SNPs or 554 SNP haplotypes, and SSRs provided more resolution in measuring genetic distance based on allele-sharing. Except for closely related pairs of individuals, measures of distance based on SSRs were only weakly correlated with measures of distance based on SNPs. Our results suggest that 1) large numbers of SNP loci will be required to replace highly polymorphic SSRs in studies of diversity and relatedness and 2) relatedness among highly-diverged maize lines is difficult to measure accurately regardless of the marker system. PMID- 18159251 TI - Shotgun lipidomics identifies a paired rule for the presence of isomeric ether phospholipid molecular species. AB - BACKGROUND: Ether phospholipids are abundant membrane constituents present in electrically active tissues (e.g., heart and the brain) that play important roles in cellular function. Alterations of ether phospholipid molecular species contents are associated with a number of genetic disorders and human diseases. METHODOLOGY/PRINCIPAL FINDINGS: Herein, the power of shotgun lipidomics, in combination with high mass accuracy/high resolution mass spectrometry, was explored to identify a paired rule for the presence of isomeric ether phospholipid molecular species in cellular lipidomes. The rule predicts that if an ether phospholipid A'-B is present in a lipidome, its isomeric counterpart B' A is also present (where the ' represents an ether linkage). The biochemical basis of this rule results from the fact that the enzymes which participate in either the sequential oxidation of aliphatic alcohols to fatty acids, or the reduction of long chain fatty acids to aliphatic alcohols (metabolic precursors of ether lipid synthesis), are not entirely selective with respect to acyl chain length or degree of unsaturation. Moreover, the enzymatic selectivity for the incorporation of different aliphatic chains into the obligatory precursor of ether lipids (i.e., 1-O-alkyl-glycero-3-phosphate) is also limited. CONCLUSIONS/SIGNIFICANCE: This intrinsic amplification of the number of lipid molecular species present in biological membranes predicted by this rule and demonstrated in this study greatly expands the number of ether lipid molecular species present in cellular lipidomes. Application of this rule to mass spectrometric analyses provides predictive clues to the presence of specific molecular species and greatly expands the number of identifiable and quantifiable ether lipid species present in biological samples. Through appropriate alterations in the database, use of the paired rule increases the number of identifiable metabolites in metabolic networks, thereby facilitating identification of biomarkers presaging disease states. PMID- 18159252 TI - Neuregulin-1 regulates cell adhesion via an ErbB2/phosphoinositide-3 kinase/Akt dependent pathway: potential implications for schizophrenia and cancer. AB - BACKGROUND: Neuregulin-1 (NRG1) is a putative schizophrenia susceptibility gene involved extensively in central nervous system development as well as cancer invasion and metastasis. Using a B lymphoblast cell model, we previously demonstrated impairment in NRG1alpha-mediated migration in cells derived from patients with schizophrenia as well as effects of risk alleles in NRG1 and catechol-O-methyltransferase (COMT), a second gene implicated both in schizophrenia susceptibility and in cancer. METHODOLOGY/PRINCIPAL FINDINGS: Here, we examine cell adhesion, an essential component process of cell motility, using an integrin-mediated cell adhesion assay based on an interaction between ICAM-1 and the CD11a/CD18 integrin heterodimer expressed on lymphoblasts. In our assay, NRG1alpha induces lymphoblasts to assume varying levels of adhesion characterized by time-dependent fluctuations in the firmness of attachment. The maximum range of variation in adhesion over sixty minutes correlates strongly with NRG1alpha induced migration (r(2) = 0.61). NRG1alpha-induced adhesion variation is blocked by erbB2, PI3K, and Akt inhibitors, but not by PLC, ROCK, MLCK, or MEK inhibitors, implicating the erbB2/PI3K/Akt1 signaling pathway in NRG1-stimulated, integrin-mediated cell adhesion. In cell lines from 20 patients with schizophrenia and 20 normal controls, cells from patients show a significant deficiency in the range of NRG1alpha-induced adhesion (p = 0.0002). In contrast, the response of patient-derived cells to phorbol myristate acetate is unimpaired. The COMT Val108/158Met genotype demonstrates a strong trend towards predicting the range of the NRG1alpha-induced adhesion response with risk homozygotes having decreased variation in cell adhesion even in normal subjects (p = 0.063). CONCLUSION/SIGNIFICANCE: Our findings suggest that a mechanism of the NRG1 genetic association with schizophrenia may involve the molecular biology of cell adhesion. PMID- 18159253 TI - Cryptococcus neoformans is resistant to surfactant protein A mediated host defense mechanisms. AB - Initiation of a protective immune response to infection by the pathogenic fungus Cryptococcus neoformans is mediated in part by host factors that promote interactions between immune cells and C. neoformans yeast. Surfactant protein A (SP-A) contributes positively to pulmonary host defenses against a variety of bacteria, viruses, and fungi in part by promoting the recognition and phagocytosis of these pathogens by alveolar macrophages. In the present study we investigated the role of SP-A as a mediator of host defense against the pulmonary pathogen, C. neoformans. Previous studies have shown that SP-A binds to acapsular and minimally encapsulated strains of C. neoformans. Using in vitro binding assays we confirmed that SP-A does not directly bind to a fully encapsulated strain of C. neoformans (H99). However, we observed that when C. neoformans was incubated in bronchoalveolar fluid, SP-A binding was detected, suggesting that another alveolar host factor may enable SP-A binding. Indeed, we discovered that SP-A binds encapsulated C. neoformans via a previously unknown IgG dependent mechanism. The consequence of this interaction was the inhibition of IgG-mediated phagocytosis of C. neoformans by alveolar macrophages. Therefore, to assess the contribution of SP-A to the pulmonary host defenses we compared in vivo infections using SP-A null mice (SP-A-/-) and wild-type mice in an intranasal infection model. We found that the immune response assessed by cellular counts, TNFalpha cytokine production, and fungal burden in lungs and bronchoalveolar lavage fluids during early stages of infection were equivalent. Furthermore, the survival outcome of C. neoformans infection was equivalent in SP-A-/- and wild type mice. Our results suggest that unlike a variety of bacteria, viruses, and other fungi, progression of disease with an inhalational challenge of C. neoformans does not appear to be negatively or positively affected by SP-A mediated mechanisms of pulmonary host defense. PMID- 18159254 TI - Sterile protection against malaria is independent of immune responses to the circumsporozoite protein. AB - BACKGROUND: Research aimed at developing vaccines against infectious diseases generally seeks to induce robust immune responses to immunodominant antigens. This approach has led to a number of efficient bacterial and viral vaccines, but it has yet to do so for parasitic pathogens. For malaria, a disease of global importance due to infection by Plasmodium protozoa, immunization with radiation attenuated sporozoites uniquely leads to long lasting sterile immunity against infection. The circumsporozoite protein (CSP), an important component of the sporozoite's surface, remains the leading candidate antigen for vaccines targeting the parasite's pre-erythrocytic stages. Difficulties in developing CSP based vaccines that reproduce the levels of protection afforded by radiation attenuated sporozoites have led us to question the role of CSP in the acquisition of sterile immunity. We have used a parasite transgenic for the CSP because it allowed us to test whether a major immunodominant Plasmodium antigen is indeed needed for the induction of sterile protective immunity against infection. METHODOLOGY/MAIN FINDINGS: We employed a P. berghei parasite line that expresses a heterologous CSP from P. falciparum in order to assess the role of the CSP in the protection conferred by vaccination with radiation-attenuated P. berghei parasites. Our data demonstrated that sterile immunity could be obtained despite the absence of immune responses specific to the CSP expressed by the parasite used for challenge. CONCLUSIONS: We conclude that other pre-erythrocytic parasite antigens, possibly hitherto uncharacterised, can be targeted to induce sterile immunity against malaria. From a broader perspective, our results raise the question as to whether immunodominant parasite antigens should be the favoured targets for vaccine development. PMID- 18159255 TI - A ribosomal S-6 kinase-mediated signal to C/EBP-beta is critical for the development of liver fibrosis. AB - BACKGROUND: In response to liver injury, hepatic stellate cell (HSC) activation causes excessive liver fibrosis. Here we show that activation of RSK and phosphorylation of C/EBPbeta on Thr217 in activated HSC is critical for the progression of liver fibrosis. METHODOLOGY/PRINCIPAL FINDINGS: Chronic treatment with the hepatotoxin CCl(4) induced severe liver fibrosis in C/EBPbeta(+/+) mice but not in mice expressing C/EBPbeta-Ala217, a non-phosphorylatable RSK inhibitory transgene. C/EBPbeta-Ala217 was present within the death receptor complex II, with active caspase 8, and induced apoptosis of activated HSC. The C/EBPbeta-Ala217 peptides directly stimulated caspase 8 activation in a cell-free system. C/EBPbeta(+/+) mice with CCl(4)-induced severe liver fibrosis, while continuing on CCl(4), were treated with a cell permeant RSK-inhibitory peptide for 4 or 8 weeks. The peptide inhibited RSK activation, stimulating apoptosis of HSC, preventing progression and inducing regression of liver fibrosis. We found a similar activation of RSK and phosphorylation of human C/EBPbeta on Thr266 (human phosphoacceptor) in activated HSC in patients with severe liver fibrosis but not in normal livers, suggesting that this pathway may also be relevant in human liver fibrosis. CONCLUSIONS/SIGNIFICANCE: These data indicate that the RSK C/EBPbeta phosphorylation pathway is critical for the development of liver fibrosis and suggest a potential therapeutic target. PMID- 18159256 TI - Fever as a cause of hypophosphatemia in patients with malaria. AB - Hypophosphatemia occurs in 40 to 60% of patients with acute malaria, and in many other conditions associated with elevations of body temperature. To determine the prevalence and causes of hypophosphatemia in patients with malaria, we retrospectively studied all adults diagnosed with acute malaria during a 12-year period. To validate our findings, we analyzed a second sample of malaria patients during a subsequent 10-year period. Serum phosphorus correlated inversely with temperature (n = 59, r = -0.62; P<0.0001), such that each 1 degrees C increase in body temperature was associated with a reduction of 0.18 mmol/L (0.56 mg/dL) in the serum phosphorus level (95% confidence interval: -0.12 to -0.24 mmol/L [-0.37 to -0.74 mg/dL] per 1 degrees C). A similar effect was observed among 19 patients who had repeat measurements of serum phosphorus and temperature. In a multiple linear regression analysis, the relation between temperature and serum phosphorus level was independent of blood pH, PCO2, and serum levels of potassium, bicarbonate, calcium, albumin, and glucose. Our study demonstrates a strong inverse linear relation between body temperature and serum phosphorus level that was not explained by other factors known to cause hypophosphatemia. If causal, this association can account for the high prevalence of hypophosphatemia, observed in our patients and in previous studies of patients with malaria. Because hypophosphatemia has been observed in other clinical conditions characterized by fever or hyperthermia, this relation may not be unique to malaria. Elevation of body temperature should be added to the list of causes of hypophosphatemia. PMID- 18159258 TI - A guide to contraindications to childhood vaccinations. PMID- 18159257 TI - Rotavirus vaccine withdrawal in the United states; the role of postmarketing surveillance. PMID- 18159259 TI - Febrile illness in a toddler: the diagnostic clue lays skin deep. PMID- 18159260 TI - Establishing priorities for national communicable disease surveillance. PMID- 18159261 TI - Clostridium difficile-associated diarrhea - the new scourge of the health care facility. PMID- 18159262 TI - Long term trends in the occurrence of nosocomial blood stream infection. AB - OBJECTIVE: To determine trends in the occurrence of nosocomial blood stream infection at the University of Alberta Hospital. METHODS: A prospective survey of nosocomial blood stream infection was conducted; cases from August 1986 to December 1996 were reviewed. Cases were detected by a review of positive blood cultures reported by the microbiology laboratory. Centers for Disease Control and Prevention definitions of nosocomial infection were used to categorize isolates as nosocomial, community acquired or contaminant. RESULTS: There were 2389 cases; primary bacteremia was the most common source (57%), followed by urinary tract, respiratory tract and surgical site sources (10% each). The nosocomial blood steam infection rate rose progressively from 6.0/1000 admissions and 4.59/10,000 patient days in 1986 to 11.2/1000 admissions and 14.31/10,000 days in 1996 (P<0.01); 48% of the total increase in rate occurred between 1995 and 1996. Significant increases occurred between 1986 and 1996 in primary infections (from 3.2 to 7.5/1000 admissions, P<0.01) and infections from all secondary sources (from 2.5 to 3.8/1000 admissions, P=0.01). Coagulase-negative staphylococci (27%), Staphylococcus aureus (19%) and enterococci (9%) were the most common microbial causes. Aerobic Gram-negative bacilli accounted for 28% and candida for 6%. Coagulase-negative staphylococci, enterococci and candida all became more prevalent as causes of infection over the study period. CONCLUSIONS: The nosocomial blood stream infection rate in the hospital has nearly doubled in the past 10 years, largely due to increased primary bacteremia. PMID- 18159263 TI - The impact of health care restructuring on nosocomially acquired blood stream infections. AB - OBJECTIVE: To assess the impact of the health care restructuring, which occurred in Alberta in 1995, on the occurrence of nosocomial blood stream infection and risk factors for these infections at the University of Alberta Hospital. PATIENTS AND METHODS: Changes in patient population, hospital bed numbers, admissions and hospital days for 1993 and 1994 (1993/94) were compared with those for 1996 and 1997(1996/97). Central venous catheter (CVC) use in intensive care units (ICU), days of total parenteral nutrition (TPN) and hemodialysis were compared for the two time periods. Prospectively collected data obtained by monitoring blood culture results on nosocomial blood stream infections in 1993/94 were compared with those obtained in 1996/97. RESULTS: Hospital bed number fell by 10% between 1993/94 and 1996/97. Annual admissions fell by 19% and patient days by 17%. Some services markedly increased patient days (neurosurgery 49%, nephrology 30%, orthopedic surgery 24%), and others markedly reduced patient days (obstetrics and gynecology 99%, ophthalmology 100%, adult medicine 41%, general paediatrics 38%). ICU use of CVCs increased by 41%, TPN days increased by 25% and hemodialysis runs increased by 9%. Annual nosocomial blood stream infections increased by 31% and the annual rate per 10,000 patient days increased by 60%. TPN-related blood stream infection rates and ICU CVC infection rates did not change from 1993/94 to 1996/97. CONCLUSIONS: Hospital restructuring has been associated with a 31% increase in nosocomial blood stream infection number and a 60% increase in rate. The increase has been associated with a change in patient populations and increases in risk factors for blood stream infection. PMID- 18159264 TI - Low prevalence of VRE gastrointestinal colonization of hospitalized patients in Manitoba tertiary care and community hospitals. AB - OBJECTIVE: To determine the prevalence of vancomycin-resistant enterococci (VRE) bowel colonization in hospitalized patients in Manitoba who had stool specimens collected for Clostridium difficile toxin and/or culture testing. DESIGN: Two tertiary care and five community hospitals in Winnipeg and three rural Manitoba community hospitals participated in this study. From January 1 to December 31, 1997 stool specimens, one per patient, submitted to hospital microbiology laboratories for C difficile toxin and/or culture testing were screened for VRE on colistin-nalidixic acid-vancomycin (6 microg/mL) (CNAV) agar plates. The study was divided into six, eight-week intervals. Stool specimens received in the first two weeks of each eight week interval were screened for VRE. MAIN RESULTS: A total of 1408 stool specimens were submitted over the 48-week study period. Sixty seven (4.8%) patients with VRE colonization of their lower gastrointestinal tract were identified. Three of the 67 (4.5%) VRE isolates were Enterococcus faecium, with the remaining 64 (95.5%) were Enterococcus gallinarum. The three vancomycin resistant E faecium -VREF- (from two different Winnipeg hospitals) demonstrated the vanA genotype, and were resistant to vancomycin, teicoplanin and ampicillin. All three VREF isolates also demonstrated high level resistance to both gentamicin and streptomycin but were susceptible to quinuprisitin/dalfopristin and LY333328. CONCLUSION: VRE colonization in hospitalized patients in Manitoba is infrequent and most commonly due to E gallinarum. The prevalence of VREF colonization in the patients studied was 0.2% (three of 1408). PMID- 18159265 TI - Leptospira infections in trappers from Ontario. AB - BACKGROUND: Four trappers presented to the Middlesex-London Health Unit in November, 1997 with similar clinical presentations. All four complained of fever, chills and headache, and three of the four had severe muscle aches. All gave histories of trapping raccoons before the onset of illness. Three of the four men exhibited diagnostic seroconversions to Leptospira grippotyphosa. OBJECTIVE: To describe the four suspected cases of leptospira infections and to determine whether raccoons might serve as a reservoir of infection using field studies. DESIGN: Raccoon serology were undertaken using the microscopic agglutination test against eight serovars of Leptospira interrogans including L grippotyphosa. Raccoons were trapped using Tomahawk live traps, anaesthetized with intramuscular injection of ketamine and acepromazine, bled by cardiac puncture and released. RESULTS: Forty-two raccoons were trapped in Middlesex (n=36) and Kent counties (n=6) from April 25 to May 2, 1998, and 10 (23.8%) of these animals had antibodies to L grippotyphosa. CONCLUSIONS: Infections due to L grippotyphosa or a closely related serovar are a risk for trappers in Ontario, and raccoons are a likely reservoir of this bacterium. PMID- 18159266 TI - Recommendations from the national varicella consensus conference, montreal, quebec, may 5 to 7, 1999. PMID- 18159267 TI - Medical ethics: debates, dilemmas and decisions. PMID- 18159268 TI - Integrating laboratory and epidemiological techniques for population-based surveillance of HIV strains and drug resistance in Canada. PMID- 18159269 TI - What if a child has never been immunized? PMID- 18159271 TI - Forearmed and deceived: diagnosis? PMID- 18159270 TI - An outbreak of cyclosporiasis in 1996 associated with consumption of fresh berries- Ontario. AB - BACKGROUND: A large foodborne outbreak of cyclosporiasis occurred in North America in 1996. An index cluster of cases associated with a catered event on May 11, 1996, in Ontario sparked the recognition of this outbreak in Canada. OBJECTIVES: To describe the Ontario experience with the North American outbreak of cyclosporiasis in 1996. PATIENTS AND METHODS: Public health units investigated the index and subsequent event-associated clusters. Investigations included retrospective cohort studies of clusters, traceback of suspect foods and a case control study of sporadic cases. These activities, coordinated with those in the United States, were part of an international investigation. RESULTS: In Ontario, 232 cases of cyclosporiasis (20 laboratory-confirmed and 72 clinically defined cases associated with seven events plus 140 additional laboratory-confirmed sporadic cases) were identified between May 1 and July 30, 1996. For the index cluster, a strawberry flan with raspberries and blueberries was the only significant exposure (relative risk 2.16, P=0.02). Fresh berries were served at all seven events associated with clusters of cases. Raspberries were definitely served at three events, possibly served at three events, and not served at one event. Only imported berries were available in Ontario in May 1996, when initial clusters and sporadic cases were identified. The raspberries served at the two events with well documented traceback data came from Guatemala. Univariate analyses of the matched case-control study demonstrated that illness was associated with consumption of raspberries (matched odds ratio 21.0, 95% CI 3.48 to 448) and strawberries (matched odds ratio 28.5, 95% CI 4.02 to 478). Further evidence amassed by the international investigation compellingly implicated Guatemalan raspberries as the vehicle of the outbreak. CONCLUSION: Cyclosporiasis may be acquired domestically from the consumption of contaminated produce. The scope and vehicle of this international foodborne outbreak were recognized through a coordinated public health response. PMID- 18159272 TI - Outcome of tuberculosis treatment: a comparison between Alberta and Nicaragua. AB - OBJECTIVE: To measure the outcome of tuberculosis treatment in a low incidence, high income region, Alberta, and compare with an intermediate incidence, low income country with a model national tuberculosis program, Nicaragua. DESIGN: All 1992 sputum smear-positive pulmonary cases from both regions were included. Treatment outcome was assigned retrospectively to Alberta cases according to the International Union Against Tuberculosis and Lung Diseases' (IUATLD) criteria of cure, failure, transfer, absconder and death. SETTING: Alberta laboratories are required to report all Mycobacterium tuberculosis cultures to Alberta provincial tuberculosis services. Nicaragua cases are reported centrally to the Programa de control de tuberculosis in Managua using the IUATLD criteria. MAIN RESULTS: In Alberta, 222 tuberculosis cases were identified, of which 61 were smear positive. Nicaragua had 1552 smear positive cases of 2885 tuberculosis cases. Alberta's outcomes were 82% cured, no failed treatment, 5% absconded, 2% transferred and 11% died; Nicaragua's outcomes were 77% cured, 2% failed, 13% absconded, 5% transferred and 4% died. There was no significant difference in cure rates between Alberta and Nicaragua, P=0.33. CONCLUSIONS: Treatment outcomes can be measured effectively and reported in high income, low incidence settings. Alberta is achieving comparable cure rates with the Nicaraguan national tuberculosis program. PMID- 18159273 TI - Reactogenicity to a live attenuated varicella vaccine in Canadian children. AB - OBJECTIVE: To assess the reactogenicity and safety of a thermostable, high titre, varicella vaccine in healthy infants and children. DESIGN: Open study of 505 children monitored for 42 days after vaccination. SETTING: Three urban Canadian centres (Halifax, Ottawa and Vancouver). PARTICIPANTS: 505 healthy children one to 12 years of age were enrolled and 504 completed the study. All were susceptible to varicella by history. INTERVENTIONS: All participants received one dose of live attenuated varicella vaccine (1x10(4.5) plaque forming units/dose) subcutaneously. MAIN OUTCOME MEASURES: The children were monitored from the day of vaccine administration (day 0) until day 42. All local and general symptoms and signs were recorded on diary cards by the patients' parents, who were encouraged to fill in the cards on days 2 to 3 and 18 to 24 via telephone calls from study personnel. RESULTS: Most of the symptoms noted after vaccine administration were mild and transient, and all resolved within the respective follow-up periods. Injection site symptoms included pain (17.5%, 13.9% and 30.4% in centres 1, 2 and 3 respectively), redness (21.1%, 32.1% and 48.8%) and swelling (7%, 10.3% and 29.2%). The general symptoms reported were fever 37.5 degrees C or higher (3.5%, 4.8% and 3.0%) and varicella-like rashes (6.4%, 2.4% and 0%). Two subjects had severe symptoms (one with cervical lymphadenopathy, and one with a fever higher than 39 degrees C) probably related to vaccine administration. No serious adverse events were reported during the entire study. CONCLUSION: The vaccine was well tolerated. PMID- 18159274 TI - Microbiological surveillance and parenteral antibiotic use in a critical care unit. AB - OBJECTIVE: To evaluate parenteral antibiotic utilization and bacterial resistance patterns in a critical care unit (CrCU). DESIGN: Descriptive, prospective audit of infection site, culture and antimicrobial susceptibility test results, parenteral antibiotic usage and duration, total antibiotic acquisition costs, and length of stay. SETTING: A 17-bed medical-surgical CrCU in a tertiary care teaching hospital in Metropolitan Toronto. PATIENTS: Two hundred and fifty-eight patients admitted to the CrCU between May 1995 and April 1996 who received antimicrobial therapy. RESULTS: The most frequently prescribed antibiotics were cefazolin (47%, 1098 g), gentamicin (33%,141 g) and ceftriaxone (20%, 255 g). The most common indications for antimicrobial therapy included surgical prophylaxis (34%) and pneumonia (35%). The following organisms were isolated from patients treated with antibiotics: Staphylococcus aureus (26%), Pseudomonas aeruginosa (13%), enterococci (12%), Haemophilus influenzae (11%), Escherichia coli (11%), Enterobacter cloacae (8%) and other Gram-negative bacilli (19%). Only 9% of Gram negative bacilli were resistant to aminoglycosides, 3% were resistant to ciprofloxacin and no extended-spectrum beta-lactamases or imipenem-resistance were detected. No vancomycin-resistant enterococci and only two methicillin resistant Staphylococcus aureus isolates were identified. CONCLUSIONS: Antibiotic use during the audit appeared appropriate for the specific clinical indications. Low levels of bacterial resistance were detected during the audit. PMID- 18159275 TI - The first smallpox epidemic on the Canadian Plains: In the fur-traders' words. AB - William Tomison, in charge of the Hudson's Bay Company's Cumberland House on the Saskatchewan River, described the devastating smallpox epidemic of 1781 and 1782. He understood contagion, practised isolation and disinfection, and provided mortality statistics during a 'virgin soil' epidemic. Above all, he showed remarkable compassion. He and his men took dying Indians into their already crowded quarters, and provided them with food, shelter and 24 h care. This article describes the epidemic and its aftermath. PMID- 18159276 TI - Viruses without borders. PMID- 18159277 TI - Vancomycin-resistant enterococci in Canada revisited. PMID- 18159278 TI - Lyme disease vaccine. PMID- 18159279 TI - Multiorgan failure and rhabdomyolysis in a recent emigre: your diagnosis? PMID- 18159280 TI - Population-based surveillance of Hib invasive infections in children in British Columbia Alberta and Ontario - 1995 to 1997. AB - OBJECTIVE: To assess vaccine effectiveness through enhanced disease surveillance following the change in childhood immunization programs in 1995, when all provinces and territories chose to use polyribosyl ribitol phosphate-tetanus protein (PRP-T) Haemophilus influenzae type b (Hib) conjugate vaccine, generally in combination with diphtheria-pertussis-tetanus inactivated polio vaccine (DPT IPV) (as PENTA vaccine) because the protective efficacy of this regimen had not been directly measured. DESIGN: Prospective, active, laboratory-based Hib case surveillance was implemented in British Columbia and Alberta, and enhanced, stimulated laboratory surveillance in Ontario during 1995 to 1997, centred on invasive infections in children. Case details and immunization histories were uniformly collected and centrally collated. MAIN RESULTS: Thirty-eight Hib cases were detected, but only 12 cases arose among PENTA-eligible children, an attack rate of 0.85 cases/100,000 child-years of observation. Annual case totals declined from 20 in 1995 to seven in 1997, when only one to three cases were encountered in each province and the incidence rate in children under age five years was 0.6/100,000. Only four cases occurred after primary immunization with PENTA, a failure rate of 0.28 cases/100,000 child-years of observation. Three cases among PENTA-eligible children reflected parental refusal of infant vaccinations, accounting for 25% of cases in eligible children. CONCLUSIONS: PRP T conjugate vaccine was highly effective when given in combination with DPT-IPV vaccine. Provincial programs that used this regimen resulted in the near elimination of invasive Hib disease in children, but unimmunized children remain at risk. PMID- 18159281 TI - Issues around host immunity. PMID- 18159282 TI - How novel methods can help discover more information about foodborne pathogens. AB - Considerable emphasis is being placed on quantitative risk assessment modelling as a basis for regulation of trade in food products. However, for models to be accurate, information about the behaviour of potential pathogens in foods needs to be available. The question is how to obtain this knowledge in a simple and cost effective way. One technique that has great potential is the use of reporter bacteria which have been genetically modified to express a phenotype that can be easily monitored, such as light production in luminescent organisms. Bacteria carrying these (lux) genes can easily be detected using simple luminometers or more sophisticated low light imaging equipment.By monitoring light output from these bacteria over time, it can easily be determined if the organism is growing (resulting in an increase in light emission), is dead (causing a decrease in light production) or is injured (light output remains constant). The use of imaging systems allows the response of bioluminescent bacteria to be studied directly on the food, making the technique even more useful. Applications of bioluminescence are discussed below and include use as reporters of gene expression; biocide efficacy and antibiotic susceptibility; sub-lethal injury; adhesion and biofilm formation; the microbial ecology of foods; pathogenesis; and as biosensors. PMID- 18159283 TI - Everyday and exotic foodborne parasites. AB - Everyday foodborne parasites, which are endemic in Canada, include the protozoans Entamoeba histolytica, Giardia lamblia and Cryptosporidium parvum. However, these parasites are most frequently acquired through unfiltered drinking water, homosexual activity or close personal contact such as in daycare centres and occasionally via a food vehicle. It is likely that many foodborne outbreaks from these protozoa go undetected. Transmission of helminth infections, such as tapeworms, is rare in Canada because of effective sewage treatment. However, a common foodborne parasite of significance is Toxoplasma gondii. Although infection can be acquired from accidental ingestion of oocysts from cat feces, infection can also result from consumption of tissue cysts in undercooked meat, such as pork or lamb. Congenital transmission poses an immense financial burden, costing Canada an estimated $240 million annually. Also of concern is toxoplasmosis in AIDS patients, which may lead to toxoplasmosis encephalitis, the second most common AIDS-related opportunistic infection of the central nervous system. Exotic parasites (ie, those acquired from abroad or from imported food) are of growing concern because more Canadians are travelling and the number of Canada's trading partners is increasing. Since 1996, over 3000 cases of Cyclospora infection reported in the United States and Canada were epidemiologically associated with importation of Guatemalan raspberries. Unlike toxoplasmosis, where strategies for control largely rest with individual practices, control of cyclosporiasis rests with government policy, which should prohibit the importation of foods at high risk. PMID- 18159284 TI - Foodborne spread of hepatitis A: Recent studies on virus survival, transfer and inactivation. AB - Hepatitis A virus (HAV) is responsible for considerable morbidity and economic losses worldwide, and is the only reportable, foodborne viral pathogen in Canada. Outbreaks caused by it occur more frequently in settings such as hospitals, daycare centres, schools, and in association with foods and food service establishments. In recent years, the incidence of hepatitis A has increased in Canada. Many factors, including changing lifestyles and demographics, faster and more frequent travel, and enhanced importation of foods from hepatitis A-endemic regions, may be behind this increase. Despite its increasing significance as a human pathogen, not much was known until recently about the survival and inactivation of HAV, and even less was understood about the effectiveness of measures to prevent and control its foodborne spread. Studies conducted in the past decade have shown that HAV can survive for several hours on human hands and for several days on environmental surfaces indoors. The virus can also retain its infectivity for several days on fruits and vegetables which are often consumed raw, and such imported items have already been incriminated in disease outbreaks. Casual contact between contaminated hands and clean food items can readily lead to a transfer of as much as 10% of the infectious virus. HAV is also relatively resistant to inactivation by heat, gamma irradiation and chemical germicides. In view of these findings, better approaches to prevent the contamination of foods with HAV and more effective methods for its inactivation in foods, on environmental surfaces and on the hands of food handlers are needed. PMID- 18159285 TI - Bar coding vaccines - more than a 'check-out' issue. PMID- 18159286 TI - West Nile virus - where did it come from and where might it go? PMID- 18159287 TI - When your child needs a blood transfusion. PMID- 18159288 TI - Surveillance of antimicrobial resistance in salmonella, shigella and Vibrio cholerae in Latin America and the Caribbean: a collaborative project. PMID- 18159289 TI - Use of antiviral prophylaxis in influenza outbreaks in long term care facilities. AB - Influenza is a major cause of illness and death in residents of long term care facilities for the elderly, in part because residents' age and underlying illness increase the risk of serious complications, and in part because institutional living increases the risk of influenza outbreaks. The administration of antiviral medications active against influenza to persons exposed to influenza has been shown to protect them effectively from illness, and mass antiviral prophylaxis of residents is an effective means of terminating influenza A outbreaks in long term care facilities. The only antiviral currently licensed in Canada for influenza prophylaxis is amantadine, a medication active against influenza A but not influenza B. The National Advisory Committee on Immunization recommends that amantadine prophylaxis be offered to residents when influenza A outbreaks occur in long term care facilities. However, there remain a number of unanswered questions about how best to use amantadine for controlling influenza A outbreaks in long term care facilities. In addition, two members of a new class of antivirals called neuraminidase inhibitors have recently been licensed in Canada for the treatment of influenza, and are effective in prophylaxis. Issues in the use of amantadine in the control of outbreaks of influenza A in long term care facilities for the elderly are reviewed, and the potential uses of neuraminidase inhibitors in this setting are discussed. PMID- 18159290 TI - Hospitalization due to adverse drug reactions and drug interactions before and after HAART. AB - OBJECTIVE: To characterize and compare the rates of adverse drug reactions (ADRs) and interactions on admission in two, one-year periods: pre-highly active antiretroviral therapy (HAART) (phase 1) and post-HAART (phase 2). DESIGN: Retrospective chart review. SETTING: University-affiliated tertiary care centre. POPULATION STUDIED: HIV-positive patients admitted to hospital. MAIN RESULTS: In phase 1, 436 of 517 admissions, and, in phase 2, 323 of 350 admissions were analyzed. Over 92% of patients were male, with a mean age of 38 years. Significant differences (P<0.05) in the mean length of stay (12.08 versus 10.02 days), the CD4 counts (99.25 versus 129.45) and the number of concurrent diseases (4.20 versus 3.63) were found between phase 1 and 2, respectively. The mean number of medications taken (5.52 versus 5.94) and the rates of hospitalization with ADRs (20.4% versus 21.4%) or interactions (2.5% versus 2.16%) were similar between the two phases. Antiretrovirals were more common in ADR admissions post HAART (21.3% versus 36.2%), while antiparasitics, psychotherapeutics and antineoplastics were more common pre-HAART. Other classes of drugs involved in both phases were sulphonamides, narcotics, ganciclovir, foscarnet, antimycobacterials and antifungals. ADR causality was possible or probable in more than 80% of cases. Over 60% of ADRs were grades 3 to 4, and about 85% were either the main or contributing reason for admission. About 65% of patients had at least partial recovery at the time of discharge. In phases 1 and 2, 8.9% and 2.9% of admissions,respectively, with ADRs were fatal. CONCLUSIONS: Although hospitalizations with ADRs and interactions were similar in both phases, HAART therapy has had a significant impact on the incidence and nature of ADRs at St Michael's Hospital, Wellesley Central Site, Toronto, Ontario. PMID- 18159291 TI - Is methicillin-resistant Staphylococcus aureus an emerging community pathogen? A review of the literature. AB - OBJECTIVES: To discuss the historical epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) and review the literature suggesting that MRSA has become a community pathogen. DATA SOURCES: A search of the MEDLINE database was performed, encompassing all English or French language citations from 1966 to 1999 and containing the subjects and/or text words: 'Staphylococcus aureus', 'methicillin resistance', 'endocarditis', 'cellulites', 'pneumonia' and 'community-acquired'. Articles published in other languages that provided English or French abstracts were included. All relevant references cited in articles obtained from the MEDLINE database and book chapters were also included. DATA EXTRACTION: All articles obtained from the above sources were examined and were included in the review if a laboratory or epidemiological study of community acquired MRSA was presented. DATA SYNTHESIS AND CONCLUSIONS: MRSA has emerged over the past 30 years to become a worldwide nosocomial pathogen and has recently been reported as a cause of community-acquired infections. The changing epidemiology of MRSA is likely because of two mechanisms: the movement of nosocomial MRSA strains into the community and the de novo appearance of community strains resulting from the transfer of genetic material from methicillin-resistant Gram-positive organisms to sensitive S aureus strains. The emergence of MRSA as a community pathogen has occurred at a slower rate than it did for penicillin-resistant S aureus (PRSA) in the 1950s and 1960s, possibly because the mechanism of methicillin resistance does not exhibit the same ease of transferability as that of penicillin resistance. Four case reports, seven case series, 10 case-control studies and two cohort studies on community-acquired MRSA were analyzed. Determining whether these reports involve new community-acquired strains rather than previously acquired nosocomial strains can be problematic. It appears, however, that MRSA strains of both nosocomial and community origin are now endemic in certain communities in different parts of the world. Few surveillance studies of nonhospitalized patient populations have been performed to date; thus, the true prevalence of MRSA in the community at large is essentially unknown, although it appears to be low. At present, the empirical treatment of community-acquired S aureus infections with a beta-lactamase-stable beta-lactam antibiotic is appropriate for most populations. However, empirical vancomycin therapy for serious S aureus infections should be strongly considered for patients with significant risk factors for previously-acquired nosocomial MRSA or for patients belonging to outpatient populations with a proven high prevalence of MRSA. Increasing vancomycin use will likely have a significant impact on the development of resistance in Gram-positive organisms. PMID- 18159292 TI - Influenza vaccination for health care workers: A duty of care. PMID- 18159293 TI - When your best friend bites: A note on dog and cat bites. PMID- 18159294 TI - Back pain and immunosuppression: What is at the root of it all? PMID- 18159295 TI - Antibiotic resistance in Canada at the dawn of the new millennium - a model for the developed world? PMID- 18159296 TI - Summary of Canadian guidelines for the initial management of community-acquired pneumonia: an evidence-based update by the Canadian Infectious Disease Society and the Canadian Thoracic Society. AB - Community-acquired pneumonia (CAP) is a serious illness with a significant impact on individual patients and society as a whole. Over the past several years, there have been significant advances in the knowledge and understanding of the etiology of the disease, and an appreciation of problems such as mixed infections and increasing antimicrobial resistance. The development of additional fluoroquinolone agents with enhanced activity against Streptococcus pneumoniae has been important as well.It was decided that the time had come to update and modify the previous CAP guidelines, which were published in 1993. The current guidelines represent a joint effort by the Canadian Infectious Diseases Society and the Canadian Thoracic Society, and they address the etiology, diagnosis and initial management of CAP. The diagnostic section is based on the site of care, and the treatment section is organized according to whether one is dealing with outpatients, inpatients or nursing home patients. PMID- 18159297 TI - Varicella susceptibility in a Canadian population. AB - OBJECTIVE: To determine susceptibility to varicella zoster virus (VZV) infection among children, pregnant women and health care workers in Newfoundland. DESIGN: Cohort and cross-sectional, province-wide, population-based seroprevalence study. STUDY POPULATION AND METHODS: A cohort of 586 children aged one year, a cross sectional group of 1135 school children aged five to 15 years, 3643 pregnant women aged 15 to 45 years, and 5386 persons in health care settings aged 15 to 55 years. Susceptibility to varicella was determined by enzyme immunoassay based on serum antibody to VZV. RESULTS: Among the cohort of 586 children, 565 (96.4%) did not have detectable VZV antibody at one year of age. The proportion with VZV antibody increased thereafter to 12.8% and 33.9%, respectively, at age two and four years, indicating the extent of exposure to VZV at these ages. Among the 1135 school-age children, the proportion testing positive for VZV antibody increased from 44% at five years of age to 88.9% at 15 years of age, indicating the cumulative incidence of varicella in this age group. Among pregnant women, 92.1% tested positive for VZV antibody, and the corresponding figure for the health care group was 93.1%. In both groups, the proportion testing positive for VZV antibody increased with advancing age, from 89.6% for the 15- to 19-year age group to 96.5% for those over the age of 40 years. CONCLUSIONS: The risk of VZV infection increases steadily from one year of age, reaching a peak during school years. The study data support the recent Canadian recommendation to vaccinate any person older than 12 months of age who is susceptible to VZV. Among the adult population, the proportion susceptible will be under 10% for the foreseeable future, and for those at risk, selective vaccination based on their immune status would be a cost effective approach. PMID- 18159298 TI - Pharmacokinetic interaction between zidovudine and trimethoprim/sulphamethoxazole in HIV-1 infected children. AB - OBJECTIVE: To evaluate the effect of the antimicrobial agent trimethoprim/sulphamethoxazole (TMP/SMX) on the pharmacokinetic properties of the antiretroviral drug zidovudine (ZDV). DESIGN: This single dose, open label, crossover study involved the oral administration of ZDV (150 mg/m2) alone and in combination with oral TMP/SMX (2.5 mg/kg) on two separate occasions. Serial blood samples (0 to 8 h) were collected, and concentrations of ZDV and its glucuronide metabolite were quantified using a radioimmunoassay. ZDV pharmacokinetics were determined by noncompartmental analysis. PATIENTS AND SETTING: Six HIV-1 infected children aged four months to five years were recruited from the HIV clinic at The Hospital for Sick Children, Toronto, Ontario. Only three patients completed both study phases and were included in the pharmacokinetic analysis. MAIN RESULTS: With TMP/SMX therapy, no statistically significant changes were observed in ZDV pharmacokinetic parameters. However, there was a trend towards increased ZDV half life and area under the concentration versus time curve, as well as decreased apparent oral clearance. Similarly, a trend towards an increased half-life of the ZDV-glucuronide metabolite was also observed. CONCLUSION: The changes in ZDV pharmacokinetics in the presence of TMP/SMX did not reach statistical significance, most likely due to the limited number of patients involved. Despite the limited data, a possible interaction between ZDV and TMP/SMX in young HIV-1 infected children should be considered, and patients may require close clinical monitoring. PMID- 18159299 TI - Persistently HIV-1 seronegative Nairobi sex workers are susceptible to in vitro infection. AB - OBJECTIVE: To evaluate whether resistance to HIV-1 infection in a subset of highly exposed sex workers correlates with resistance at the cellular level. DESIGN: In vitro evaluation of susceptibility to infection by Kenyan HIV-1 isolates and cellular production of potential mediators of resistance. SETTING: Samples were collected in a primary care clinic in Nairobi. PATIENTS: Thirteen individuals from a cohort of sex workers with a similar risk of acquiring HIV infection and six unexposed controls. INTERVENTIONS: Subjects were provided with appropriate primary care and counselling on the prevention of sexually transmitted diseases. RESULTS: No inherent cellular resistance to infection was identified. CD8+ cells from a subset of subjects strongly inhibited viral replication. CONCLUSIONS: Lack of infection in this cohort was not attributable to factors inherent to CD4+ cells. Resistance to HIV infection is likely to be multifactorial, and products of CD8+ cells and unique features of mucosal sites probably contribute to this state. PMID- 18159301 TI - Writing clinical guidelines with evidence-based medicine. PMID- 18159300 TI - Double-blind, randomized study of the effects of influenza vaccination on the specific antibody response and clinical course of patients with chronic fatigue syndrome. AB - OBJECTIVE: To determine whether influenza immunization is associated with early side effects, a deleterious impact on the illness course and depressed antibody response in patients with chronic fatigue syndrome (CFS). DESIGN: Prospective, randomized, double-blind, placebo controlled trial. CFS patients and healthy volunteers filled out a questionnaire on immunization side effects and had hemagglutination-inhibiting (HI) antibody titres measured pre- and three weeks after immunization. CFS patients completed symptom and function questionnaires before and during the six-week, postimmunization period. SETTING: Ambulatory care. POPULATION STUDIED: Convenience sample of 40 CFS patients fulfilling the Centers for Disease Control and Prevention criteria and 21 demographically matched healthy volunteers. INTERVENTIONS: CFS patients were randomly selected to receive commercially available whole virus influenza vaccine (n=19) or an injection of saline placebo (n=21). Healthy volunteers received vaccine only. MAIN RESULTS: As a group, immunized CFS patients had lower geometric mean HI antibody rises than healthy volunteers (P<0.001). However, there was no difference in the rates of fourfold titre rises, and immunization did achieve a probably protective titre (1:32 or greater) in most CFS patients. No difference could be detected between immunized and placebo CFS patients in immunization side effects, although CFS patients as a group reported four times as many side effects as healthy volunteers. Further, in the six weeks following immunization, placebo and immunized CFS patients did not demonstrate any differences in terms of functioning, symptom severity and sleep disturbance. CONCLUSIONS: In patients with CFS, influenza immunization is safe, not associated with any excess early reactions, and stimulates an immunizing response comparable with that of healthy volunteers. PMID- 18159302 TI - Application of DNA array technology for diagnostic microbiology. PMID- 18159303 TI - The battle against influenza: The role of neuraminidase inhibitors in children. PMID- 18159304 TI - Guidelinitis: A new syndrome? PMID- 18159305 TI - How Nova Scotia general practitioners choose antibiotics for the empirical treatment of community-acquired pneumonia. AB - OBJECTIVE: To gain an understanding of how physicians in general practice choose antibiotics for the empirical treatment of community-acquired pneumonia (CAP). DESIGN: Questionnaire with three sample cases of CAP and a knowledge assessment (mailed to half of the physicians). POPULATION STUDIED: Nova Scotia family physicians. RESULTS: One hundred and eighty-four of the 841 (21.9%) physicians who were mailed a questionnaire responded. A knowledge assessment showed satisfactory knowledge except in two areas - an overestimation of the prevalence of penicillin-resistant Streptococcus pneumoniae in Nova Scotia and the view that ciprofloxacin was an effective antibiotic for the treatment of CAP (42% of physicians). As the complexity of the case increased, there was decreasing consensus regarding the choice of antibiotic therapy and a decline in prescribing according to guidelines for the treatment of CAP. Also, as the complexity of the cases increased, it became increasingly difficult to discern a decision-making strategy. For the simplest case - a 17-year-old male with presumed Mycoplasma pneumoniae pneumonia - physician factors (age, family practice training), desire to target specific pathogens, and concern with resistance and side effects affected the choice of antibiotic. However, for the most complex case - a 45-year old female with severe pneumonia - familiarity with such a case was the only significant factor and led to treatment with a combination of antibiotics designed to treat both typical and atypical pathogens. CONCLUSIONS: For uncomplicated cases of CAP, physician factors, desire to treat specific pathogens and concern with resistance affect the choice of antibiotic therapy. For complex cases, familiarity with such cases was the only factor that influenced choice of antibiotic therapy. PMID- 18159306 TI - Hemoptysis and fever post-transplant: Diagnosis? PMID- 18159307 TI - Evaluation of the rubella surveillance system in Quebec. AB - OBJECTIVE: To evaluate the validity of information in the rubella surveillance system in Quebec. DATA AND METHODS: Cases of rubella in the provincial registry of notifiable diseases, "Maladies a declaration obligatoire" (MADO), from 1994 to 1996 were matched with laboratory-identified cases and with cases in a reference file created from all case investigation records of regional departments of public health for the same period. Sensitivity and the proportion of cases in agreement were calculated. RESULTS: Compared with laboratories, the sensitivity of the provincial registry was 56%. Compared with the reference file, global sensitivity (confirmed cases plus clinical cases) was 58% and the positive predictive value was 50%. Of the 356 cases reported to regional public health departments, 65% were classified in the same diagnostic category (confirmed case, clinical case, excluded case) by public health professionals and a group of experts (weighted kappa=0.32). Information on rubella vaccination status was missing in 25% of cases in the MADO file for rubella. CONCLUSIONS: Notification of positive results for immunoglobulin M antibodies and viral cultures should be required of all laboratories. Uniform procedures should be adopted and applied for the validation of cases that are reported to regional departments of public health. In the context of the rarefaction of rubella, any immunoglobulin M positive result should be interpreted using all available epidemiological information. PMID- 18159308 TI - Medical decision analysis in infectious diseases. AB - Medical decision analysis (MDA) has played an important role in assisting infectious disease physicians make decisions associated with varying levels of complexity. Clinicians are often uncomfortable with some aspects of MDA, particularly when utilities are used as outcome measures. However, as the present paper outlines, MDA may use other outcome variables, including costs and disease complications. In this context, this explicit, reproducible analytic framework is an important tool in the area of infectious diseases, and is frequently applied to many situations, including cost effectiveness analyses, studies involving assessment of risks versus benefits of preventive and treatment strategies, and other situations. The objective of this paper is to assist infectious diseases clinicians to understand better the role of MDA in clinical practice. In this regard, the principles of MDA are reviewed and a common clinical example is used for illustrative purposes. PMID- 18159309 TI - Prosthetic knee septic arthritis due to Pseudomonas stutzeri. AB - Prosthetic joint infection is usually caused by Staphylococcus aureus, coagulase negative staphylococci and, less commonly, by Gram-negative bacilli and anaerobes. A case of prosthetic joint infection due to Pseudomonas stutzeri in a 73-year-old female with acute promyelocytic leukemia is presented, and the pertinent literature is reviewed. Although the patient had prolonged neutropenia, the infection was successfully treated with antibiotics and without artificial joint replacement. PMID- 18159310 TI - Immunization: Must be safe or we will all be sorry. PMID- 18159311 TI - Nevirapine use to reduce mother-to-child transmission of HIV in Canada. PMID- 18159312 TI - The role of the acellular pertussis vaccine and the demise of 'Pertussis Pete'. PMID- 18159313 TI - Severe complications associated with varicella: Province of Quebec, April 1994 to March 1996. AB - OBJECTIVE: To determine the frequency and severity of serious complications associated with varicella in Quebec; the frequency and severity of cases of congenital varicella; and hospital costs associated with hospitalizations for varicella. STUDY DESIGN: All hospitalizations related to varicella were identified through the use of a hospital data bank and pertinent data were collected from hospital records. SETTING: Province of Quebec with a population of 6,895,960 people. STUDY POPULATION: All cases with a principal or secondary diagnosis of varicella hospitalized in Quebec between April 1, 1994 and March 31, 1996. OUTCOME MEASURES: Types of complications and reason for hospitalization, risk of complications and calculation of associated costs were studied. RESULTS: Nine hundred nine eligible hospitalizations were identified between April 1, 1994 and March 31, 1996. In all, 583 (64.1%) hospitalizations were for the treatment of complications, 127 (14.0%) for administration of intravenous acyclovir and 199 (21.9%) for supportive care. Healthy people accounted for 644 (70.8%) hospitalizations and immunosuppressed individuals for 136 (15.0%). Among children, one-half of the principal complications were skin infections, while 13.5% and 8.4% of principal complications were pneumonia and neurological complications, respectively. Among adults, the most common complication was pneumonia, with a rate of 43.5%, followed by thrombocytopenia and skin infections, with rates of 22.2% and 14.8%, respectively. The complication rate was 29.2 cases/10,000 cases of varicella. CONCLUSIONS: Although perceived as a benign childhood disease by the general population, varicella may be accompanied by severe complications. Morbidity associated with varicella is one of the elements that must be considered when evaluating the usefulness of varicella vaccine. PMID- 18159314 TI - Burden of chickenpox on families: A study in Quebec. AB - OBJECTIVE: To estimate the nonhospital costs of treating chickenpox and to ascertain the opinion of parents regarding the usefulness of vaccination. DESIGN: Retrospective postal survey. SETTING: Province of Quebec. PARTICIPANTS: Random sample of 3333 families with children aged six months to 12 years. OUTCOME MEASURES: For cases of chickenpox that occurred between September 1, 1997 and August 31, 1998, the use of health services, time away from school or work, patient care required, direct and indirect costs for the families and the health care system, and the opinion of parents regarding chickenpox and the vaccine were evaluated. RESULTS: The response rate was 64.7%, and 18.8% of households reported a history of chickenpox, a total of 693 cases. A physician was consulted in 45.8% of these cases, and medication was used in 91.7%. The frequency of hospitalizations was 0.6%. Time away from work or school caused by the disease was 4.1 days on average, with 46.5% of absences being attributed to the risk of contagion. The total average cost of a case of chickenpox was $225. Direct expenses for households accounted for 11% of the total cost, public sector direct costs 7%, indirect costs related to absence from work 38% and caregiving time 45%. A majority of parents (70%) were in favour of a systematic childhood immunization program. CONCLUSIONS: Chickenpox without complications is disruptive for families, but the direct costs for families and the public sector are relatively small. PMID- 18159315 TI - Evaluation of potential factors contributing to microbiological treatment failure in Streptococcus pyogenes pharyngitis. AB - BACKGROUND: A cohort study of children with pharyngitis aged two to 16 years was conducted to assess the role of microbial and host factors in group A beta hemolytic streptococcus (GABHS) microbiological treatment failure. METHODS: GABHS infected children had pharyngeal swabs repeated two to five days after completing a 10-day course of penicillin V. M and T typing, and pulsed field gel electrophoresis were performed on the isolates, and the isolates were evaluated for tolerance. Patient characteristics and clinical features were noted and nasopharyngeal swabs for respiratory viruses were taken at enrolment. RESULTS AND CONCLUSIONS: Of 286 patients enrolled, 248 (87%) could be evaluated. GABHS was cultured from 104 patients (41.9%), of whom 33 (33.7%) had microbiological treatment failures on follow-up. Although there was a trend toward failure for younger children (mean 6.5+/-2.4 years versus 7.3+/-2.4 years, P=0.07) and M type 12 (24% versus 10%, P=0.08), no factors were associated with treatment failure. PMID- 18159316 TI - Appearances can be deceiving: What is the diagnosis for this community-acquired pneumonia? PMID- 18159317 TI - Orbital myositis complicating sinusitis. AB - Orbital myositis is a common cause of extraocular muscle enlargement. It is characterized by nonspecific inflammation of one or more extraocular muscles. Although often idiopathic in origin, orbital myositis has been associated with various noninfectious diseases. Several cases have also been reported as occurring after upper respiratory tract infections. The present report describes a case of orbital myositis together with subclinical sinusitis and its rapid resolution after antibiotic treatment. The literature on this clinical entity is also reviewed. PMID- 18159318 TI - Of Microsoft and mycobacteria. PMID- 18159319 TI - Pertussis immunization for adolescents: What are we waiting for? PMID- 18159320 TI - Bioterrorism in 2001 - How ready are we? PMID- 18159321 TI - N-CDAD in Canada: results of the Canadian Nosocomial Infection Surveillance Program 1997 N-CDAD Prevalence Surveillance Project. AB - BACKGROUND: A 1996 preproject survey among Canadian Hospital Epidemiology Committee (CHEC) sites revealed variations in the prevention, detection, management and surveillance of Clostridium difficile-associated diarrhea (CDAD). Facilities wanted to establish national rates of nosocomially acquired CDAD (N CDAD) to understand the impact of control or prevention measures, and the burden of N-CDAD on health care resources. The CHEC, in collaboration with the Laboratory Centre for Disease Control (Health Canada) and under the Canadian Nosocomial Infection Surveillance Program, undertook a prevalence surveillance project among selected hospitals throughout Canada. OBJECTIVE: To establish national prevalence rates of N-CDAD. METHODS: For six weeks in 1997, selected CHEC sites tested all diarrheal stools from inpatients for either C difficile toxin or C difficile bacteria with evidence of toxin production. Questionnaires were completed for patients with positive stool assays who met the case definitions. RESULTS: Nineteen health care facilities in eight provinces participated in the project. The overall prevalence of N-CDAD was 13.0% (95% CI 9.5% to 16.5%). The mean number of N-CDAD cases were 66.3 cases/100,000 patient days (95% CI 37.5 to 95.1) and 5.9 cases/1000 patient admissions (95% CI 3.4 to 8.4). N-CDAD was found most frequently in older patients and those who had been hospitalized for longer than two weeks in medical or surgical wards. CONCLUSIONS: This national prevalence surveillance project, which established N-CDAD rates, is useful as 'benchmark' data for Canadian health care facilities, and in understanding the patterns and impact of N-CDAD. PMID- 18159322 TI - Initial drug regimen for active tuberculosis cases in Montreal, 1995 to 1998. AB - OBJECTIVES: To evaluate the proportion of tuberculosis (TB) cases initially treated with the recommended four-drug regimen of isoniazid (INH), rifampin (RIF), pyrazinamide (PZA) and ethambutol (EMB) or streptomycin; and to identify factors associated with the choice of initial therapy. DESIGN: Descriptive analysis of surveillance data obtained by TB case notifications from physicians and microbiology laboratories. SETTING: The island of Montreal (with a population of 1,854,435 people). STUDY POPULATION: All TB cases reported between January 1, 1995 and December 31, 1998. OUTCOME MEASURE: The proportion of TB cases initially treated with a four-drug regimen by sex, age, country of birth, site of disease and year of reporting. MAIN RESULTS: Seven hundred forty-one cases were reported during the study period. Among the 687 analyzed cases, 406 (59.1%) received the recommended initial four-drug regimen (INH-RIF-PZA-EMB), 187 (27.2%) received an INH-RIF-PZA regimen, 61 (8.9%) received an INH-RIF-EMB regimen and 33 (4.8%) received an INH-RIF regimen only. In a logistical regression model, a four-drug regimen was significantly associated with respiratory disease (odds ratio [OR] 4.48; 95% CI 3.15 to 6.39), age younger than 65 years (OR 2.32; 95% CI 1.55 to 3.45), being foreign-born (OR 1.62; 95% CI 1.06 to 2.48) and later year of reporting (OR 1.27; 95% CI 1.09 to 1.47). CONCLUSIONS: The proportion of TB cases initially treated with a four-drug regimen has increased steadily since 1995, reaching 65% in 1998. However, given the rate of INH resistance in Montreal, efforts to promote the use of the initial four-drug regimen must continue. PMID- 18159323 TI - Beliefs and behaviours of parents regarding antibiotic use by children. AB - OBJECTIVE: To determine parents' beliefs and behaviours about antibiotic use by their children in the ambulatory setting. DESIGN: Cross-sectional survey, where a self-administered questionnaire was completed by adult caregivers of children before the medical assessment of the child. SETTING: Three paediatric acute care settings (paediatric emergency department [PED], group paediatric practice and after hours walk-in medical clinic). POPULATION STUDIED: Adult caregivers (n=114; 76% mothers, 19% fathers and 4% other caregivers) of children brought for acute care were surveyed. MAIN RESULTS: Forty-one caregivers completed the survey in the PED, 37 in the paediatric office and 36 in the walk-in clinic. They believed that antibiotics were appropriate for ear infections (86%), pharyngitis (77%), bronchitis (49%), sinus colds (20%), cough (12%), colds (8%) and influenza (8%). Sixty-eight per cent of children had received antibiotics in the previous year. Thirteen per cent of caregivers reported previously requesting an antibiotic for their child, 18% believed a previous antibiotic prescription had been unnecessary and 19% had not complied with prescriptions in the past. Concerns about antibiotic use included antibiotic resistance (50%), harm to the immune system (40%), adverse effects (28%) and lack of efficacy (19%). CONCLUSIONS: In this population, parental knowledge and understanding of indications for antibiotics and their adverse effects were good; however, incorrect beliefs and disagreements with physicians did occur. To improve patterns of antibiotic use by children, it will be necessary to understand parents' beliefs, behaviours and information sources better so that misconceptions and disagreements with caregivers can be addressed appropriately. PMID- 18159324 TI - Infectious diseases training in Canada: one size does not fit all. AB - PURPOSE: To evaluate training in infectious diseases, determining which components of the training program best prepare residents for their career choices and where improvements are needed. METHOD: A cross-sectional survey was mailed to all 14 physicians who had graduated from both the Adult and Paediatric Infectious Diseases Training Program at the University of Calgary from 1985 to 1998. Responses about the adequacy of training were measured using a Likert-type scale and a qualitative questionnaire. RESULTS: Of 14 mailed questionnaires, nine responses were received (64%). Two-thirds of respondents were in an academic setting, and seven (78%) graduates obtained postfellowship training. The specialists in academic settings were all engaged in multiple nonclinical activities. The clinical and diagnostic microbiological components of training received the highest scores in terms of adequacy of training. CONCLUSION: Graduates of the University of Calgary training program indicated an overall satisfaction with their training. However, improvements are needed in career counselling, health administration, antibiotic utilization, infection prevention and specialized outpatient clinics. Potential strategies for addressing these issues include didactic lectures, enhanced exposure to clinical outpatient settings and provision of designated faculty mentors. PMID- 18159325 TI - Explosive pleuritis. AB - The objective of the present paper is to describe the clinical and computed tomography features of 'explosive pleuritis', an entity first named by Braman and Donat in 1986, and to propose a case definition. A case report of a previously healthy, 45-year-old man admitted to hospital with acute onset pleuritic chest pain is presented. The patient arrived at the emergency room at 15:00 in mild respiratory distress; the initial chest x-ray revealed a small right lower lobe effusion. The subsequent clinical course in hospital was dramatic. Within 18 h of admission, he developed severe respiratory distress with oxygen desaturation to 83% on room air and dullness of the right lung field. A repeat chest x-ray, taken the morning after admission, revealed complete opacification of the right hemithorax. A computed tomography scan of the thorax demonstrated a massive pleural effusion with compression of pulmonary tissue and mediastinal shift. Pleural fluid biochemical analysis revealed the following concentrations: glucose 3.5 mmol/L, lactate dehydrogenase 1550 U/L, protein 56.98 g/L, amylase 68 U/L and white blood cell count 600 cells/mL. The pleural fluid cultures demonstrated light growth of coagulase-negative staphylococcus and viridans streptococcus, and very light growth of Candida albicans. Cytology was negative for malignant cells. Thoracotomy was performed, which demonstrated a loculated parapneumonic effusion that required decortication. The patient responded favourably to the empirical administration of intravenous levofloxacin and ceftriaxone, and conservative surgical methods in the management of the empyema. This report also discusses the patient's rapidly progressing pleural effusion and offers a potential case definition for explosive pleuritis. Explosive pleuritis is a medical emergency defined by the rapid development of a pleural effusion involving more than 90% of the hemithorax over 24 h, which causes compression of pulmonary tissue and mediastinal shift to the contralateral side. PMID- 18159326 TI - Pondering parotid masses. PMID- 18159327 TI - Infection control in acute care facilities: Evidence-based patient safety. PMID- 18159328 TI - Rare but not so rare: The evolving spectrum of Whipple's disease. PMID- 18159329 TI - Flesh-eating disease: A note on necrotizing fasciitis. PMID- 18159330 TI - Tuberculosis drug resistance in Canada, 1998 to 2000. PMID- 18159331 TI - Mumps caused by an inadequately attenuated measles, mumps and rubella vaccine. AB - PROBLEM: Reports of mumps following measles, mumps and rubella (MMR) immunization. OBJECTIVE: To determine whether mumps was caused by immunization or whether there was a concurrent epidemic of a wild strain of mumps. DESIGN AND PARTICIPANTS: Analysis of surveillance data and a cohort study of three schools that participated in the campaign. OUTCOME MEASURES: Cases of clinical mumps and orchitis, and immunization history and records were reviewed. The MMR vaccine was produced by the Serum Institute of India and contained the Leningrad-Zagreb strain of mumps virus. Four lots were used in the specific immunization campaign. RESULTS: Sentinel health facility surveillance showed an increase in mumps after two school immunization campaigns in western Suriname and a mass immunization campaign in the same region. There was also an increase in a geographically separate region following a mass campaign with the same vaccine. Three hundred fifteen children from three schools that were targeted in the immunization campaign were interviewed. The attack rate for mumps in those immunized was 15.1%; in those not immunized, the attack rate was 4.7%. In the affected males, the attack rate for orchitis was five of 19 (21%). Assuming 90% protection by the MMR vaccine, the incidence ratio (observed to expected) was 32. CONCLUSIONS: The mumps outbreak was caused by an inadequately attenuated MMR vaccine. Because this vaccine had not been used in these populations before in Suriname, it was not possible to determine wether the outbreak was due the virulence of the Leningrad Zagreb mumps strain or due to production problems with one or more specific lots of vaccine. The vaccine was withdrawn from further use. PMID- 18159332 TI - Community-based cluster of methicillin-resistant Staphylococcus aureus in Manitoba. AB - A community-based, single strain cluster of methicillin-resistant Staphylococcus aureus (MRSA) is described. The cluster of cases was identified in a small rural community in southwestern Manitoba, although some spread of the MRSA strain to neighbouring communities was observed. The majority of people were otherwise healthy, had no contact with the hospital system and did not fit the profile of those at risk for MRSA infection. PMID- 18159333 TI - Opinions of Quebec parents and vaccinators on the usefulness of chickenpox vaccine. AB - BACKGROUND: A chickenpox vaccine was recently licensed in Canada. Because this vaccine has caused some controversy within the health care profession, studies among Quebec parents and vaccine providers were carried out, surveying their opinions concerning chickenpox vaccination. METHODS: Three studies among parents of preadolescents, parents of two-year-old children completely or incompletely vaccinated and vaccinators were completed. The studies asked for opinions concerning the usefulness of vaccinating children against chickenpox. RESULTS: The majority of parents of preadolescents (56%), and parents of two-year-old children completely (64%) and incompletely vaccinated (60%) favoured chickenpox vaccination. Among vaccinators, 53% of paediatricians, 37% of general practitioners and 33% of nurses considered universal vaccination of young children to be useful. A greater proportion of health care professionals were in favour of a policy of vaccinating groups at risk, such as susceptible adolescents (86%, 75% and 58%, respectively). There was a positive association between the perceived severity of chickenpox and the potential usefulness of the vaccine. CONCLUSION: Quebec parents are more favourably disposed to chickenpox vaccine than vaccine providers. In contrast, strategies targeting susceptible groups would be generally well received by health care professionals. A considerable amount of work will be needed to convince vaccinators of the benefits of a universal childhood vaccination against chickenpox. PMID- 18159334 TI - Characteristics of hepatitis C virus infection in HIV-infected people. AB - Knowledge pertaining to hepatitis C virus (HCV)/human immunodeficiency virus (HIV) co-infection is currently incomplete or conflicting. Several points are well studied, however. Plasma HCV RNA levels are higher in matched HIV-infected people than in HIV-seronegative control subjects and are inversely correlated with CD4(+) T lymphocyte counts. HCV genotype does not appear to influence this value. Co-infected individuals develop histological and clinical features of HCV liver disease more rapidly than HIV-seronegative patients. Co-infected individuals appear to respond to interferon-alpha therapy equally as well as HIV seronegative HCV-infected adults, but minimal information exists regarding the efficacy and toxicity of combination HCV therapy (interferon-alpha plus ribavirin) in this population. Adverse consequences of highly active antiretroviral therapy in co-infected patients include hepatic toxicity and, in a minority of patients, an 'immune restoration syndrome'. It is unclear whether long term, highly active antiretroviral therapy positively or negatively influences the natural history of HCV infection. PMID- 18159335 TI - Hantavirus pulmonary syndrome in Manitoba. AB - The first confirmed case of hantavirus pulmonary syndrome in Manitoba was diagnosed in 1999. To define better the risk of exposure to hantaviruses in this area, the clinical features and epidemiological factors pertaining to this case were described, and a serological survey of rodents collected near the patient's residence was undertaken. Small mammals were collected using live traps, were anesthetized via inhalation of isoflurane and were bled. Human and mouse serologies were undertaken using an ELISA to detect hantavirus-specific immunoglobulin G and/or immunoglobulin M antibodies. In addition, a full medical and epidemiological assessment, as well as individual risk factor and exposure analysis, were conducted. A 27-year-old Manitoba woman presented with severe respiratory distress and diffuse bilateral air space disease radiologically. Despite extremely aggressive measures, including mechanical ventilation, antibiotics, fluid management and inotropic support, the patient's condition rapidly deteriorated, and she died 8 h after admission. Hantavirus pulmonary syndrome was confirmed by the detection of immunoglobulin M and immunoglobulin G antibodies to the Sin Nombre virus (SNV) in her sera and by the demonstration of SNV genomic sequences in her lung tissue. Exposure to hantavirus likely occurred in and around the home or in the rural area in which she resided. A total of 252 small mammals, primarily deer mice (Peromyscus maniculatus), were collected from 17 different sites at or near where the patient lived. Antibodies to SNV were detected in 28 of 244 (11.5%) deer mice, which were collected within 9 km of the residence of the fatal case, indicating that these rodents are a significant reservoir for SNV in this area. PMID- 18159336 TI - Syncope on steroids. PMID- 18159337 TI - Canadian-acquired hydatid disease: A case report. AB - Echinococcal cysts are unusual in Canada, and most cases seen are in immigrants. In northern Canadian communities, Echinococcus granulosis infection occasionally is acquired from dogs that feed on the entrails of caribou or moose. Seventeen patients with Canadian-acquired hydatid cysts were seen over an 11-year period. One challenging case is described in detail. An 18-year-old aboriginal woman presented with jaundice, pain, lower extremity edema and coagulopathy from a 26 cm echinococcal hepatic cyst. She was successfully treated with a combination of oral albendazole, percutaneous drainage and surgery. One-year follow-up showed no recurrence of disease. The management options for echinococcal cysts are extensively reviewed. PMID- 18159338 TI - Wasting away: To sludge or not to sludge? PMID- 18159339 TI - A perspective on biosolids management. PMID- 18159340 TI - The case against land application of sewage sludge pathogens. PMID- 18159341 TI - It's time for a national immunization strategy. PMID- 18159342 TI - Re-examining treatment of latent tuberculosis infection. PMID- 18159343 TI - Antimicrobial resistance in Canada: An update on activities of the Canadian Committee on Antibiotic Resistance. PMID- 18159344 TI - Macrolides: A Canadian Infectious Disease Society position paper. AB - Since the introduction of erythromycin in 1965, no new compounds from the macrolide antimicrobial class were licensed in Canada until the 1990s. Clarithromycin and azithromycin, since their introduction, have become important agents for treating a number of common and uncommon infectious diseases. They have become prime agents in the treatment of respiratory tract infections, and have revolutionized the management of both genital chlamydial infections, by the use of single-dose therapy with azithromycin, and nontuberculous mycobacterial infections, by the use of clarithromycin. The improvement of clarithromycin and azithromycin over the gastrointestinal intolerability of erythromycin has led to supplanting the use of the latter for many primary care physicians. Unfortunately, the use of these agents has also increased the likelihood for misuse and has raised concerns about a resultant increase in the rates of macrolide resistance in many important pathogens, such as Streptococcus pneumoniae. This paper reviews the pharmacology and evidence for the current indications for use of these newer agents, and provides recommendations for appropriate use. PMID- 18159345 TI - Population-based surveillance of invasive group A streptococcal disease in British Columbia: 1996 to 1998. AB - OBJECTIVE: To identify and describe all cases of invasive group A streptococcal (GAS) infection occurring in British Columbia during a two-year period. DESIGN: Active, laboratory-based surveillance with supplemental case description. SETTING: Forty community and regional hospitals and the provincial laboratory participated, encompassing all health regions. POPULATION STUDIED: Entire provincial population from April 1, 1996 to March 31, 1998. MAIN RESULTS: Over the 24-month surveillance period, 182 eligible cases were identified, yielding a mean annual incidence rate of 2.3/100,000. Patients ranged in age from two to 91 years, with a mean of 39.1 years. Soft tissue infections accounted for 89 of 130 cases (68.5%) with a defined clinical syndrome, 20 of which were necrotizing fasciitis. Injection drug use was described in 55 patients, who, as a group, were younger, more likely to have soft tissue infections and less likely to die of infection than nondrug users. Other risk factors for infection included HIV infection (19 patients); skin damage (26 patients, damage independent of injection drug use); chronic illness (27 patients); and immunosuppresion (three patients). Death from GAS infection occurred in 15 of 131 (11.5%) cases with known outcome, yielding an annual case fatality rate of 1.9/million population. Among necrotizing faciitis cases, the mortality rate was 30%. CONCLUSIONS: Invasive GAS infections are rare in British Columbia and tend to involve persons with chronic illness or prior skin trauma, especially injection drug abuse, which accounted for nearly half of the cases. PMID- 18159346 TI - Reduced susceptibility to penicillin among pneumococci causing invasive infection in children - Canada, 1991 to 1998. AB - OBJECTIVE: To determine, over time, the rate and serotypes of pneumococci with reduced penicillin susceptibility obtained from children with invasive infection. DESIGN: Active, hospital-based, multicentre surveillance spanning from 1991 to 1998. SETTING: Eleven Canadian tertiary care paediatric facilities located from coast to coast. POPULATION STUDIED: 1847 children with invasive pneumococcal infection whose isolates (from a normally sterile site) were available for serotyping and standardized testing for penicillin susceptibility at the National Centre for Streptococcus. MAIN RESULTS: The prevalence of reduced penicillin susceptibility increased from 2.5% of 197 cases in 1991 to 13.0% of 276 cases in 1998. In the latter year, 8.7% of isolates had intermediate level resistance, and 4.3% had high level resistance. Since they were first detected in 1992, strains with high level resistance have been encountered only sporadically at most centres, but by 1998, all centres but two had encountered examples. Of 40 isolates with high level resistance and 101 isolates with intermediate level resistance, serotypes matched those included in new seven-valent conjugate vaccines for children in 97.5% and 79.2% of cases, respectively. CONCLUSIONS: Pneumococci with reduced susceptibility to penicillin are increasing in frequency across Canada among children with invasive infection. The Immunization Monitoring Program, Active data indicate that new conjugate vaccines could help to curb infections due to pneumococci with reduced susceptibility to penicillin but are unlikely to control completely the problem of antibiotic resistance. PMID- 18159347 TI - Erysipelothrix rhusiopathiae endocarditis and presumed osteomyelitis. AB - Erysipelothrix rhusiopathiae is known to cause infections in humans following exposure to decaying organic matter or animals colonized with the organism, such as swine and fish. Invasive infections with this organism are unusual and are manifested primarily as infective endocarditis. The present report is believed to be the first to report a case of E rhusiopathiae endocarditis and presumptive osteomyelitis. E rhusiopathiae appears to have intrinsic resistance to vancomycin. Because vancomycin is often used empirically for the treatment of endocarditis, rapid differentiation of E rhusiopathiae from other Gram-positive organisms is critical. In patients with endocarditis caused by a Gram-positive bacillus and epidemiological risk factors for E rhusiopathiae exposure, empirical treatment with vancomycin should be reconsidered. PMID- 18159348 TI - Reflections. PMID- 18159349 TI - The impact of health care restructuring on nosocomial infections and transmission of antimicrobial resistant organisms. PMID- 18159350 TI - Bioterrorism in Canada: An economic assessment of prevention and postattack response. AB - The present paper calculates the human and economic consequences of a bioterrorist attack on Canadian soil using aerosolized Bacillus anthracis and Clostridium botulinum. The study assumed that 100,000 people in a Canadian suburban neighbourhood were exposed over a 2 h period to an infectious dose of one of the agents. Using an epidemic curve based on the epidemiology and management of anthrax and botulinum poisoning, the costs of intervention and treatment after an attack were compared with the costs of preparedness before a bioterrorist attack. The results show that an investment in planning and preparedness to manage the consequences of an attack can reduce morbidity, mortality and economic costs. The sooner that an intervention program is instituted, the more significant are the health and economic benefits. The greatest benefits were realized when postattack intervention was initiated before day 3 after the event. The economic impact of a bioterrorist attack in Canada could range from $6.4 billion/100,000 exposed to B anthracis to $8.6 billion/100,000 exposed in an attack using C botulinum. Without the benefit of an effective consequence management program, predicted deaths totalled 32,875 from anthrax and 30,000 from botulinum toxin. Rapid implementation of a postattack prophylaxis program that includes the stockpiling of antibiotics, vaccines and antitoxins; training of first responders in the diagnosis, handling and treatment of pathogens; and the general enhancement of Canada's response capability would reduce both human and economic losses. PMID- 18159351 TI - Previous creatinine levels safely predict amantadine dose for influenza A outbreak control. AB - BACKGROUND: Amantadine, an antiviral agent, is the only drug currently approved in Canada for prophylaxis of influenza A virus infection. To minimize side effects, the amantadine dose is adjusted for age and estimated creatinine clearance (CrCl) based on plasma creatinine (Cr) levels. As amantadine is used more frequently for influenza A outbreak control in care facilities for elderly people, physicians are increasingly called on to prescribe it for residents and to consider the necessity of requesting plasma Cr levels. OBJECTIVE: To determine whether previous Cr levels are predictive in estimating current CrCl and safe amantadine dose determination. DESIGN AND SETTING: Residents' charts were reviewed in two facilities in Vancouver, British Columbia. CrCl estimated using previous or current Cr results, current weight and age, as well as recommended amantadine doses based on Canadian National Advisory Committee on Immunization guidelines, were studied. RESULTS: 165 charts with Cr results in March 1998 were included; 122 had results before March 1998, and 103 had Cr results after March 1998. Pearson's correlation coefficient for CrCl estimated from current and previous Cr values was 0.929 for results less than six months previously, 0.974 for six to 12 months previously and 0.952 for 12 to 18 months previously. The same or a more conservative dose of amantadine was predicted in 92% of cases when using a Cr result taken within the previous year and in 76% of cases when using a Cr result taken 12 to 18 months previously. CONCLUSION: In long term care facilities, Cr levels measured up to 12 months previously can usually safely be used to estimate CrCl. Using previous Cr results permits advance preparation of doctor's orders for amantadine prophylaxis and avoids repeating Cr testing on every resident when an outbreak occurs, reducing related staff time and cost. PMID- 18159352 TI - Susceptibility of community Gram-negative urinary tract isolates to mecillinam and other oral agents. AB - OBJECTIVE: To determine the susceptibility of community outpatient Gram-negative urinary tract isolates to mecillinam and other commonly used oral agents. DESIGN AND SETTING: The study was a laboratory-based study of consecutive Gram-negative urinary tract isolates. Only those isolates considered to be significant pathogens were included in the study. Susceptibility testing was performed using agar dilution methodology following guidelines published by the National Committee for Clinical Laboratory Standards. POPULATION STUDIED: Outpatients presenting to a family physician or clinic with signs or symptoms suggestive of a urinary tract infection were included in the study. MAIN RESULTS: Of 2000 consecutive community isolates (91.8% Escherichia coli, 3.9% Klebsiella species, 2.0% Proteus species, 2.3% others), in vitro susceptibilities were: mecillinam 98.8%, ampicillin 77.0%, ciprofloxacin 100%, trimethoprim/sulfamethoxazole 91.6% and nitrofurantoin 95.4%. Susceptibility to mecillinam was significantly better than all other agents except ciprofloxacin (P<0.001, McNemar's test). Organisms with reduced susceptibility to mecillinam included Citrobacter species, Pseudomonas aeruginosa and Providencia species. CONCLUSIONS: Community Gram negative urinary tract isolates remain highly sensitive to mecillinam and ciprofloxacin, but a significant number have developed resistance to trimethoprim/sulfamethoxazole. Further studies are required to determine the clinical significance of these results. PMID- 18159353 TI - Treatment of CMV retinitis with intravitral ganciclovir in the HAART era. AB - OBJECTIVE: To describe the course and outcome of cytomegalovirus (CMV) retinitis among AIDS patients treated with intravitreal ganciclovir and systemic highly active antiretroviral therapy (HAART). The secondary objective was to compare the course of CMV retinitis between patients receiving HAART and those not receiving this treatment. DESIGN: A retrospective cohort design consisting of 21 eyes from 16 patients with AIDS and CMV retinitis consecutively enrolled between January 1996 and August 1999. All patients received intravitreal ganciclovir therapy, and half of the patients began HAART as well. Duration of intravitreal therapy and ensuing disease quiescence, as well as CD4+ T cell counts at diagnosis and at cessation of ganciclovir, were calculated. Secondly, instantaneous hazards for outcomes such as CMV retinitis progression, ocular complications and mortality were compared. SETTING: Tertiary care centre in Ottawa, Ontario. RESULTS: Five of eight patients receiving HAART discontinued intravitreal ganciclovir after a mean treatment period of 428 days. During this period, their mean CD4+ count rose from 7.5 to 190microL. Subsequently, none of these patients experienced retinitis progression during follow-up periods lasting up to 820 days (mean of 617 days). Progression of CMV retinitis was 11.4 times more likely among those not receiving HAART (P=0.049). CONCLUSIONS: On initiating HAART, patients with CMV retinitis may enjoy significant recovery in CD4+ counts and sustained retinitis quiescence without specific anti-CMV therapy. Intravitreal ganciclovir injections seem well suited to offer effective CMV control during temporary periods of decreased CD4+ counts while awaiting HAART-mediated immune system reconstitution. PMID- 18159354 TI - Isolated native tricuspid valve endocarditis caused by viridans streptococcus. AB - The present report describes a case of native tricuspid valve endocarditis caused by viridans group streptococcus in a 43-year-old man who had recently undergone dental extraction. The patient had no history of intravenous drug use, heart disease or right heart catheterization. Although there have been scattered reports of unusual organisms, to the authors' knowledge, this is the first case of viridans group streptococcal endocarditis involving only the tricuspid valve after dental manipulation. PMID- 18159355 TI - Cotrimoxazole-induced hypoglycemia in an HIV-infected patient. AB - A case of cotrimoxazole-induced hypoglycemia is described in a male patient infected with HIV. Ten days after initiating high dose cotrimoxazole for suspected Pneumocystis carinii pneumonia, the patient developed neuroglycopenic symptoms and diaphoresis. Blood glucose levels were repeatedly low, with elevated insulin and C-peptide levels despite multiple intravenous bolus doses and infusions of dextrose. Hypoglycemia resolved after approximately 36 h of treatment with dextrose and discontinuation of cotrimoxazole. A review of reported cases of hypoglycemia associated with cotrimoxazole is provided, including information about onset, risk factors and possible mechanism. PMID- 18159356 TI - Gemella species endocarditis in a child. AB - Organisms of the genus Gemella can, on occasion, cause serious systemic illness. The present paper reports a successfully treated case of endocarditis in a 12 year-old girl with congenital heart disease caused by species of Gemella. The child presented with cough, fatigue and decreased appetite without fever. Echocardiogram demonstrated marked mitral insufficiency with flail posterior mitral valve leaflet, mitral valve vegetations, and an enlarged left atrium and ventricle. While being treated with vancomycin, the child initially had persistent bacteremia, which resolved after the addition of gentamycin; the course of therapy was completed with penicillin G and gentamycin once antimicrobial susceptibilities were available. Attempts to identify the species of Gemella were unsuccessful in the local laboratory, and at reference laboratories in Canada and the United States. The isolate is undergoing further evaluation to determine its taxonomic status. PMID- 18159357 TI - Hepatitis. PMID- 18159358 TI - Creutzfeldt-Jakob disease and infection control. PMID- 18159359 TI - Historical review of croup. PMID- 18159360 TI - Current hepatitis A status in Canada. AB - Hepatitis A, caused by the hepatitis A virus, occurs most frequently in developing countries, but also causes sporadic cases or outbreaks in industrialized countries. The most common route of transmission is fecal-oral. The incidence of hepatitis A varies with geography, and economic and environmental conditions. The epidemiological pattern of the disease has changed with improvements in hygiene and economic conditions. The incidence and prevalence of hepatitis A has decreased, while the average age of exposure and subsequent infection has increased. The present report describes the current status of hepatitis A in Canada. The incidence rate of reported cases in Canada varies from over 10/100,000 (1991) to 3.6/100,000 (1998), and is higher in males, 4.7/100,000 (1998), than in females, 2.5/100,000 (1998). The highest reported hepatitis A rates are in age groups 30 to 39 years and 40 to 59 years, and in British Columbia. Such information is important for assessing current immunization approaches and for decision-making about new preventive strategies against hepatitis A in Canada. PMID- 18159361 TI - Epidemiology of hepatitis B in Canada. AB - OBJECTIVE: To provide a current and comprehensive review of the epidemiology of hepatitis B virus (HBV) in Canada. DATA SOURCES: Published and unpublished epidemiological studies and surveillance reports of the past decade, primarily from Canada were studied. Fifty reports addressing HBV surveillance, incidence and prevalence, transmission-associated risk factors, co-infections, and prevention strategies were reviewed. DATA SYNTHESIS: HBV infection is an important vaccine-preventable infectious disease in Canada. The incidence rate of clinically recognized, acute HBV infection in 1998/1999 was estimated to be 2.3/100,000 people or approximately 700 cases a year. The prevalence of HBV carriers is estimated to be 0.5% to 1.0% of the population, but varies substantially according to population-specific risk factors. Most acute HBV infections are associated with injection drug use or high risk heterosexual activities, but 20% to 30% of acute cases did not report any identified risk factors. Surveillance activities such as the National Notifiable Disease Reporting system provide information regarding trends and risk factors. The primary preventive strategy for HBV consists of universal immunization for preadolescents and/or infants. Other strategies, such as the universal prenatal screening and postnatal immunization, and the prevention of nosocomial acquisition, are also important. The recently described hepatitis B surface antigen (HBsAg) escape mutants may not be detected by current HBsAg test assays, and the existing HBV vaccines may not protect vaccinees from infections by such mutants. CONCLUSION: Ongoing surveillance and research are required to assess risk factors for HBV transmission, evaluate the effectiveness of immunization programs and monitor the impact of HBsAg escape mutants. PMID- 18159362 TI - Chronic hepatitis B infection in Canada. AB - Recent developments in the treatment and prevention of hepatitis B virus (HBV) infections warrant revisiting important epidemiological questions, such as how prevalent is chronic HBV infection in Canada, in which Canadian subpopulations are HBV prevalence rates the highest, in what percentage of infected individuals is the virus actively replicating, and how many infected Canadians are candidates for antiviral therapy? Currently available data suggest the overall prevalence of HBV-infected individuals in the general population is approximately 2%, with 5% to 10% having serological evidence of previous HBV infection. In high risk groups, such as street-connected individuals, Aboriginals and immigrants from endemic areas, these rates of viral prevalence and serological evidence of previous HBV infection are approximately two to 10 and five to 10 times higher, respectively, than in the general population. Candidates for antiviral therapy range from less than 1% of infected Aboriginals to 15% to 30% of Asians with chronic HBV. From these data, it is clear that chronic HBV remains an important public health problem in this country. Hence, resources must be identified to enhance Canadians' awareness of HBV infection, maintain, if not expand, efforts to identify and implement safe and effective antiviral therapy for HBV-infected individuals, and continue programs for universal vaccination to prevent new HBV infections. PMID- 18159363 TI - Enhanced surveillance of acute hepatitis B and C in four health regions in Canada, 1998 to 1999. AB - OBJECTIVE: To assess the incidence and risk factors for acute hepatitis B and acute hepatitis C in a defined Canadian population. PATIENTS AND METHODS: An enhanced surveillance system was established in October 1998 to identify cases of acute hepatitis B and C infections in four regions in Canada, with a total population of approximately 3.2 million people. Information on demographic and clinical characteristics, laboratory results and potential risk factors was collected using predefined questionnaires. RESULTS: A total of 79 cases of acute hepatitis B and 102 cases of acute hepatitis C were identified from October 1998 to December 1999, resulting in an incidence rate of 2.3 and 2.9/100,000 person years, respectively. Males had higher incidence rates than females. The incidence of acute hepatitis B peaked at age 30 to 39 years for both males and females, whereas acute hepatitis C peaked at 30 to 39 years for males and 15 to 29 years for females. At least 34% of acute hepatitis B and 63% of acute hepatitis C were associated with injection drug use. Persons who were 15 to 39 years of age were more likely to report injection drug use as a risk factor. Heterosexual contact was reported to be a risk factor for 36.6% of acute hepatitis B cases and 3.5% of acute hepatitis C cases. CONCLUSIONS: The surveillance provides national incidence estimates of clinically recognized acute hepatitis B and C. Both hepatitis B and C are important public health threats to Canadians. Prevention efforts for both diseases should focus on injection drug use, especially for people aged 15 to 39 years. Risky sexual behaviour is also a major concern in prevention of hepatitis B in Canada. PMID- 18159364 TI - The emerging epidemiology of VRE in Canada: results of the CNISP Passive Reporting Network, 1994 to 1998. AB - OBJECTIVE: To provide a rapid and efficient means of collecting descriptive epidemiological data on occurrences of vancomycin-resistant enterococcus (VRE) in Canada. DESIGN AND METHODS: Passive reporting of data on individual or cluster occurrences of VRE using a one-page surveillance form. SETTING: The surveillance form was periodically distributed to all Canadian Hospital Epidemiology Committee members, Community and Hospital Infection Control Association members, L'Association des professionnels pour la prevention des infections members and provincial laboratories, representing 650 health care facilities across Canada. PATIENTS: Patients colonized or infected with VRE within Canadian health care facilities. RESULTS: Until the end of 1998, 263 reports of VRE were received from 113 health care facilities in 10 provinces, comprising a total of 1315 cases of VRE, with 1246 cases colonized (94.7%), 61 infected (4.6%)and eight of unknown status. (0.6%). VRE occurrences were reported in 56% of acute care teaching facilities and 38% of acute care community facilities. All facilities of more than 800 beds reported VRE occurences compared with only 10% of facilities with less than 200 beds (r2=0.86). Medical and surgical wards accounted for 51.4% of the reported VRE occurences. Sixty-five (24.7%) reports indicated an index case was from a foreign country, with 85.2% from the United States and 14.8% from other countries. Some type of screening was conducted in 50% of the sites. CONCLUSIONS: A VRE passive reporting network provided a rapid and efficient means of providing data on the evolving epidemiology of VRE in Canada. PMID- 18159365 TI - A critical review of oxazolidinones: an alternative or replacement for glycopeptides and streptogramins? AB - OBJECTIVE: To review the available data on the oxazolidinones linezolid and eperezolid. DATA SELECTION: Published reports were obtained by searching MEDLINE for articles published between 1992 and 2000, inclusive. References of published papers were also obtained and reviewed. Abstracts from scientific proceedings were reviewed. DATA EXTRACTION: Due to the limited data available regarding these agents, the criteria for study inclusion were not restrictive. DATA SYNTHESIS: The oxazolidinones (eg, linezolid) are a new antimicrobial class with a unique mechanism of action. They are active against resistant Gram-positive cocci including methicillin-susceptible and -resistant Staphylococcus aureus (MRSA), methicillin-susceptible and -resistant Staphylococccus epidermidis, vancomycin resistant enterococci (VRE) and penicillin-resistant Streptococcus pneumoniae (PRSP). Linezolid is active against anaerobes and displays modest activity against fastidious Gram-negative pathogens such as Haemophilus influenzae, but is not active against Enterobacteriaceae. Linezolid is available both orally and parenterally, and has a bioavailability of 100%. Clinical trials comparing linezolid with standard therapy have demonstrated similar bacteriological and clinical cures rates to standard therapy in community- and hospital-acquired pneumonia, uncomplicated and complicated skin and soft tissue infections, and infections caused by MRSA and VRE. Adverse effects have been minor and infrequent; however, platelets should be monitored in patients who have received more than two weeks of linezolid therapy. It is expected that these agents will have a bright future due to their excellent spectrum of activity against antibiotic-resistant Gram-positive organisms, such as MRSA, VRE and PRSP, and their excellent bioavailability. CONCLUSION: The oxazolidinones represent a new class of antimicrobials with a unique mechanism of action. They have excellent activity against susceptible and resistant Gram-positive organisms such as MRSA, methicillin-susceptible S epidermidis, VRE and PRSP, and a good adverse effect profile; they can be administered both intravenously and orally. Their potential use in Canada may be as an intravenous and oral alternative to glycopeptides and streptogramins. PMID- 18159366 TI - Is our vaccine supply secure? PMID- 18159367 TI - Macrolide resistance in Streptococcus pneumoniae: Fallacy or fact? PMID- 18159368 TI - Antimicrobial resistance in Canada: Update on activities of the Canadian Committee on Antibiotic Resistance. PMID- 18159369 TI - A typical atypical pneumonia. PMID- 18159370 TI - A national survey of antimicrobial prophylaxis in adult cardiac surgery across Canada. AB - OBJECTIVE: To characterize national and regional patterns of antimicrobial prophylaxis in adult cardiac surgery across Canada. DESIGN: Retrospective, cross sectional analysis. SETTING: Thirty-three adult cardiac surgical centres across Canada. INTERVENTIONS: A one-page questionnaire collecting information regarding institutional demographics and antimicrobial prophylaxis regimens for adult cardiac surgical procedures was mailed to all adult surgical centres across Canada. If a response was not received within one month, a second survey was mailed, followed by a telephone reminder within two weeks of the second mailing. MAIN RESULTS: The Overall response rate was 100%. Prophylactic antimicrobials were used in all the adult cardiac centres; single-agent prophylaxis was used in 97% (32 of 33) of centres; Single-dose antimicrobial prophylaxis was used in only 3% (one of 33) of centres. Preoperative and postoperative antimicrobial prophylaxis regimens varied both between provinces and within provinces across Canada. Cefazolin was the antimicrobial used in 88% (38 of 43) and 87% (33 of 38) of the reported pre-operative and post-operative prophylaxis regimens, respectively. Antimicrobial prophylaxis was initiated in the operating room 72% (26 of 36) of the time and intra-operative supplemental antimicrobial doses were administered for cardiac procedures longer than a median of 4 hours (range 4 to 8 hr). Overall, the median duration of antimicrobial prophylaxis was 36 hours (range 8 to 96 hr). CONCLUSIONS: Despite the availability of various published guidelines, our survey identified several areas for improvement with respect to antimicrobial prophylaxis in adult cardiac surgery across Canada. PMID- 18159371 TI - Introductory note on the correlation between the molecular and clinical aspects of infection. PMID- 18159372 TI - The buccale puzzle: The symbiotic nature of endogenous infections of the oral cavity. AB - The indigenous, 'normal', microflora causes the majority of localized infectious diseases of the oral cavity (eg, dental caries, alveolar abscesses, periodontal diseases and candidiasis). The same microflora also protects the host from exogenous pathogens by stimulating a vigorous immune response and provides colonization resistance. How can a microflora that supports health also cause endogenous oral disease? This paradoxical host-symbiont relationship will be discussed within the dynamic of symbiosis.Symbiosis means 'life together' - it is capable of continuous change as determined by selective pressures of the oral milieu. Mutualistic symbiosis, where both the host and the indigenous microflora benefit from the association, may shift to a parasitic symbiosis, where the host is damaged and the indigenous microflora benefit. Importantly, these are reversible relationships. This microbial dynamism, called amphibiosis, is the essential adaptive process that determines the causation of endogenous oral disease by a parasitic microflora or the maintenance of oral health by a mutualistic microflora.Complex microbial consortiums, existing as a biofilm, usually provide the interfaces that initiate and perpetuate the infectious assault on host tissue. The ecology of the various oral microhabitats is critical for the development of the appropriate selecting milieux for pathogens. The microbiota associated with dental caries progression is primarily influenced by the prevailing pH, whereas periodontal diseases and pulpal infection appear to be more dependent on redox potential. Candidiasis results from host factors that favour yeast overgrowth or bacterial suppression caused by antibiotics. Oral health or disease is an adventitious event that results from microbial adaptation to prevailing conditions; prevention of endogenous oral disease can occur only when we realize that ecology is the heart of these host-symbiont relationships. PMID- 18159373 TI - Interactions between host and oral commensal microorganisms are key events in health and disease status. AB - The oral cavity has sometimes been described as a mirror that reflects a person's health. Systemic disease such as diabetes or vitamin deficiency may be seen as alterations in the oral mucosa. A variety of external factors cause changes in the oral mucosa, thus altering mucosal structure and function, and promoting oral pathologies (most frequently bacterial, fungal and viral infections). Little is known, however, about immune surveillance mechanisms that involve the oral mucosa.There is no direct contact between specific immune cells in the basal epithelium and microorganisms in the upper layers of the oral mucosa. The author's hypothesis is that the protective immunity is conveyed through epithelial cells. The present brief review assesses the oral mucosa's role as the main defense in the interactions between the host and the oral microbial community. A unique model was used to investigate these interactions as the cause of oral disease and to develop new treatments that exploit our knowledge of the host-microorganism relationship. PMID- 18159374 TI - Safety of fluoroquinolones: An update. AB - The fluoroquinolone class of antimicrobials has been in clinical use for over 13 years. During that period, some representatives of the class have been extensively prescribed, such as ciprofloxacin and levofloxacin, while others have seen minimal use and have been restricted or withdrawn, namely, trovafloxacin and grepafloxacin. Manipulation of the fluoroquinolone structure by substituting a range of moieties around the core has yielded enhanced antibacterial activity, but in some cases this has come at a price. Specific substitutions are discussed in relation to particular recognized adverse events. In the present paper, newly introduced fluoroquinolones, such as moxifloxacin and gatifloxacin, are examined in terms of anticipated class effects and recent clinical experience. These antimicrobials are associated with reactions such as diarrhea, nausea, headache and other typical antimicrobial phenomena at rates less than 5%. New fluoroquinolone agents should be examined carefully in light of structural findings until adequate clinical data are amassed. PMID- 18159375 TI - A tropical skin eruption. PMID- 18159376 TI - Rabies: Still with us. PMID- 18159377 TI - Point-of-care testing for HIV: HIV counselling and testing. PMID- 18159378 TI - Epstein-Barr virus infection in transplant recipients: Summary of a workshop on surveillance, prevention and treatment. AB - Diseases caused by the Epstein-Barr virus are of great significance among organ transplant recipients. One of these diseases, post-transplant lymphoproliferative disease, is a major complication among organ transplant recipients. Management of this entity is problematic due to the difficulties with laboratory surveillance, diagnosis, prevention and treatment. A group of Canadian and American experts was assembled to discuss these aspects of Epstein-Barr virus diseases in Canadian organ transplant recipients. This report summarizes the relevant background literature and levels of evidence in relation to the outcomes of the deliberations and recommendations by the expert panel. PMID- 18159379 TI - Polyclonal intravenous immunoglobulin for the prophylaxis and treatment of infection in critically ill adults. AB - Infection is a major cause of morbidity and mortality in critically ill patients. Despite advances in technology, its mortality rate has changed minimally over the past two decades, and new therapies are needed. Polyclonal intravenous immunoglobulin (IVIG) has been investigated both as a preventive and a treatment modality for sepsis and septic shock in critically ill adult patients. Prophylaxis with IVIG has been shown to reduce significantly the incidence of infection, particularly pneumonia, in selected postsurgical intensive care patients. However, it does not reduce mortality. The risk-benefit and cost effectiveness of this therapeutic intervention have not been determined, and its routine use is therefore not recommended. Treatment with IVIG has been shown in a number of small trials and a meta-analysis to reduce dramatically sepsis and septic shock mortality. However, a large, unpublished randomized trial has apparently shown no mortality benefit with this therapy. Despite limited evidence, IVIG has become the standard of care for the management of group A streptococcal toxic shock syndrome. At present, clinical equipoise exists for the use of IVIG in the treatment of sepsis and septic shock, and further study is needed. PMID- 18159380 TI - Multiple sclerosis and Epstein-Barr virus. AB - OBJECTIVE: To evaluate the epidemiological evidence for an etiological role of Epstein-Barr virus in multiple sclerosis (MS). DATA SOURCES: MEDLINE and Cochrane Library searches of the medical literature identified 24 studies. DATA EXTRACTION: Studies were categorized as seroepidemiological, case-control or historical cohort, and were then classified within each group according to methodological rigour using criteria derived from published guidelines for the epidemiological study of MS. DATA SYNTHESIS: There was significant variability in the quality of evidence, and while two well-designed cohort studies found increased relative risks of MS in subjects with infectious mononucleosis, results from other studies were unconvincing. CONCLUSIONS: The evidence was insufficient to accept or reject the hypothesis that Epstein-Barr virus has an etiological role in MS. Further study, ideally using large samples of incident cases with blinded, trained interviewers using confirmatory sources for recalled data, is needed. PMID- 18159381 TI - Rabies in a nine-year-old child: The myth of the bite. AB - A nine-year-old boy died from rabies encephalitis caused by a rabies virus variant associated with insectivorous bats. The patient was most likely infected in the Laurentian Mountains of western Quebec, but neither the patient nor his parents remembered any direct contact with an animal. The diagnosis was made seven days after the start of symptoms. After examining the most recent cases of rabies in North America, it is obvious that rabies following bat exposure can occur without history of a documented bite. The present case report emphasizes that the general public and medical care providers need better information about the risks associated with exposure to bats. PMID- 18159382 TI - Diagnosis and analysis of a recent case of human rabies in Canada. AB - BACKGROUND: On September 30, 2000, staff at the Canadian Food Inspection Agency's Centre of Expertise for Rabies, located at the Animal Diseases Research Institute in Ottawa, Ontario, diagnosed rabies in a child from Quebec. This was the first case of rabies in a human in Canada in 15 years and in 36 years in the province of Quebec. After spending a week in intensive care in a Montreal hospital, the nine-year-old boy succumbed to this nearly always fatal disease. The boy had been exposed to a bat in late August 2000, while vacationing with his family in the Quebec countryside. METHODS: Antemortem specimens taken from the patient were sent to the Canadian Food Inspection Agency laboratory for rabies diagnosis. Samples included saliva, eye secretions, corneal impressions, cerebral spinal fluid and skin. Specimens were examined by direct immunofluorescence microscopy, and results were confirmed using molecular biological techniques. Virus strain identification was performed by both genetic methods and phenotypic analysis with monoclonal antibodies. RESULTS: Initial results from direct immunofluorescence staining indicated that rabies virus was present in the skin biopsy specimen but not in the corneal impressions. This diagnosis of rabies was confirmed by polymerase chain reaction product analysis from several of the submitted specimens. Virus isolation from postmortem samples was not possible because fresh brain tissue was not available. Rabies virus was isolated from saliva and was determined to be similar to a variant that circulates in populations of silver haired bats. INTERPRETATION: Intravitam diagnosis of rabies in humans is very dependent on the samples submitted for diagnosis and the method used for testing. Upon first examination, only skin specimens were positive for rabies virus antigen; using polymerase chain reaction analysis, only saliva yielded positive results for rabies virus genome. Extensive testing and retesting of specimens submitted for rabies diagnosis in humans must be done to avoid false negative results. PMID- 18159383 TI - Colonic tuberculosis clinically misdiagnosed as anorexia nervosa, and radiologically and histopathologically as Crohn's disease. AB - A case of colonic tuberculosis presenting with severe wasting was misdiagnosed as anorexia nervosa at another institution. Double contrast barium enema showed strictures, and ulcerations of the cecum and ascending colon with a skip area. The radiologist believed that these findings were due to Crohn's disease. Colonoscopic biopsies from the involved area revealed histopathological changes typical of Crohn's disease and not tuberculosis. The patient completely recovered with antituberculous therapy without steroids. The case demonstrated the clinical, radiological and histopathological difficulties and misdiagnoses that may be associated with the diagnosis of colonic tuberculosis. PMID- 18159384 TI - Food for thought. PMID- 18159385 TI - Mupirocin - Are we in danger of losing it? PMID- 18159386 TI - Allergy to antibiotics in children: Perception versus reality. PMID- 18159387 TI - Seroepidemiology of Coxiella burnetii infection and its frequency as a cause of community-acquired pneumonia in Canada. AB - The present study tested acute and convalescent serum samples from 788 patients hospitalized for community-acquired pneumonia in seven Canadian provinces for antibodies to Coxiella burnetii. One hundred nine patients (13.8%) had antibodies to this microorganism, and seven patients had acute Q fever. Serological evidence of infection with C burnetii was present in patients from all seven provinces. Three of the seven cases of acute Q fever were from Manitoba, suggesting that there may be unrecognized cases of Q fever in this province. In addition, a case of acute Q fever in Newfoundland, where there had previously been no reported cases, was noted, although subsequently, an outbreak of Q fever on goat farms has been reported. PMID- 18159388 TI - Economic analysis of rotavirus-associated diarrhea in the metropolitan Toronto and Peel regions of Ontario. AB - OBJECTIVE: To measure the economic cost of rotavirus-associated diarrhea for a selected group of families, in a nonexperimental setting, and to estimate the factors that influence these costs. DESIGN: Use and other socioeconomic data from a family survey (the Pediatric Rotavirus Epidemiology Study for Immunization) of children who tested positive for rotavirus were collected for the metropolitan Toronto and Peel regions of Ontario during the rotavirus season of 1997-1998. Service costs were estimated from provider data. A statistical regression analysis was used to explain the variances of provincial health care costs, prescription drug costs and indirect (work-loss) costs. SETTING: Data were collected in hospitals, emergency rooms, paediatric practices, primary care clinics and licensed daycare centres. Hospital coverage was wide, but community coverage was not. PATIENTS AND OTHER PARTICIPANTS: Children with diarrhea were tested for rotavirus. Those who tested positive and whose parents consented for their children to participate were included in the study. INTERVENTIONS: None MAIN OUTCOME MEASURES: The main outcomes were provincial health care costs, drug costs, nonmedical costs and the number of days of work missed by parents per child, as well as factors that determine these costs. RESULTS: Children in the most severe category incurred costs of $2,663/person, and those in the least severe categories incurred costs of approximately $350/person. The most important determinant to explain provincial health care costs was the number of health problems that the child had before having rotavirus. Costs due to work loss of parents were considerable for children in all severity groups and were influenced by family working conditions. CONCLUSIONS: When considering the economic implications of rotavirus, prior health status should be considered and indirect costs should be recognized for their importance. PMID- 18159389 TI - Outbreak of Escherichia coli 0157:H7 related to animal contact at a petting zoo. AB - OBJECTIVE: To determine the cause of an outbreak of Escherichia coli 0157:H7 related to animal exposures so that further transmission could be prevented. DESIGN: Description of laboratory investigations and a case control study. SETTING: Agricultural pavilion at an annual fair in Ontario. POPULATION: People with laboratory evidence of E coli 0157:H7 (seven people) and others with diarrhea (155 people) who called the health unit following a media release were interviewed. Animals that were accessed most frequently by the public in the agriculture pavilion were tested for E coli 0157:H7. In the case control study, a case was defined as someone with laboratory confirmed E coli 0157:H7, or someone who developed severe or bloody diarrhea two to eight days after attending the agricultural pavilion at the fair (61 people). A convenience sample of people who attended the agricultural pavilion but did not develop diarrhea was selected as the control group (89 people). INTERVENTIONS: Human and animal E coli 0157:H7 specimens were subtyped. Cases and controls were interviewed using a standardized questionnaire. RESULTS: Subtyping of the seven human isolates of E coli 0157:H7 revealed five that were of an extremely uncommon phage type. Three samples from goats and one from sheep at the petting zoo in the agricultural pavilion were of this same phage type. The case control study also implicated goats (odds ratio [OR] 3.65; 95% CI 1.63 to 8.52) and sheep (OR 2.94; 95% CI 1.33 to 6.57) from the petting zoo. CONCLUSIONS: Results of this investigation suggest strongly that the goats and sheep from the petting zoo were the source of this outbreak of E coli 0157:H7. PMID- 18159390 TI - The 'farm to plate' approach to food safety - Everyone's business. AB - Food safety is growing in importance as a public health concern for health practitioners and the general public. The Canadian public is exposed more extensively than ever before to exotic foods and pathogens via international travel, changing lifestyles and domestic contact with fresh foodstuff that is imported from faraway lands. Global warming, changing microbial ecology and resistance, and reduced host immunity are also having their effect in increasing the risk. To manage the risk adequately, interventions must be implemented at every point of the food supply chain, from production and processing to distribution, preparation and consumption, at home and in retail food service establishments. The role of government is explained, and the roles of other stakeholders, including physicians, are reviewed briefly. PMID- 18159392 TI - Assessing immunization programs. PMID- 18159391 TI - Risk factors for genital chlamydial infection. AB - OBJECTIVE: To discuss the occurrence of genital chlamydia in developed countries and review the literature assessing the potential risk factors for this sexually transmitted disease. DATA SOURCES: A MEDLINE search was performed for all English citations from 1985 to 2000 that contain the keywords "Chlamydia trachomatis", "chlamydial infections", "risk factors" and "sex behaviour". All relevant references cited in articles that were obtained from the search were also included. DATA EXTRACTION: ALL ARTICLES OBTAINED FROM THE ABOVE SOURCES WERE EXAMINED, AND WERE INCLUDED IN THE REVIEW IF THEY MET THE FOLLOWING CRITERIA: primary study examining sociodemographic or behavioural risk factors associated with genital chlamydial infection using multivariate analysis; study subjects 12 years of age and older; and study setting in a developed country. DATA SYNTHESIS AND CONCLUSIONS: Genital chlamydial infection has become the most commonly reported bacterial infection in North America over the past decade. Thirty-eight cross-sectional studies and six cohort studies were included in the present review. Most studies demonstrated that young men and women are at higher risk of being infected with chlamydia than older subjects. Chlamydia seems to be found in a diverse group of people, and unlike gonorrhea, is not concentrated in low income, minority core groups with high rates of partner change. However, a number of studies have shown that communities with well-established control programs are beginning to demonstrate this pattern. There is no clear evidence that chlamydia is associated with type of partners, contraceptive use, or age at first intercourse. Future research should follow this sexually transmitted disease as it evolves through the epidemiological stages to ensure that preventive and treatment services are reaching those people who are most likely to be infected. PMID- 18159393 TI - Routine adult immunization in Canada: recommendations and performance. PMID- 18159395 TI - Antimicrobial resistance in Canada: update on activities of the Canadian Committee on Antibiotic Resistance. PMID- 18159394 TI - Preventing group B streptococcal infections: new recommendations. PMID- 18159396 TI - Hospitalizations for diarrhea in Quebec children from 1985 to 1998: estimates of rotavirus-associated diarrhea. AB - OBJECTIVE: To characterize the incidence and duration of hospitalization due to diarrhea and to assess the proportion of hospitalizations that are attributed to rotavirus-associated diarrhea. DESIGN: Retrospective study of hospitalization data. SETTING: Hospitals located in Quebec. POPULATION STUDIED: Children from one to 59 months of age who were discharged from hospital from April 1, 1985 to March 31, 1998. MAIN RESULTS: There were 63,827 hospitalizations for diarrhea over the study period, for an average of 4910 hospitalizations/year. The epidemic curve showed a periodicity with regular alternation of high and low annual peaks. The number of hospitalizations for rotavirus-associated diarrhea was estimated according to three different methods. The estimates varied between 1353 and 1849 hospitalizations due to rotavirus-associated diarrhea/year over the 13-year period, with good agreement between the results of the three methods for a one month to five years of age incidence of 320 hospitalizations for rotavirus associated diarrhea/100,000 children. The average duration of hospital stay decreased from 5.2 days in 1985 to 3.3 days in 1998. CONCLUSIONS: The present article shows the importance of diarrhea hospitalizations among children and the alternating peak-year periodicity. PMID- 18159397 TI - Epidemiology and clinical features of methicillin-resistant Staphylococcus aureus in the University Hospital, Jeddah, Saudi Arabia. AB - OBJECTIVE: To describe the prevalence, demography and clinical characteristics of patients who were colonized or infected with methicillin-resistant Staphylococcus aureus (MRSA) in 1998 at King Abdulaziz University Hospital, Jeddah, Saudi Arabia. PATIENTS AND METHODS: Results of MRSA-positive cultures of clinical specimens obtained as part of investigations for suspected infections were retrieved from the King Abdulaziz University Hospital Infection Control Department's records. Charts of patients were reviewed. RESULTS: Of 292 S aureus isolates identified, 111 (38%) were MRSA, or 6.0 MRSA isolates/1000 admissions, which represented a marked increase over MRSA prevalence in 1988 (less than 2%). Nosocomial acquisition occurred in 74.8% of isolates. All age groups were affected, but 45.9% of patients were in the 'extremes of age' group (younger than one or older than 60 years of age). The prevalence was highest in the medical ward (27%), followed by the paediatrics combined medical and surgical ward (20.7%), the outpatient department (18%), the adult surgical ward (17.1%) and the intensive care units (17.1%). Two-thirds (66.7%) of cases represented infection and the remainder represented colonization. Surgical wounds (31.1%), the chest (27%) and endovascular catheters (20.3%) were the most common sites of infection. Bacteremia occurred in 27% of patients. Local signs (68.9%) and fever (60.8%) were the most common clinical manifestations. Respiratory distress and septic shock occurred in 28.4% and 6.8% of cases, respectively. Of 74 patients with MRSA infection and 37 patients with MRSA colonization, 91.9% and 56.8% received antibiotics in the preceding six weeks, respectively (P<0.0001). The total mortality of patients with MRSA infection was 60.8%; 37.8% of deaths were the result of MRSA infection and 23% were the result of other diseases. CONCLUSIONS: The prevalence of MRSA is high and rapidly increasing at King Abdulaziz University Hospital, as it is worldwide. Control measures to prevent the spread of MRSA in hospitals should continue with reinforcement of hygienic precautions and development of policies to restrict the use of antibiotics. PMID- 18159398 TI - Review of the use of cephalosporins in children with anaphylactic reactions from penicillins. AB - OBJECTIVE: It is a widely accepted practice that children with anaphylaxis from penicillins should avoid cephalosporins. The purpose of the present study was to determine whether there is evidence in the literature to support this practice. DATA SOURCES: MEDLINE, EMBASE, Toxline, International Pharmaceutical Abstracts and PubMed were used to search the literature published from 1966 to 2001. The Canadian Medical Protective Association, Health Canada and the Boston Collaborative Drug Surveillance Program were also contacted to determine whether there were any unpublished cases of cross-reactivity between penicillins and cephalosporins. DATA EXTRACTION: Cases describing the use of cephalosporins in adults and children with positive penicillin skin tests or anaphylaxis from penicillin were evaluated. Case reports of anaphylaxis from cephalosporins in paediatric patients were identified. DATA SYNTHESIS: There have been five reported cases of serious reactions from cephalosporins in patients with a history of anaphylaxis from penicillins. All cases occurred in adults; three developed anaphylaxis from the older, first-generation cephalosporins, cephalothin and cephaloridine; one developed anaphylaxis from cefamandole; and one developed anaphylaxis from cefaclor. There have been 12 other published reports of anaphylaxis from cephalosporins in adults with a history of penicillin allergy or a positive penicillin skin test, but with no history of anaphylaxis from penicillin. In seven studies, in which a total of 158 patients with positive penicillin skin tests were administered cephalosporins, seven had apparent immunoglobulin E-mediated reactions when they were given a cephalosporin. When the class of cephalosporin was able to be determined, none of the reports of reactions from cephalosporins in patients with allergies to penicillin involved third-generation cephalosporins. There have been 13 case reports of anaphylaxis from cephalosporins in paediatric patients. CONCLUSION: There are no published case reports of anaphylaxis from cephalosporins in children with anaphylaxis from penicillin, and there are only a small number of such reports in adults. Anaphylaxis from cephalosporins appears to be incredibly rare in children. There is minimal evidence in the literature to support the avoidance of cephalosporins in children with anaphylaxis from penicillins. PMID- 18159399 TI - Blastomycosis acquired by three children in Toronto. AB - Three paediatric cases of blastomycosis, apparently acquired in or near Toronto, Ontario, a region not known to be endemic for this disease, are described. Blastomycosis was not suspected clinically in any of the three cases, and the diagnosis was established only when the diagnostic net was broadened to include fungal and mycobacterial cultures. All three patients were diagnosed after significant delays, which is consistent with the rarity of the disease in children and its acquisition outside previously accepted geographical boundaries. Pulmonary involvement was present in all three children, while one also had multifocal osteomyelitis. Drug therapy was successful in all three cases, either with amphotericin B followed by itraconazole, or itraconazole alone. Blastomycosis should be included in the differential diagnosis of a patient from the Toronto area who presents with a compatible history despite a negative travel history to known endemic zones. PMID- 18159400 TI - Fever and a rat bite. PMID- 18159401 TI - A new society. PMID- 18159402 TI - VISA, hetero-VISA and VRSA: the end of the vancomycin era? PMID- 18159403 TI - From soap and water, to waterless agents: update on hand hygiene in health care settings. PMID- 18159404 TI - Combined topical and oral antimicrobial therapy for the eradication of methicillin-resistant Staphylococcus aureus (MRSA) colonization in hospitalized patients. AB - OBJECTIVE: How to eradicate methicillin-resistant Staphylococcus aureus (MRSA) colonization in hospitalized patients is uncertain. We reviewed our experience with MRSA decolonization therapy in hospitalized patients. SETTING: An 1100-bed, university-affiliated tertiary care teaching hospital in Toronto, Ontario. DESIGN: Retrospective chart review of 207 adult inpatients with MRSA colonization hospitalized between February 1996 and March 1999. INTERVENTIONS: All patients with MRSA colonization were assessed for possible decolonization therapy with a combination of 4% chlorhexidine soap for bathing and washing, 2% mupirocin ointment applied to the anterior nares three times/day, rifampin (300 mg twice daily) and either trimethoprim/sulfamethoxazole (160 mg/800 mg twice daily) or doxycycline (100 mg twice daily). This treatment was given for seven days. RESULTS: A total of 207 hospitalized patients with MRSA colonization were identified and 103 (50%) received decolonization therapy. Patients who received decolonization therapy were less likely than untreated patients to have intravenous (P=0.004) or urinary catheters (P<0.001), or extranasal sites of colonization (P=0.001). Successful decolonization was achieved in 90% of the 43 patients who were available for at least three months of follow-up. CONCLUSIONS: Combined topical and oral antimicrobial therapy was found to be effective in eradicating MRSA colonization in selected hospitalized patients, especially those without indwelling medical devices or extranasal sites of colonization. PMID- 18159405 TI - Oral fluoroquinolones in the treatment of pneumonia, bronchitis and sinusitis. AB - BACKGROUND: Despite a relatively large number of clinical studies comparing oral fluoroquinolones to one antibiotic class comparator, there is limited information on the relative efficacy of different fluoroquinolones. OBJECTIVE: To examine the efficacy and tolerability of oral fluoroquinolones in the treatment of mild to moderate community-acquired pneumonia, acute exacerbations of chronic bronchitis and sinusitis. METHODS: A systematic review was undertaken with a MEDLINE search for antibiotics and indications. Included studies met the following criteria: original study; random allocation to treatment groups; treatment with one of the following oral antibiotics - moxifloxacin, levofloxacin, ciprofloxacin, gatifloxacin; controlled by either placebo or an active comparator medication; double-blind, single-blind or open treatment; men and women (18 years of age and older); diagnosis of one of the three indications; and treatment duration of at least three days. Outcome measures included efficacy and safety. Comparative and single arm meta-analyses were conducted. Statistical differences in antibiotic success rates were evaluated. Pooled point estimates and 95% CIs for the comparative statistics (z-scores, P-values) and the single-arm analysis were examined to evaluate equivalence. RESULTS: The results of the comparative and single meta-analyses revealed no major differences between the new fluoroquinolones. This is not surprising because the clinical studies were designed to show equivalence versus their comparators. Few comparative evaluations were conducted due to a paucity of studies. In relation to other competitors, small differences were seen. CONCLUSIONS: Results indicate that, in general, fluoroquinolones had similar efficacy, overall safety and dropout rates. PMID- 18159406 TI - Patterns of susceptibility in an outbreak of Bordetella pertussis: evidence from a community-based study. AB - OBJECTIVE: To describe an outbreak of Bordetella pertussis and to assess which factors were associated with the development of clinical pertussis in children and adults during the outbreak. DESIGN: A case series was described to define the epidemiology of the pertussis outbreak. A school-based survey of children was used to measure the incidence of clinical pertussis over the previous six months. Vaccination records from the local public health facility were used to look at the relationship between age and vaccination parameters, and susceptibility to clinically diagnosed pertussis. A cross-sectional survey of teachers, parents and some hospital workers was used to assess these associations in adults. SETTING: An outbreak of pertussis in an isolated northern community in British Columbia. POPULATION STUDIED: All children in the community who attend daycare, kindergarten or school, and their parents were surveyed. In addition, some health care workers and mothers of preschool children were surveyed. MAIN RESULTS: A total of 31 suspected cases of pertussis were identified over a three-month period. Ninety per cent of the affected children who had available vaccination records had received four or five doses of pertussis vaccine. Sixty per cent of the town's 209 children returned completed surveys. Of these, 69% had available vaccination records. Thirty-six children (28%) reported symptoms that fit the case definition for pertussis over the previous three months. Attack rates were highest for the group of children aged 10 to 14 years. In a multivariate logistic regression analysis, receiving prophylactic medication and an increased number of years from the last vaccine dose were found to be significant predictors for developing pertussis. Thirty-four per cent of the estimated 291 adults in the community returned completed surveys. The attack rate of pertussis in the adults was only 9%. Being a member of the school staff and/or having a household contact with pertussis were significant predictors of developing pertussis. CONCLUSIONS: Immunity to pertussis appears to wane during childhood. Peak susceptibility appears to be during early adolescence. Adults do not seem to be at greater risk than adolescents for developing the disease, but it seems unlikely that this is due to better immunity. Rather, it is probably related to a lower risk of exposure to pertussis and a lower rate of progression to symptomatic disease when adults are infected. PMID- 18159407 TI - Demographic, clinical and microbiological characteristics of maternity patients: a Canadian clinical cohort study. AB - OBJECTIVE: To determine the demographic, clinical and microbiological characteristics of a representative Canadian obstetrical population. DESIGN: A one-year cohort study of all maternity patients who were followed to delivery, using detailed patient questionnaires containing more than 60 demographic and clinical variables, and three microbiological evaluations during gestation - first trimester, 26 to 30 weeks, and labour and delivery. Outcome measurements included birth weight and gestational age. SETTING: Labour and delivery suites of all office obstetrical practices affiliated with a single hospital. POPULATION STUDIED: A consecutive sample of pregnant women in the study practices during one year were eligible for enrolment; 2237 consecutive patients were approached for consent, 2047 enrolled and 1811 completed the study through delivery. RESULTS: The average patient was white, married and 29 years of age. Slightly more than half of the patients had postsecondary education, but 10% fell below the national poverty line for income. Frequency of factors linked to adverse pregnancy outcomes included cigarette smoking (19%), alcohol ingestion (18%), previously having had a premature infant (7%), and maternal diabetes (2%). Overall prevalence of genital microbes variously implicated in prematurity was 37% for ureaplasma, 11% for group B streptococcus and 4% for Mycoplasma hominis. Prevalence of bacterial vaginosis was 14%. The median gestational age for the cohort was 39 weeks, with 7% of infants born less than 37 weeks' gestation. Mean birth weight was 3415 g. CONCLUSIONS: The present clinical cohort represents demographic and medical characteristics of the Canadian obstetrical population. The birth outcomes are consistent with national data. This database provides valuable information about a general obstetrical population that is managed by a universal health care system. PMID- 18159409 TI - Imported from Cuba? PMID- 18159408 TI - Verotoxigenic Escherichia coli (VTEC): a major public health threat in Canada. AB - BACKGROUND: Verotoxigenic Escherichia coli (VTEC) was first described in Canada during the 1980s as an emerging foodborne disease in association with morbidity and mortality in outbreaks of hemorrhagic colitis caused by E coli O157:H7. OBJECTIVE: To describe the surveillance activities and epidemiological laboratory markers of VTEC that are used at the National Laboratory for Enteric Pathogens (NLEP) to investigate sporadic cases and outbreaks of E coli O157:H7 and non-O157 VTEC in Canada. METHODS: Passive surveillance was conducted by obtaining data on laboratory confirmed cases of VTEC from the Provincial Laboratories of Public Health across Canada. The laboratory epidemiological markers generated for isolates of VTEC included biotyping, serotyping, phage typing, toxin detection and characterization, and molecular typing using pulsed-field gel electrophoresis. RESULTS: Major outbreaks of VTEC O157:H7 disease have been associated with ground beef, unpasteurized apple juice, salami and untreated water. In 1999 and 2000, a total of 46 outbreaks of E coli O157:H7 disease were investigated. Among those, one outbreak was associated with contact at a petting zoo and a second with the consumption of salami. An outbreak in 2000 in Ontario was associated with water and resulted in more than 1000 cases of human illness, with six deaths. The NLEP has also identified more than 100 non-O157 VTEC serotypes from cattle and meat products. At least 23 VTEC serotypes found in humans were also identical to those found in cattle and meat products. CONCLUSIONS: The laboratory-based information that is generated is used to define the incidence, sources of infection, risk factors, trends, distribution and transmission of VTEC to humans from food, water and animal sources. Prevention and control of outbreaks are high-priority health concerns. PMID- 18159410 TI - Antimicrobial resistance: A continuing Canadian tale. PMID- 18159411 TI - Immunization for bone marrow transplant recipients. PMID- 18159412 TI - Vaccine schedules. PMID- 18159413 TI - Guidance on patient identification and administration of recombinant human activated protein C for the treatment of severe sepsis. AB - Approximately one-third of cases of severe sepsis result in death. Endogenous activated protein C (APC) plays a key role in the regulation of the inflammation, fibrinolysis and coagulation associated with severe sepsis. In a recently published phase III trial, Protein C Worldwide Evaluation in Severe Sepsis (PROWESS), intravenous administration of recombinant human APC (rhAPC) 24 mug/kg/h for 96 h to patients with severe sepsis resulted in a 6.1% reduction in absolute mortality and a 19.4% reduction in the relative risk of death from any cause within 28 days (number needed to treat = 16). This dose is now being applied in clinical practice.rhAPC is recommended for the treatment of severe sepsis (sepsis associated with acute organ dysfunction) occurring as a result of all types of infection (Gram-negative bacterial, Gram-positive bacterial and fungal). A panel of Canadian clinicians experienced in the treatment of severe sepsis and the management of critical care patients has developed this consensus document to assist clinicians in appropriate patient selection and management of potential challenges associated with rhAPC therapy. PMID- 18159414 TI - Outcome of bacteremia and fungemia in paediatric oncology patients. AB - OBJECTIVE: To determine the outcome of paediatric oncology patients with positive blood cultures. DESIGN: Retrospective chart review. SETTING: Tertiary care hospital. POPULATION STUDIED: Oncology patients up to 17 years of age with positive blood cultures from January 1, 1994 to March 31, 1999. MAIN RESULTS: There were 121 episodes of positive blood cultures in 76 patients. Seventeen episodes were excluded because blood cultures were contaminated. Of the organisms grown from the remaining episodes, 63% were Gram-positive organisms, 23% were Gram-negative organisms, 3% were fungal and 11% were mixed. There were 13 episodes with pure or mixed isolates of Staphylococcus aureus, of which nine occurred within 14 days of the placement of a new central venous tunnelled catheter. Central venous tunnelled catheters were retained in 76 of the 102 episodes when they were present. There were two relapses, and four children were admitted to the intensive care unit with septic shock, but all survived. CONCLUSIONS: The outcome was excellent with the current management of possible bacteremia in paediatric oncology patients, but the high incidence of S aureus bacteremia suggests that empirical antibiotics should be altered if sepsis is suspected within 14 days of the placement of a central venous catheter. PMID- 18159415 TI - Three-year follow-up of protection rates in children given varicella vaccine. AB - OBJECTIVE: To determine the rate and severity of subsequent varicella and zoster among children given a varicella vaccine. DESIGN: Retrospective survey conducted three years after vaccination, using standardized telephone interviews. SETTING: Three urban Canadian centres (Halifax, Ottawa and Vancouver). PARTICIPANTS: Of 475 eligible children, 431 aged three to 15 years participated. INTERVENTION: Participants had received one dose of a live, attenuated varicella vaccine (Oka/GSK) an average of 36 months earlier. MAIN OUTCOME MEASURES: Interviewers asked about known varicella exposures and obtained descriptions of any illnesses reported as varicella or zoster. Reported varicella was classified as definite (vesicles present, physician confirmation), probable (vesicles reported), suspected (nonvesicular rash, recent contact with varicella) or unlikely (nonvesicular rash, no known contact). RESULTS: Parents reported 257 within-home exposures to varicella and alleged that 80 subjects had had varicella rash after exposure. Of these, 40 cases were classified as definite or probable (vesicles present; rate 9.3% or 3.1% per year on average), 31 as suspected (no vesicles; rate 2.4% per year) and nine as unlikely. All rash illnesses were mild: among children with vesicles, 75% were said to have up to 10 and none had more than 100. The outcome of 76 exposures to household members with varicella was evaluable: 25 (32.9%) resulted in a rash, and 13 (17.1%) of those involved vesicles (maximum 40 lesions). Parents alleged that three subjects had zoster, but only one illness was definite (six vesicles, physician confirmed). CONCLUSIONS: Varicella vaccination completely prevented the development of typical chickenpox during the three years of observation. Breakthrough infections were infrequent (about 5% per year) and mild. Zoster was rare. PMID- 18159416 TI - Distribution of serogroups of Neisseria meningitidis and antigenic characterization of serogroup Y meningococci in Canada, January 1, 1999 to June 30, 2001. AB - The relative frequency of serogroups of Neisseria meningitidis associated with meningococcal disease in Canada during the period January 1, 1999 to June 30, 2001 was examined. Of the 552 strains of N meningitidis collected from clinical specimens of normally sterile sites, 191 (34.6%), 276 (50.0%), 61 (11.1%) and 23 (4.2%) were identified by serological and molecular methods as serogroups B, C, Y and W135, respectively. About half (50.8%) of the serogroup Y isolates were isolated in the province of Ontario. The two most common serotypes found were 2c and 14. Most of the serogroup Y strains isolated from patients in Ontario were serotype 2c, while serotype 14 was the most common serotype associated with disease in the province of Quebec. The two most common serosubtypes found among the serogroup Y meningococci were P1.5 and P1.2,5. Laboratory findings, based on antigenic analysis, did not suggest that these serogroup Y strains arise by capsule switching from serogroups B and C strains. This study documented a higher incidence of finding serogroup Y meningococci in clinical specimens from patients in Ontario compared to the rest of Canada, and parallels the increase in serogroup Y meningococcal disease reported in some parts of the United States. PMID- 18159417 TI - A case of endocarditis: When three equals one. PMID- 18159418 TI - Access to vaccines: A call to action. PMID- 18159419 TI - Norwalk virus - Off and running. PMID- 18159420 TI - Antimicrobial resistance in Canada: Update on activities of the Canadian Committee on Antibiotic Resistance. PMID- 18159421 TI - Contemporary antiviral drug regimens for the prevention and treatment of orolabial and anogenital herpes simplex virus infection in the normal host: Four approved indications and 13 off-label uses. AB - Herpes simplex virus (HSV) orolabial and anogenital infection causes substantial and recurring disease in healthy individuals due directly to infection of these sites and, indirectly, due to its complications. These complications include eczema herpeticum plus erythema multiforme and neonatal HSV infection, respectively. Four drugs: acyclovir, famciclovir, valacyclovir and penciclovir, are currently licensed by the Therapeutics Products Directorate of Health Canada for the management of HSV infections. Although these drugs are only approved for four orolabial and anogenital infections in healthy persons, their efficacy and safety for 13 other related uses in this population have been demonstrated in controlled clinical trials, so called off-label uses. In this review, the evidence supporting these 17 uses, the drugs and regimens evaluated, and their current costs, are described. PMID- 18159422 TI - Choice of antibiotics in late neonatal sepsis in the extremely low birth weight infant. AB - OBJECTIVE: To review the choice of antibiotics in treating suspected late neonatal sepsis in infants weighing 1000 g or less in a neonatal intensive care unit. METHODS: Retrospective review of medical records. RESULTS: Ninety-six infants weighing 1000 g or less were admitted to the neonatal intensive care unit during the study period. Sixty-two infants survived beyond four days of life and had at least one sepsis workup done to exclude late neonatal infection. Of the 62 study patients, 42 (68%) were started on ampicillin and netilmicin (A/N) and 20 (32%) were started on vancomycin and ceftizoxime (V/C) as the antibiotics of choice, pending culture results. Of the patients started on A/N, 17 of 42 had a positive blood culture compared with 11 of 20 on V/C (40% versus 55%, P=0.40). The mean (+/-SD) birth weight of infants started on A/N was 793+/-133 g compared with a mean of 728+/-153 g in the group that received V/C (P=0.09). Seven patients died in the A/N group compared with three in the V/C group (16.7% versus 15%, P=0.84). In addition to the sepsis episode studied, before they were discharged from hospital, 21 of 42 (50%) infants in the A/N group had further workups for suspected sepsis, compared with 16 of 20 (80%) (P=0.048) infants initially given V/C. CONCLUSIONS: Ampicillin and netilmicin is a safe antibiotic combination for neonates suspected of late sepsis. This, in turn, may be important in reducing vancomycin overuse and the potential for bacterial resistance to this antimicrobial agent. PMID- 18159423 TI - Reasons for testing women for genital Chlamydia trachomatis infection in the Calgary region. AB - OBJECTIVE: To determine the clinical reason(s) for screening women with varying degrees of risk for genital Chlamydia trachomatis (CT) in the Calgary region. DESIGN: Women aged 15 to 75 years were enrolled at various patient care locations. Pertinent risk factors for genital CT infection were recorded and a gynecological examination was performed. Two endocervical swabs and a first-void urine sample were collected for CT detection using two different nucleic acid amplification test methods. SETTING: Calgary is an urban region that provides healthcare services to a population of almost one million people. Microbiology services are provided by Calgary Laboratory Services through a centralized regional laboratory service. MAIN RESULTS: 504 women with a mean age of 28.1 +/ SD 8.22 years were enrolled. Two hundred ninety-one women (57.8%) were at high risk for acquiring genital CT infection. Twenty-eight (5.6%) tested positive for CT infection and almost all of these women (26 of 28, 93%) had risk factors for acquiring infection. Of the high-risk women, 9.8% were CT positive versus only 1.3% of women at low risk (P=0.0001). Only two of 152 (1.3%) women older than 30 years had genital CT infections. Although most women were asymptomatic, those with laboratory-confirmed CT infection were more likely to have genitourinary symptoms. Three hundred forty-three of 476 (72%) women who did not have genital CT infection had no risk factors, and screening was done as part of a routine gynecological examination for other purposes (prenatal visit, Pap smear). CONCLUSION: Women without risk factors are being screened routinely for genital CT infection as part of a routine gynecological examination done for other reasons. Elimination of the routine screening of low-risk women older than 30 years of age would decrease the current regional utilization of CT tests by as much as one-third. PMID- 18159424 TI - Canada's first universal varicella immunization program: Lessons from Prince Edward Island. AB - A live attenuated varicella vaccine was licensed in Canada in December 1998. The first universal varicella immunization program in Canada was initiated in Prince Edward Island in 2000. Students in grades one to six without a history of varicella were offered the vaccine, administered by Public Health Nurses, in school clinics during February and March 2000. The acceptance rate ranged from 29.1% of all grade one students to 9.8% of all grade six students; overall, 18.8% of students received the vaccine. A universal program for children 12 months of age was introduced on April 1, 2000 and catch-up clinics for those between 12 months of age and those in grade one were introduced in June 2000. Repeated media announcements and the cooperation of staff in schools and daycare facilities assisted in informing parents about the availability of the vaccine. Vaccine associated adverse events have been uncommon and the vaccine has been well accepted. PMID- 18159425 TI - Mycobacterium neoaurum bacteremia in a hemodialysis patient. AB - Bacteremia due to Mycobacterium neoaurum, a rapidly growing mycobacterium, is described in a diabetic woman on hemodialysis. This is the first reported case of M neoaurum bacteremia in Canada. The organism initially grew on standard BacT/Alert SA aerobic blood cultures, and was subsequently positively identified using 16S rRNA sequence analysis. The present case serves to reinforce the need for a high index of clinical suspicion of infections caused by unusual microorganisms in the context of an immunocompromised host. PMID- 18159426 TI - Experiencing West Nile virus. PMID- 18159427 TI - Diagnosis of ventilator-acquired Pneumonia: Where Do We Go From Here? PMID- 18159428 TI - Transfusion and risk of infection in Canada. PMID- 18159429 TI - Clinical utility of a Legionella pneumophila urinary antigen test in a large university teaching hospital. AB - OBJECTIVE: To determine the clinical utility of diagnosing Legionella pneumonia by urinary antigen testing (LPUAT) in a low prevalence centre. DESIGN: The results of LPUATs were abstracted and analyzed from the authors' laboratory information system. Medical records were reviewed in detail for all positive tests and a random sample of 50 negative tests. SETTING: The Queen Elizabeth II Health Sciences Centre, a large university hospital complex. POPULATION STUDIED: Patients who were admitted from the emergency room with pneumonia or who had developed pneumonia in hospital and who had a LPUAT performed between April 1998 and October 2000. MAIN RESULTS: One thousand one hundred fifty-four tests were performed on 1007 patients. Seven patients had nine positive LPUATs. Three of these patients had confirmed Legionella pneumophila pneumonia. Three others had probable or possible L pneumophila pneumonia. There was one probable false positive. Six of the seven patients were already on empirical anti-L pneumophila therapy. Of the 50 negative tests reviewed in detail, 31 patients were on one of the antibiotics of choice for L pneumophila at the time the test was ordered; in 21 (68%) of these patients the negative result did not lead to a change in therapy. CONCLUSIONS: The cost to diagnose each case of Legionella pneumonia by LPUAT was approximately $5,770 and most patients were already on appropriate antibiotics. In patients with negative tests, antibiotics were often not changed in response to the test result. Rigorous screening of patients is required to increase pretest probability for LPUAT to be justified. PMID- 18159430 TI - Response to a protease-inhibitor (ritonavir)-containing combination antiretroviral regimen in HIV-infected children. AB - INTRODUCTION: The number of antiretroviral agents available for children who are failing existing therapy is limited. Data are lacking on the use of various combination regimens and the resulting viral load dynamics in such children. METHODS: Between March 1998 and March 2000, HIV-infected children younger than 18 years of age were studied in an open trial. The study regimen included ritonavir, with at least two drugs to which the virus was known or presumed to be sensitive. Subjects were ritonavir-naive and were included if they had high viral loads while receiving antiretroviral therapy. Patients had clinical assessments, CD4 counts and viral load monitoring. RESULTS: Fifteen antiretroviral-experienced HIV infected children were enrolled. Approximately 87% (13 of 15) had perinatally acquired HIV; median age was 7.9 years (range 1.6 to 14.8). At enrolment, the median CD4 count was 557 cells/mm(3) (range 57 to 1702) and the median viral load was 72,600 copies/mL (range 3626 to 796,440). The majority of children (73.3%) had increases in CD4 counts within 12 weeks. During this period, the median increase in CD4 counts over baseline was 30.0%. Approximately 73% (eight of 11) of subjects with initial improvements in CD4 counts had sustained increases at 32 to 48 weeks. Over the first 12 weeks, 60% (nine of 15) had greater than 0.5 log(10) decreases in viral load. The improvement was sustained in 88.9% (eight of nine) of these patients at 32 to 48 weeks. Three patients discontinued therapy due to taste aversion. CONCLUSIONS: Among pediatric patients with high viral loads while on existing therapy, the ritonavir-containing regimen was generally well tolerated. In a significant proportion of patients, modification of therapy was associated with sustained improvements in viral loads and CD4 counts over 32 to 48 weeks. PMID- 18159431 TI - Predominance of herpes simplex virus type 1 from patients with genital herpes in Nova Scotia. AB - The epidemiology of genital herpes is changing with evidence to suggest an increasing incidence of herpes simplex virus type 1 (HSV-1) infections. The results of 6529 HSV genital cultures taken between April 1998 and December 2001 were reviewed. overall, HSV-1 was recovered more often than HSV-2; 1213 versus 1045. This trend was particularly striking in young women 30 years of age or less, in whom 70.8% of isolates were HSV-1. In men of the same age range, 45% of isolates were HSV-1. The proportion of women with HSV-1 declined from 73.7% in those younger than 31 years of age to 4.5% in those older than 60 years of age.These observations have important implications. The decline in the relative proportion of HSV-1 isolates from young adults may be the result of changing sexual practices, changing susceptibility or increased exposure to HSV-1 during vaginal intercourse. In this setting HSV-2 vaccines may be less likely to produce the desired reduction in the overall prevalence of genital herpes infections. PMID- 18159432 TI - Coxiella burnetii seroprevalence of shepherds and their flocks in the lower Saint Lawrence River region of Quebec, Canada. AB - OBJECTIVE: To determine the seroprevalence of Coxiella burnetii among the shepherds and their sheep in the lower Saint-Lawrence River region (LSLRR) of Quebec, Canada. DESIGN: A prospective human-animal comparative study was conducted with 81 shepherds from 46 farms and a control group matched for sex and age. All participants answered a standardized questionnaire to evaluate their risk factors for Q fever, including a specific section on the work practices of the shepherds. All human subjects had a blood sample taken for serology to phase I and phase II antigens of C burnetii performed by indirect immunofluorescence assay. At each participating farm, seven to nine sheep had blood samples taken for C burnetii serology to be assessed by the complement fixation test. RESULTS: The seroprevalence to C burnetii was higher in the group of shepherds (28.4%) than the control group (1.2%) (P<0.005). Among the group of shepherds, spending more than 5 h/week in the sheep barn (P=0.06) and buying and/or trading sheep within the past six months (P=0.004) were associated with positive C burnetii serology. A total of 137 of 334 sheep (41%) were seropositive for C burnetii. These positive sheep were distributed in 41 of the 46 flocks (89%). No correlation could be demonstrated between a serology for C burnetii in the herds and the shepherds. CONCLUSION: Q fever is highly prevalent in the LSLRR of Quebec, affecting 89% of the flocks and 28% of the shepherds. Shepherds in this region are at increased risk for C burnetii infection in comparison to the general population. PMID- 18159433 TI - West Nile virus surveillance and diagnostics: A Canadian perspective. AB - A surveillance program has been in place since 2000 to detect the presence of West Nile virus (WNV) in Canada. Serological assays are most appropriate when monitoring for human disease and undertaking case investigations. Genomic amplification procedures are more commonly used for testing animal and mosquito specimens collected as part of ongoing surveillance efforts. The incursion of WNV into this country was documented for the first time in 2001 when WNV was demonstrated in 12 Ontario health units during the late summer and fall. In 2002 WNV activity was documented by avian surveillance in Ontario by mid-May with subsequent expansion of the virus throughout Ontario and into Quebec, Manitoba, Saskatchewan and Nova Scotia. Human cases were recorded in both Ontario and Quebec in 2002 with approximately 800 to 1000 probable, confirmed and suspect cases detected. The possible recurrence and further spread of WNV to other parts of Canada in 2003 must be anticipated with potential risk to public health. The continued surveillance and monitoring for WNV-associated human illness is necessary and appropriate disease prevention measures need to be in place in 2003. PMID- 18159434 TI - Xylohypha bantiana multiple brain abscesses in a patient with systemic lupus erythematosus. AB - Xylohypha bantiana is a rare cause of cerebral fungal infection (phaeohyphomycosis). We report on a 72-year-old man who, while taking several immunosuppressive medications for systemic lupus erythematosus, presented with multiple bilateral cerebral abscesses caused by X bantiana. The lesions were not surgically amenable and the patient died two months after discontinuing antifungal therapy. PMID- 18159435 TI - Cepacia-like syndrome caused by Burkholderia multivorans. AB - The variable severity of Burkholderia cepacia complex infections in cystic fibrosis (CF) has recently been ascribed to differences in the virulence between genomovars. Specifically, genomovar III isolates have been associated with higher transmission rates and adverse outcomes compared to other B cepacia genomovars, and consequently further segregation between genomovar III and non-genomovar III B cepacia infected patients is advocated in some centres. The important role of non-genomovar III isolates is presented in the context of a clinical case whereby a patient with long-standing pulmonary infection with B multiovorans developed bacteremic infection reminiscent of the fatal 'cepacia syndrome'. PMID- 18159436 TI - Residential water heater temperature: 49 or 60 degrees Celsius? PMID- 18159437 TI - Influenza - Expect the unexpected. PMID- 18159438 TI - Routine immunization schedule: Update 2004. PMID- 18159439 TI - Blood cultures in ambulatory patients who are discharged from emergency with community-acquired pneumonia. AB - OBJECTIVES: To determine the factors that predict whether or not ambulatory patients with community-acquired pneumonia (CAP) treated in an emergency room (ER) setting will have blood cultures drawn and the factors that predict a positive blood culture. METHODS: Prospective observational study of all patients with a diagnosis of CAP, as made by an ER physician, who presented to any of seven Edmonton-area ERs over a two-year period. RESULTS: Seven hundred ninety three (19.2%) of 4124 patients with CAP had blood cultures drawn. The site specific blood culture rates ranged from 7.8% to 25% (P<0.001); 41 of 793 (5.1%) were positive. Streptococcus pneumoniae accounted for 58.5% of the isolates while Staphylococcus aureus and Escherichia coli each accounted for 14.6%, or six patients each. Only two of the 24 patients with S pneumoniae bacteremia were subsequently admitted to hospital while all six of the patients with S aureus were admitted. Only one of the six patients with E coli bacteremia was treated at home. No factors were predictive of positive blood cultures on multivariate analysis. CONCLUSIONS: Physicians are selective in ordering blood cultures on patients with ambulatory pneumonia who present to an ER, and the positivity rate of 5.1% is quite high. No factors are predictive of positive blood cultures on multivariate analysis, thus clinical judgment has to prevail in the decision to perform blood cultures. Breakthrough bacteremia can occur with microorganisms susceptible to the antibiotics that the patient is receiving. PMID- 18159440 TI - Should university students be vaccinated against meningococcal disease in Canada? AB - OBJECTIVE: To evaluate the benefit and costs of vaccination of university students against invasive meningococcal disease (IMD) in Canada. METHODS: Published studies were reviewed and a simulation model was used. RESULTS: IMD risk seems to be of low magnitude, but consequences can be dramatic. Over a 10 year period, IMD risk reduction would be slightly greater using a monovalent C conjugate vaccine than a quadrivalent polysaccharide vaccine. From a societal perspective, costs per quality-adjusted life-years gained would be between $135,000 and $698,000, according to epidemiological scenarios and with vaccine purchase prices between $35 and $50 per dose. CONCLUSIONS: Economic indices exceed proposed criteria for cost effective public health programs, but from the perspective of students and parents, the cost of vaccination might be worth the benefit. PMID- 18159441 TI - Measurement of antibiotic consumption: A practical guide to the use of the Anatomical Thgerapeutic Chemical classification and Definied Daily Dose system methodology in Canada. AB - Despite the global public health importance of resistance of microorganisms to the effects of antibiotics, and the direct relationship of consumption to resistance, little information is available concerning levels of consumption in Canadian hospitals and out-patient settings. The present paper provides practical advice on the use of administrative pharmacy data to address this need. Focus is made on the use of the Anatomical Therapeutic Chemical classification and Defined Daily Dose system. Examples of consumption data from Canadian community and hospital settings, with comparisons to international data, are used to incite interest and to propose uses of this information. It is hoped that all persons responsible for policy decisions regarding licensing, reimbursement, prescribing guidelines, formulary controls or any other structure pertaining to antimicrobial use become conversant with the concepts of population antibiotic consumption and that this paper provides them with the impetus and direction to begin accurately measuring and comparing antibiotic use in their jurisdictions. PMID- 18159443 TI - Myiasis in an urban setting: A case report. AB - Myiasis is considered to be a condition only found in tropical, developing countries. However, this paper reports a case identified in an urban, North American setting. The clinical presentation is discussed along with the underlying comorbidities and social determinants. PMID- 18159442 TI - The management of infection and colonization due to methicillin-resistant Staphylococcus aureus: A CIDS/CAMM position paper. AB - Methicillin-resistant Staphylococcus aureus (MRSA) is being seen with greater frequency in most hospitals and other health care facilities across Canada. The organism may cause life-threatening infections and has been associated with institutional outbreaks. Several studies have confirmed that MRSA infection is associated with increased morbidity and mortality compared with infections caused by susceptible strains, even when the presence of comorbidities is accounted for. Treatment of MRSA infection is complicated by the fact that these organisms are resistant to multiple antimicrobial agents, so treatment options are limited. The effectiveness of decolonization therapy (attempting to eradicate MRSA carriage) is also uncertain. This paper reviews the medical management of MRSA infections, discusses the potential role of decolonization and provides an overview of evidence to support recommended infection control practices. PMID- 18159444 TI - Headache, fever and back pain in a 16-year-old boy. PMID- 18159445 TI - Tropical travel and life-threatening disease: preventing malaria misadventures. PMID- 18159446 TI - A SARS commentary. PMID- 18159447 TI - SARS : A Tale of Two Epidemics. PMID- 18159448 TI - West Nile virus - Mosquitoes no longer just an annoyance! PMID- 18159449 TI - Legionnaires' disease - Results of a multicentre Canadian study. AB - BACKGROUND: There has never been a cross-Canada surveillance project to determine the rate of Legionella species as a cause of community-acquired pneumonia requiring hospitalization and to determine whether there are any regional differences in the rates of Legionnaires' disease in Canada. Anecdotally, Legionnaires' disease is thought to be uncommon in Western Canada. METHODS: From January, 1996 through to October 31, 1997, a prospective study of the etiology of community acquired pneumonia requiring admission to 15 tertiary care hospitals in eight Canadian provinces was conducted. A urine sample from each patient was tested for Legionella pneumophila serogroup 1 antigen using a commercially available ELISA assay. A culture of sputum or other respiratory specimens for Legionellaceae was carried out at the discretion of the attending physician. Two hundred thirty-four patients had acute and 6-week convalescent serum samples tested for antibodies to L pneumophila serogroups 1 through 6 using an ELISA method. RESULTS: 28 of the 850 patients (3.2%) had Legionnaires' disease; 18 of 823 (2.1%) were positive for L pneumophila serogroup 1 by urinary antigen testing. The rate of Legionnaires' disease, based on urinary antigen, at the Halifax site was higher than that at the other sites (seven of 163 patients versus 11 of 660 [P=0.04]). Of the 28 cases of Legionnaires' disease identified using all methods, 11 of 277 patients (3.9%) were enrolled from Western provinces versus 17 of 573 patients (2.9%) from Eastern provinces (P=nonsignificant). CONCLUSIONS: Legionnaires' disease is just as common in Western as in Eastern Canada. L pneumophila serogroup 1 may be more common in Halifax than at the other sites studied. PMID- 18159450 TI - A cross-sectional study of intestinal parasitic infections in a rural district of west China. AB - BACKGROUND: Parasitic infections are widespread in rural areas of West China. The remote and humid environment, traditional ways of life, contaminated potable water and limited health services all contribute to the transmission and persistence of fecal parasites. OBJECTIVE: To describe the prevalence of intestinal parasitic infections in an area of West China, including its associations with demographic variables. METHODS: Single fecal specimens were collected using a previously validated kit. Parasites were detected microscopically by iodine-stained smear, modified Kato-Katz thick smear, simple saline smear, test tube filter paper culture and adhesive cellophane tape anal swab. RESULTS: The prevalence of enteric parasites in the population was 51.7%. The proportion of individuals infected with one parasite was 36.5%, two parasites 12.7%, three parasites 3.0% and four parasites 0.08%. Parasites identified included Ascaris lumbricoides in 41.4% of all individuals tested, Ancylostoma duodenale in 17.7%, Trichurias trichuria in 8.25%, Enterobius vermicularis in 0.43%, Blastocystis hominis in 1.37%, Entamoeba coli in 0.47%, Entamoeba histolytica in 0.16%, Giardia lamblia in 0.04%, Strongyloides stercoralis in 0.04% and Clonorchis sinensis in 0.04%. The prevalence was significantly increased in women, people aged 15 to 19 years and over 80 years, the rural farm population, farmers, preschool and primary school groups, residents of a hilly rather than mountainous or plains terrain, and in intermediate income groups. The prevalence was significantly negatively associated with educational level, but not with age or income. CONCLUSIONS: This study supports the use of simple fecal examinations to monitor the parasite burden in rural areas of developing countries. Enteric parasites remain common in this population in rural China. PMID- 18159451 TI - Necrotizing lung infection caused by the protozoan Balantidium coli. AB - Balantidium coli, a ciliated protozoan, is well known to cause intestinal infection in humans. Extraintestinal spread to the peritoneal cavity and genitourinary tract has rarely been reported. There have also been a few cases of lung involvement from this parasite. A case of B coli causing a thick-walled right upper lobe cavity in an organic farmer who had contact with aerosolized pig manure is reported. Bronchoalveolar lavage fluid examined for ova and parasite revealed trophozoites of B coli in large numbers. Treatment with doxycycline hyclate led to marked improvement. Necrotizing lung infection caused by the protozoan B coli should be considered in individuals who report contact with pigs. PMID- 18159452 TI - Chest wall hydatidosis as the unique location of the disease: Case report and review of the literature. AB - The chest wall is a rare location of secondary hydatidosis, but secondary hydatidosis may occur from the rupture of a lung cyst, from a liver cyst invading the diaphragm into the pleural cavity, following previous thoracic surgery for hydatidosis, or by hematogenous spread. This report describes a case of chest wall hydatidosis, which was the primary disease site in the patient, who had no previous history or current disease (hydatidosis) at other sites. The cyst invaded and partially destroyed the 9th and 10th ribs and the 10th thoracic vertebra, and protruded outside the pleural cavity through the 9th intercostal space. Preoperative albendazole administration for 10 days, surgical resection of the disease through a posterolateral thoracotomy incision, and postoperative albendazole treatment resulted in a cure with no evidence of local recurrence or disease at other sites in four years of follow-up. PMID- 18159453 TI - Pyo-mediastinitis: A complication of pulmonary coccidioidomycosis. AB - A rare fatal case of pulmonary coccidioidomycosis complicated by mediastinal and visceral abscesses treated with antifungal medications is described. The case report discusses the potential need for early surgical intervention to drain mediastinal and visceral abscesses as a primary mode of therapy in disseminated coccidioidomycosis for a successful control of infection and clinical outcome. PMID- 18159454 TI - An opinion paper: Strengthening the weakest link in food safety. PMID- 18159455 TI - Nosocomial transmission of bloodborne viruses from infected health care workers to patients. PMID- 18159456 TI - Current management of Herpes simplex infection in pregnant women and their newborn infants: What's hot and what's not. PMID- 18159457 TI - Nelfinavir and non-nucleoside reverse transcriptase inhibitor-based salvage regimens in heavily HIV pretreated patients. AB - OBJECTIVE: To assess the efficacy of nelfinavir mesylate (NFV) in combination with delavirdine mesylate (DLV) or efavirenz (EFV) and other antiretroviral agents following virological failure on other protease inhibitor (PI)-based regimens. DESIGN: Multicentre, retrospective chart review. METHODS: One hundred one patients who were naive to both NFV and non-nucleoside reverse transcriptase inhibitors (NNRTIs) and who initiated NFV plus DLV or EFV-based salvage regimens were reviewed. Response to treatment was defined as a reduction in HIV ribonucleic acid (RNA) levels to unquantifiable levels (less than 50 copies/mL, less than 400 copies/mL, less than 500 copies/mL) on at least one occasion after the initiation of salvage therapy. Baseline correlates of response, including prior duration of HIV infection, prior number of regimens, viral load and CD4 cell counts were also evaluated. RESULTS: Patients had a mean duration of HIV infection of 10 years, a mean duration of prior therapy of four years, a median of four prior nucleoside reverse transcriptase inhibitors and a median of two prior PIs. At the time of review the mean duration of salvage therapy was 63.4 weeks. Virological suppression was achieved in 59 (58.4%) patients within a mean of eight weeks and maintained for a mean of 44.9 weeks (the mean follow-up was 78 weeks). Of the non-responders, 16 (38%) achieved a less than 1 log(10) decrease in HIV RNA levels. Although there was no association between baseline correlates, response rate (75.7%) was significantly higher in patients with HIV RNA levels of 50,000 copies/mL or lower and CD4 counts greater than 200 cells/mm(3). CONCLUSION: NFV/NNRTI-based highly active antiretroviral therapy regimens are an effective therapy in many patients who have experienced virological breakthroughs on at least one prior PI-based regimen. PMID- 18159458 TI - An outbreak of foodborne botulism in Ontario. AB - Botulism is a rare paralytic illness resulting from a potent neurotoxin produced by Clostridium botulinum. Botulism in Canada is predominately due to C botulinum type E and affects mainly the First Nations and Inuit populations. The most recent outbreak of botulism in Ontario was in Ottawa in 1991 and was caused by C botulinum type A. We report an outbreak of foodborne type B botulism in Ontario, which implicated home-canned tomatoes. The outbreak was characterized by mild symptoms in two cases and moderately severe illness in one case. The investigation shows the importance of considering the diagnosis of botulism in patients presenting with cranial nerve and autonomic dysfunction, especially when combined with gastrointestinal complaints; it also highlights the importance of proper home canning technique. PMID- 18159459 TI - Predictors of a positive duplex scan in patients with a clinical presentation compatible with deep vein thrombosis or cellulitis. AB - BACKGROUND: Deep vein thrombosis (DVT) and cellulitis are common conditions whose symptoms lead patients to seek medical attention in the emergency department (ED). Distinguishing between these two conditions quickly and accurately is important. OBJECTIVES: To determine the yield of duplex scanning among ED patients whose clinical presentation is compatible with DVT or cellulitis. In addition, to determine whether baseline clinical variables are predictive of the final diagnosis among ED patients with an initial clinical impression of 'DVT versus cellulitis' who underwent duplex scanning. METHODS: In this historical cohort study, patients with a final diagnosis of DVT (positive duplex) were compared on several baseline variables with patients with a final diagnosis of cellulitis (negative duplex and antibiotics prescribed) . RESULTS: One hundred nine of 542 ED patients referred for a duplex scan were initially diagnosed as 'DVT versus cellulitis', 17% of whom had DVT confirmed by a positive duplex scan. Comparing patients with DVT versus those with cellulitis, 0% versus 15.3% had rigors (P=0.06); 0% versus 8.3% had distinct margins of erythema (P<0.01); 5.3% versus 22.2% were currently on antibiotics (P=0.09); and 50% versus 21.3% had an elevated white blood cell count (P=0.04). CONCLUSION: There are differences in a number of baseline characteristics of 'DVT versus cellulitis' patients who went on to have either positive or negative duplex scans, some of which were statistically significant despite the limited sample size. These findings should be confirmed prospectively in a larger study sample since they may have the potential to aid in the clinical differentiation between DVT and cellulitis. PMID- 18159460 TI - Epidemiological and economic burden of pneumococcal diseases in Canadian children. AB - BACKGROUND: With the arrival of a new conjugate pneumococcal vaccine, it is important to estimate the burden of pneumococcal diseases in Canadian children. The epidemiological data and the economic cost of these diseases are crucial elements in evaluating the relevance of a vaccination program. METHODS: Using provincial databases, ad hoc surveys and published data, age-specific incidence rates of pneumococcal infections were estimated in a cohort of 340,000 children between six months and nine years of age. The costs of these diseases to the health system and to families were also evaluated using data from Quebec and Manitoba. RESULTS: Cumulative risks were one in 5000 for pneumococcal meningitis, one in 500 for bacteremia and one in 20 for pneumonia, leading to 16 deaths in the cohort. About 262,000 otitis media episodes and 32,000 cases of myringotomy with ventilation tube insertion were attributable to Streptococcus pneumoniae. Societal costs were estimated at $125 million, of which 32% was borne by the health system and 68% was borne by families. Invasive infections represented only 2% of total costs, while 84% were generated by otitis media. CONCLUSION: Pneumococcal infections represent a significant burden for Canadian children and society that could be significantly reduced through immunization. PMID- 18159461 TI - T cell costimulatory molecules in anti-viral immunity: Potential role in immunotherapeutic vaccines. AB - T lymphocyte activation is required to eliminate or control intracellular viruses. The activation of T cells requires both an antigen specific signal, involving the recognition of a peptide/major histocompatibility protein complex by the T cell receptor, as well as additional costimulatory signals. In chronic viral diseases, T cell responses, although present, are unable to eliminate the infection. By providing antigens and costimulatory molecules together, investigators may be able to increase and broaden the immune response, resulting in better immunological control or even elimination of the infection. Recent progress in understanding the function of costimulatory molecules suggests that different costimulatory molecules are involved in initial immune responses than are involved in recall responses. These new developments have important implications for therapeutic vaccine design. In this review the authors discuss the function of T cell costimulatory molecules in immune system activation and their potential for enhancing the efficacy of therapeutic vaccines. PMID- 18159462 TI - Case report: Acute hepatitis E infection with coexistent glucose-6-phosphate dehydrogenase deficiency. AB - Hepatitis E virus is one of the leading causes of acute viral hepatitis in India but usually manifests as a mild self-limiting illness. Viral hepatitis in the presence of glucose-6-phosphate dehydrogenase (G6PD) deficiency may be associated with complications such as severe anemia, hemolysis, renal failure, hepatic encephalopathy and even death. The incidence of G6PD deficiency in the general population of northern India is reported to be between 2.2% and 14%. Despite both hepatitis E infection and G6PD deficiency being common, their impact on patient illness has only recently been reported. The present study reports a case of severe hemolysis in a patient with G6PD deficiency and hepatitis E infection. PMID- 18159463 TI - Smallpox 2003. PMID- 18159465 TI - The Red Book is coming to town. PMID- 18159464 TI - The emergence of methicillin-resistant Staphylococcus aureus as a community acquired pathogen in Canada. PMID- 18159466 TI - Antimicrobials in acute exacerbations of chronic obstructive pulmonary disease - An analysis of the time to next exacerbation before and after the implementation of standing orders. AB - OBJECTIVE: To compare the mean time to next exacerbation in patients with acute exacerbations of chronic obstructive pulmonary disease (COPD) before and after the implementation of standing orders. SETTING: Tertiary care hospital, Halifax, Nova Scotia, Canada. POPULATION STUDIED: The records of 150 patients were analyzed, 76 were in the preimplementation group, 74 in the postimplementation group. INTERVENTION: The management and outcomes of patients admitted with an acute exacerbation of COPD before and after the implementation of standing orders were compared. DESIGN: A retrospective chart review. MAIN RESULTS: THERE WAS NO DIFFERENCE IN THE MEAN TIME TO NEXT EXACERBATION BETWEEN TREATMENT GROUPS (PREIMPLEMENTATION GROUP: 310 days, postimplementation group: 289 days, P=0.53). Antibiotics were used in 90% of the cases (preimplementation group: 87%, postimplementation group: 93%). The postimplementation group had a 20% increase in the use of first-line agents over the preimplementation group. Overall, first line agents represented only 37% of the antibiotic courses. CONCLUSIONS: The implementation of standing orders encouraged the use of first-line agents but did not influence subsequent symptom resolution, length of hospital stay, or the infection-free interval in patients with acute exacerbations of COPD. PMID- 18159467 TI - Epidemiology, antibiotic susceptibility, and serotype distribution of Streptococcus pneumoniae associated with invasive pneumococcal disease in British Columbia - A call to strengthen public health pneumococcal immunization programs. AB - BACKGROUND: This study examined the epidemiology, antibiotic susceptibility and serotype distribution of Streptococcus pneumoniae associated with invasive pneumococcal disease (IPD) in British Columbia. METHODS: Six hospitals and one private laboratory network participated in a prospective, sentinel laboratory based surveillance study of IPD, between October 1999 and October 2000. At each site, S pneumoniae isolates were collected and epidemiological data were gathered using a structured questionnaire, for all cases of IPD meeting the study case definition. Isolates were serotyped and tested for antimicrobial susceptibility. Bivariate associations were analyzed and multivariate logistic regression was used to identify independent risk factors associated with hospitalization or death. RESULTS: One hundred three reports and isolates were collected. Seventy nine per cent of cases were community-acquired, 64% required hospitalization and 5% died. Cases with one or more assessed risk factor for IPD and of female sex were independent variables associated with hospitalization or death. One-third of isolates had reduced penicillin susceptibility and 96% of these represented serotypes contained in the 23-valent pneumococcal polysaccharide vaccine (PPV 23). Overall, 89% of serotypes identified are included in the PPV-23 vaccine and 88% of isolates from children under five years of age are found in the 7-valent pneumococcal conjugate vaccine (PCV-7). Forty-one per cent of cases qualified for publicly funded pneumococcal vaccine and 34% of eligible persons were vaccinated. CONCLUSIONS: Overall, pneumococcal serotypes associated with IPD in this study closely matched serotypes included in PPV-23 products currently licensed in Canada. Most serotypes associated with IPD in children under five years of age are included in a recently licenced PCV-7. One third of isolates demonstrated reduced penicillin susceptibility, most involving serotypes included in PPV-23. Effective delivery of current public health immunization programs using PPV-23 and extending protection to infants and young children using the PCV-7 will prevent many cases of IPD. PMID- 18159468 TI - A descriptive study of human Salmonella serotype typhimurium infections reported in Ontario from 1990 to 1998. AB - BACKGROUND: Salmonella infections cause gastrointestinal and systemic diseases worldwide and are the leading causes of food-borne illnesses in North America (1 4). Salmonella serotype typhimurium (ST), in particular, is increasingly becoming a major public health concern because of its ability to acquire multiple resistant genes (5,6). OBJECTIVE: To describe demographic, temporal and geographical distributions, and reported risk factors of nonoutbreak cases of ST reported to a surveillance system in Ontario. METHODOLOGY: Descriptive analyses were performed on data on salmonellosis cases reported in Ontario between 1990 and 1998. Direct age- and sex-standardized rates were computed, and temporal trend analyses were performed using simple linear regression and a general additive model with a locally weighted regression (LOESS) smoother. RESULTS: The mean annual rates of infections with all Salmonella serotypes and with ST were 27 cases per 100,000 persons and 3.7 cases per 100,000 persons, respectively. Males and children under five years of age had significantly higher rates of both ST and ST definitive type 104 (DT104) infections. There was also evidence of temporal clustering of all strains of Salmonella, with significantly more cases being reported during the summer. Significantly higher rates of ST DT104 were observed in urban areas compared with rural areas, suggesting potential differences in the geographical distribution of risk factors. CONCLUSIONS: Information on demographic, temporal and geographical distributions, and risk factors is critical in planning disease control strategies. Further prospective analytical observation studies are needed to gain a better understanding of the epidemiology of ST and ST DT104 in Ontario, which will better guide disease control decisions. PMID- 18159469 TI - Food safety and foodborne disease in 21st century homes. AB - Over the past decade there has been a growing recognition of the involvement of the home in several public health and hygiene issues. Perhaps the best understood of these issues is the role of the home in the transmission and acquisition of foodborne disease. The incidence of foodborne disease is increasing globally. Although foodborne disease data collection systems often miss the mass of home based outbreaks of sporadic infection, it is now accepted that many cases of foodborne illness occur as a result of improper food handling and preparation by consumers in their own kitchens. Some of the most compelling evidence has come from the international data on Salmonella species and Campylobacter species infections.By its very nature, the home is a multifunctional setting and this directly impacts upon the need for better food safety in the home. In particular, the growing population of elderly and other immnocompromised individuals living at home who are likely to be more vulnerable to the impact of foodborne disease is an important aspect to consider. In addition, some developed nations are currently undergoing a dramatic shift in healthcare delivery, resulting in millions of patients nursed at home. Other aspects of the home that are unique in terms of food safety are the use of the home as a daycare centre for preschool age children, the presence of domestic animals in the home and the use of the domestic kitchen for small-scale commercial catering operations. At the global level, domestic food safety issues for the 21st century include the continued globalization of the food supply, the impact of international travel and tourism, and the impact of foodborne disease on developing nations.A number of countries have launched national campaigns to reduce the burden of foodborne disease, including alerting consumers to the need to practice food safety at home. Home hygiene practice and consumer hygiene products are being refined and targeted to areas of risk, including preventing the onward transmission of foodborne illness via the inanimate environment. It has been said that food safety in the home is the last line of defense against foodborne disease, and it is likely that this will remain true for the global population in the foreseeable future. PMID- 18159470 TI - The changing pattern of nontuberculous mycobacterial disease. AB - Nontuberculous mycobacteria are human opportunistic pathogens whose source of infection is the environment. These include both slow-growing (eg, Mycobacterium kansasii and Mycobacterium avium) and rapid-growing (eg, Mycobacterium abscessus and Mycobacterium fortuitum) species. Transmission is through ingestion or inhalation of water, particulate matter or aerosols, or through trauma. The historic presentation of pulmonary disease in older individuals with predisposing lung conditions and in children has been changing. Pulmonary disease in elderly individuals who lack the classic predisposing lung conditions is increasing. Pulmonary disease and hypersensitivity pneumonitis have been linked with occupational or home exposures to nontuberculous mycobacteria. There has been a shift from Mycobacterium scrofulaceum to M avium in children with cervical lymphadenitis. Further, individuals who are immunosuppressed due to therapy or HIV-infection are at a greatly increased risk for nontuberculous mycobacterial infection. The changing pattern of nontuberculous mycobacterial disease is due in part to the ability of these pathogens to survive and proliferate in habitats that they share with humans, such as drinking water. The advent of an aging population and an increase in the proportion of immunosuppressed individuals suggest that the prevalence of nontuberculous mycobacterial disease will increase. PMID- 18159471 TI - Shoulder pain, paralysis and fever: Dog attack or infection? PMID- 18159472 TI - Witnesses. PMID- 18159473 TI - Staphylococcus aureus: The persistent pathogen. PMID- 18159474 TI - A retrospective analysis of practice patterns in the treatment of methicillin resistant Staphylococcus aureus skin and soft tissue infections at three Canadian tertiary care centres. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) infections are increasingly being encountered and pose an increasing burden to the health care system in Canada. OBJECTIVE: To elucidate and characterize the factors influencing the current MRSA treatment patterns in patients with skin and soft tissue infections (SSTIs) before linezolid became available on the Canadian market. METHODS: A retrospective study collected demographic, treatment and resource use data on patients hospitalized at one of three geographically distinct tertiary care facilities, where MRSA SSTI treatment was initiated with intravenous (IV) vancomycin. Analysis of opportunities for IV-to-oral switch therapy was based on eligibility criteria. RESULTS: Of 89 patients identified over a 43-month period, the mean (+/-SD) durations of anti-infective treatment and hospitalization were 22.4+/-21 days and 28.9+/-20.8 days, respectively. An infected surgical wound was most common, representing 62.9% of infections. The mean duration of vancomycin treatment was 19.5 days and the mean number of 1 g doses received was 29.0+/-32.9. The majority of patients (55.1%) initiated vancomycin therapy a mean of 5.4+/-8.9 days after confirmation of MRSA. Of the 70% of patients meeting criteria for IV-to-oral switch therapy, only 10% received oral treatment. The most common reason cited for not switching was lack of an effective oral alternative. Analysis of switch therapy criteria found that IV treatment continued for a mean of 13 days despite the appropriateness of the oral route. CONCLUSIONS: Considerable variation exists in treatment patterns for MRSA infections. Improvements in the initiation of therapy and the use of IV-to-oral switch therapy may improve care and reduce the duration of hospitalization for MRSA SSTIs. PMID- 18159475 TI - Assessment of two alternative sample transport and fixation methods in the microbiological diagnosis of bacterial vaginosis. AB - BACKGROUND: The standard method for specimen collection and transport for microbiological diagnosis of bacterial vaginosis is an air-dried smear of vaginal secretions, promptly heat- or alcohol-fixed, Gram-stained and scored by Nugent's criteria. OBJECTIVE: TWO ALTERNATIVE METHODS ARE EVALUATED: sending a swab in transport medium to be smeared and Gram-stained in the laboratory two days later; and sending a smear of vaginal secretions sprayed with cytological fixative to the laboratory for Gram staining seven days later. METHODS: One hundred fifty-two women aged 18 years and older who attended a hospital colposcopy clinic or a community healthy sexuality clinic were studied. This was a prospective study: three vaginal swabs were taken from each patient and handled as described above. Each slide was blindly and independently interpreted by two microbiology technologists. The sensitivity, specificity and coefficient of agreement of the transported swab and cytologically fixed methods were compared with the air-dried smear method. RESULTS: Smears from swabs in transport medium and cytologically fixed smears both had 90% sensitivity and 97% specificity for bacterial vaginosis compared with diagnosis from air-dried smears. Cohen's kappa was 0.88 (95% CI 0.79 to 0.97) for each method. Inter-rater reliability assessed over all slides (all sampling techniques) was excellent (kappa 0.94). CONCLUSIONS: For the diagnosis of bacterial vaginosis, both alternative techniques provide results equivalent to air-dried direct smears. A vaginal smear sprayed with cytological fixative provides immediate fixation of material to the slide, permits delays in swab transport and avoids the requirement for transport at a controlled temperature imposed by swabs. PMID- 18159476 TI - Three generations of experience and thought in microbiology and infection. AB - Views and comments were sought from Brian Lacey, who was professionally active from the 1930s to the 1970s, Alain Dublanchet, active from the 1960s to the 2000s, and Mark Pallen, active from the 1990s to 2000 and beyond. Professor Lacey was professor of microbiology at the Westminster Medical School, University of London, United Kingdom, for many years and is now retired. Docteur Dublanchet is the long time head of the laboratory of microbiology and virology at the Centre Hospitalier Intercommunal de Villeneuve-Saint-Georges in the greater Paris area. Professor Pallen is currently professor of bacterial genetics at the Medical School, University of Birmingham (United Kingdom); he is a keen enthusiast of genomic studies in the interest of molecular pathogenesis research. All three are medically qualified. Four questions were posed to each:What was the situation like in the infectious disease field when you first started your career?What do you feel have been the most important accomplishments with regard to problems of infectious disease during your period of activity?What do you foresee as the vital matters that still need to be addressed for countering infectious disease?Can infectious disease ever, practically, be eradicated and, if so, how would this be accomplished? PMID- 18159477 TI - Invasive Streptococcus pneumoniae infection causing hemolytic uremic syndrome in children: Two recent cases. AB - INTRODUCTION: Streptococcus pneumoniae is an uncommon cause of hemolytic uremic syndrome (HUS) with a unique pathophysiology that differs from Shiga toxin related HUS. METHODS: Case descriptions for each patient are provided. Each strain of S pneumoniae was subjected to a pulsed-field gel electrophoresis (PFGE) analysis, Shiga toxin assay and polymerase chain reaction to detect Shiga toxin genes. A review of the current literature was conducted. CASE PRESENTATIONS: Two patients with S pneumoniae-related HUS that presented to the Alberta Children's Hospital, Calgary, Alberta, within four weeks of each other in 2001 are described. Both presented with pneumonia and empyema with associated HUS. Both patients required dialysis, one patient for 10 days and the other for 18 days. Neither patient demonstrated evidence of Shiga toxin-related disease. S pneumoniae isolated from blood or pleural fluid was penicillin susceptible. One isolate was serotype 3 and the other was serotype 14. The two strains had different PFGE patterns. Both patients recovered well with no persistent renal dysfunction. CONCLUSIONS: S pneumoniaecontinues to be an uncommon but important cause of HUS. Most cases can be confirmed or at least considered probable without performing a renal biopsy. PMID- 18159478 TI - Immunization and autism links: Ethics in research. PMID- 18159479 TI - Patient safety: What does it all mean? PMID- 18159480 TI - Transfusion and risk of infection in Canada: UPDATE 2004. PMID- 18159481 TI - Translocation of Clostridium difficile toxin B across polarized Caco-2 cell monolayers is enhanced by toxin A. AB - Clostridium difficile is the etiological agent of antibiotic-associated diarrhea; the most common form of nosocomial infectious diarrhea. The basis for the shock like systemic symptoms observed in severe cases of this infection are not known. It is hypothesized that the invasion of C difficile toxins A and/or B from the gut mucosa may contribute to these symptoms.A polarized tissue culture model employing Caco-2 cells grown on transwell inserts was established to study the translocation of purified C difficile toxins A and B. C difficile toxins were (125)I labelled and inoculated onto confluent polarized Caco-2 cell monolayers to study translocation dynamics. Electrical resistance measurements were used to monitor monolayer confluence and tight junction integrity. Samples were taken from the apical and basal sides of the insert, as well as the insert itself, and tested using the human foreskin fibroblasts cell cytotoxicity assay to monitor partitioning of the radiolabelled toxins. Toxin A produced a 50% reduction in electrical resistance in 3 h whereas the same concentration of toxin B required at least 7 h to achieve the same effect. Both toxins A and B were able to translocate across confluent monolayers of Caco-2 cells. The combination of toxin A and B together was synergistic with respect to promoting the translocation of toxin B. Although the addition of toxin A resulted in a 100% increase in the amount of toxin B able to translocate, no increases in toxin A translocation were observed. These findings suggest a model of pathogenesis in which C difficile toxin A facilitates the translocation of toxin B from the gut into submucosal areas where it may play a role in inflammatory damage. PMID- 18159482 TI - Risk and prevention of meningococcal disease among education workers: A review. AB - The aims of the present study were to review the risk of invasive meningococcal disease (IMD) among education workers, particularly pregnant women, and to evaluate preventive measures, in a context of endemicity, outbreak or epidemic as observed in the province of Quebec. The literature was reviewed and persons in charge of IMD surveillance in France, Quebec, the United Kingdom and the United States were interviewed. Surveys of asymptomatic carriage of Neisseria meningitidis show that transmission among students is higher than transmission between students and teachers. IMD incidence among education workers was analyzed in Cheshire (United Kingdom) in the period from 1997 to 1999, and the results indicated a risk six times higher than that in the general population. Overestimation of the magnitude of the risk is possible because the analysis focused on a cluster. None of the population-based studies of IMD mentioned a risk of secondary cases among education workers. Six IMD cases in education workers were identified in five clusters in schools in the United Kingdom, but not in the other countries. There is no epidemiological study on IMD risk among pregnant women, and this factor was not mentioned in any published review of IMD. Immunization of education workers at the beginning of their employment, using serogroup C glycoconjugate vaccine or a combined A, C, W-135, and Y conjugate vaccine (still under development), could reduce IMD risk, but the cost effectiveness of this measure should be evaluated. The societal benefit of excluding pregnant women from the work place during an outbreak seems to be very low, even if disease risk could be decreased for this specific group. When chemoprophylaxis is indicated for the control of an outbreak in an educational setting, treatment should be offered both to students and teachers in the group at risk. PMID- 18159483 TI - Clinical outcome with oral linezolid and rifampin following recurrent methicillin resistant Staphylococcus aureus bacteremia despite prolonged vancomycin treatment. AB - Drug-resistant Gram-positive bacteria, especially Staphylococcus aureus, are emerging as the predominant organisms involved in both nosocomial and community acquired infections. Since the 1980s, vancomycin has been the first-line antibiotic used to treat methicillin- resistant S aureus. However, allergy and intolerance to vancomycin, the increasing number of vancomycin clinical failures and the existence of vancomycin intermediate-susceptible isolates of S aureus suggest that new antibiotics are needed. This paper reports the only known case of a successful clinical outcome with long term oral linezolid and rifampin therapy in the management of recurrent and persistent methicillin-resistant S aureus bacteremia with metastatic infections despite prolonged vancomycin use. More than two years since the initiation of linezolid and rifampin, the study patient has been clinically well with no evidence of adverse drug reactions including cytopenia and hepatic toxicities. Physicians must be aware of the novel developments in antibiotic therapy to treat drug-resistant bacterial infections. PMID- 18159484 TI - The hemophagocytic syndrome in an immunocompromised patient: A diagnostic challenge. PMID- 18159485 TI - HIV: A journey just begun. PMID- 18159486 TI - Coming full circle: From antibiotics to probiotics and prebiotics. PMID- 18159487 TI - West Nile update - Preparing for summer 2004. PMID- 18159488 TI - Innovation and challenges in funding rapid research responses to emerging infectious diseases: Lessons learned from the outbreak of severe acute respiratory syndrome. AB - Although the local public health response to the severe acute respiratory syndrome outbreak in Canada was critical to the diagnosis, management and treatment of patients, such a rapid research response required a national effort to engage the research and stakeholder communities. The Canadian research effort, coordinated through the Institute of Infection and Immunity of the Canadian Institutes of Health Research and the Michael Smith Foundation for Health Research, has provided insight into the mechanisms required to ensure the rapid development of strategical initiatives in response to emerging infectious diseases. It has also provided a rational basis to set up a national network to be engaged if needed in the future. PMID- 18159489 TI - Recurrent chronic ambulatory peritoneal dialysis-associated infection due to rothia dentocariosa. AB - Rothia dentocariosa is a commensal organism of the human oropharynx. Clinical infection due to this organism is rare. A case of recurrent peritoneal dialysis related peritonitis caused by R dentocariosa and a review of the literature is reported. Isolation of R dentocariosa from dialysate fluid should not be dismissed as a contaminant. Although there are no interpretive criteria for antimicrobial susceptibility testing, R dentocariosa appears to be susceptible to a variety of antibiotics including beta-lactams, vancomycin and aminoglycosides. Optimal therapy of peritoneal dialysis peritonitis caused by this organism may also require removal of the catheter. PMID- 18159490 TI - An unusual intracerebral hemorrhage. PMID- 18159491 TI - Antimicrobial resistance in food. PMID- 18159492 TI - Severe acute respiratory syndrome: Did quarantine help? PMID- 18159493 TI - Intracardiac device and prosthetic infections: What do we know? PMID- 18159494 TI - Bag urine specimens still not appropriate in diagnosing urinary tract infections in infants. PMID- 18159495 TI - The costs and consequences of methicillin-resistant Staphylococcus aureus infection treatments in Canada. AB - BACKGROUND: A multinational randomized controlled trial has shown a trend toward early discharge of patients taking oral linezolid versus intravenous vancomycin (IV) in the treatment of methicillinresistant Staphylococcus aureus (MRSA) infections. Infection treatments resulting in shorter hospitalization durations are associated with cost savings from the hospital perspective. OBJECTIVE: To determine whether similar economic advantages are associated with oral linezolid, the costs and consequences of linezolid use following vancomycin IV versus the existing practice in the treatment of infections caused by MRSA were compared. METHODS: The charts of all patients admitted to one of three tertiary care teaching hospitals between January 1, 1997 and August 31, 2000 and treated with vancomycin IV for an active MRSA infection (skin and soft tissue only) were reviewed. Based on the vancomycin IV chart review data set and a simulated linezolid data set, the clinical consequences and the associated costs of MRSA treatment with vancomycin IV, and oral and IV forms of linezolid were quantified and compared within the framework of a cost-consequence analysis. RESULTS: Patients treated with oral and IV forms of linezolid compared with the existing practice had a shorter length of stay and required fewer home IV care services, which resulted in a cost savings of $750 (2001 values) to the Canadian health care perspective. CONCLUSIONS: The estimated cost savings associated with linezolid use not only offset the higher acquisition cost of the anti-infective, but may be substantial to health care systems across Canada, especially as the incidence of MRSA continues to rise. PMID- 18159496 TI - HIV and hepatitis C virus testing and seropositivity rates in Canadian federal penitentiaries: A critical opportunity for care and prevention. AB - BACKGROUND: Incarcerated persons experience high rates of HIV and hepatitis C virus (HCV) infection, but little is known about the burden of these bloodborne viruses among federal penitentiary inmates in Canada. OBJECTIVE: The present study investigates rates of testing and seropositivity for HIV and HCV among inmates in all 53 Canadian federal penitentiaries. METHODS: A cross-sectional design using surveillance data on voluntary HIV and HCV antibody testing in 2002 were applied to estimate the rate of testing uptake and the rate of incident seropositive tests among new admissions to federal penitentiaries and resident inmates. Rates of testing and infection were further examined by sex and region. Seroprevalence of HIV and HCV was estimated from the number of cumulative positive tests to year-end. RESULTS: Of 7670 new admissions during 2002, 30% were tested for HIV and HCV. Test seropositivity rates in this group were 0.7% for HIV and 10% for HCV. Of the 12,426 resident inmates, 28% were tested for HIV and 27% for HCV. Seropositivity rates in this group were 0.3% for HIV and 7% for HCV. Seroprevalence rates at yearend for 2002 were 2.0% for HIV and 26% for HCV and were substantially higher among women offenders (HIV: 3.7% of women, 1.9% of men; HCV: 34% of women, 26% of men). Variations in testing uptake and test seropositivity were observed across regions. CONCLUSIONS: The present study underscores the value of continued monitoring and evaluation of trends in HIV and HCV infection, which remain prevalent in federal penitentiaries. Higher rates of testing are warranted for at-risk inmates to improve early detection of infection and provide infected inmates with timely care and treatment. For those who remain free of infection, testing can provide the additional benefits of exposing inmates to health counselling and for the reinforcement of prevention messages. The period of incarceration is also a critical opportunity to link inmates with outside resources in preparation for release to the community. PMID- 18159497 TI - Recovery of cephalosporin-resistant Escherichia coli and Salmonella from pork, beef and chicken marketed in Nova Scotia. AB - BACKGROUND: Antimicrobial use in farm animals is a potentially important contributor to the emergence of antimicrobial resistance. Resistant Salmonella may lead to serious human infections and resistant Escherichia coli may transfer plasmid-encoded resistance genes to other pathogens. OBJECTIVE: To determine the prevalence of E coli and Salmonella species resistant to the third generation of cephalosporins in retail meat products in Halifax, Nova Scotia in 2002. METHODS: Ground beef, ground pork and chicken wings were tested for E coli and Salmonella. E coli were selected on ceftriaxonecontaining media. Beta-lactamases were characterised by isoelectric focusing, polymerase chain reaction and sequencing. Pulsed field gel electrophoresis was performed to determine the relationship of strains. The transferability of plasmids and location of resistance genes was also determined. RESULTS: Forty-three of 75 packages of chicken wings contained ceftriaxone-resistant E coli; 42 of these contained beta-lactamases with isoelectric points at approximately 8.7. Six of seven CMY primer amplicons that were sequenced contained plasmid-mediated Citrobacter freundii-derived blaCMY-2; the other contained a CMY-2- like beta-lactamase. Pulsed field gel electrophoresis patterns demonstrated that strains were not clonal in nature. Four chicken samples contained Salmonella, one of which contained bla CMY-2 mediated resistance and an E coli bearing the same gene, but on different plasmids. Four of 100 beef samples contained blaCMY-2-bearing E coli; none contained Salmonella. Two of 75 pork samples contained ceftriaxone resistant E coli, one of which encoded for CMY-2. One susceptible Salmonella strain was recovered from pork. CONCLUSIONS: Chicken from retail outlets located in Halifax, Nova Scotia, commonly contained blaCMY-2-bearing E coli. The relationship antibiotics used in food-producing animals and its effect on resistance of commensals and pathogens needs to be determined. PMID- 18159498 TI - Microscopy and polymerase chain reaction detection of Leishmania chagasi in the pleural and ascitic fluid of a patient with AIDS: Case report and review of diagnosis and therapy of visceral leishmaniasis. AB - Atypical visceral leishmaniasis is increasingly reported in immunocompromised patients, including patients with AIDS. A case of visceral leishmaniasis in an HIV-infected Brazilian patient with pulmonary and peritoneal involvement is reported. Histological evaluation of pleural fluid and ascites aspirate revealed macrophages with intracellular Leishmania. Polymerase chain reaction analysis was positive for Leishmania in the pleural and ascitic fluid with use of primers specific for Leishmania chagasi. In addition to classical methods for diagnosing leishmaniasis, such as microscopy and culture, polymerase chain reaction detection and identification of Leishmania species in pleural effusions and ascites are important diagnostic tools that should be considered by clinicians evaluating HIV-infected patients from endemic areas of visceral leishmaniasis. The authors review the clinical manifestations, diagnostic and therapeutic aspects of visceral leishmaniasis in immunocompetent and HIV-infected patients. PMID- 18159499 TI - Antimicrobial resistance - Judicious use is the key. PMID- 18159500 TI - Avian influenza - The next pandemic? PMID- 18159501 TI - Occupational head-butting and skin nodules. PMID- 18159502 TI - Canadian Committee on Antibiotic Resistance report. PMID- 18159503 TI - Mycophenolic acid inhibits replication of Type 2 Winnipeg, a cerebrospinal fluid derived reovirus isolate. AB - BACKGROUND: The role of reoviruses in human disease is uncertain. Most identified cases are sporadic and asymptomatic or produce minor upper respiratory or gastrointestinal symptoms. In November 1997, a reovirus was isolated from the cerebrospinal fluid of a severe combined immune deficient infant in Winnipeg, Manitoba. RNA characterization and sequencing studies demonstrated this reovirus isolate to be unique. Thus, the virus was named Type 2 Winnipeg (T2W). OBJECTIVE: Mycophenolic acid (MPA), a drug primarily used as an immunosuppressive agent, was assessed in the capacity to inhibit T2W viral growth. METHODS: The effects of MPA on viral growth were determined by plaque reduction assays. Cells were treated with different MPA concentrations, infected with T2W and incubated at 37 degrees C for 0 h to 72 h. Virus titres were determined and compared with untreated controls. RESULTS: Production of infectious T2W progeny decreased more than 99% at 3 microg/mL MPA compared with untreated controls. Inhibition was not caused by cell toxicity because there was no difference in cell viability. The 50% cell toxic dose was 30 microg/mL MPA. CONCLUSIONS: MPA was able to inhibit viral growth of the novel reovirus T2W. Although MPA is usually used as an immunosuppressive agent, and despite the fact that T2W was isolated from an immunocompromised patient, these results suggest that MPA could have been used as a possible treatment at subimmunosuppressive doses. Animal studies to better define the antiviral and immunosuppressive activities of MPA (and its prodrug mycophenolate mofetil) appear warranted. PMID- 18159504 TI - Uropathogen antibiotic resistance in adult women presenting to family physicians with acute uncomplicated cystitis. AB - BACKGROUND: Increasing rates of trimethoprim-sulfamethoxazole (TMP-SMX) resistance among uropathogens have raised concerns about its continued role in empirical treatment of acute uncomplicated cystitis in adult women. OBJECTIVE: To determine current rates of antibiotic resistance among uropathogens in the community. METHODS: Urine culture reports from adult women with symptoms of cystitis attending the offices of family physicians from across Canada were examined. Antibiotic sensitivities and the total number of antibiotics an organism was resistant to was determined. RESULTS: In 446 women, 235 (61.4%) positive urine cultures were identified. Of these, 38.2% were resistant to at least one antibiotic and 21.5% were resistant to two or more antibiotics. The rate of ampicillin resistance was 34.1%. For TMP-SMX, resistance was reported in 10.8% of samples. Antibiotic resistance was higher in British Columbia (55%) and western provinces (48%), compared with Ontario (33.3%) and the eastern provinces (26.3%, P=0.04, Fisher's exact test). Multidrug resistance was also higher in western Canada (33.9%) than in eastern Canada (16.6%, P=0.007). CONCLUSIONS: TMP SMX resistance in Canada remains within current recommended guidelines, allowing for its continued use as a first line empirical treatment for acute cystitis in adult women. The reasons for higher rates of antibiotic resistance in western Canada merit further study. PMID- 18159505 TI - Serological studies of West Nile virus in a liver transplant population. AB - BACKGROUND: Solid organ transplant populations are at increased risk for serious clinical manifestations of West Nile virus (WNV) infection. OBJECTIVE: To monitor liver transplant recipients during the 2003 WNV season in Manitoba and to identify incidence, clinical presentation and serology. METHODS: Serial blood specimens were obtained from adult patients followed at the liver transplant outpatient clinic between May 2003 and October 2003. Studies for WNV infection included immunoglobulin (Ig) G and IgM enzyme immunoassay (EIA), hemagglutination inhibition (HI), plaque reduction neutralization test and reverse transcriptase polymerase chain reaction. RESULTS: None of the 79 patients had clinical presentations suggestive of WNV infection. On testing of the final serum specimen obtained, 14 patients (18%) had positive IgG anti-WNV by EIA and six patients (7%) had indeterminate IgG anti-WNV by EIA, although all were negative by IgM EIA. Four (20%) of the EIA-positive samples were reactive by HI, but all of these were negative by WNV plaque reduction neutralization test; this is consistent with the presence of non-West Nile flavivirus antibody in these sera. Blood specimens obtained throughout the season from EIA- and HI-positive individuals were uniformly negative for WNV-RNA by reverse transcriptasepolymerase chain reaction. Age, sex, hematology and biochemistry findings, hepatitis B or C virus status, immunosuppressive regimen (cyclosporin or tacrolimus) and pretransplant diagnosis of liver disease were similar for EIA-positive and EIA-negative patients. For the 10 patients with a positive IgG EIA maintained on cyclosporin, the cyclosporin level was 129.1+/-28.6 microg/L compared with 85.6+/-36.7 microg/L in 26 patients who were EIA-negative (P=0.002). CONCLUSIONS: False positive IgG EIA serology for WNV was common in this cohort of liver transplant recipients, and was associated with elevated serum cyclosporin levels. PMID- 18159506 TI - An economic evaluation of voriconazole versus amphotericin B for the treatment of invasive aspergillosis in Canada. AB - BACKGROUND: Invasive aspergillosis (IA) is a serious fungal infection that affects immunocompromised patients. The Global Comparative Aspergillosis study demonstrated that voriconazole, a new broad-spectrum triazole, had better responses and improved survival compared with conventional amphotericin B deoxycholate (CAB) and other licensed antifungal therapy (OLAT) for the treatment of definite or probable aspergillosis. OBJECTIVES: To compare costs and outcomes of voriconazole and CAB for the treatment of definite or probable aspergillosis in Canada. METHODS: A cost-consequence decision tree model was designed to reflect the treatment pathways used in clinical practice when using voriconazole or CAB as primary therapy for IA. Therapy included initial treatment with either voriconazole or CAB and then switched to an OLAT in the event of an inadequate response, severe toxicity or intolerance. The principal data source used was the Global Comparative Aspergillosis study. RESULTS: The total cost of voriconazole when compared with CAB as initial therapy for IA was $38,319 versus $42,495 per patient, respectively, representing a 9.8% cost reduction for each patient treated with voriconazole. The higher mean cost in the CAB arm was primarily due to the high proportion of patients (73.7%) who were switched to an OLAT due to severe side effects or an inadequate response. Treating with voriconazole was a dominant strategy. The number of patients that had to be treated with voriconazole instead of CAB to save one additional life was eight. CONCLUSIONS: Voriconazole as primary treatment for IA increased the chances of successful treatment, improved survival and may represent a potential cost saving strategy in Canada. PMID- 18159507 TI - Screening donors of solid organs for West Nile virus: First, do no harm! PMID- 18159508 TI - Severe acute respiratory syndrome: What have we learned two years later? PMID- 18159509 TI - The management of chronic viral hepatitis: A Canadian consensus conference 2004. AB - Several government and nongovernment organizations held a consensus conference on the management of acute and chronic viral hepatitis to update previous management recommendations. The conference became necessary because of the introduction of new forms of therapy for both hepatitis B and hepatitis C. The conference issued recommendations on the investigation and management of chronic hepatitis B, including the use of lamivudine, adefovir and interferon. The treatment of hepatitis B in several special situations was also discussed. There were also recommendations on the investigation and treatment of chronic hepatitis C and hepatitis C-HIV coinfection. In addition, the document makes some recommendations about the provision of services by provincial governments to facilitate the delivery of care to patients with hepatitis virus infection. The present document is meant to be used by practitioners and other health care providers, including public health staff and others not directly involved in patient care. PMID- 18159510 TI - Canadian consensus guidelines for the management of cytomegalovirus disease in HIV/AIDS. AB - BACKGROUND: The management of HIV-infected patients with cytomegalovirus (CMV) disease has changed significantly with the availability of highly active antiretroviral therapy (HAART). OBJECTIVES: These updated guidelines are intended to provide practical help to physicians managing HIV-positive patients with or at risk for CMV disease. METHODS: The 10 members of the Canadian CMV Disease in HIV/AIDS Consensus Group were infectious disease specialists, a primary care physician and ophthalmologists with expertise in HIV and CMV infection. Financial support by Hoffmann-La Roche Canada Ltd was unrestricted, and was limited to travel expenses and honoraria. The consensus group met in June and October 2002. Key areas to be considered were identified, and group members selected, reviewed and presented relevant recent literature for their assigned section for the group's consideration. Evidence was assessed based on established criteria, which were expert opinions of the members. Draft documents were circulated to the entire group and modified until consensus was reached. The final guidelines represent the group's consensus agreement. The guidelines were approved by the Canadian Infectious Disease Society. RESULTS AND CONCLUSIONS: The guidelines address symptom monitoring, screening for early detection and prevention, and treatment using oral, intravenous and intraocular anti-CMV therapies in conjunction with HAART. PMID- 18159511 TI - A case of spontaneous methicillin-resistant Staphylococcus aureus meningitis in a health care worker. AB - Meningitis caused by Staphylococcus aureus is an unusual illness that is often associated with bacteremia, contiguous infection and/or a post-neurosurgical state. Until recently, most cases were caused by methicillin-susceptible strains, but there are a few reports of methicillinresistant S aureus strains causing meningitis. A new case of meningitis caused by methicillin-resistant S aureus in a neonatal intensive care nurse who did not have bacteremia or neurosurgery is reported. The patient made a full recovery after treatment with vancomycin and rifampin. PMID- 18159512 TI - Perspectives on emerging zoonotic disease research and capacity building in Canada. AB - Zoonoses are fundamental determinants of community health. Preventing, identifying and managing these infections must be a central public health focus. Most current zoonoses research focuses on the interface of the pathogen and the clinically ill person, emphasizing microbial detection, mechanisms of pathogenicity and clinical intervention strategies, rather than examining the causes of emergence, persistence and spread of new zoonoses. There are gaps in the understanding of the animal determinants of emergence and the capacity to train highly qualified individuals; these are major obstacles to preventing new disease threats. The ability to predict the emergence of zoonoses and their resulting public health and societal impacts are hindered when insufficient effort is devoted to understanding zoonotic disease epidemiology, and when zoonoses are not examined in a manner that yields fundamental insight into their origin and spread.EMERGING INFECTIOUS DISEASE RESEARCH SHOULD REST ON FOUR PILLARS: enhanced communications across disciplinary and agency boundaries; the assessment and development of surveillance and disease detection tools; the examination of linkages between animal health determinants of human health outcomes; and finally, cross-disciplinary training and research. A national strategy to predict, prevent and manage emerging diseases must have a prominent and explicit role for veterinary and biological researchers. An integrated health approach would provide decision makers with a firmer foundation from which to build evidence-based disease prevention and control plans that involve complex human/animal/environmental systems, and would serve as the foundation to train and support the new cadre of individuals ultimately needed to maintain and apply research capacity in this area. PMID- 18159513 TI - Community-acquired methicillin-resistant Staphylococcus aureus: Getting over it. PMID- 18159514 TI - Lyme disease: Is it or is it not? PMID- 18159515 TI - Enhanced surveillance of non-O157 verotoxin-producing Escherichia coli in human stool samples from Manitoba. AB - BACKGROUND: Relatively few enhanced surveillance studies have been undertaken to investigate the extent to which verotoxin-producing non-O157 serotypes of Escherichia coli occur in stool samples received for the detection of verotoxin producing organisms. OBJECTIVES: To describe the prevalence, molecular and epidemiological characteristics, and geographical patterns associated with non O157 verotoxin-producing E coli (VTEC) in Manitoba. RESULTS: Thirty-two VTEC isolates consisting of 10 serogroups and 13 different serotypes were isolated over a 22-month period. Twenty-three isolates (71.8%) possessed verotoxin encoding gene stx1 only, five isolates (15.6%) possessed stx2 only, two isolates (6.3%) possessed both stx1 and stx2, and two isolates (6.3%) possessed stx2c. Only three instances of indistinguishable pulsed-field gel electrophoresis patterns were identified. The age of the individuals from whom non-O157 VTEC were isolated ranged from eight months to 87 years. Mean and median ages were 30 and 22 years of age, respectively. Some areas of the province appeared to experience a higher than expected number of non-O157 E coli in comparison with the number of stools that were received from these areas. CONCLUSIONS: The present study demonstrated a large number of infections associated with non-O157 VTEC in Manitoba. Most non-O157 cases appear to result from sporadic infections, and these occur typically in rural areas. Continued enhanced surveillance is necessary to understand the temporal patterns of non-O157 VTEC and the underlying epidemiological factors driving these patterns. PMID- 18159516 TI - The role of the cytomegalovirus antigenemia assay in the detection and prevention of cytomegalovirus syndrome and disease in solid organ transplant recipients: A review of the British Columbia experience. AB - BACKGROUND: The pp65 cytomegalovirus (CMV) antigenemia assay has been used as a means of guiding the pre-emptive therapy of CMV disease in solid organ transplant (SOT) recipients. Recently, concerns have been raised regarding the utility of the test to accurately and precisely detect viral activity early enough to reduce the morbidity and mortality associated with CMV OBJECTIVE: To determine the performance characteristics of the method of antigenemia testing of SOT recipients used at Vancouver General Hospital, Vancouver, British Columbia. METHODS: All SOT recipients between January 1, 1999, and June 30, 2000, were retrospectively reviewed for six months following transplantation. Physical examination results, laboratory parameters, antigenemia results and treatment information were reviewed. RESULTS: A total of 134 kidney, liver, lung and kidney pancreas transplant recipients were included in the analysis. The overall performance characteristics of the antigenemia assay in predicting CMV disease included a sensitivity of 64%, a specificity of 81%, a positive predictive value of 76% and a negative predictive value of 71%. A mean of 18 days passed between the onset of signs and symptoms of CMV disease/syndrome and the first recorded positive antigenemia result, and only 26% of patients had a positive test result before the onset of symptoms. It was found that an antigenemia test breakpoint of at least one positive cell for defining a positive test provided the most sensitive and specific prediction, with increased odds of developing CMV disease. CONCLUSIONS: Based on performance characteristics, the Vancouver General Hospital's current method of antigenemia testing to guide pre-emptive ganciclovir therapy in SOT patients is not optimal for the early detection of disease. Further study is needed on new molecular testing methods to determine if our ability to predict CMV disease can be improved. PMID- 18159517 TI - Outbreaks of infection caused by community-acquired methicillin-resistant Staphylococcus aureus in a Canadian correctional facility. AB - BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has been identified in prison settings in the United States. The present study investigated two clusters of skin and soft tissue infection caused by community acquired (CA) MRSA in a correctional facility in southern Ontario. METHODS: Outbreak investigations were conducted by the responsible public health authority. Strain relatedness was assessed through comparison of pulsed-field gel electrophoresis and antibiograms. RESULTS: Two distinct outbreaks of CAMRSA associated disease occurred in 2002 and 2004. Most patients presented with abscesses in the lower extremities. All isolates had identical DNA banding patterns on pulsed-field gel electrophoresis. One-half of the affected inmates resided in a cellblock with one other affected inmate. No other risk factors were identified. CONCLUSIONS: One of the first outbreaks of CAMRSA infections in a correctional facility in Canada is documented. Taken in conjunction with outbreaks elsewhere, this suggests that residence in correctional facilities may be a risk factor for CAMRSA infection. PMID- 18159519 TI - A case of atypical croup. PMID- 18159518 TI - Complicated urinary tract infection in adults. AB - BACKGROUND: Complicated urinary tract infection occurs in individuals with functional or structural abnormalities of the genitourinary tract. OBJECTIVE: To review current knowledge relevant to complicated urinary tract infection, and to provide evidence-based recommendations for management. METHODS: The literature was reviewed through a PubMed search, and additional articles were identified by journal reference review. A draft guideline was prepared and critically reviewed by members of the Association of Medical Microbiology and Infectious Disease Canada Guidelines Committee, with modifications incorporated following the review. RESULTS: Many urological abnormalities may be associated with complicated urinary infection. There is a wide spectrum of potential infecting organisms, and isolated bacteria tend to be more resistant to antimicrobial therapy. Morbidity and infection outcomes in subjects with complicated urinary infection are principally determined by the underlying abnormality rather than the infection. Principles of management include uniform collection of a urine specimen for culture before antimicrobial therapy, characterization of the underlying genitourinary abnormality, and nontreatment of asymptomatic bacteriuria except before an invasive genitourinary procedure. The antimicrobial regimen is determined by clinical presentation, patient tolerance, renal function and known or anticipated infecting organisms. If the underlying abnormality contributing to the urinary infection cannot be corrected, then early post-treatment recurrence of infection is anticipated. CONCLUSIONS: The management of complicated urinary infection is individualized depending on patient variables and the infecting organism. Further clinical investigations are necessary to assist in determining optimal antimicrobial regimens. PMID- 18159520 TI - Distinguishing between acute and serous otitis media. PMID- 18159522 TI - Syndromes associated with sexually transmitted infections. PMID- 18159521 TI - Sexually transmitted infections. PMID- 18159523 TI - The laboratory diagnosis of Neisseria gonorrhoeae. AB - The present article describes the laboratory diagnosis of Neisseria gonorrhoeae by culturing of the organism from different types of clinical specimens followed by confirmatory tests. The success of culture methods requires good quality collection and transport of clinical specimens. The present guide describes the media requirements and cultural conditions for N gonorrhoeae growth and the characteristics for a presumptive identification of N gonorrhoeae. Confirmatory tests include biochemical tests, chromogenic enzyme substrate tests, immunoassays and nucleic acid methods. Nucleic acid detection methods include either amplification-based methods or nonamplification tests, and are increasingly used in clinical laboratories where a viable culture is not possible to obtain. Nucleic acid methods can also be used to detect the presence of low numbers in a specimen. Nucleic acid detection methods need confirmation with another amplification method or gene target. Controls must be included to ensure true positive and negative results, and to rule out nucleic acid contamination. Monitoring of antimicrobial susceptibilities of N gonorrhoeae is important to investigate treatment failure and to evaluate the efficacy of currently recommended therapies. Many methods for the characterization of N gonorrhoeae require cultures. The useful typing methods for determining strain relatedness include auxotyping, serotyping, plasmid profile analysis, DNA sequencing of the porB gene and pulsed-field gel electrophoresis. Quality assurance programs for diagnostic testing and antimicrobial susceptibility testing is reviewed. PMID- 18159524 TI - The laboratory diagnosis of HIV infections. AB - HIV diagnostic testing has come a long way since its inception in the early 1980s. Current enzyme immunoassays are sensitive enough to detect antibody as early as one to two weeks after infection. A variety of other assays are essential to confirm positive antibody screens (Western blot, polymerase chain reaction [PCR]), provide an adjunct to antibody testing (p24 antigen, PCR), or provide additional information for the clinician treating HIV-positive patients (qualitative and quantitative PCR, and genotyping). Most diagnostic laboratories have complex testing algorithms to ensure accuracy of results and optimal use of laboratory resources. The choice of assays is guided by the initial screening results and the clinical information provided by the physician; both are integral to the laboratory's ability to provide an accurate laboratory diagnosis. Laboratories should also provide specific information on specimen collection, storage and transport so that specimen integrity is not compromised, thereby preserving the accuracy of laboratory results. Point of Care tests have become increasingly popular in the United States and some places in Canada over the past several years. These tests provide rapid, on-site HIV results in a format that is relatively easy for clinic staff to perform. However, the performance of these tests requires adherence to good laboratory quality control practices, as well as the backup of a licensed diagnostic laboratory to provide confirmation and resolution of positive or indeterminate results. Laboratory quality assurance programs and the participation in HIV proficiency testing programs are essential to ensure that diagnostic laboratories provide accurate, timely and clinically relevant laboratory results. PMID- 18159525 TI - The laboratory diagnosis of Haemophilus ducreyi. AB - Chancroid is a sexually transmitted infection caused by Haemophilus ducreyi. This fastidious, Gram-negative coccobacilli dies rapidly outside the human host, making diagnostic testing using culture methods difficult. This genital ulcer infection is not common in Canada and, therefore, can often be misdiagnosed. The objective of the present paper is to provide practical approaches for the diagnosis of chancroid in Canadian patients where the prevalence of this infection is low. Issues related to sample collection, sample transport and available diagnostic tests are reviewed, and several alternative approaches are outlined. Although antigen detection, serology and genetic amplification methods have all been reported for H ducreyi, none are commercially available. Culture is still the primary method available to most laboratories. However, the special media necessary for direct bedside inoculation is often not available; therefore, communication with the diagnostic laboratory and rapid specimen transport are essential when chancroid is suspected. PMID- 18159526 TI - The laboratory diagnosis of Trichomonas vaginalis. AB - Trichomonas vaginalis, a parasitic protozoa that causes the sexually transmitted infection trichomoniasis, is the sexually transmitted infection with the largest annual incidence, exceeding 170 million cases per year. The disease can be difficult to diagnose due to its heterogeneous presentation and problems with diagnostic testing. All diagnostic tests are fraught with imperfections, but the old, reliable wet mount examination (in trained hands), and the newer InPouch method may be advantageous due to simplicity in technology and cost. The present article reviews the pros and cons of culture, antibody and nucleic acid-based technologies that may point to future diagnostic advances. PMID- 18159527 TI - The laboratory diagnosis of Chlamydia trachomatis infections. AB - Lower genital tract infections with Chlamydia trachomatis are predominantly asymptomatic in men and women. Diagnostic technology has provided several approaches to the diagnosis of C trachomatis. Outside of cells, Chlamydia can die or degrade without optimal storage and transportation. Because some of the other assays perform better on certain specimen types, it is important for laboratories to recognize these differences and provide advice to physicians and nurses collecting patient specimens, with the objective of diagnosing lower genital tract infections to prevent transmission and upper tract damage. Most invasive specimens, such as cervical or urethral swabs, may be collected for culture, antigen or nucleic acid detection. Noninvasive samples such as first-void urine and vaginal swabs can be easily collected by the patient; these samples must be tested by more sensitive nucleic acid amplification tests. These newer investigative strategies should enable implementation of screening programs to identify and treat partners. Serology has not been particularly useful for the diagnosis of acute C trachomatis infections in adults. Presently, it appears that antibiotic-resistant C trachomatis is not a clinical problem. Laboratories providing C trachomatis diagnosis require participation in continuous quality improvement programs. PMID- 18159528 TI - The laboratory diagnosis of syphilis. AB - Syphilis has several clinical manifestations, making laboratory testing a very important aspect of diagnosis. In North America, many unsuspected cases are discovered by laboratory testing. The etiological agent, Treponema pallidum, cannot be cultured, and there is no single optimal alternative test. Serological testing is the most frequently used approach in the laboratory diagnosis of syphilis. The present paper discusses the various serological and alternative tests currently available along with their limitations, and relates their results to the likely corresponding clinical stage of the disease. The need to use multiple tests is discussed, and the importance of quality control is noted. The complexity of syphilis serology means that the services of reference laboratories and clinical experts are often needed. PMID- 18159529 TI - The laboratory diagnosis of sexually transmitted infections in cases of sexual assault and abuse. AB - Laboratory staff dealing with samples from victims must be aware that such patients have been psychologically traumatized and deserve special care. The help of a sexual assault care team should be sought if available, and appropriate specimens should be collected two to 10 days after an incident, preferably in a single visit. Specimens should be clearly labelled, and the laboratory should be informed. In the laboratory, all procedures need to be clearly documented. There are special requirements for the collection of forensic specimens and associated records, which may later be required for legal proceedings. The laboratory must know what the current legal status is for any test being used in that community. The present article serves as a guideline to more detailed practice standards for the investigation of individual sexually transmitted infections in assault and abuse situations. PMID- 18159530 TI - The laboratory diagnosis of hepatitis B virus. AB - Hepatitis B virus (HBV) chronically infects approximately 250,000 Canadians and 350 million people worldwide. Without intervention, approximately 15% to 40% of chronically infected individuals will eventually develop cirrhosis, end-stage liver disease or hepatocellular carcinoma, or require liver transplantation. The availability and extensive use of the HBV vaccine has dramatically reduced the number of incident infections in Canada and worldwide. Effective therapeutic agents have been and continue to be developed to treat chronic infection. The present review provides a comprehensive overview of diagnostic tests for HBV infection and immunity, and elaborates on HBV risk factors, vaccine prevention and therapeutic monitoring. HBV diagnosis is accomplished by testing for a series of serological markers of HBV and by additional testing to exclude alternative etiological agents such as hepatitis A and C viruses. Serological tests are used to distinguish acute, self-limited infections from chronic HBV infections and to monitor vaccine-induced immunity. Nucleic acid testing for HBV-DNA is increasingly being used to quantify HBV viral load and measure the effectiveness of therapeutic agents. Given the multitude of available tests and the complexity of clinical management, there is a critical need for greater coordination among clinicians, diagnostic laboratory personnel and researchers to define optimal laboratory diagnostic and monitoring assays so that the appropriate tests are used to maximize prevention and optimize treatment outcomes. PMID- 18159531 TI - Screening for sexually transmitted infection pathogens in semen samples. AB - The transmission of sexually transmitted infection (STI) pathogens from an infected donor to the recipient of a semen donation in assisted conception may result not only in acute infection but also in long-term reproductive complications or adverse outcomes of pregnancy, including infection of the offspring. Screening for bacterial STI pathogens, Chlamydia trachomatis and Neisseria gonorrhoeae is strongly recommended because these pathogens can cause serious reproductive complications in the recipients of semen donations and infection in their offspring. Screening for these pathogens should be performed using the most sensitive methods, such as nucleic acid amplified tests. False negative results due to inhibitory substances in the semen sample should be monitored using amplification controls. Where specimen transport is not a problem and culture facilities are available, N gonorrhoeae can also be detected by culture. Laboratories performing screening should subscribe to proficiency programs and have strict quality controls. Although Trichomonas vaginalis, group B streptococcus and genital mycoplasmas have been associated with adverse outcomes of pregnancy, the frequent finding of these organisms in healthy individuals brings into question the validity of mandatory inclusion of these organisms in the screening panel. Although viral STI pathogens and Treponema pallidum - the causative agent of syphilis - may be detected in semen, their presence may be more sensitively detected through antibody testing of the donor. Screening donors for HIV, hepatitis B and syphilis by serology is uniformly recommended in all of the guidelines, but the value of screening either donors or semen samples for cytomegalovirus, herpes simplex viruses and human papilloma viruses is less clear. PMID- 18159533 TI - The evaluation of diagnostic tests for sexually transmitted infections. AB - Diagnostic tests should receive method- and use-effectiveness evaluations. Method effectiveness evaluations determine sensitivity, specificity and predictive values for new tests. Use-effectiveness evaluations determine how practical or convenient a new test will be in a specific setting and may not be performed in a formal way in North American laboratories. To perform a clinical method evaluation of diagnostic tests, a good relationship between laboratory and clinical personnel is essential. Studies are usually conducted separately on populations of men and women, and should include sampling from different prevalence groups. Test performance comparisons may be made on a single specimen type or on more than one specimen from the same patient, which allows for the expansion of a reference standard and includes the ability of a particular assay, performed on a specimen type to diagnose an infected individual. The following components of the evaluation should be standardized and carefully followed: specimen identification; collection; transportation; processing; quality control; reading; proficiency testing; confirmatory testing; discordant analysis - sensitivity, specificity and predictive value calculations; and record keeping. Methods are available to determine whether sample results are true or false positives or negatives. Use-effectiveness evaluations might determine the stability or durability of supplies and equipment; the logistics of shipping, receiving and storing supplies; the clarity and completeness of test instructions; the time and effort required to process and read results; the subjectivity factors in interpretation and reporting; and the costs. These determinations are usually more apparent for commercial assays than for homemade tests. PMID- 18159532 TI - The laboratory diagnosis of bacterial vaginosis. AB - Bacterial vaginosis (BV) is an extremely common health problem for women. In addition to the troublesome symptoms often associated with a disruption in the balance of vaginal flora, BV is associated with adverse gynecological and pregnancy outcomes. Although not technically a sexually transmitted infection, BV is a sexually associated condition. Diagnostic tests include real-time clinical/microbiological diagnosis, and the current gold standard, the standardized evaluation of morphotypes on Gram stain analysis. The inappropriate use of vaginal culture can be misleading. Future developments into molecular based diagnostics will be important to further understand this complex endogenous flora disruption. PMID- 18159534 TI - The laboratory diagnosis of genital human papillomavirus infections. AB - Human papillomaviruses (HPVs) are the etiological agents of several genital cancers, including cancer of the uterine cervix. The detection of HPV infection in genital samples may increase the sensitivity of primary and secondary screenings of cervical cancer. HPV testing may also improve the specificity of screening programs, resulting in the avoidance of overtreatment and cost savings for confirmatory procedures. The major determinants of clinical progression of HPV infection include persistence of HPV infection, involvement of high-risk HPV types, high HPV viral load, integration of viral DNA and presence of several potential cofactors. Signal amplification HPV-DNA detection techniques (Hybrid Capture II, Digene Corporation, USA) are standardized, commercially available, and capable of detecting several high-risk HPV types. They also increase the sensitivity of screening for high-grade lesions in combination with cytology. The sensitivity of these techniques to detect high-grade lesions is higher than that of cytology, but the referral rate for colposcopy is greater. These techniques are approved for the triage to colposcopy of women with cervical smears interpreted as atypical squamous cells of undetermined significance. Triage and screening for cervical cancer using HPV will probably be restricted to women aged 30 years or older because of the high prevalence of infection in younger women. Amplification techniques are ideal for epidemiological studies because they minimize the misclassification of HPV infection status. These techniques can detect low HPV burden infections. Consensus primers amplify most genital types in one reaction, and the reverse hybridization of amplicons with type-specific probes allows for the typing of HPV-positive samples. Consensus PCR assays are currently under evaluation for diagnostic purposes. HPV testing is currently implemented for the clinical management of women. PMID- 18159535 TI - The laboratory diagnosis of herpes simplex virus infections. AB - Herpes simplex virus (HSV) types 1 and 2 cause genital herpes infections and are the most common cause of genital ulcer disease in industrialized nations. Although these infections are very common, the majority of them remain underdiagnosed because they are asymptomatic or unrecognized. A clinical diagnosis of genital herpes should always be confirmed by laboratory testing; this can be accomplished through the use of direct tests for viral isolation, the detection of antigen or, more recently, the detection of HSV DNA using molecular diagnostic techniques. Testing for serotypes is recommended because of the different prognostic and counselling implications. Type-specific HSV serology is becoming more readily available and will enhance the ability to make the diagnosis and guide clinical management in select patients. PMID- 18159536 TI - Where are all the new antibiotics? The new antibiotic paradox. PMID- 18159537 TI - Transfusion and risk of infection in Canada: Update 2005. PMID- 18159538 TI - An evaluation of the management of asymptomatic catheter-associated bacteriuria and candiduria at The Ottawa Hospital. AB - BACKGROUND: Asymptomatic catheter-associated urinary tract infections (CAUTIs) are common in hospitalized patients. They are associated with a low incidence of sequelae and morbidity, and in most patients resolve spontaneously on removal of the catheter. As a result, it is not recommended that asymptomatic catheter associated bacteriuria or candiduria be treated with antimicrobial agents while the catheter remains in place because it may lead to the evolution of resistant flora. OBJECTIVE: To assess the current management of patients with CAUTIs with respect to antimicrobial therapy at The Ottawa Hospital and the University of Ottawa Heart Institute, Ottawa, Ontario. METHODS: A prospective observational study over a period of 26 consecutive days was conducted at The Ottawa Hospital (General and Civic campuses) and the University of Ottawa Heart Institute. Inpatients with an indwelling catheter, a positive urine culture and the absence of UTI signs or symptoms were assessed. Patients were followed for five days to determine whether antimicrobials were prescribed. RESULTS: From March 3 to March 28, 2003, 29 of 119 patients screened met inclusion criteria. Of these 29 patients, 15 (52%) were prescribed antimicrobials and were therefore considered to be inappropriately managed. Differences were observed between the appropriate and inappropriate management groups in terms of duration of stay to positive urine culture and whether yeast or bacteria were isolated from the culture. CONCLUSION: Antimicrobial agents were prescribed in over one-half of CAUTI cases, contrary to recommendations from the literature. Education is required to bring this strongly supported recommendation into clinical practice. PMID- 18159539 TI - Potential capsule switching from serogroup Y to B: The characterization of three such Neisseria meningitidis isolates causing invasive meningococcal disease in Canada. AB - Three group B Neisseria meningitidis isolates, recovered from meningococcal disease cases in Canada and typed as B:2c:P1.5, were characterized. Multilocus sequence typing showed that all three isolates were related because of an identical sequence type (ST) 573. Isolates typed as 2c:P1.5 are common in serogroup Y meningococci but rare in isolates from serogroups B or C. Although no serogroup Y isolates have been typed as ST-573, eight isolates showed five to six housekeeping gene alleles that were identical to that of ST-573. This suggested that the B:2c:P1.5 isolates may have originated from serogroup Y organisms, possibly by capsule switching. PMID- 18159540 TI - The prevalence of hepatitis A in children in British Columbia. AB - BACKGROUND: The risk of hepatitis A virus (HAV) infection during childhood is difficult to estimate without population serosurveys because HAV-related symptoms are often mild at this age. Few serosurveys have been conducted in Canada. The present study surveyed teenagers in two nonurban regions of British Columbia where the historical rate of reported HAV either exceeded (region A) or was less than (region B) the historical provincial rate. METHODS: A point prevalence survey of salivary HAV-specific immunoglobulin G was conducted in high schools among grade 9 students in regions A and B. A questionnaire was used to gather sociodemographic data. The survey was extended to grade 1 and grade 5 students in community 1 of region B. Associations between risk factors and prior infection were evaluated by logistic regression. RESULTS: Eight hundred eleven grade 9 students were tested. Antibody to HAV was detected in 4.7% of students in region A (95% CI 2.9% to 7.2%) and 9.6% of students in region B (95% CI 6.9% to 12.9%). The region B figure reflected HAV antibody prevalence rates of 19.5% in community 1 and 2.5% in the remainder of the region. Younger students in community 1 had low HAV antibody to HAV prevalence rates (3.9% for grade 1 and 3.1% for grade 5), and positive tests in this community were associated with a particular school, foreign travel and brief residence. The risk factors for HAV infection in grade 9 students were not determined. CONCLUSIONS: Children in nonurban areas of British Columbia are generally at low risk of HAV infection during the first decade of life regardless of the reported population rates, thereby permitting the consideration of school-based HAV immunization programs. PMID- 18159541 TI - Cardiac device infections due to Mycobacterium fortuitum. AB - Two cases of cardiac device infection due to Mycobacterium fortuitum are reported along with a discussion of their clinical management. Long-term therapy and removal of the infected device is needed. The slow progression and absence of systemic signs and symptoms suggest a low pathogenicity of M fortuitum. PMID- 18159542 TI - Cerebral Mycobacterium avium abscesses: Late immune reconstitution syndrome in an HIV-1-infected patient receiving highly active antiretroviral therapy. AB - A patient who developed an atypical manifestation of Mycobacterium avium complex (MAC) infection almost two years after starting effective highly active antiretroviral therapy is described. The recurrence, manifested as brain abscesses in the central nervous system, was an uncommon form of MAC disease usually reported postmortem. An increased CD4 cell count, localized and suppurative infection, and the absence of systemic evidence of infection were consistent with a late immune reconstitution syndrome. The present case report adds to the understanding of MAC disease in HIV-infected patients. PMID- 18159543 TI - Illicit drugs, infectious disease and public health: A historical perspective. AB - The present report outlines a presentation by Professor Virginia Berridge at the Second Stanier Lecture held in Halifax, Nova Scotia, on November 5, 2002. The relationship among public health concepts, illicit drug use prevention and policy, and infection control strategies in England and other locations is paralleled over the course of two centuries. This historical journey analyzes changes in public health and demonstrates how history and public health have intersected at various times to result in the public health approaches used today. PMID- 18159544 TI - Avian influenza: Myth or mass murder? AB - The purpose of the present article was to determine whether avian influenza (AI) is capable of causing a pandemic. Using research from a variety of medical journals, books and texts, the present paper evaluates the probability of the AI virus becoming sufficiently virulent to pose a global threat.Previous influenza A pandemics from the past century are reviewed, focusing on the mortality rate and the qualities of the virus that distinguish it from other viruses. Each of the influenza A viruses reviewed were classified as pandemic because they met three key criteria: first, the viruses were highly pathogenic within the human population; second, the viruses were easily transmissible from person to person; and finally, the viruses were novel, such that a large proportion of the population was susceptible to infection. Information about the H5N1 subtype of AI has also been critically assessed. Evidence suggests that this AI subtype is both novel and highly pathogenic. The mortality rate from epidemics in Thailand in 2004 was as high as 66%. Clearly, this virus is aggressive. It causes a high death rate, proving that humans have a low immunity to the disease.To date, there has been little evidence to suggest that AI can spread among humans. There have been cases where the virus has transferred from birds to humans, in settings such as farms or open markets with live animal vending. If AI were to undergo a genetic reassortment that allowed itself to transmit easily from person to person, then a serious pandemic could ensue, resulting in high morbidity and mortality. Experts at the World Health Organization and the United States Centers for Disease Control and Prevention agree that AI has the potential to undergo an antigenic shift, thus triggering the next pandemic. PMID- 18159545 TI - The Editorial Board is changing. PMID- 18159546 TI - What do central venous catheter-associated bloodstream infections have to do with bundles?g. PMID- 18159547 TI - Immunization update 2005: Stepping forward. PMID- 18159548 TI - Necrotizing fasciitis resulting from human bites: A report of two cases of disease caused by group A streptococcus. AB - Necrotizing fasciitis is a serious and potentially life-threatening condition. Although bite wounds are common, they are not frequently reported as a cause of necrotizing fasciitis. In the present article, two cases of bite-associated necrotizing fasciitis caused by group A streptococcus are reported. Previously published cases are also reviewed. PMID- 18159549 TI - The prevalence and incidence of sexually transmitted infections in a prospective cohort of injection drug users in Vancouver, British Columbia. AB - BACKGROUND: While several studies have reported on sexual risk behaviours and the prevalence of sexually transmitted infections (STIs) among injection drug users (IDUs), there are fewer prospective studies that have been able to examine populations of IDUs with no history of STIs. Therefore, the authors examined prevalence, correlates and factors associated with time to first STI infection in a prospective cohort of IDUs in Vancouver, British Columbia. METHODS: The authors examined the prevalence and correlates of STIs among IDUs at the time of recruitment into a prospective cohort study. The authors also evaluated the cumulative rate of time to first STI among IDUs with no history of STIs at baseline using the Kaplan-Meier method, and modelled factors independently associated with first STI using Cox regression. RESULTS: Between May 1996 and November 2003, 1560 individuals were recruited into the cohort; of these individuals, 745 reported a history of STI at baseline. Among the 815 who did not report an STI at baseline, 671 (82%) had at least one follow-up visit and were eligible for the analysis of time to first STI. After 36 months of follow-up, the cumulative rate of first STI was 8.2% for men and 15.9% for women (log-rank P<0.001), whereas the cumulative rate of first STI was 8.0% for IDUs who did not report sex trade involvement versus 19.8% for IDUs who reported sex trade involvement (log-rank P<0.001). In multivariate analyses, the risk of first STI remained independently associated with unprotected sex with regular partners (relative hazard=2.04, 95% CI 1.29 to 3.23; P=0.001) and unprotected sex with sex trade clients (relative hazard=2.36, 95% CI 1.46 to 3.82; P=0.005). CONCLUSIONS: In the present study, the authors found that STIs were associated with both regular sex partnerships and sex trade involvement. These findings are of particular concern because both unprotected sex with regular partners and sex trade involvement is common among IDUs. Interventions to encourage condom use among IDUs, particularly those with regular sex partners and those involved in the sex trade, should be further developed. PMID- 18159550 TI - Risk factors for new-onset diabetes mellitus in patients receiving protease inhibitor therapy. AB - BACKGROUND: Metabolic complications including diabetes mellitus (DM) have been associated with protease inhibitor (PI) therapy. Risk factors for the development of DM are not well-defined. OBJECTIVES: To determine risk factors for the development of new-onset DM in patients initiated on PI therapy. METHODS: A retrospective cohort study was conducted to identify predictors of developing DM in subjects started on PI therapy between January 1997 and January 2003. Diabetes cases were defined as physician documentation of DM in the outpatient medical chart and/or those subjects receiving an antidiabetic agent. Logistic regression was used to examine the relationship between new-onset DM and demographic characteristics, and between new-onset DM and total treatment days with PI therapy. Body mass index could not be entered into the model due to missing height measurements. RESULTS: A total of 496 subjects on PI therapy were included, of which 18 (3.6%) developed DM. The mean age of the subjects was 43.4+/-9.4 years (range 19 to 77) and the mean duration of therapy was 3.0+/-1.9 years (range 0.17 to 7.9). In the multivariate model, older subjects were more likely to develop DM (OR 1.12, 95% CI 1.05 to 1.19; P=0.001). This corresponds to a 12% increased risk of DM for each one-year increase in age. Subjects that weighed more had an increased risk (OR 1.06, 95% CI 1.03 to 1.10; P=0.001), as did those belonging to a non-Aboriginal minority group when compared with Caucasians (OR 6.67, 95% CI 1.56 to 28.41; P=0.01). A longer duration of PI therapy was also significantly associated with developing DM (OR 1.52, 95% CI 1.07 to 2.17; P=0.02). CONCLUSION: A longer duration of PI therapy is associated with an increased risk of developing DM. As with HIV-negative subjects, demographic characteristics such as age, weight and ethnicity were important predictors of developing DM in the present study. PMID- 18159552 TI - Francisella philomiragia adenitis and pulmonary nodules in a child with chronic granulomatous disease. AB - Francisella philomiragia is a rare and opportunistic pathogen capable of producing invasive infection in patients with compromised neutrophil function and in patients that have survived a near-drowning. A case of F philomiragia adenitis and lung nodules, refractory to cephalosporin therapy, is reported in a 10-year old boy with chronic granulomatous disease following a facial abrasion from a saltwater crab. To the authors' knowledge, this is the first Canadian clinical isolate to be reported. Genus and species identification was confirmed via 16S ribosomal RNA sequence analysis. A literature review revealed three groups at risk of F philomiragia infection: young patients with chronic granulomatous disease; adults with hematogenous malignancy; and near-drowning patients. Pneumonia, fever without an apparent source and sepsis are the main clinical presentations. Invasive procedures may be required to isolate this organism and ensure appropriate antimicrobial therapy. Limited awareness of F philomiragia has led to delayed identification, patient death and misidentification as Francisella tularensis - a biosafety level three pathogen and potential bioterrorism agent. PMID- 18159551 TI - HIV-associated lipodystrophy syndrome: A review of clinical aspects. AB - Approximately two years after the introduction of highly active antiretroviral therapy for the treatment of HIV infection, body shape changes and metabolic abnormalities were increasingly observed. Initially, these were ascribed to protease inhibitors, but it is now clear that nucleoside reverse transcriptase inhibitors also contribute to lipodystrophy syndrome. The syndrome groups together clinical conditions describing changes in body fat distribution that include lipoatrophy, lipoaccumulation or both. However, there does not appear to be a direct link between lipoatrophy and lipoaccumulation that would support a single mechanism for the redistribution of body fat. Currently, there is no clear definition of lipodystrophy, which explains the difficulty in determining its prevalence and etiology. There are no current guidelines for the treatment of fat distribution abnormalities that occur in the absence of other metabolic complications. The present article reviews the current state of knowledge of the definition, symptoms, risk factors, pathogenesis, diagnosis and treatment of the morphological changes associated with lipodystrophy syndrome. PMID- 18159553 TI - Bacillary angiomatosis in an HIV-positive man with multiple risk factors: A clinical and epidemiological puzzle. AB - A 36-year-old man with advanced AIDS presented with skin lesions, fever and weight loss. Physical examination revealed skin lesions suggestive of bacillary angiomatosis. Diagnostic imaging identified bone lesions, soft tissue masses and a single hepatic lesion. His clinical presentation was most consistent with Bartonella quintana; however, his risk for infection with B quintana was limited to a remote, brief period of homelessness. In contrast, his frequent contact with cats presented substantial potential exposure to Bartonella henselae.Diagnosing a Bartonella infection presents challenges. The detection of bacteremia is hindered by slow growth in BACTEC media (BD Diagnostic Systems, Canada). In the present case, recovery of the organism required blind subcultures after at least eight days of incubation. Speciation by 16s ribosomal RNA analysis confirmed B quintana as the pathogen.Because the patient presented with skin lesions, the diagnosis was straightforward; however, skin lesions are nondiagnostic in some cases. Microbiological confirmation of the diagnosis rests on sufficient clinical suspicion to apply labour-intensive laboratory protocols. PMID- 18159554 TI - Overwhelming pulmonary infection after a tobogganing accident. PMID- 18159555 TI - Chlamydia pneumoniae and atherosclerosis: The end? PMID- 18159556 TI - Natural disasters, corpses and the risk of infectious diseases. PMID- 18159557 TI - A comparison of pertussis rates in the Northwest Territories: Pre- and postacellular pertussis vaccine introduction in children and adolescents. AB - INTRODUCTION: During the past decade, a trend toward increasing cases of Bordetella pertussis in older children and adults has been witnessed in Canada. The National Advisory Committee on Immunization now recommends that the adult formulation of the acellular pertussis (adult dTap) vaccine combined with diphtheria and tetanus toxoids be substituted for diphtheria and tetanus toxoids alone for the 14- to 16-year-old booster dose. In October 2000, the government of the Northwest Territories was one of the first to adopt adult dTap into their territorial immunization program free of charge. OBJECTIVE: To evaluate the effect of the acellular pertussis vaccine in children and adolescents on the epidemiology of pertussis in the Northwest Territories. METHODS: Pertussis is a reportable disease in the Northwest Territories, and data on the incidence rates of pertussis are available from 1989 to 2004. The present study reviews pertussis cases during three four-year periods: the whole-cell vaccine era (1993 to 1996); the preadult dTap era (1997 to 2000); and the postadult dTap era (2001 to 2004). RESULTS: The incidence of pertussis decreased from 18.0 cases per 10,000 population in 1993 to 0.2 cases per 10,000 population in 2004. The number of cases decreased from 186 to 129 to 19 cases in the three chronological time periods (ie, whole-cell vaccine era, preadult dTap era and postadult dTap era, respectively), with the most substantial reduction coming with the introduction of postadult dTap. CONCLUSIONS: There appears to be a decrease in the incidence of pertussis with the targeted introduction of adult dTap in the Northwest Territories. PMID- 18159558 TI - Enhanced surveillance for acute and likely acute hepatitis B in Canada: 1999 to 2002. AB - OBJECTIVE: To assess the incidence of clinically identified hepatitis B cases, as well as the demographic and risk behaviour profiles of these cases in a defined Canadian population. METHODS: An enhanced hepatitis surveillance system was established in October 1998 to identify acute hepatitis B and C cases in Canada. Acute and likely acute cases, as determined by laboratory testing, collectively defined incident clinical hepatitis B cases. Data from 1999 to 2002 on incidence, demographic characteristics and risk behaviour characteristics were collected and analyzed. RESULTS: During the 1999 to 2002 surveillance period, 379 cases were identified in a target population ranging from 3,128,179 to 8,576,071 individuals. The observed hepatitis B incidence rate was 1.93/100,000 person years in the surveillance area. The incidence rate was 2.74 times higher in men. The observed incidence rates decreased for all age groups over the surveillance period but remained high in the 20- to 29-year-old and 30- to 39-year-old age groups, as well as in men. Of the incident cases, 55.9% self-identified as being born in Canada and 18.5% as being born in Asia, while 18.7% did not identify a birth area. Of the Canadian-born cases, 61.3% identified themselves as Caucasian, 11.3% as Aboriginal and 23.6% as no ethnic category. Injection drug use was the most reported risk behaviour (19.1%), followed closely by sex-associated risk behaviours. A large proportion, 24%, indicated none of the known risk behaviours. CONCLUSIONS: The hepatitis B incidence rate has shown some decrease in the surveillance population from 1999 to 2002 and this may be due, in part, to past intervention programs (eg, vaccination programs and health promotion campaigns). The authors' results identify some high-risk groups that would benefit from additional prevention and control programs, and further targeted research and intervention. PMID- 18159559 TI - Key research issues in Clostridium difficile. AB - Clostridium difficile is an emerging pathogen that causes C difficile-associated diarrhea, an important nosocomial infection. Control of this infection remains a challenge, and much needs to be determined about the antimicrobial resistance of the organism, antibiotic stewardship, contamination of the patient environment, and various host factors that determine susceptibility or resistance to infection. A national symposium focusing on C difficile infections, the Clostridium difficile Symposium on Emerging Issues and Research, was hosted on November 23, 2004, by the Department of Medical Microbiology and Infectious Diseases at the University of Manitoba, Winnipeg, Manitoba, in partnership with the Canadian Institutes of Health Research. This symposium, which aimed to summarize key research issues regarding C difficile infections in Canada, had the following objectives: to provide a forum for learning and discussion about C difficile and its impact on the health of Canadians; to identify the key research issues that should be addressed; and to explore potential research funding opportunities and collaboration. The present report summarizes key research issues identified for C difficile infections in Canada by addressing four major themes: diagnosis and surveillance, infection prevention and control, antibiotic stewardship, and clinical management. PMID- 18159560 TI - Probable linezolid-induced pancytopenia. AB - A 75-year-old male outpatient with cardiac disease, diabetes, chronic renal insufficiency and iron deficiency anemia was prescribed linezolid 600 mg twice daily for a methicillin-resistant Staphylococcus aureus diabetic foot osteomyelitis. After one week, his blood counts were consistent with baseline values. The patient failed to return for subsequent blood work. On day 26, he was admitted to hospital with acute renal failure secondary to dehydration, and was found to be pancytopenic (erythrocytes 2.5x10(12)/L, leukocytes 2.9x10(9)/L, platelets 59x10(9)/L, hemoglobin 71 g/L). The patient was transfused, and linezolid was discontinued. His blood counts improved over the week and remained at baseline two months later.The patient's decline in blood counts from baseline levels met previously established criteria for clinical significance. Application of the Naranjo scale indicated a probable relationship between pancytopenia and linezolid.Clinicians should be aware of this rare effect with linezolid, and prospectively identify patients at risk and emphasize weekly hematological monitoring. PMID- 18159561 TI - Listeria meningitis complicating infliximab treatment for Crohn's disease. AB - Infliximab, a monoclonal antibody directed against tumour necrosis factor-alpha, is an effective therapy for Crohn's disease. Though uncommon, serious opportunistic infections, including reactivation of tuberculosis, have occurred in patients after infliximab administration.Meningitis caused by Listeria monocytogenes developed in a 37-year-old man six days after the second infusion of infliximab. The patient, who also was treated with azathioprine and corticosteroids, had an uneventful recovery after a course of antibiotics. Several other recent reports have implicated infliximab therapy in the development of severe Listeria infections, particularly meningitis and sepsis. With the increasing use of tumour necrosis factor-alpha-neutralizing agents, clinicians should be aware of the risk of opportunistic infections caused by L monocytogenes in patients with Crohn's disease following infliximab treatment. PMID- 18159562 TI - Cardiobacterium hominis endocarditis: A case report and review of the literature. AB - The present case report describes the clinical course of a patient who presented with Cardiobacterium hominis endocarditis. A review of the literature follows the case presentation. C hominis, a fastidious Gram-negative bacillus, is a member of the HACEK group of microorganisms (Haemophilus species, Actinobacillus actinomycetemcomitans, C hominis, Eikenella corrodens and Kingella kingae). Endocarditis caused by C hominis is uncommon and generally follows a subacute course. Patients may present with constitutional symptoms, symptoms related to valvular destruction or symptoms secondary to embolic events. Diagnosis requires identification of the pathogen from blood or vegetation by either culture or molecular techniques. Blood cultures may require prolonged incubation, highlighting the importance of incubating blood cultures for at least two to three weeks in patients with suspected endocarditis. In the past, C hominis was generally sensitive to penicillin. However, reports of beta-lactamase-producing C hominis have appeared in the literature over the past decade. The current recommendation for first-line treatment is a third-generation cephalosporin (ceftriaxone) for four weeks (six weeks if a prosthetic valve is in place). PMID- 18159563 TI - Chlamydial bacteriophage: No role in acute coronary events? AB - BACKGROUND: A relationship between Chlamydia pneumoniae infection and acute coronary syndromes has not been consistently found in published studies. It has been hypothesized that a bacteriophage-infected subset of C pneumoniae may be uniquely equipped to promote atherosclerosis and acute coronary syndromes through the expression of phage genes. METHODS: The authors performed a pilot case control study of acute coronary events. Case and control subjects were characterized demographically and according to recognized coronary risk factors. These subjects also provided serum for the detection of antibody to the elementary bodies of C pneumoniae and antibody to the Vp1 protein coded by the phage. Bivariate and multivariate comparisons were performed using statistics appropriate for paired analyses. RESULTS: Antibodies to C pneumoniae, Vp1 protein or both were not associated with acute coronary events by bivariate or multivariate analysis. As expected, case subjects were significantly more likely to have hypertension, hypercholesterolemia or diabetes mellitus. CONCLUSION: The present study adds to a growing body of literature that does not support the hypothesized relationship between C pneumoniae (or a phage-infected subset of C pneumoniae) and acute coronary syndromes. PMID- 18159564 TI - Fever and shock in a child: How 'good' is a good blood test? PMID- 18159565 TI - The risk of tuberculosis transmission in solid organ transplantation: Is it more than a theoretical concern? AB - The present case report describes a therapeutic dilemma regarding the transmission of Mycobacterium tuberculosis during transplantation of solid organs, places this phenomenon within the context of the literature, and makes recommendations for screening and therapy. PMID- 18159566 TI - Antimicrobial resistance: An update from the Canadian Committee on Antibiotic Resistance. PMID- 18159567 TI - A simultaneous onset of organizing pneumonia and rheumatoid arthritis, along with a review of the literature. AB - Organizing pneumonia (OP) is a specific type of interstitial pneumonia that has been noted as one of the pulmonary manifestations during the course of rheumatoid arthritis (RA). In this study, we report a case with a simultaneous development of OP and RA. The patient presented with concurrent flu-like symptoms and arthralgia of multiple joints, and antibiotic therapy was not effective. The rheumatoid factor (RF) and anti-cyclic citrullinated antibodies were both high. Multiple air-space opacities on chest radiographs and bilateral peripheral consolidations on high-resolution computed tomography films were evident. The histology of transbronchial lung biopsy samples was characterized by intra alveolar buds of granulation tissue consisting of intermixed myofibroblasts and connective tissues. Treatment with prednisolone induced a complete recovery from OP without relapses. Our review of previous reports about RA-associated OP (RA OP) suggested that the high titer of RF and increased disease activity of RA indicate a great risk of developing OP. This condition may represent a lung's reaction in the RA-associated inflammatory and/or immune process. We should be aware of RA-OP cases in which pulmonary manifestations precede articular symptoms. In these cases, respiratory manifestations are the main evidence of RA activity. In most cases of steroid-resistant RA-OP, the use of immunosuppressants was effective. Since OP may progress to fibrotic lung disease during the course of RA, we may consider performing a second lung biopsy for steroid-resistant patients, even if they have once been diagnosed as OP. PMID- 18159568 TI - A case of pulmonary asbestosis presenting with temporal arteritis involving multiple medium-sized vessels. AB - A 76-year-old woman with pulmonary asbestosis was admitted with fever and polymyalgia. She subsequently developed a visual disorder, hemoptysis, and hemoperitoneum. A biopsy of the temporal artery revealed the presence of giant cell arteritis. CT and angiography showed hemorrhaging from the bronchial and abdominal arteries. These observations suggested temporal arteritis in which medium-sized vessels were involved. This case implies the association between vasculitis and asbestosis, and suggests a problem in the classification of vasculitides. PMID- 18159569 TI - A case of lupus nephritis improved after appropriately adjusting the dosage of mizoribine. AB - A 29-year-old male presenting nephrotic syndrome and facial skin erythema was admitted to our hospital in September of 2000. We diagnosed him as having systemic lupus erythematosus (SLE) accompanied by lupus nephritis (WHO class V). The disease activity had decreased after treatment with methylprednisolone (m PSL) pulse therapy, which was followed by oral PSL. Thereafter, when tapering the dosage from 60 to 30 mg/day, the lupus nephritis flared up and he was re hospitalized in February of 2001. After successful retreatment with m-PSL pulse therapy followed by the tapering of the dosage from 60 to 30 mg/day, we used mizoribine (MZR) as a combination therapy. The lupus nephritis flared up again after tapering down to 17.5 mg/day of PSL. Then, we changed the MZR dosage from 150 mg/day in three divided daily doses to 200 mg/day in two divided daily doses. This modification increased the peak blood concentration (Cmax) of MZR from 0.63 to 1.55 microg/ml. At present, we have been able to successfully taper the dosage to 7.5 mg/day of oral PSL and the patient has achieved a state of remission without any side effects. Monitoring of the serum concentration of MZR is thus considered to be important for achieving effective therapy of SLE, especially for steroid-resistant lupus nephritis. If the serum concentration of MZR does not reach an effective level, then the dosage of MZR should be adjusted appropriately in order to maintain an adequate serum concentration of MZR. PMID- 18159570 TI - Effect of estrogen replacement therapy on arthritis and bone mineral density in estrogen-replete rats with collagen-induced arthritis. AB - The influence of estrogen therapy on changes in arthritis and bone mineral density (BMD) was evaluated using an estrogen-replete collagen-induced arthritis (CIA) rat model. Seven-month-old female Sprague-Dawley rats (n = 30) were divided into the three groups; control (CONT), collagen sensitization (CIA), and CIA + 17beta-estradiol administration for 7 weeks (CIA + E). BMD was measured by peripheral quantitative computed tomography in the proximal tibia every 2 weeks. Eight weeks after the initial sensitization the rats were killed and histomorphometry of tibia was performed. The hind paw thickness increased with time in CIA rats and there was a significant difference between CONT and CIA at 8 weeks after the initial sensitization. Estrogen tended to make the development of arthritis milder. In CIA, BMD at metaphyseal cancellous bone began to decrease with the onset of arthritis and became significantly lower than in CONT after 8 weeks. Compared with the CIA, the deterioration in BMD was inhibited in CIA + E. Histomorphometrical parameters of bone resorption were increased in CIA compared with CONT, and those elevations were reduced by estrogen treatment, but estrogen had no effect on bone formation parameters. In conclusion, estrogen could partially suppress arthritis and bone loss in estrogen-replete rats as well as estrogen-deplete ones. PMID- 18159571 TI - A propos time and autoimmunity. AB - The integrated defense system has been shaped over eons showing noteworthy robustness by surviving a million-year prehistory, a comparatively short evolving history and current transformation. Self-identification being part of it, so are deviations manifold expressed in autoimmunity. Epidemiological incidence and intensity, both being subject of change, are focused in the light of the time factor. Furthermore, it is stressed that there is no bi-univocal mutual relationship between immunity and defense and the origins of autoimmunity still remain mysterious. We question whether the present transforming events have occurred within too short a time to be attributed to genetic predisposition exclusively. PMID- 18159572 TI - Level of operant training rather than cocaine intake predicts level of reinstatement. AB - RATIONALE: Extended cocaine self-administration has been shown to potentiate reinstatement. This increased vulnerability to relapse could be attributed not only to extended cocaine exposure but also to extended operant training. OBJECTIVE: This study was aimed at determining the influence of different operant training histories on cocaine-induced reinstatement when cocaine intake is kept constant. MATERIALS AND METHODS: Cocaine intake and operant training were dissociated by using experimental procedures generating different histories of operant training but almost identical histories of cocaine intake. Rats were first trained to self-administer cocaine at a classical unit dose (250 microg/inf, FR1), then in independent groups, the level of operant response was changed for the next 20 sessions by changing either the unit dose available (83, 250, or 750 microg/inf, Experiment 1) or the fixed ratio required (FR-1, FR-3, or FR-10, Experiment 2). Then, all rats were tested for reinstatement with different priming doses of cocaine (0, 5, 10, and 15 mg/kg; i.p.) at an early and late stage of an extinction period. RESULTS: Level of responding during training predicts the level of reinstatement later on, independently of the amount of cocaine consumed. High FR requirement and low unit dose access led to higher level of reinstatement at early and late stage of the extinction period, respectively. CONCLUSIONS: This study shows that the level of operant responding required to maintain optimal cocaine intake directly influences later levels of reinstatement. This finding suggests that environmental constrains that make drug taking demanding and effortful may increase the vulnerability to relapse. PMID- 18159573 TI - Integrated strategies for identification of selenometabolites in animal and plant samples. PMID- 18159574 TI - [The experience of chronic pain and pain beliefs]. AB - Chronic pain is well appreciated as a universal problem in the sense that it causes serious impairment in the individuals' physical and psychosocial status of functioning. Research, psychological assessment and psychotherapeutic process towards chronic pain is most frequently based on the principles of cognitive behavioral approach (CBA). However, Sharp criticised CBA for its ongoing dependency on the operant premises of the pure behavioral approach and proposed a "reformulated CBA" where the emphasis is on specific cognitive factors, primarily the pain beliefs. According to current studies in chronic pain, it is becoming increasingly apparent that pain beliefs play an important role in the maintainance and management of chronic pain. In Turkey, there are no studies and tools on pain beliefs. In this specific review, the literature on pain beliefs is discussed in terms of necessity of research and material development on pain beliefs that tab factors specific to Turkish culture. PMID- 18159575 TI - [The effects of various doses of intrathecal morphine in addition with ropivacaine in combined spinal-epidural technique for labour analgesia]. AB - We evaluated the duration of analgesia, side effects and neonatal effects of intrathecal ropivacaine combined with two different doses of morphine using combined spinal epidural (CSE) technique for labour analgesia. Fourty term parturients were enrolled. Group I received 0,2% ropivacaine 3 mg + morphine 50 microg and Group II received 0,2% ropivacaine 3 mg + morphine 100 microg intrathecally. When VAS for pain was equal or above 30 mm, PCEA(Patient controlling epidural analgesia) was started using 0,1% ropivacaine. There wasn't any statistical significant difference considering the duration of analgesia between two groups. The first stage of labour was significantly shorter in Group II then Group I, and the epidural local anaesthetic volume was significantly lower in Group II. The most common side effect in both groups was pruritus. There was not any side effect of local anaesthetic and opioid on the newborns. Adequate analgesia was obtained during labour with CSE technique, using ropivacaine combined with 50 and 100 microg morphine intrathecally. However cervical dilatation was faster and epidural local anaesthetic volume was lower in the group in which 100 microg morphine was used. Also considering the unchanged incidence of the side effects, 3 mg 0,2% ropivacaine + 100 microg morphine may be a useful combination for labour analgesia. PMID- 18159576 TI - [Comparison of tramadol, tramadol-metamizol and tramadol-lornoxicam administered by intravenous PCA in management of postoperative pain]. AB - Comparison of tramadol, tramadol-metamizol and tramadol-lornoxicam administered by intravenous PCA in management of postoperative pain. The aim of the present study was to compare of the postoperative analgesic effects of tramadol, combinations of tramadol-metamizol and tramadol-lornoxicam administered by intravenous Patient Control Analgesia (PCA) in lower abdominal surgery. Sixty adult, female patients who undergoing lower abdominal surgery, were included in this study. Patients were randomized to three groups. The solutions were prepared containing 500 mg tramadol in 50 ml saline (10 mg/ml tramadol) for Group I, 250 mg tramadol+3000 mg metamizol in 50 ml saline (5 mg/ml tramadol+60 mg/ml metamizol) for Group II and 250 mg tramadol+20 mg lornoxicam in 50 ml saline (5mg/ml tramadol + 0.4 mg/ml lornoxicam) for Group III. Loading dose 10 ml was administrated within 30 min 30 to 40 min before the end of the surgery. PCA was started at the first complaint of pain. Pain was evaluated by VAS in every 15 minute intervals at the first hour and later at 2nd, 4th, 8th, 12th, 18th and 24th hours postoperatively. Vital parameters, side-effects, sedation scores and total analgesic consumptions were also recorded concurrently. Total tramadol and anti-emetic consumption, the incidence of postoperative nausea and vomiting (PONV) were significantly higher in group I than the other groups (p<0.05). In conclusion; Tramadol-metamizol and tramadol-lornoxicam combinations administered by intravenous PCA provide efficient postoperative analgesia with less side effects. PMID- 18159577 TI - The effect of opioid administration by different routes on the psychological functions of elderly patients. AB - Psychological status of patients during epidural and intravenous (i.v.) morphine administration are not known enough. It's known that after administration of morphine epidurally and intravenously plasma concentrations are similar but the concentrations in cerebrospinal fluide are different. The effects of two postoperative analgesia treatment regimens on the psychological and mental functions of the elderly after major orthopedic surgery were investigated in this randomized, blinded study. Morphine was used epidurally in Group E and intravenously in Group IV with patient controlled analgesia method for postoperative analgesia treatment. All patients were assessed for psychological status the day before surgery and 2 days after surgery using the Brief Symptom Inventory (BSI). The results of the BSI were assessed with the Global Severity Index (GSI). Nine subscales were also derived: depression, somatization, obsessive-compulsive, anxiety, interpersonal sensitivity, hostility, phobic anxiety, paranoid ideation, and psychoticism. There was not significant difference between the groups for preoperative and postoperative GSI results (p>0.05). Only interpersonal sensitivity subscale results in Group i.v. were decreased compared to the baseline (p<0.001). Preoperative BSI and submeasurement test scores revealed no correlation with total morphine consumption in both groups (p>0.05). Mental changes like agitation, time and place disorientation were seen equally in both groups. This study showed that after epidural and intravenouse morphine administration, no psychologycal changes were observed in the postoperative period compared with baseline. Patients who received morphine intravenously were showed less interpersonal sensitivity in the postoperative period. PMID- 18159578 TI - Prevalence of patent foramen ovale in patients with migraine. AB - Recent evidence supports that the prevalence of patent foramen ovale is higher in patients with migraine with aura. We conducted a case-control study and searched for intra-atrial right to left shunt in 53 patients with migraine. PFO was detected by means of transthoracic echocardiography with administration of contrast medium during valsalva maneuver and the results were compared with age and sex matched 27 healthy controls. Patent foramen ovale was more frequent in the migraine group (p<.01). The percentages of PFO in migraine patients with aura, without aura and the control group were 66.7%, 47.4% and 22.2%, respectively. Our results are supportive of an association between PFO and migraine, especially with aura. PMID- 18159579 TI - [Eagle's syndrome: a case presentation]. AB - Eagle's syndrome occurs when an elongated styloid process or calcified stylohyoid ligament causes recurrent throat pain or foreign body sensation, dysphagia. Diagnosis can usually be made on physical examination by digital palpation of the styloid process in the tonsiller fossa. Three-dimensional computed tomography can utilized for supporting diagnosis. The treatment of Eagle's syndrome is primarily surgical. The styloid process can be shortened through an intraoral oe external approach. We report a 46- year-old woman with the seymptomatology of Eafle's syndrome and literature review. PMID- 18159580 TI - Rethinking statistical approaches to evaluating drug safety. AB - PURPOSE: The current methods used to evaluate the efficacy of drug products are inadequate. We propose a non-inferiority approach to prove the safety of drugs. MATERIALS AND METHODS: Traditional hypotheses for the evaluation of the safety of drugs are based on proof of hazard, which have proven to be inadequate. Therefore, based on the concept of proof of safety, the non-inferiority hypothesis is employed to prove that the risk of new drugs does not exceed a pre specified allowable safety margin, hence proving that a drug has no excessive risk. The results from papers published on Vioxx and Avandia are used to illustrate the difference between the traditional approach for proof of hazard and the non-inferiority approach for proof of safety. RESULTS: The p-values from traditional hypotheses were greater than 0.05, and failed to demonstrate that Vioxx and Avandia are of cardiovascular hazard. However, these results cannot prove that both Vioxx and Avandia are of no cardiovascular risk. On the other hand, the non-inferiority approach can prove that they are of excessive cardiovascular risk. CONCLUSION: The non-inferiority approach is appropriate to prove the safety of drugs. PMID- 18159582 TI - Lamivudine therapy for korean children with chronic hepatitis B. AB - PURPOSE: Lamivudine is known to be very effective in suppressing hepatitis B virus replication and virus induced necroinflammation. The aim of this study was to evaluate lamivudine therapy efficacy, predictive factors, breakthrough, prevalence of YMDD mutation, and relapse rate in Korean children with chronic hepatitis B. MATERIALS AND METHODS: Between August 1999 and February 2005, 60 children on lamivudine therapy for chronic hepatitis B were enrolled. Treatment response was defined as alanine aminotransferase (ALT) normalization, and HBeAg and HBV-DNA disappearance. RESULTS: Seroconversion rates of HBeAg and HBV- DNA were 42% and 53%, respectively, and ALT normalization rate was 88%. Seroconversion rates of HBeAg (60.0%) and anti-HBe (60.0%) were higher in patients younger than 6 years. Seroconversion rate of HBV-DNA (68.4%) and normalization rate of serum ALT (94.7%) were highest in patients between 6 and 12 years. Seroconversion rates of all HBV markers were lowest in patients older than 12 years. Predicted 3 year cumulative seroconversion rates, were 70%, 68% for HBeAg, HBV-DNA, respectively. These were calculated by Kaplan-Meier method. Cox proportional hazard regression model showed that pre-treatment ALT was a positive predictive factor for seroconversion of HBeAg and HBV-DNA. Breakthrough phenomenon was noted in 6 patients, and 3 had a YMDD mutation. CONCLUSION: Lamivudine therapy had a significant effect on HBeAg seroconversion and HBV-DNA disappearance, and ALT normalization for Korean children with chronic hepatitis B. PMID- 18159581 TI - Antiphospholipid syndrome and vascular ischemic (occlusive) diseases: an overview. AB - Antiphospholipid syndrome (APS) is primarily considered to be an autoimmune pathological condition that is also referred to as "Hughes syndrome". It is characterized by arterial and/or venous thrombosis and pregnancy pathologies in the presence of anticardiolipin antibodies and/or lupus anticoagulant. APS can occur either as a primary disease or secondary to a connective tissue disorder, most frequently systemic lupus erythematosus (SLE). Damage to the nervous system is one of the most prominent clinical constellations of sequelae in APS and includes (i) arterial/ venous thrombotic events, (ii) psychiatric features and (iii) other non- thrombotic neurological syndromes. In this overview we compare the most important vascular ischemic (occlusive) disturbances (VIOD) with neuro psychiatric symptomatics, together with complete, updated classifications and hypotheses for the etio-pathogenesis of APS with underlying clinical and laboratory criteria for optimal diagnosis and disease management. PMID- 18159583 TI - Incomplete colonoscopy in patients with occlusive colorectal cancer: usefulness of CT colonography according to tumor location. AB - PURPOSE: We sought to evaluate the clinical usefulness of CT colonography (CTC) after incomplete conventional colonoscopy (CC) for occlusive colorectal cancer (CRC) according to the tumor location. MATERIALS AND METHODS: Seventy-five patients with occlusive CRC underwent subsequent CTC immediately after incomplete CC. Fifty-nine patients had distal CRC and 16 had proximal colon cancer. Experienced radiologists prospectively analyzed the location, length, and TNM staging of the main tumor. The colorectal polyps in the remaining colorectum and additional extraluminal findings were also recorded. Sixty-seven patients underwent colorectal resection. We retrospectively analyzed the surgical outcome and correlated CTC and CC findings. RESULTS: The overall accuracies of tumor staging were: T staging, 86%; N staging (nodal positivity), 70% (80%); and intra abdominal M staging, 94%. Additional colonic polyps were found in 23 patients. Six synchronous carcinomas were detected (9%); three in the proximal colon and three in the distal colon of occlusion. Clinically significant localization errors at CC were noted in 8 patients (12%, 5 proximal colon cancers and 3 distal CRCs) and were corrected by CTC. After CTC, the surgeons modified the initial surgical plan in 11 cases (16%). CONCLUSION: In occlusive CRC, CTC is not only useful in the evaluation of the proximal bowel, but can also provide surgeons with accurate information about staging and tumor localization. CTC is recommended when endoscopists encounter occlusive CRC, regardless of tumor location. PMID- 18159584 TI - Therapeutic comparison of chemotherapy and surgery for early stage diffuse large B-cell gastric lymphoma. AB - PURPOSE: The use of surgery versus stomach-preserving treatment for primary gastric lymphoma has caused controversy among doctors. This retrospective, single center study aims to evaluate the efficacy and benefit of stomach-preserving treatment against surgery for early stage diffuse large B-cell lymphoma of stomach. MATERIALS AND METHODS: From August 1991 to January 2006, 43 cases of early-stage diffuse large B-cell gastric lymphoma were reviewed. RESULTS: Eleven cases were treated with chemotherapy or chemotherapy plus radiation (CT +/- RT), 17 were treated with surgery alone (OP), and 15 were treated with surgery plus adjuvant chemotherapy (OP + CT). The complete remission and response rates were 63.6% and 90.9% in those treated with CT +/- RT (7 complete responders, 3 partial responders, 1 non-responder), 100% and 100% in those treated with OP, and 100% and 100% in those treated with OP + CT, respectively. Five-year overall survival rates were 85.7%, 87.5%, and 100% in those treated by CT +/- RT, OP, and OP + CT, respectively (p=0.76). The five-year disease free survival rates were 100%, 87.5% and 100% in those treated by CT +/- RT, OP, and OP + CT, respectively (p=0.99). There was no significant difference in overall survival and disease free survival between modalities. Even though there are no definite differences in the number of complications between those treated by CT +/- RT or OP, these facts reflect little concern on complications after surgery. CONCLUSION: In preventing morbidity arising from early or late complications from surgery and promoting quality of life, chemotherapy should be a primary consideration for early stage diffuse large B-cell lymphoma of the stomach. PMID- 18159585 TI - IGF-1 counteracts TGF-beta-mediated enhancement of fibronectin for in vitro human lens epithelial cells. AB - PURPOSE: To determine whether insulin-like growth factor (IGF-1) affects transforming growth factor (TGF-beta)- mediated fibronectin accumulation in human lens epithelial cell line (HLE B-3) cells. MATERIALS AND METHODS: HLE B-3 cells were incubated for 24 hours with TGF-beta (10 ng/ mL), IGF-1 (10 ng/mL), or both. Expression of the fibronectin gene was determined using a real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Fibronectin levels were examined using western blot analysis and immunofluorescence staining. RESULTS: Expression of the fibronectin gene was not different between the TGF-beta/IGF-1 treated group and the TGF-beta treated group (p= 0.116). However, western blot analysis demonstrated decreased fibronectin levels in human lens epithelial cells treated with TGF-beta and IGF-1 compared to those treated with TGF-beta only (p < 0.01). Immunofluorescence staining disclosed inhibition of TGF-beta-induced fibronectin in the presence of IGF-1. CONCLUSION: This study suggests that IGF-1 counteracts TGF-beta-mediated fibronectin accumulation in human lens epithelial cells. PMID- 18159586 TI - Conjunctival expansion using a subtenon's silicone implant in New Zealand white rabbits. AB - PURPOSE: In the field of ophthalmology, the conjunctival autograft is a useful therapeutic material in many cases, but the small size of the autograft is a disadvantage. Therefore, we evaluated the feasibility of taking an expanded sample of conjunctival tissue using a subtenon's silicone implant. MATERIALS AND METHODS: We included a total of nine rabbits; eight rabbits were operative cases, and one was a control. A portion of conjunctival tissue from the control rabbit, which did not undergo surgery, was dissected and examined to determine whether it was histologically different from the experimental group. The surgical procedure was performed on eight rabbits via a subtenon's insertion of a silicone sponge in the left superior-temporal portion; after surgery, we dropped antibiotics into the eyes. We sacrificed a pair of rabbits every three days (on days 3, 6, 9, and 12) after surgery, removed the expanded conjunctival tissues with the silicone sponge implants, and measured their sizes. RESULTS: The mean size of the expanded conjunctival tissues was 194.4 mm2. On the third day, we were able to harvest a 223.56 mm2 section of conjunctival tissue, which was the most expanded sample of tissue in the study. On the twelfth day, we removed a 160.38 mm2 section of conjunctival tissue, which was the least expanded sample of tissue. Statistically, there were no significant differences in the mean dimensions of the expanded conjunctival tissues for each time period. Microscopic examinations showed no histological differences between the expanded conjunctival tissues and the normal conjunctival tissues. CONCLUSION: The results reveal that this procedure is a useful method to expand the conjunctiva for grafting and transplantation. PMID- 18159587 TI - Correlation between MRI and operative findings in Bell's palsy and Ramsay Hunt syndrome. AB - PURPOSE: To investigate the correlation between gadolinium enhanced magnetic resonance image (MRI) results and surgical findings of facial nerves in Bell's palsy and Ramsay Hunt syndrome. MATERIALS AND METHODS: From 1995 to 2004, MRI was performed on 13 patients with Bell's palsy or Ramsay Hunt syndrome, who were offered with surgical decompression of the facial nerve through the middle cranial fossa approach. Gadolinium enhanced MRI was performed on all patients and the enhancement of the facial nerve was evaluated by radiology specialists. Operative findings including the degree of the facial nerve segment swelling were examined. Furthermore, the time interval from the onset of palsy to surgery was evaluated. RESULTS: Swelling of facial nerve segments was found in patients with enhanced facial nerves from MRI. The swelling of the facial nerve in the labyrinthine segment in particular was identified in all patients with enhanced labyrinthine segments in MRI. The intraoperative swelling of geniculate ganglion of facial nerve was found in 78% of patients with enhanced facial segment in MRI (p=0.01). The intraoperative swelling of tympanic segment was observed from fourth to ninth weeks after the onset of palsy. CONCLUSION: MRI enhancement of facial nerves in Bell's palsy and Ramsay Hunt syndrome is associated with the extent of intratemporal lesions of facial nerves, especially in the labyrinthine segment. PMID- 18159588 TI - The value of routinely culturing for tuberculosis during bronchoscopies in an intermediate tuberculosis-burden country. AB - PURPOSE: Many medical centers routinely culture bronchoscopy samples for Mycobacterium tuberculosis, even when tuberculosis is not strongly suspected. The value of this practice, however, is controversial. We evaluated the role of that procedure in the diagnosis of pulmonary tuberculosis in an intermediate tuberculosis-burden country. PATIENTS AND METHODS: A prospective, observational study was conducted in a tertiary referral center and included 733 consecutive patients who underwent bronchoscopy examination. RESULTS: M. tuberculosis was isolated in 47 patients (6.4%). According to radiographic features, the rate of positive culture for M. tuberculosis was relatively high in patients with atelectasis (5/33, 15.2%) and those with pulmonary infiltrations of suspicious infections (26/183, 14.2%). M. tuberculosis was isolated even in patients with pulmonary masses (9/266, 3.4%) and those with pulmonary nodules (5/175, 2.9%). In 16/47 (34.0%) patients with positive cultures for M. tuberculosis, active pulmonary tuberculosis was not suspected at the time of bronchoscopy. CONCLUSION: These results suggest that routinely culturing for M. tuberculosis during bronchoscopy is still useful in the diagnosis of pulmonary tuberculosis in an intermediate tuberculosis-burden country. PMID- 18159589 TI - Isolated and complex scimitar vein anomalies and their differentiation from the meandering right pulmonary vein. AB - PURPOSE: Four pediatric patients with isolated (the adult form) and complex (the infantile form) scimitar vein anomalies were reviewed and compared with patients with meandering right pulmonary veins. MATERIALS AND METHODS: From January 1990 to December 2006, 4 female patients, aged 2 days to 3.5 years, with isolated and complex scimitar vein anomalies were retrospectively studied. The clinical features, chest radiographs, echocardiography, magnetic resonance imaging, magnetic resonance angiography, computer tomography, bronchography, cardiac catheterization with angiography, surgery, and autopsy were reviewed to substantiate the diagnosis of isolated and complex scimitar vein anomalies. RESULTS: Clinical manifestations were lung infections with radiographic scimitar signs in 4 patients (3 right; 1 left), respiratory distress, congestive heart failure, pulmonary hypertension and/or cyanosis in 3 patients, dextrocardia, heterotaxy, and/or right atrial isomerism in 2 patients, as well as dextroversion, right bronchial isomerism, bronchial stenosis, and/or sepsis in 1 patient. Two patients with right atrial isomerism expired. CONCLUSION: The clinical discrimination between scimitar vein anomaly with and without cardiovascular and bronchopulmonary malformations is crucial for the outcomes vary. PMID- 18159590 TI - Surgical outcome of synchronous second primary cancer in patients with gastric cancer. AB - PURPOSE: In order to improve the likelihood of curative and safe gastric surgery, this study investigated the clinical features and surgical outcomes of gastric cancer with a synchronous cancer. PATIENTS AND METHODS: The clinicopathological data of 10,090 gastric cancer patients at Samsung Medical Center from September 1994 to December 2006 were retrospectively analyzed. Of them, 90 patients with gastric cancer and a synchronous second primary cancer underwent simultaneous surgery for gastric cancer and second primary cancer. The clinicopathological characteristics of the patients, surgical outcome, and prognosis were examined. RESULTS: The most common synchronous second primary cancer was colorectal cancer (37 patients), followed by hepatocellular carcinoma (13 patients), renal cell carcinoma (11 patients), and pancreatic carcinoma (5 patients). The incidence of a second primary cancer in the gastric cancer patients was higher than the incidence in the general population. Stage I gastric cancer patients had more synchronous cancers than stage II patients (59 vs. 31). Postoperative complications were encountered in 7 patients. Four patients underwent reoperation. Two patients died from hepatic failure and leakage of esophagojejunal anastomosis. The 5-year survival rate of stage I and II gastric cancer was 61% and 39%, respectively. CONCLUSION: Since gastric cancer patients with a synchronous second primary cancer are not rare, the possibility of synchronous cancers in gastric cancer patients should be considered. The prognosis of early stage gastric cancer patients with a synchronous second primary cancer was influenced more by the presence of the second primary cancer than by the gastric cancer itself. PMID- 18159591 TI - Spinal cord tumors of the thoracolumbar junction requiring surgery: a retrospective review of clinical features and surgical outcome. AB - PURPOSE: A retrospective review of medical records and imaging studies. To investigate characteristic clinical features and surgical outcomes of spinal cord tumors (SCTs) of the thoracolumbar junction (TLJ). The spinal cord transitions to the cauda equina in the TLJ. The TLJ contains the upper and lower motor neurons of the spinal cord and cauda equina. As a result, the clinical features of lesions in the TLJ vary, and these anatomical characteristics may affect surgical outcome. MATERIALS AND METHODS: Pathological diagnosis, clinical features, neurological signs, and surgical outcomes were investigated in 76 patients surgically treated at our institute for SCTs arising from T11 to L2. The patients were divided into epiconus (T11-12, n=18) and conus groups (L1-2, n=58). RESULTS: Patients in the epiconus group had hyperactive deep tendon reflexes (DTRs), while those in the conus group had hypoactive DTRs (p < 0.05). Nine patients were misdiagnosed with intervertebral disc diseases (IVDs) before correct diagnoses were made. It was impossible to definitively determine the exact cause of symptoms in four patients who had both SCTs and IVDs. CONCLUSION: Among SCTs of the TLJ, the epiconus group displayed upper motor neuron syndrome and the conus group displayed lower motor neuron syndrome. SCTs of the TLJ were frequently misdiagnosed as IVDs due to symptomatic similarities. SCTs of the TLJ should be included in differential diagnosis of back and leg pain, and it is highly recommended that routine lumbar magnetic resonance imaging include the TLJ. PMID- 18159592 TI - High-dose terazosin therapy (5mg) in Korean patients with lower urinary tract symptoms with or without concomitant hypertension: a prospective, open-label study. AB - PURPOSE: We determined the efficacy and safety of a relatively high dose of terazosin (5mg) in Korean patients with lower urinary tract symptoms (LUTS), with or without concomitant hypertension. MATERIALS AND METHODS: From July to December 2006, 200 men who consecutively presented with LUTS were prospectively studied. Eight weeks after treatment, blood pressure (BP), uroflowmetry, and International Prostate Symptom Score (I-PSS) were assessed. For analysis purposes, patients were stratified according to concomitant hypertension. Of the 200 patients, 173 completed the scheduled eight-week treatment period. RESULTS: At baseline, no differences were evident in the two groups in terms of I-PSS, Qmax, PVR and BP. After eight weeks of treatment-although I-PSS and uroflowmetry parameters were not significantly different in the two groups-systolic and diastolic BP in the non-hypertensive control group were higher than in the hypertensive group (p= 0.001 and p=0.0100, respectively). Changes in I-PSS, uroflowmetry parameters, and BPs measured at week eight post- treatment commencement did not significantly differ between the two groups. Moreover, the addition of 5mg of terazosin to antihypertensives did not cause a significant reduction in either systolic or diastolic BP in either group. CONCLUSION: Adding terazosin to existing antihypertensive regimens did not seem to increase the incidence of adverse events. Our findings suggest that 5mg terazosin is effective and that it has an acceptable safety profile as an add-on therapy for patients with LUTS and concomitant hypertension. PMID- 18159593 TI - A prospective, multicenter, open-label trial of zoledronic acid in patients with hormone refractory prostate cancer. AB - PURPOSE: The short-term safety and efficacy of zoledronic acid for the treatment of skeletal metastasis was evaluated in patients with hormone-refractory prostate cancer. PATIENTS AND METHODS: A total of 19 hormone-refractory prostate cancer patients with bone metastases were enrolled. All patients received up to six infusions of zoledronic acid (4 mg, given intravenously over 15 minutes, every 3 4 weeks). Safety was assessed by monitoring adverse events and serum creatinine levels. Efficacy was assessed by monitoring skeletal-related events, brief pain inventory score, quality of life score, type of pain medication, and analgesic score. Mean age of patients was 67.3 years (46-86 years), mean time from diagnosis of bone metastases was 27.6 months (0-117 months), and mean time from diagnosis of hormone-refractory disease was 7.5 months (0-26 months). RESULTS: There was no clinically significant change in serum creatinine levels. Eleven adverse events (musculoskeletal disorders and systemic disorders) in 8 patients were classed as having a possible relationship to study drug. Fifteen patients completed six courses of zoledronic acid infusion. There were no significant changes in the brief pain inventory composite scores, quality of life questionnaire scores or analgesic score. No new skeletal-related events developed during the treatment period. CONCLUSION: Zoledronic acid administered in this study as a 15-minute infusion demonstrated an acceptable and well-known safety profile in patients with refractory prostate cancer with bone metastases. However, prospective placebo- controlled clinical trials are required to elucidate the efficacy of zoledronic acid. PMID- 18159594 TI - Identification of enhancer of zeste homolog 2 expression in peripheral circulating tumor cells in metastatic prostate cancer patients: a preliminary study. AB - PURPOSE: Enhancer of zeste homolog 2 (EZH2), a kind of transcriptional repressor, is reportedly over-expressed in metastatic prostate cancer. In this study, we analyzed EZH2 mRNA in circulating tumor cells (CTCs) in peripheral blood as a biomarker in patients with metastatic prostate cancer. PATIENTS AND METHODS: Ber EP4 coated immunomagnetic beads were used to harvest CTCs, and mRNA was isolated by oligo- dT conjugated immunomagnetic beads. Reverse transcriptase- polymerase chain reaction for EZH2 mRNA was performed and the expression density was measured. The sensitivity of this test for detection of EZH2 mRNA was determined by serial dilutions of a human prostate cancer cell line. Blood samples were collected from 20 patients each with metastatic or localized prostate cancer and 10 healthy volunteers. RESULTS: Sensitivity experiments showed that the test was highly sensitive as it could detect 10 tumor cells per 5 mL. EZH2 mRNA expression was obtained from peripheral blood samples of patients and control subjects. EZH2 mRNA expression density in the metastatic prostate cancer group was significantly higher than in the control (p=0.023) and localized prostate cancer groups (p=0.019). There was no difference between the control and localized prostate cancer groups (p > 0.05). CONCLUSION: EZH2 mRNA expression in circulating epithelial cells represents a promising marker for detecting early metastasis in prostate cancer. However, more specific and sensitive techniques for detection of CTCs are needed to avoid mononuclear cell contamination. PMID- 18159595 TI - A relationship of sex hormone levels and erectile dysfunction: which tests should be done routinely? AB - PURPOSE: In this study, the relationship between sex hormone levels and erectile dysfunction (ED), as well as the necessity of routinely measuring sex hormone levels were evaluated. MATERIALS AND METHODS: This study included one hundred patients admitted to a urology clinic for sexual dysfunction. To determine the hormone levels, following the history (included IIEF-5 score) and physical examination, triple blood samples were collected at intervals of 15 minutes between 08:00 and 10:00 am. Total and free testosterone, prolactin, follicle stimulating hormone and luteinizing hormone levels were studied. RESULTS: Mean age was 43 (23-80) years. IIEF-5 score was less than 21 [9.8-4.3 (3-19)] in all study groups. There was a statistically significant correlation between tT and FSH, as well as between LH and FSH in Pearson (r =-0.513, p < 0.001, respectively) and also in Spearman tests (r=-0.224, p=0.042 and r=-0.459, p < 0.001, respectively). However, there was no correlation between age and serum hormone levels (p > 0.05). Of the 100 patients, 18 (18%) had low tT, 77 (77%) had normal and 5 (5%) had high tT levels. No statistically significant correlation was found between decreased libido and tT levels (p > 0.05). Twelve (66.6%) of the 18 patients with low tT had normal libido. CONCLUSION: Analyzing the medical history in detail and performing a thorough physical examination can reduce the need for excessive studies and consultations, and enables patients to save time and costs. PMID- 18159596 TI - Sulindac prevents esophageal adenocarcinomas induced by gastroduodenal reflux in rats. AB - PURPOSE: It is known that cyclooxygenase (COX)-2 expression is increased in Barrett's esophagus and esophageal adenocarcinomas. We studied COX-2 expression and the effect sulindac has on the genesis of Barrett's esophagus and adenocarcinoma in rats undergoing esophagogastroduodenal anastomosis (EGDA). MATERIALS AND METHODS: Fifty-one rats were divided into a control group (n=27), a 500 ppm sulindac-treated group (n=15) and 1000 ppm sulindac-treated group (n=9). Randomly selected rats were killed by diethyl ether inhalation at 20 and 40 weeks after surgery. RESULTS: At 40 weeks, rats treated with 1000 ppm sulindac showed narrower esophageal diameter and milder inflammation than the control rats. At 40 weeks, the incidence of Barrett's esophagus was similar between control and sulindac-treated groups, but the incidence of adenocarcinoma was significantly lower in the 1000 ppm sulindac-treated group than either the control or 500 ppm sulindac-treated groups. COX-2 was significantly increased in the lower esophagus of control rats killed at 40 weeks. Cyclin D1 expression was negligible in the sulindac- treated group compared with the control group. CONCLUSION: We suggest that the chemopreventive effect of sulindac is related to decreased COX-2 and cyclin D1 expression, which may be influenced by reduced inflammation. PMID- 18159597 TI - Ascorbic acid alleviates pancreatic damage induced by dibutyltin dichloride (DBTC) in rats. AB - PURPOSE: Because previous studies have reported depleted antioxidant capacity in patients with chronic pancreatitis (CP), prevention of free radical production has gained importance in antifibrotic treatment strategies for CP. The aim of this study was to investigate the effects of ascorbic acid on oxidative capacity and pancreatic damage in experimental CP. MATERIALS AND METHODS: CP was induced in male Sprague-Dawley rats by infusion of dibutyltin dichloride (DBTC) into the tail vein. Ascorbic acid was given intraperitoneally at a daily dose of 10 mg/kg body weight. The treatment groups were as follows: group 1, DBTC plus intraperitoneal physiologic saline; group 2, DBTC plus intraperitoneal ascorbic acid; group 3, solvent plus intraperitoneal physiologic saline; group 4, no operation plus intraperitoneal physiologic saline. Each group contained 15 animals. Treatment was started after CP was established. After 4 weeks of treatment, serum hyaluronic acid and laminin levels were determined by radioimmunoassay, pancreatic tissue oxidative stress was analyzed, and the degree of pancreatic damage was determined. RESULTS: Ascorbic acid treatment markedly increased superoxide dismutase (SOD) activity and decreased malondialdehyde (MDA) concentrations in pancreatic tissue (p < 0.01 for both). Significant serum hyaluronic acid and laminin reductions were observed in group 2 as compared with group 1 (p < 0.05). However, the serum hyaluronic acid and laminin levels remained elevated when compared with those of groups 3 and 4 (p < 0.05). Histopathologic scores were also lower in animals with CP that underwent ascorbic acid-treatment (p < 0.05). CONCLUSION: Ascorbic acid treatment alleviated the degree of oxidative stress and pancreatic damage in rat CP. Antioxidant treatment might be considered a potential option to improve the pathologic process in CP. PMID- 18159598 TI - A case of successful intrapleural chemotherapy with Cisplatin plus cytarabine for intractable malignant pleural effusion. AB - When conventional treatments of malignant pleural effusion, such as repeated thoracentesis, closed thoracotomy and pleurodesis by instilled sclerosing agents, are ineffective, there are few alternative therapies available. Our case involves a 47-year-old woman with uterine cervical carcinoma suffering from malignant pleural effusion. She presented with a chief complaint of severe dyspnea, and was classified as an Eastern Cooperative Oncology Group (ECOG) performance status of 4. Her underlying cervical carcinoma progressed despite various systemic chemotherapy regimens. In addition, pleural effusion persisted in spite of 4 weeks of drainage through the thoracotomy tube and talc pleurodesis. Under such circumstances, we attempted intrapleural chemotherapy with cisplatin plus cytarabine, which resulted in significant decrease of the pleural effusion. No serious systemic toxicities, including myelosuppression, were observed. As a result, the patient's dyspnea was relieved, and her ECOG performance status improved from 4 to 2. However, the thoracotomy tube was not removed due to subsequent iatrogenic pneumothorax. Pleural effusion did not recur for the 4 weeks leading up to her death. PMID- 18159599 TI - Two separated ileal adenocarcinomas in neurofibromatosis type 1. AB - Patients with neurofibromatosis-1 (NF-1) have increased susceptibility to a variety of malignancies. Here, we document a rare case of two separated ileal adenocarcinomas in NF-1. The adenocarcinomas were surrounded by a diffuse tubular adenomatous lesion of the mucosa, and ganglion cells were scattered in the NF background. We found this case meaningful for several reasons: two separated adenocarcinomas arising in an unusual ileal segment, the association with precancerous tubular adenoma, and the presence of ganglion cells, which suggests ganglioneuromatosis in NF-1. PMID- 18159600 TI - A case of cardiac lymphangioma presenting as a cystic mass in the right atrium. AB - A 44-year-old woman underwent surgery for an asymptomatic primary tumor of the heart located in the right atrium. The tumor was detected incidentally during follow-up computed tomography for a resected breast cancer. The mass, lying along the lower portion of the right atrial septum, was homogenous and cystic in nature, as detected by transthoracic and transesophageal echocardiography. Complete resection was performed via a median sternotomy under cardiopulmonary bypass. The postoperative course was uneventful. However, the histological result was surprising: the mass was a cardiac lymphangioma. PMID- 18159601 TI - Catheter ablation of a left free-wall accessory pathway via the radial artery approach. AB - Catheter ablation of the left free-wall accessory pathways (APs) is normally performed by the retrograde transaortic approach via a femoral artery or the transseptal approach. Here we report a case of an overt left free-wall AP, which was successfully ablated with a retrograde transaortic approach via the radial artery without any vascular complications. The patient has remained free of any symptoms or pre-excitation observed on the ECG during a 10-month post- ablation follow-up. PMID- 18159602 TI - A case of noncompaction of the ventricular myocardium combined with situs ambiguous with polysplenia. AB - A 33-year-old man was admitted to our hospital with chest pain and exertional dyspnea. Two-dimensional echocardiography showed prominent trabeculations and deep intertrabecular recesses, findings consistent with noncompaction of the ventricular myocardium. Thoracoabdominal CT and cardiac magnetic resonance imaging (CMR) revealed situs ambiguous with polysplenia and noncompaction of the left ventricular myocardium. CMR also demonstrated delayed enhancement of the trabeculations located at the apical portion of the left ventricle. The coronary angiogram was normal. This is the first case of noncompaction of the ventricular myocardium associated with situs ambiguous with polysplenia. PMID- 18159603 TI - Solitary fibrous tumor arising from stomach: CT findings. AB - Solitary fibrous tumors are spindle-cell neoplasms that usually develop in the pleura and peritoneum, and rarely arise in the stomach. To our knowledge, there is only one case reporting a solitary fibrous tumor arising from stomach in the English literature. Here we report the case of a 26-year-old man with a large solitary fibrous tumor arising from the stomach which involved the submucosa and muscular layer and resembled a gastrointestinal stromal tumor in the stomach, based on what was seen during abdominal computed tomography. A solitary fibrous tumor arising from the stomach, although rare, could be considered as a diagnostic possibility for gastric submucosal tumors. PMID- 18159604 TI - Synchronous adenocarcinoma and mantle cell lymphoma of the stomach. AB - Synchronous occurrence of mantle cell lymphoma (MCL) and gastric cancer in the same patient has not yet been reported in the English literature. MCL comprises 2.5-7% of non-Hodgkin's lymphomas and is characterized by a poor prognosis with a median survival probability of 3-4 years in most series. A 62-year-old man was referred to our hospital for evaluation of an abnormal gastric lesion. The endoscopic finding was compatible with type IIc early gastric cancer (EGC) in the middle third of the stomach, and a biopsy of the lesion proved to be carcinoma. Radical total gastrectomy with splenectomy and Roux-en-Y esophagojejunostomy were performed. The resected specimen revealed two grossly separated lesions. Postoperative histological examination reported both adenocarcinoma and MCL. Immunohistochemical staining showed positivity for CD5, CD20, and cyclin D1 in the infiltrated lymphoid cells. MCL is an aggressive non-Hodgkin's lymphoma, and the current treatment approach is still unsatisfactory. Further advancements in the understanding of the synchronous occurrence of both diseases, and more efforts on investigations of treatment are needed. PMID- 18159605 TI - Extrapulmonary small cell carcinoma of the liver: clinicopathological and immunohistochemical findings. AB - Patients with primary small cell carcinoma of the liver have rarely been described in medical literature. Knowledge of clinical, pathological and immunohistochemical properties remains limited. We described an 82-year-old female patient with primary small cell carcinoma of the liver. Histologically, the tumor showed typical morphology of a pulmonary small cell carcinoma. Immunohistochemically, the tumor revealed neuroendocrine differentiation; positive reaction for chromogranin, synaptophysin, CD56, and neuron specific enolase. The tumor was also positive for TTF-1 and c-kit but completely negative for hepatocyte, carcinoembryonic antigen, cytokeratin 7; 19; and 20. Herein, we discussed the clinical, pathological and immunohistochemical findings of extrapulmonary small cell carcinoma of the liver and reviewed the relevant literature. PMID- 18159606 TI - Value of manganese-enhanced T1- and T2-weighted MR cholangiography for differentiating cystic parenchymal lesions from cystic abnormalities which communicate with bile ducts. AB - We present a case report to show how manganese-enhanced T1- and T2-weighted MR cholangiography could differentiate cystic parenchymal lesions from cystic abnormalities which communicate with the bile ducts. PMID- 18159607 TI - A case of more abundant and dysplastic adenomas in the interposed colon than in the native colon. AB - We report a 60-year-old woman with intramucosal adenocarcinoma arising in the interposed colon, 40 years after the esophageal reconstruction for lye induced esophageal stricture. Although synchronous adenomas were also found in the native colon where the graft was taken, the number of adenomas was greater in the interposed colon and more dysplastic, even progressed to adenocarcinoma, than that of the native colon. The microsatellite instability-testing performed in the intramucosal carcinoma from interposed colon showed absence of microsatellite instability. Changing of location and functional demand of colonic segment, and the exposure to different intraluminal contents might have facilitated the adenoma- carcinoma transformation in the interposed colon. PMID- 18159608 TI - Association of beta 2-adrenergic receptor polymorphism with the phenotype of aspirin-intolerant acute urticaria. AB - The genetic mechanism of aspirin intolerant acute urticaria (AIAU) is unknown. To demonstrate an association between the beta 2 adrenergic receptor (ADRB2) polymorphism and the phenotype of AIAU, one hundred fourteen patients with AIAU, 110 patients with aspirin intolerant chronic urticaria (AICU), and 498 normal healthy controls (NC) based on a Korean population were enrolled. The genotype of ADRB2 at 46 A > G was analyzed using a direct sequencing method. The ADRB2 polymorphism at 46 A > G showed a significant difference between AIAU and NC; the frequency of the major genotype was significantly higher in the AIAU group (p= 0.017 in recessive model), while no differences were noted in allele and genotype frequencies between AICU and NC. In conclusion, the ADRB2 (46 A > G) gene polymorphism may contribute to the development of the phenotype of AIAU. PMID- 18159609 TI - Retraction: Suat Canbaz et al. Bilateral popliteal artery aneurysms with rupture and pseudoaneurysm formation on the left. Yonsei Med J 2003;44:159-62. PMID- 18159610 TI - Comparison of the Atkins, Zone, Ornish, and LEARN diets for change in weight and related risk factors among overweight premenopausal women. PMID- 18159611 TI - Atkins is an easy learning curve. PMID- 18159613 TI - Fail to test the impressive ability of diet to favorably affect long-term health and body weight loss. PMID- 18159612 TI - The A TO Z weight loss study is a flawed attempt to evaluate different nutritional life styles. PMID- 18159614 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159615 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159616 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159617 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159618 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159620 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159619 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159621 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159622 TI - Why don't the doctors attend the funerals of their patients who die? PMID- 18159623 TI - The Supreme Court and the Partial-birth Abortion Ban Act of 2003: a political procedure replaces woman-centered care. PMID- 18159624 TI - The Supreme Court and the Partial-birth Abortion Ban Act of 2003: a political procedure replaces woman-centered care. PMID- 18159625 TI - The Supreme Court and the Partial-birth Abortion Ban Act of 2003: a political procedure replaces woman-centered care. PMID- 18159626 TI - Medical student 'hazing' is unhealthy and unproductive. PMID- 18159627 TI - Medical student "hazing" is unhealthy and unproductive. PMID- 18159628 TI - Medical student "hazing" is unhealthy and unproductive. PMID- 18159630 TI - I am not like your father's pharmacist. PMID- 18159629 TI - I am not like your father's pharmacist. PMID- 18159631 TI - To be a great physician, you must understand the whole story. PMID- 18159632 TI - Peripheral neurotoxicity of platinum-based chemotherapy. PMID- 18159633 TI - An interview with... Sydney Brenner. Interview by Errol C. Friedberg. PMID- 18159635 TI - CO2 emissions: getting bang for the buck. PMID- 18159634 TI - Cancer filter deja vu. PMID- 18159636 TI - [Transcription of the KLRB1 gene is suppressed in human cancer tissues]. AB - The KLRB1 gene encodes the CD161 receptor of natural killer cells (NK-cells). The gene is also expressed in the NKT-cells. The two cell types are cytotoxic and capable of recognizing and eliminating various cell species, e.g. tumorous, virus infected, and allotransplants. The biological function of human CD161 is still insufficiently understood; probably it is involved in regulation of the cytotoxic functions of the cells and in regulation of cytokine production. Because immune system, in particular, the activity of cytotoxic cells, is suppressed in most cancer patients, it was suggested that the KLRB1 expression might be suppressed in cancerous cells. The results of this work demonstrated that the transcription of the KLRB1 was suppressed in tumor tissues in 68% patients with nonsmall-lung cancer (p < 0.0001) and 57% patients with esophageal squamous-cell carcinoma (p = 0.0003). High frequency of the KLRB1 transcription suppression upon cancers makes it possible to use this parameter as a highly informative marker of lung and esophageal cancers. PMID- 18159637 TI - [Theory of functional systems: postulates and principles of human body construction in health and pathology]. AB - It is shown that many functional systems with different level of organization harmoniously interact in healthy humans and animals. Early stress discoordinates information links of functional systems which can be easily corrected by nonpharmacological methods. Long-term and intensive stress disturbs mechanisms of self-regulation of the weakest functional systems. This converts a pathological process to a molecular tissue level. Principles of systemic functional human organization in pathology and compensation of impaired functions are considered. PMID- 18159638 TI - [Possible causes of temperature-dependent disorders of regional circulation in cryoglobulinemia]. AB - We studied effects of changed blood theology in cryoglobulinemia on temperature dependent disorders of peripheral circulation. The rheological status was assessed by aggregation characteristics of blood at temperature 37 degrees C and 4 degrees C at nephelometry of reverse light diffusion. Blood cryoglobulins concentration was determined spectrophotometrically. Turbidimetry estimated the size of cryocomplexes at different temperatures and energetic characteristics of their formation. It was found that blood of healthy donors in cooling does not change its rheological properties significantly. Blood of patients with cryoglobulinemia cooled to 4 degrees accelerated formation of linear erythrocyte aggregates and increased their strength. The results agreed with estimated values of molecular connection energy in cryocomplexes. The greatest difference between the patients' and donors' blood was in strength of the largest aggregates. Sharp strengthening of both large and small aggregates in blood cooling makes vascular red cell desaggregation impossible. This may entail microcirculatory arrest. PMID- 18159639 TI - [An antistress action of a biologically active additive "nervine tea"]. PMID- 18159640 TI - [Modification of an antitumor action of photodynamic therapy with anticoagulant drug fenylin]. AB - Noninbred rat experiments with M-1 tumor have found that combination of fenylin with photodynamic therapy produced a 1.5-fold slowing down in the growth of the tumor compared to the control animals on day 15 and 21 after the treatment. PMID- 18159641 TI - [The superficial architectonics of peripheral blood lymphocytes in patients with pulmonary tuberculosis]. AB - Peripheral blood lymphocytes were electron microscopically studied in patients with disseminated drug-sensitive and drug-resistant pulmonary tuberculosis before and during specific antituberculous chemotherapy. There was a tendency for an increase in the count of lymphocytes with membranous morphostructural changes during the therapy, which was more clearly defined in the drug-resistant type of the infectious process. It is suggested that antituberculous drugs have a damaging effect on the membrane of immunocompetent blood cells. PMID- 18159642 TI - [Morphometric characteristics of eosinophils in Opisthorchis invasion]. PMID- 18159643 TI - [The diagnostic value of macroprolactin in hyperprolactinemia]. AB - Macroprolactinemia (MP) in which there is a discrepancy between prolactin (P) levels and clinical symptoms has recently generated great interest among physicians of various specialties since not the whole of P is biologically active. The diagnosis of this condition is a topical problem. The purpose of the investigation was to detect high molecular-weight or big big P in patients with hyperprolactinemia (HP) of various genesis and to define its clinical importance. A hundred and one females with HP (P > 700 mU/l) were examined. In each patient, biologically active monomeric P was measured via treatment of polyethylene glycol (PEG) 6000. MP was detected in 23% of the examinees. Among them, only 11 females had clinical symptoms of the disease: 72.7% complained about menstrual irregularities; galactorrhea and headache were observed in 18.3 and 9.0%, respectively. The remaining 12 females had no clinical symptoms of HP. MP in which the clinical symptoms are feebly marked has been found to be not a rare condition. The determination of big big P in HP is of diagnostic value. It is recommended that the serum separation diagnostic technique using PEG should be made routine in the examination of patients with HP, particularly in the discrepancy of biochemical and clinical data and in idiopathic HP. PMID- 18159644 TI - [Markers of Fas-mediated apoptosis in patients with heart failure]. PMID- 18159645 TI - Early identification and treatment of insulin resistance: impact on subsequent prediabetes and type 2 diabetes. AB - Insulin resistance (IR) is characterized by decreasing sensitivity of target tissues to the action of insulin, elevated blood glucose concentration, and increased hepatic production of atherogenic lipids. IR is associated with declining insulin production by the pancreas, the emergence of type 2 diabetes, and increasing risk of cardiovascular disease (CVD). Clinical markers of IR include elevated plasma glucose concentration under fasting conditions or following ingestion of an oral glucose challenge. IR and hyperinsulinemia produce a number of effects that promote CVD, including adverse effects on blood pressure, endothelial cell function, lipid profile, platelet function, and blood coagulation. Glucose dysregulation often occurs in combination with other cardiovascular risk factors, including hypertension, obesity, and dyslipidemia. Clinical trials have shown that lifestyle changes to promote weight loss and medical therapy with insulin-sensitizing agents can reduce the likelihood of progression from early stages of IR to type 2 diabetes. However, it is important to recognize that obesity is a chronic condition that needs strategies beyond a diet plan to maintain sufficient weight loss over time. Pharmacologic therapies that are currently in development may help not only to promote weight loss but also to improve the symptoms of cardiometabolic risk in patients with and without diabetes. PMID- 18159646 TI - The benefits of tight glycemic control in type 2 diabetes mellitus. AB - Type 2 diabetes mellitus (DM) is associated with significantly increased risk of microvascular and macrovascular disease. Although most studies have focused on the microvascular complications of diabetes (eg, nephropathy, neuropathy, retinopathy), most patients with type 2 DM die from causes that are related to macrovascular disease (eg, myocardial infarction). Poor glycemic control increases the risk of future cardiovascular events, and prospective studies of patients with type 1 and type 2 DM have demonstrated that the incidence of vascular complications is reduced by lifestyle modifications or medications that reduce blood glucose concentrations. PMID- 18159647 TI - Clinical management of EGFRI dermatologic toxicities: the European perspective. AB - Dermatologic treatment of epidermal growth factor receptor inhibitor (EGFRI) skin toxicity is supportive and aims at maintaining quality of life while continuing EGFRI therapy. Despite the lack of randomized controlled trials or evidence-based guidelines, most cases of acneiform eruption are well controlled by topical metronidazole and oral minocycline 100 mg qd. For severe reactions, the minocycline dose is doubled and saline compresses are used. For superinfection with Staphylococcus aureus, oral cefuroxime axetil can be added for a short term. Emollients and topical steroids can be administered for skin dryness or eczema. Paronychia is the hardest to treat but antiseptic soaks and a corticosteroid paste can alleviate symptoms to some degree. PMID- 18159648 TI - Clinical management of EGFRI dermatologic toxicities: the Japanese perspective. AB - Epidermal growth factor receptor inhibitors (EGFRIs) have demonstrated clinical activity in patients with non-small-cell lung cancer, pancreatic cancer, and colorectal cancer. EGFRIs are generally well tolerated, but reversible dermatologic toxicities are commonly associated with their use. Limited clinical evidence has characterized these adverse reactions as a class effect. For panitumumab (Vectibix), mild-to-moderate dermatologic toxicities are the most common associated adverse reactions. This report details the Japanese experience in the management of dermatologic toxicities associated with panitumumab use. Treatment selection for skin toxicity in Japan is also detailed, with a flowchart depicting strategies to treat various stages of dermatologic toxicities. Panitumumab was well tolerated in Japanese patients with advanced solid tumors, with a safety profile similar to that seen in non-Japanese patients. PMID- 18159649 TI - The future of IT? Aggressive educational reform. TIGER initiative preps nurses for healthcare's digital era. PMID- 18159650 TI - A "most-wired" hospital targets information sharing. Process improvements include faster communication of key patient indicators. PMID- 18159652 TI - Ostomy care and radiation therapy. PMID- 18159651 TI - Over-the-counter vaginal contraceptive and spermicide drug products containing nonoxynol 9; required labeling. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule establishing new warning statements and other labeling information for all over-the-counter (OTC) vaginal contraceptive drug products (also known as spermicides, hereinafter referred to as vaginal contraceptives or vaginal contraceptives/spermicides) containing nonoxynol 9 (N9). These warning statements will advise consumers that vaginal contraceptives/spermicides containing N9 do not protect against infection from the human immunodeficiency virus (HIV), the virus that causes acquired immunodeficiency syndrome (AIDS), or against getting other sexually transmitted diseases (STDs). The warnings and labeling information will also advise consumers that use of vaginal contraceptives and spermicides containing N9 can irritate the vagina and rectum and may increase the risk of getting the AIDS virus (HIV) from an infected partner. This final rule is part of FDA's ongoing review of OTC drug products. FDA is issuing this final rule after considering public comments on its proposed regulation, and all relevant data and information on N9 that have come to our attention. PMID- 18159654 TI - Inside the Nobel committee on medicine: prize competition procedures 1901-1950 and the fate of Carl Neuberg. PMID- 18159655 TI - A review of development of cemeteries in Hong Kong: 1841-1950. PMID- 18159656 TI - Breaking bounds: Alice Profe, radical and emancipationist. AB - Alice Prof was the first female sport physician in Germany and she influenced ideas on female bodies and female sports for more than 40 years. Her dream to become a doctor could be realized only in Switzerland because women were not admitted to universities in Prussia before 1908. After her examination and her PhD she established herself as one of the first female doctors in Berlin in 1905 and she worked there until her death in 1946 as general practitioner and medical specialist for pediatrics and sport medicine. As an expert on the female body she was active in many committees and organizations. Alice Prof worked her whole life for the improvement of the situation of girls and women. The focus of her work was physical education and sport and she was always active in different types of sports herself. Her first article on this topic was published in 1906. In the following years over and over again she 'took the floor' criticizing traditional stereotypes of women and the female body, demanding resources for the education of girls and women. Her efforts also helped to reduce anxieties about women in sport and to encourage women to participate in sport. In her articles and in her lectures she encouraged women to decide for themselves about their involvement in sport. She rejected all attempts to patronize female athletes and refused to accept their exclusion from many sports. Prof propagated ideas which were not popular and she never swam with the tide. In one obituary it is stated: 'you never changed your ideas about life for material rewards or professional advancement. You stayed yourself'. Her ideas are relevant even today. PMID- 18159657 TI - Selling and saving "mother Iran": gender and the Iranian press in the 1940s. PMID- 18159658 TI - Neonatal intensive care utilisation by infants born to mothers older than 40 years of age: a 10-year review. AB - BACKGROUND: Increased maternal age is associated with increased rates of maternal hypertension, diabetes, growth restriction, and Caesarean section in pregnancy. However, there are few data on outcome and utilisation of neonatal resources by this group of infants. AIM: The primary aim was to determine how many infants born to women 40 years old or over were admitted for neonatal care at National Women's Hospital in Auckland, New Zealand during the period 1995-2004 inclusive. Secondary aims were to evaluate trends in admission, neonatal mortality, morbidity, and service utilisation. METHOD: Infants admitted for neonatal care and eligible for registration with the Australian New Zealand Neonatal Network were identified from the National Women's Neonatal Intensive Care Unit (NICU) database, trends were evaluated and resource utilisation estimated. RESULTS: For the study period, 203 infants were admitted following birth to women over 40 years. The median gestation was 33 weeks. Admissions increased to a peak in 2001 2002. A similar increase was seen in the total days in NICU and the percentage of total respiratory support used. Changes in referral practice from 2004 may have affected the infant numbers thereafter. CONCLUSION: Infants born to mothers over the age of 40 currently represent about 3% of total births but 5% of the infants requiring neonatal intensive care. PMID- 18159659 TI - Wives and mothers like ourselves? Exploring white women's intervention in the politics of race, 1920s-1940s. AB - This paper takes the issue of the removal of Aboriginal children, and the broader white anxiety over the 'half-caste problem' which underpinned the policy, to explore white women reformers' intervention in the politics of race in the years 1920-40. In these years middle-class women's citizenship was based on maternalism and the defence of motherhood. At the same time the national feminist lobby, the Australian Federation of Women Voters, joined the public debate about the 'Aboriginal problem'. In this context it is necessary to ask: What was the feminist view of Aboriginal women's status? Were they considered 'wives and mothers' like themselves, as Louisa Lawson suggested in the 1890s? What was their view of the 'half-caste problem' and the 'absorption proposal' formulated to solve it? By asking such questions I hope to modify the current feminist historiographical view of white women reformers as 'pro-Aboriginal' and 'radical' and their policies as challenging White Australia in these years. PMID- 18159660 TI - African voices from two world wars. PMID- 18159661 TI - Physical fitness and economic opportunity in the Bechuanaland Protectorate in the 1930s and 1940s. PMID- 18159662 TI - Development and death: reinterpreting malaria, economics and ecology in British India. PMID- 18159663 TI - Lymphoedema garment hosiery available on FP10 prescription. PMID- 18159664 TI - Compression hosiery: donning aids and garment removal. PMID- 18159665 TI - Best practice in managing scrotal lymphoedema. PMID- 18159666 TI - Dispelling myths concerning the humble compression stocking. PMID- 18159667 TI - Morphologic description of infectious salmon anaemia virus (ISAV)-induced lesions in rainbow trout Oncorhynchus mykiss compared to Atlantic salmon Salmo salar. AB - In the present study the pathogenesis of experimental infectious salmon anaemia virus (ISAV) infection in rainbow trout Oncorhynchus mykiss (Walbaum, 1972) and Atlantic salmon Salmo salar was compared. The virus infection in the 2 species demonstrated different mortality patterns and pathology characteristics. Atlantic salmon showed a typical acute mortality pattern peaking at 8 to 16 d post infection (dpi) depending on virus dose, whereas in rainbow trout, only the highest virus dose (10(7.13-7.8) TCID50/200 microl) showed a similar pattern. The middle (10(4.13) TCID50/200 microl) and lowest virus doses (10(2.13) TCID50/200 microl) in rainbow trout induced only sporadic protracted mortality, lasting up to 46 dpi. Infected rainbow trout that were live-sampled and those that died demonstrated increased erythrophagia, clusters of cellular degeneration in the haematopoietic portion of the kidney, and occasionally epicarditis, endocarditis and myocarditis. These lesions are very different from the typical necrosis in liver and kidney that occur in infected Atlantic salmon, and some of them may be indicative of an antiviral response by a resistant host to the ISAV infection. Virus was detected in the endothelium of the rainbow trout tissues using in situ hybridization, supporting our conclusions of the ISAV-induced lesions in this report. PMID- 18159668 TI - Pathogenesis of Herpesvirus anguillae (HVA) in juvenile European eel Anguilla anguilla after infection by bath immersion. AB - A clinical infection in post-larval (glass) European eels Anguilla anguilla was successfully induced after artificial bath immersion with Herpesvirus anguillae (HVA), isolated from diseased European eel. HVA caused a clinical infection after 7 d post-inoculation (pi); virus was detected by polymerase chain reaction (PCR) from Day 1 pi; virus isolation was positive from Day 7 pi, and HVA antigen was detected by immunohistochemistry in gills and stomach from Day 4 pi. Tissue changes were found by histological examination in gills and skin from Day 4 pi. In general, there was good correlation in the timing of the clinical signs, PCR, virus isolation, immunohistochemistry and histopathology results, although PCR, histopathology and immunohistochemistry were the first positive tests. HVA was first detected in skin and stomach, followed by gills, and later heart and intestine, whereas HVA was detected persistently in gills and skin. Koch's postulates were fulfilled. For diagnosis of HVA infections, clinical pathology combined with virus isolation and/or PCR are recommended. PMID- 18159669 TI - Detection of Cyprinid herpesvirus-3 (CyHV-3) DNA in infected fish tissues by nested polymerase chain reaction. AB - Diagnostic tests with high analytical sensitivity are required to detect Cyprinid herpesvirus-3 (CyHV-3) in carriers that have been implicated in the dissemination of this important disease of koi and common carp Cyprinus carpio. A nested polymerase chain reaction (PCR) test was developed to detect CyHV-3 DNA in tissues of infected fish. The 2-round PCR amplified 529 and 379 bp segments of the CyHV-3 gene coding for the major capsid protein using specific external and internal primers, respectively. The PCR test did not amplify genomic DNA from either Cyprinid herpesvirus-1 (CyHV-1) or Cyprinid herpesvirus-2 (CyHV-2). The nested PCR test detected CyHV-3 DNA at a 10-fold lower concentration than a routinely used, 1-round test. Hence, the new method should provide more sensitive diagnosis of CyHV-3 infection among koi and carp populations. PMID- 18159670 TI - Effect of an acute necrotic bacterial gill infection and feed deprivation on the metabolic rate of Atlantic salmon Salmo salar. AB - In this study, experiments were conducted to examine the effect of an acute necrotic bacterial gill infection on the metabolic rate (M(O2)) of Atlantic salmon Salmo salar. Fed and unfed Atlantic salmon smolts were exposed to a high concentration (5 x 10(12) CFU ml(-1)) of the bacteria Tenacibaculum maritimum, their routine and maximum metabolic rates (M(O2rout) and M(O2max), respectively) were measured, and relative metabolic scope determined. A significant decrease in metabolic scope was found for both fed and unfed infected groups. Fed infected fish had a mean +/- standard error of the mean (SEM) decrease of 2.21 +/- 0.97 microM O2 g(-1) h(-1), whilst unfed fish a mean +/- SEM decrease of 3.16 +/- 1.29 microM O2 g(-1) h(-1). The decrease in metabolic scope was a result of significantly increased M(O2rout) of both fed and unfed infected salmon. Fed infected fish had a mean +/- SEM increase in M(O2rout) of 1.86 +/- 0.66 microM O2 g(-1) h(-1), whilst unfed infected fish had a mean +/- SEM increase of 2.16 +/- 0.72 microM O2 g(-1) h(-1). Interestingly, all groups maintained M(O2max) regardless of infection status. Increases in M(O2rout) corresponded to a significant increase in blood plasma osmolality. A decrease in metabolic scope has implications for how individuals allocate energy; fish with smaller metabolic scope will have less energy to allocate to functions such as growth, reproduction and immune response, which may adversely affect the efficiency of fish growth. PMID- 18159671 TI - Metabolic effects of amoebic gill disease (AGD) and chloramine-T exposure in seawater-acclimated Atlantic salmon Salmo salar. AB - Our aim was to determine possible metabolic effects amoebic gill disease (AGD) on Atlantic salmon Salmo salar. Standard (R(S)) and routine (R(ROU)) metabolic rates were evaluated by continually measuring oxygen consumption in 2 independent tanks of fish (18.69 +/- 1.01 kg m(-3), mean +/- SE). Active metabolic rate (R(ACT)) and metabolic scope (R(ACT) - R(S)) were assessed using a chasing protocol and determined at 3 time periods: (1) pre-infection, (2) 3 d post-infection, and (3) 2 d post-treatment. On Day 3 of the study, the fish were infected with amoebae isolated from the gills of AGD-affected salmon (2300 cells l(-1)). No significant elevations in R(ACT) or metabolic scope were detected 3 d post-infection and 2 d post-treatment; however, significant elevations in R(S) and R(ROU) were detected 3 d post-infection and 2 d post-treatment. Assessment of R(ROU) data, especially for the light period, also indicated a rise in oxygen consumption rate over the course of the experiment. Treatment of AGD-affected Atlantic salmon with chloramine-T (CL-T) appeared to briefly mitigate the rise in R(S), as there was a 30% drop (though non-significant) in R(S) following treatment. Despite this, R(S) continued the upward trend 1 d following treatment. These results suggest that over the course of AGD development, R(S) in Atlantic salmon increases. Therefore, considering the physical conditions which constrain R(ACT), we expect that metabolic scope would become compromised in fish more heavily affected with AGD. Treatment with CL-T shows promise for mitigating the respiratory effects of AGD and potentially minimising the loss of metabolic scope. PMID- 18159672 TI - Site preference of myxozoans in the kidneys of Hungarian fishes. AB - Myxozoans are common parasites of fish kidneys, with most having specific sites of development. Five specific sites of development include (1) the lumen of renal tubules, (2) the renal corpuscles followed by location in renal tubules, (3) intracellular location within the tubular epithelium followed by a stage in the lumen of the ducts, (4) haematopoietic tissue with dispersed trophozoites, and (5) haematopoietic tissue with large, localized plasmodia. A coelozoic development preceded by presporogonic multiplication characterises most Sphaerospora spp. Early plasmodial stages of Myxidium and Chloromyxum spp. are frequently found in the renal glomerules, while spores develop in the urinary channels in plasmodia released from the renal corpuscles. In Hoferellus and Myxobilatus spp., spores are formed in small plasmodia inside the lumen of the urinary ducts after several internal cleavages in the epithelium of renal tubules. The presence of dispersed trophozoites among haematopoietic tissue cells of the renal interstitium characterises the development of Sphaerospora tincae and Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD). Spores of S. tincae are formed at the place of plasmodial development, while spore formation of PKD is in the renal tubules. A large mass of spores, often surrounded by a connective tissue capsule, can appear in the renal interstitium during infections by several Myxobolus spp.; furthermore, a large number of these spores formed in plasmodia in distant tissues can also accumulate in melano-macrophage centres. PMID- 18159673 TI - Non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on ELISA plate wells. AB - Enzyme-linked immunosorbent assay (ELISA) is a popular technique for quantifiable detection of specific antibodies in warm-blooded animals, but it has not been accepted for detection of fish antibodies because of its low reproducibility, which is due in part to high background optical density (OD) measurements. In the present study, we report that the high background of a fish antibody-detection ELISA resulted from non-specific adsorption of fish immunoglobulin M (IgM) to blocking reagents on the ELISA plate wells. Four fish sera (from rainbow trout Oncorhynchus mykiss, masu salmon O. masou, Japanese flounder Paralichthys olivaceus and koi Cyprinus carpio) were poured into ELISA plate wells pre-blocked with several blocking reagents (skim milk, soybean milk, bovine serum albumin, fetal bovine serum, gelatin and Roche BlockingReagent) and then washed out in order to measure the remaining fish IgM on the ELISA plate wells. Significant amounts of fish IgMs (OD absorbance at 492 nm: 0.3 to 1.1) remained on the ELISA plate wells with no antigenic protein except blocking reagents. The amount of remaining fish IgMs on the ELISA plate wells decreased significantly following treatment of fish sera with skim milk. However, the specific immuno-reactivity of fish IgM was not reduced by such treatment. Thus, we conclude that treatment of fish sera with skim milk is useful in reducing the high background OD often observed in fish IgM detection ELISA. PMID- 18159674 TI - Disease associated with integumentary and cloacal parasites in tadpoles of northern red-legged frog Rana aurora aurora. AB - A total of 6830 northern red-legged frog Rana aurora aurora tadpoles were examined under a dissecting microscope for oral disc, integumentary, and cloacal abnormalities in 13 ponds in and near Redwood National Park in northern California. Of these, 163 tadpoles were collected for histopathological investigation, including 115 randomly collected individuals, 38 collected with oral disc abnormalities, and 10 collected due to severe morbidity of unknown etiology. The tadpoles were infected with 8 parasites, including Batrachochytrium dendrobatidis (the amphibian chytrid), trematodes, leeches, and protozoa. Chytridiomycosis was detected at an overall prevalence of 6.4%, but prevalence was higher in tadpoles with oral disc lesions than in those with normal oral discs (43.5% versus 6.1%). Interestingly, infection was associated with some environmental and co-infection risk factors. Individual tadpoles possessed 0 to 5 species of parasites in varying intensities. Apiosoma sp. was the most prevalent (66%) and widespread. Tadpoles infected with B. dendrobatidis had a lower diversity of oral parasites than those uninfected. During the field portion of the study, a large number (approximately 500) of moribund and dead tadpoles was seen occurring at multiple locations within and surrounding Redwood National Park. Ten animals were collected for histological examination and a diverse protozoal infection was discovered, including some known pathogens of fish. This study is the first reporting parasitism and disease in natural populations of northern red-legged frogs. PMID- 18159675 TI - Histopathology of the mangrove land crab Ucides cordatus (Ocypodidae) affected by lethargic crab disease. AB - Lethargic crab disease (LCD) has caused extensive epizootic mortality of the mangrove land crab Ucides cordatus (Linnaeus 1763) (Brachyura: Ocypodidae) along the Brazilian coast. Direct culture of tissue samples from sick crabs and subsequent isolation and purification identified the causative agent as an Exophiala species of fungus. The histopathology of crabs with variable signs of LCD indicates that the most affected tissues are the epidermis, connective tissue, heart, hepatopancreas, nervous system, and gills. Gonads, somatic muscles, and digestive system are less affected by the fungus. The observed pathology is compatible with the clinical signs of LCD. Necrosis, tissue degeneration, and congestion of hemal sinuses and vessels are present in heavily infected organs. Nerve fibers may be compressed by accumulations of yeast-like cells. In heavy infections the tissue of gill lamellae is destroyed with subsequent dilation or compression. Cellular immune responses include hemocytic infiltration, agglutination and encapsulation, and phagocytosis. Phagocytosis of yeast-like cells is abundant in the connective tissue associated with the exoskeleton. These results indicate that LCD is the result of a systemic phaeohyphomycosis caused by a species of Exophiala. The present study also suggests that dispersal of the fungus within the crab occurs through the hemal system. PMID- 18159676 TI - Meagre Argyrosomus regius (Osteichthyes) as host of a gonad-infecting species of Philometra (Nematoda: Philometridae) off the Atlantic coast of Portugal. AB - Subgravid females (up to 439 mm long) of the nematode Philometra sp. were found in meagre Argyrosomus regius (Asso, 1801) (Sciaenidae: Perciformes) off the southern Atlantic coast of Portugal in 2006. The general morphology of these nematodes somewhat resembles that of Philometra lateolabracis (Yamaguti, 1935), but the gravid females of the species from A. regius are apparently much longer. This is the first documented record of a gonad-infecting species of Philometra in marine fishes off the Atlantic coast of Europe. The possible importance of the gonad-parasitizing Philometra spp. as pathogens of marine fishes is stressed. PMID- 18159677 TI - [*In Italy, a law now permits the opening up of the nursing profession"]. PMID- 18159678 TI - [Giving space in the hospital and developing a territorial organization for nurses]. PMID- 18159679 TI - [An exciting stage in the Parkinson unit]. PMID- 18159680 TI - [Polarity of organization or how the contemplate public hospitals]. PMID- 18159681 TI - [Conducting change in the hospital]. PMID- 18159682 TI - [The contract of polarity in practice]. PMID- 18159683 TI - [Polyvalence in the network of polarity, wager or reality?]. PMID- 18159684 TI - [Thoughts of nurses on work in poles]. PMID- 18159685 TI - [Reflection of the function of activity poles]. PMID- 18159686 TI - [Activity poles, an opportunity to seize]. PMID- 18159687 TI - [Urinary tract sounding: position and surveillance]. PMID- 18159688 TI - [Choosing an instructor on blood glucose control in the hospital milieu]. PMID- 18159689 TI - ["Full of back" or how to manage patients]. PMID- 18159690 TI - [Dany Guillou: companion of health without permanent housing]. PMID- 18159691 TI - [Religious laxity in the hospital]. PMID- 18159692 TI - [The nurse and the network of work]. PMID- 18159693 TI - [Louise de Marillac (1591-1660). ]. PMID- 18159694 TI - [Anxiolytics]. PMID- 18159695 TI - [Heparinemia (anti-factor Xa activity]. PMID- 18159696 TI - [Driver fatigue]. PMID- 18159697 TI - [Pulmonary embolism: initial management]. AB - Pulmonary embolism is a frequent disease with well known acquired or hereditary risk factors. The first diagnostic step is to determine a pre-test probability of the disease based on the revised Geneva score. Further work up with D-Dimers, blood gases and ECG, will ascertain the diagnostic of the condition. The prognosis will be better evaluated with BNP and troponine. Negative D-Dimers can rule out pulmonary embolism when pre test probability of the disease is low or intermediate. Radiological work-up includes computed tomographic pulmonary angiography when D-Dimers are positive and when the pre test probability is high, or ventilation/perfusion scan in case of iodine allergy or renal failure. Finally, guidelines will help choosing which patients suffering of pulmonary embolism have to undergo a hematological assessment or oncologic screening. PMID- 18159698 TI - [Long-term oxygen therapy]. AB - Two studies showed that continuous long-term oxygen therapy (> 15 h/d) undoubtedly confers a significant benefit on survival in hypoxemic patients with chronic obstructive pulmonary disease. This treatment should be prescribed to stable patients with a PaO2 < 7.3 kPa or a PaO2 < 8 kPa and either previous episodes of ankle edema, pulmonary hypertension or a haematocrit > 55%. Patients who are hypoxemic only during exercise can also benefit from mobile oxygen therapy. However, the Swiss Society of Pneumology does not recommend treating patients with isolated nocturnal hypoxemia, unless they suffer from central apnea and are shown to be responders. The characteristics of the three methods of oxygen delivery for domiciliary treatment are discussed in this review. PMID- 18159699 TI - [Current approach to bronchial carcinoid tumors]. AB - Neuroendocrine tumors of the lung comprise typical and atypical carcinoids, as well as large cell neuroendocrine carcinomas and small cell carcinomas. However carcinoid tumours differ from the latter two by their clinical presentation, radiographic, histological appearances, and treatment modalities as well as prognosis. In this paper we review these features and discuss the role of new imaging modalities such as PET scan. PMID- 18159700 TI - [Phenotypes of severe persistent asthma in adults]. AB - The clinical features of asthma may vary very widely between individuals. Among those with severe persistent asthma, one can also distinguish different phenotypes. This distinction is important both for the understanding of the disease and for the treatment. The present review will briefly discuss corticoid resistant asthma, occupational asthma, aspirin-induced asthma, and the association of asthma with obesity. PMID- 18159701 TI - [Obstructive sleep apnea: a risk for general anesthesia?]. AB - There are many case reports of serious complications and death among obstructive sleep apnea patients (OSA) during general anesthesia or postoperative analgesia. Sedatives and anesthetic agents, pharyngeal anatomy of these patients, opiates given for analgesia, and post operative REM sleep rebound represent potential hazards for general anesthesia in OSA patients. Ideally these patients should be treated with continuous positive airway pressure (CPAP) during premedication, directly after extubation and during postoperative analgesia. Unfortunately, only about 20% of these patients are diagnosed before surgery. A special attention should be given to the symptoms and signs suggestive of OSA during preoperative visits. Screening tests should be performed in patients with suspected OSA and, if positive, a treatment should be initiated. PMID- 18159702 TI - [The antisynthetase syndrome: a cause of rapidly progressive interstitial lung disease]. AB - The anti-synthetase syndrome is associated with inflammatory myopathy, interstitial lung disease, polyarthritis, Raynaud's phenomenon and typical skin lesions known as "mechanic's hands". Anti-Jo1 antibodies are frequently present. Interstitial lung disease is the most important feature, as it determines survival and the therapeutic attitude. We report here two cases of anti synthetase syndrome with acute respiratory failure. Immunosuppressive treatment combining corticosteroids and cyclosporine lead to clinical improvement, regression of radiological lesions and improvement in pulmonary functions. PMID- 18159703 TI - [The establishment of great medicine: Louis-Ferdinand Celine]. PMID- 18159704 TI - [Assisted suicide or a slippery slope]. PMID- 18159705 TI - [Hospitals, ethics and money: health services accessibility]. PMID- 18159706 TI - [The fight against the AIDS-tuberculosis co-epidemic (2)]. PMID- 18159707 TI - [Judgment of legal insanity]. PMID- 18159708 TI - [Obesity and global warming]. PMID- 18159709 TI - [Compact MRI]. PMID- 18159710 TI - [Management of osteoporosis by using biochemical markers for bone turnover]. PMID- 18159711 TI - [HIP study--how to prevent hip fractures?]. PMID- 18159712 TI - [Where do SERMs stand in the light of clinical application?]. PMID- 18159713 TI - [First choice drug for treatment of osteoporosis and the strategic points for choice of drugs]. PMID- 18159714 TI - [Diagnosis and treatment of immobilization osteoporosis]. PMID- 18159715 TI - [Serum vitamin D as an indicator of fall-related physical function among the elderly]. PMID- 18159716 TI - [How to increase bone mass during childfood]. PMID- 18159717 TI - [Treatment guideline for breast cancer and SERM-treatment]. PMID- 18159718 TI - Oral-health-related quality of life: an overview for the general dental practitioner. PMID- 18159719 TI - Mandibular overdentures: a review of treatment philosophy and prosthodontic maintenance. AB - Root overdentures and implant overdentures are two similar treatment options for the nearly-edentulous or edentulous mandible. The purpose of this literature review was to specifically compare their prosthodontic treatment philosophies and follow-up maintenance requirements. Critical comparison of these two prosthodontic treatment philosophies revealed that the foundation for overdentures which is provided by oral implants is more predictable than that provided by the roots of natural teeth. The two treatment modalities have both similarities and differences in their associated prosthodontic maintenance. The findings of this literature review suggest the future possibility that mandibular root overdentures may become obsolete as a treatment approach. PMID- 18159720 TI - A case of recurrent pyogenic granuloma. AB - Dental practitioners are usually the first health professionals to examine and diagnose patients with oral pyogenic granuloma. Usually of traumatic origin, the lesion results from an overly aggressive immune response. Despite several accepted methods of treatment, recurrence is common. This paper reports on a case of recurrent pyogenic granuloma and reviews the relevant literature. PMID- 18159721 TI - Natural rubber latex allergy and dental practice. AB - Natural rubber latex (NRL) allergy is a significant clinical problem with potentially life-threatening complications. Oral health care professionals must be able to screen for NRL allergy and refer patients or staff to a specialist for definitive diagnosis. Protocol for its management must be developed and incorporated into daily practice. Practitioners must be able to recognize and treat NRL exposure emergencies. Knowledge of the availability of substitute products and an adequate fresh stock of such products in dental practice can minimise the risk of adverse NRL sensitivity. PMID- 18159722 TI - Eighty years ago... PMID- 18159723 TI - Time to go. PMID- 18159724 TI - Support needed for nurses offering HIV counselling. PMID- 18159725 TI - Australian health care reform: a place for nurse practitioners? AB - Nurses and health professionals across the board have argued for many years that the current health care system is not sustainable and reform is necessary if access to quality health care for all Australians is to continue. PMID- 18159727 TI - Working holiday inspires career in public health. PMID- 18159726 TI - A busy year promoting nursing and health. AB - The recent federal election has been a central and consuming focus of the ANF and the professional team this year. Advocating for professional nursing and midwifery issues at a national level has involved collaboration with nursing colleagues, other health professionals and consumers. PMID- 18159728 TI - Improving retention in the NT. PMID- 18159729 TI - Topical skin care in aged care facilities. PMID- 18159730 TI - [Chlorine dioxide as an agent for optimization of drinking water preparation]. AB - The paper deals with the important urgent problem in the provision of epidemiological drinking water. It considers the chemical, biological, toxicological, technological, environmental, and economic aspects of the use of chlorine dioxide as a highly effective disinfecting and oxidizing agent in water purification and decontamination processes. The authors present the results of their own studies. They conclude that chlorine dioxide is an epidemiologically, ecologically, and toxicologically safe and technologically adequate and economically acceptable agent for water treatment in the domestic water supply systems of localities. PMID- 18159731 TI - [The need for developing a methodology of spatial rationale for a sanitary protective zone]. PMID- 18159732 TI - [Assessment of the risk of influence of environmental factors on the population's health under the conditions of a large center of chemical industry]. PMID- 18159733 TI - [Hygienic diagnosis of abiotic microbiocenotic changes under the present conditions]. PMID- 18159734 TI - [Ambient air pollution and respiratory diseases in the population of Novodvinsk]. PMID- 18159735 TI - [Prevalence of cancer in the Primorye Territory: ecological and hygienic aspects]. AB - By using the system analysis, the authors estimate the prevalence of cancer diseases upon exposure to many environmental factors. They describe a formula of the integrated exposure index. Application of the information-aentropion analysis identifies habitat modules that affect cancer morbidity. The findings may be used when developing a regional "Oncology" program. PMID- 18159736 TI - [Bioindicator role of plants]. PMID- 18159737 TI - [Problem in the planting of greenery in residential areas]. PMID- 18159738 TI - [Global hydroclimatic changes and the problems of the population's health in the coastal towns]. PMID- 18159739 TI - [Provision of the rural population with high-quality drinking water: hygienic aspects]. PMID- 18159740 TI - [Medicoecological studies in the assessment of biogeochemical province in the area of a toxic waste ground]. AB - The paper presents the results of complex sanitary-and-epidemiological, toxicological-and-hygienic, and medical diagnostic studies of health and the environment, by taking into account the activity of a toxic industrial waste ground. Toxic waste burning on primitive unfiltered apparatuses was shown to lead to the formation of a biogeochemical province that is characterized by pollution of soil, bottom sediment, subsoil well water and snow with heavy metals, the components of toxic waste. Burning of waste and its storage in the open trenches resulted in ambient air pollution with organic solvents, nitric oxide and sulfur oxide had a negative impact on the health of children living at a distance of 3 km from the ground. PMID- 18159741 TI - [Control of industrial waste consumption residues: ecological and hygienic aspects]. AB - The problem in the provision of safe handling of industrial waste and consumption residues is relatively current. According with the United Nations Organization's data, 25 to 33% of the world's notified diseases are directly associated with the low quality of the human environment. Up to now, a list of chemicals encountered in the waste and residues is unavailable in Russia and foreign countries. By keeping in mind the ubiquitous spread of industrial waste and consumption resides due to human vital activity, their huge formations and their very wide diversity in composition, type, and pattern of a possible dangerous effect, it is important to consider the problem associated with waste handling, by evaluating their environmental and hygienic hazard. PMID- 18159742 TI - [The functional status of the hepatobiliary system in workers upon occupational exposure to methylene chloride]. PMID- 18159743 TI - [Hygienic evaluation of the effects of petroleum hydrocarbons on crops]. AB - Experience in standardizing the soil levels of chemicals suggests that the translocation hazard index is limiting for most standardized substances. The current unavailability of scientifically grounded maximum allowable concentration of soil petroleum generates a need for studies using this hazard index. The studies were made to examine different soil petroleum levels, by employing various test crops. PMID- 18159744 TI - [Comparison of the acute toxicity of a class of alkylphenols]. PMID- 18159745 TI - [Hygienic evaluation of the effect of petroleum hydrocarbons on the soils of megapolises]. AB - The paper gives the results of studying the quantity of petroleum hydrocarbons in the Moscow soil, as well as the latter's hygienic evaluation by the changes in acidity (pH) and redox potential and by the sanitary state (the levels of Escherichia coli and viable helminthic eggs). The study has ascertained that the petroleum-polluted soils show changes in medium pH and redox conditions that determine the biological activity of the substances and the trend in their disintegration processes and revealed a relationship of thy sanitary state of Moscow soils to their level of petroleum hydrocarbons. PMID- 18159746 TI - [Buccal epithelial micronuclear test in the comprehensive assessment of children's ecological well-being in Moscow]. AB - In the buccal epithelial micronuclear test, only the higher frequency of cells with any nuclear degenerative changes is considered to be attributable to the adverse influence of environmental factors. Comprehensive examination of 223 children of 5-7 years of age, going to kindergartens and schools has indicated that the frequency of cells with nuclear chromatin condensation and incomplete nuclear lysis decreased when ambient air contamination was increased in the territories of children's establishments and when the child's throat was contaminated with Staphylococcus aureus or the pharyngeal resident microflora inhibited. Some poor conditions of the nasal and buccal mucosae were also associated with the increase or decrease of cell frequencies. It is suggested that nuclear chromatin condensation and nuclear lysis (rather than other signs of cell degeneration and death) reflect natural cell aging. A moderate toxic effect causes epitheliocytic physiological development retardation and a more intensive effect accelerates aging of cells and induces their death. PMID- 18159747 TI - [Characteristics of congenital morphogenetic variants in hygienic studies]. AB - The study was undertaken to examine the basic frequencies of congenital morphogenetic variants (CMVs) among preschool children. The basic frequencies were valid if the population frequency was more than 2-4 %. The least valid frequencies were characteristic of rare signs at a frequency of less than 1%. It is suggested that rare signs be integrated into an individual group and then their average be compared. PMID- 18159748 TI - [Track membranes in the filtration technique of determination of the quality of water from bacteriological values]. AB - The paper describes a new class of microfiltration track membranes (TMs) proposed for laboratories to carry out a sanitary water bacteriological test. TMs have a strictly reference pore structure, a low sorption capacity do not contain the components that able to migrate into the filtrate, they are noted for a screening mechanism of holding microparticles onto the surface. The TMs made in Russia, the reticular membranes "Vladipore", and the foreign Millipore membranes certified by the international standards were comprehensively assessed. TMs are superior to the membranes manufactured in Russia at the moment in a number of basic properties that are most essential for the study of water quality and to foreign certified membranes in physical parameters, the specific features of growth of coliform bacteria, the rate of performance of an oxidase test, and the efficiency of retention of various microorganisms. PMID- 18159749 TI - [Problem in compensation for socioecological damage]. AB - The paper considers theoretical approaches to and rules of thumb for compensation for socioenvironmental damage at the municipal level. There is evidence for the necessity of designing a uniform present-day package of methods to calculate the damage with a uniform complex of evaluation of all factors, both quantitative and non-quantitative ones. The authors propose to finish off early developed procedures to specify and quantify a health-exposure relationship, and to have a more adequate market economic interpretation of the above phenomena. PMID- 18159750 TI - [Alcoholic hepatosis in pathogenesis of hepatic encephalopathy]. AB - The problem of chronic alcoholism has a great social importance. In the literature there are not methodological grounds for verification of causes and mechanisms of unexpected death caused by chronic alcoholic intoxication. Given literature data show the reasonability of study of hepatic encephalopathy in combination with pathomorphological, histochemical and biochemical analyses in comparison with forensic chemical analysis of alcohol concentration in human organism. That will make possible to determine the mechanisms of tanatogenesis of alcoholic intoxication. PMID- 18159751 TI - [Morphofunctional interaction of adrenal zones in acute lethal carbon monoxide poisoning in people with alcoholic intoxication]. AB - Morphological examination of human epinephroses at acute lethal poisoning with carbon monoxide against a background of alcoholic intoxication was done. Acute lethal poisoning with carbon monoxide decreases effect of alcoholic intoxication upon morphofunctional state of epinephroses and disorders characteristics of interzonal interaction. Interaction between zones of one epinephros recedes but interaction between both glands increases. PMID- 18159752 TI - [Criteria of dimensional typology of the face and body constitution of young men]. AB - On the grounds of results of combined anthropometrical examination of 1108 Russian juveniles 17-21 years old the relationship between face dimensions and various complex of somatic characteristics and constitutional types from Shtefko Ostrovsky scheme was studied. The medium-sized relationship (correlation coefficient less 0.6) between parameters of various morphological subsystems "face-body" was established. Two integrative indices describing general face size and shape were calculated with the method of factor analysis. The tendency to macroprosopy in the row from asthenoid to digestive constitutional type from Shtefko-Ostrovsky scheme (p < 0.001) was revealed with the help of various statistic methods. Estimating equations to determine individual variant of general face size and shape and male somatotype by V. V. Bunak and Shtefko Ostrovsky schemes were done for practical use the received results. PMID- 18159753 TI - [Detection of antimony topography using diffuse-contact method on the skin targets of bio-manikins after shooting with Makarov gun from short distance]. AB - The authors examined sedimentation of stibium in the area of experimental skin entry holes generated by Makarov gun shots from different distances. Regularities of allocation of stibium subject to the distance of the shot were determined. Practical recommendations for expert conclusion about the distance of the shot were given on the grounds of the results of the study. PMID- 18159754 TI - [Possibility of the species identification using blood stains located on the material evidences and bone fragments with the method of solid phase enzyme immunoassay with "IgG general-EIA-BEST" kit and human immunoglobulin G]. AB - The authors give the comparative analysis of Russian and foreign forensic medical methods of species character identification of the blood from the stains on the material evidences and bone fragments. It is shown that for this purpose it is feasible to apply human immunoglobulin G (IgG) and solid phase enzyme immunoassay (EIA) with the kit "IgG general-EIA-BEST". In comparison with the methods used in Russia this method is more sensitive, convenient for objective registration and computer processing. The results of experiments shown that it is possible to use the kit "IgG general-EIA-BEST" in forensic medicine for the species character identification of the blood from the stains on the material evidences and bone fragments. PMID- 18159755 TI - [Applicability of elemental composition of bone and dental tissue for human identification]. AB - The authors give the information about elemental composition (levels and concentration ranges of chemical elements) of human bone and dental tissue in the range from sodium to uranium, compare elemental composition of bones and teeth, make conclusion about applicability of bone and dental tissue for of human attribute diagnostics. PMID- 18159756 TI - [Criteria of expert assessment of professional errors in dentistry]. AB - The most frequent errors in dental practice are analysed. Questionnaires, schemes and formula for objective assessment of medical documentation quality in dentistry are proposed. PMID- 18159757 TI - [Forensic-medical evaluation of medical care errors in obstetric-gynecological practice]. AB - The authors show the necessity of elaboration and practical application of normative documents (standards) and forensic-medical criterions of assessment of defects in obstetric-gynecological medical aid. The examples of the above assessment are given in the article. PMID- 18159758 TI - [Toxicological characteristic of neuroleptics--substituted benzamides]. AB - At present in the medical practice the group of neuroleptics, substituted benzamides is widely used. According to literature data they cause acute intoxications under particular conditions. The group of substituted benzamides includes drugs such as amisulpride, tiapride, sulpiride and sultoprid. Substituted benzamides intake causes toxic actions: psychotropic and neurotoxic. The neuroleptic intoxication is an effect of overdosage, abuse and hypersensitization. Severe intoxications with drugs of this group may cause lethal outcome. PMID- 18159759 TI - [Use of special medical knowledge in civil court practice]. PMID- 18159760 TI - [Determination of identification signs--tattooing on decomposed exhumed corpse]. PMID- 18159761 TI - [Innovation technologies in teaching of discipline "Forensic Medicine" in I. M. Sechenov Moscow Medical Academy]. PMID- 18159762 TI - [Results of crush-tests and their forensic-medical significance]. PMID- 18159763 TI - [Contribution of Military Medical Academy to formation of St. Petersburg Forensic Medical Expert service]. PMID- 18159764 TI - [Quantitative positioning of facial nerve based on three-dimensional CT image reconstruction of temporal bone]. AB - OBJECTIVE: To explore a set of quantitative methods to determine the position of the facial nerve based on three-dimensional CT reconstruction of temporal bone structures on personal computer, which can provide a series of important parameters for ear and the lateral skull base surgery. METHOD: The internal structures of temporal bone were reconstructed based on a set of axial CT images of adult patients, the complicated relationship and their morphologic characteristics were clearly presented by using Able Software 3D-DOCTOR. The precise measurement of some parameters between facial nerve and its adjacent structures could easily be processed with the software. Based on all obtained data, the relationship of facial nerve and its adjacent structures were effectively summarized and analyzed. RESULT: Three-dimensional images, including the facial nerve, tympanic anulus, auditory ossicles, chochleariform process, pyramidal eminence, internal auditory, the cochlea, semicircular canal, jugular fossa and carotid artery in the temporal bone, were reconstructed. Some parameters obtained from measuring the distance or angle between the facial nerve and its adjacent structures in the three-dimensional models had some extent regularity, which were benefit to design surgical approach and determine the position of facial nerve during relevant operation. CONCLUSION: CT 3D reconstruction can accurately display the detailed internal structures anatomy of the temporal bone and their quantitative spatial relationships. PMID- 18159765 TI - [The study on the microsurgical anatomy of maxillary sinus approach pterygopalatine fossa]. AB - OBJECTIVE: To provide morphologic data for the operation of pterygopalatine fossa (PPF) region by measuring the bony landmarks and dissecting vessel and nerve. METHOD: The shape, size and adjacent important structures of PPF were observed in 20 dry skulls (forty sides). Ten (twenty sides) skulls were fixed in 4% paraformaldehyde and their arteries were perfused with red lacteprene. The pterygopalatine fossa was exposed from the middle sagittal section dissected to inner wall of maxillary sinus and PPF and observed the course of trigeminal nerve and arteria maxillaris under the operating microscope. To investigate the related surgical anatomy approaches of the maxillary sinus and pterygopalatine fossa. RESULT: The pterygopalatine fossa shows a long and narrow cleft. PPF communicate with nasal cavity, oral cavity, pharynx, orbit, infratemporal fossa and middle cranial fossa. The pterygopalatine segment of maxillary artery and accompanying vein, the maxillary nerve and sphenopalatine ganglion are the most important structures. There are two layers of tissue in the pterygopalatine fossa, they are nervous layer and vessel layer. CONCLUSION: (1) To deal with lesion, the approach of maxillary sinus can de used, the cross point of the upper wall, the inner wall and the back wall can be thought as important landmark. (2) Safe scope could be acquired in endoscopic transnasal surgery in pterygopalatine fossa, pterygoid canal and foramen rotundum could be thought as an important landmark. PMID- 18159766 TI - [A clinical study of the influence on the drying of the ear canal by cleaning the focus of infection in the posterior tympanum]. AB - OBJECTIVE: To investigate the influence of drying the ear canal by cleaning the focus of infection in the posterior tympanum in middle ear operations. METHOD: Retrospectively analyze 168 cases of middle ear operations, which were divided into two groups by whether the posterior tympanum was open or not in the operation process. Then comparing the drying ear rate. RESULT: The drying ear rate (DEC) were 81.7% (67/82) and 91.9% (79/86) in the posterior tympanum unopened and opened group, respectively. In the posterior tympanum unopened group, the DEC rate were 78.9% (30/38) and 84.1% (37/44) in 38 cases of chronic otitis media and mastoiditis with cholesteatoma and in 44 cases of chronic otitis media and mastoiditis with granulation tissue, respectively. In the posterior tympanum opened group, the DEC rate were 88.2% (37/42) and 94.7% (42/44) in 42 cases of chronic otitis media and mastoiditis with cholesteatoma and in 44 cases of chronic otitis media and mastoiditis with granulation tissue, respectively. In the posterior tympanum unopened group, the DEC rate were 81.7% (36/44) and 84.6% (34/38) in 44 cases of radical mastoidectomy and in 38 cases of tympanoplasty; compared with the posterior tympanum opened group, the DEC rate were 91.3% (42/46) and 92.5% (37/40) correspondingly. CONCLUSION: In the middle ear operation, open the posterior tympanum in convention can not only have profits to clear the insidious focus of infection, but also have profits to skeletonized to lower facial ridge to the utmost limits and make the drainage of the ear canal easy and smooth. At last, the rate of DEC was raised. PMID- 18159767 TI - [Application of auditory brainstem response and distortion product otoacoustic emission in the clinical detection of hearing loss in patients with diabetes mellitus]. AB - OBJECTIVE: To study the characteristic of hearing loss and the changes of the auditory brainstem response and distortion product otoacoustic emission in diabetics. METHOD: General information of 136 diabetics were investigated whether had the complication of capillary vessel and hearing loss with self-made questionnaire at random. 136 cases (272 ears) were tested with ABR and DPOAE. They were divided into A, B, C, D four groups according to the age of per ten years (30-70 years). Other 120 (240 ears) matched healthy adult were enrolled in the control group. The pure tone threshold (PTT), acoustic impedance, ABR and DPOAE results were compared between the four groups and the control group, and then made statistical analysis. RESULT: When comparing to the control group, the interpeak latency I-V and ABR response were no significant difference between patients at the stage of 30 years. Not only the threshold of wave V was elevated with aging in patients over 40 years, but the amplitude of the peak latencies of waves III and V were lessening, and the interpeak latency III-V and I-V were prolonged between patients at the stage of 60 years, which had significant difference compared with age-matched patients (P < 0.05). The inducing rate of DPOAE were 100% in the test and control group, but the amplitude of DPOAE declined ranged from 1 to 8 kHz in patients with early diabetes mellitus and were significantly reduced at 4 kHz, which had significant difference compared with age-matched patients (P < 0.05). CONCLUSION: The majority of patients with diabetes mellitus had slight and moderate hearing loss gradually. The ABR and the threshold of ABR response were varied with the age and sex of patients, and whether had the complication of capillary vessel, but were not related to its type and disease course. The DPOAE can comprehend the cochlear disorder in patients with early diabetes mellitus. PMID- 18159768 TI - [Image date and clinicopathologic features of myoepithelial carcinoma in parapharyngeal space]. AB - OBJECTIVE: To study the image data and clinicopathologic features of myoepithelial carcinoma in parapharyngeal space. METHOD: Based on one patient with myoepithelial carcinoma of parapharyngeal space treated in our hospital, image data and clinicopathologic findings were analyzed. The clinical characteristics of myoepithelial carcinoma in head and neck region were discussed. RESULT: CT obviously showed the mass in parapharyngeal space with destruction of temporal bone. Immunohistochemical stain showed positive staining for S-100 protein, Smooth muscle actin, Cytokeratin and Vimentin. There were otalgia, bloody pus, headache and signs of cranial nerve lesion. CONCLUSION: Myoepithelial carcinoma was a rare malignant epithelial neoplasm and had a poor prognosis. Myoepithelial carcinoma often occurrence in salivary gland of maxillofacial region. The clinical signs were otalgia and headache in patient with myoepithelial carcinoma in parapharyngeal space. CT image showed malignant neoplasm and surrounding tissues were destroyed. Diagnosis depends on pathology and immunohistochemistry. PMID- 18159769 TI - [The audiograms of 462 tinnitus victims who never perceived hearing loss]. AB - OBJECTIVE: To investigate the risk factors of tinnitus and provide evidence for tinnitus prevention. METHOD: Retrospectively analyzed the audiograms and the risk factors of 462 tinnitus victims who never perceived hearing loss, find the relationship between every risk factor and tinnitus. RESULT: The rate of tinnitus victims with high frequency hearing loss was the 46.1%, with low frequency hearing loss was 15.8%, with single middle frequency notch was 11.04%, with an audiogram like upset-basin was 7.58%, with normal audiogram was 14.07%, with other type of audiograms was 5.41%. The age distribution in the high frequency hearing loss group was mainly between 30 and 40 years, in the upset-basin hearing loss group was mainly older than 50 years, and in the other groups, mainly between 20 and 30 years. In the high frequency hearing loss group, male to female ratio was 158/55, in the low frequency hearing loss group and single middle frequency notch group ,male to female ratio was 14/59 and 12/39 respectively. In another three groups, the male to female ratio was nearly 1. Risk factors for high frequency hearing loss group, low frequency hearing loss, single middle frequency notch group and in the upset-basin hearing loss group were noise, fatigue, long-time musical instrument contact and ageing respectively. CONCLUSION: Among the tinnitus victims who never perceived hearing loss, 86% of them have hearing impairment in high frequency area, low frequency area or single middle frequency. The rate of high frequency hearing loss was 46.01%, the major risk factor was noise, most of victims were men. The rate of low frequency hearing loss was 15.8%, the major risk factor was fatigue and nervous, most of the victims were women. The rate of single middle frequency notch was 11.04%, the major risk factor was long-time musical instrument contact. The rate of upset basin hearing loss was 7.58%, ageing was the major risk factor. PMID- 18159770 TI - [The management of the stoma in endoscopic dacryocystorhinostomy]. AB - OBJECTIVE: To explore the better management of the stoma in endoscopic dacryocystorhinostomy. METHOD: To review the 102 chronic dacryocystitis patients (109 eyes), who underwent the endoscopic dacryocystorhinostomy surgery with silver clips used to maintain the stoma. They were given combined therapy after the surgery, and were followed up for a period range from 3-73 months. RESULT: 99/109 eyes (91%) were cured, 5/109 eyes (4.5%) were improved, and the total effective rate reached to 104/109 (95.5%). CONCLUSION: The application of silver clip in endoscopy dacryocystorhinostomy surgery and combined therapy after the surgery can effectively prevent the stoma stenosis or atresia. PMID- 18159771 TI - [Effect of cisplatin on the potassium channels of spiral ganglion neurons from neonatal rats]. AB - OBJECTIVE: To investigate the effect of cisplatin on outward delayed rectifier potassium currents (IKDR currents) in isolated spiral ganglion neurons cells and to analyze possible intracellular mechanism of this effects by using the patch clamp technique. METHOD: The IKDR was recorded and measured before and after the application of 10 micromol/L cisplatin in external solution by using the whole cell patch-clamp technique. RESULT: Cisplatin could inhibit SGNs voltage dependent potassium channels, the zero current potential and reversal potential of IKDR currents were both shift to polarizing directions by 10 micromol/L cisplatin. Moreover, the maximal magnitude of IKDR currents were decreased by 23.3% when giving the test pulse at +50 mV. Cisplatin's inhibition on IKDR has dose-dependence with cisplatin-concentration in extracellular fluid and the currents recovered completely after cisplatin being washed out. CONCLUSION: This research explained the toxic mechanism of cisplatin through its action on keeping from spiral ganglion neuron's IKDR from the electrophysiological aspect and set a foundation for further research. PMID- 18159772 TI - [Inhibition of salicylate on voltage-gated sodium channels in rat inferior colliculus neurons]. AB - OBJECTIVE: To investigate the mechanism of the tinnitus inducer, sodium salicylate, on voltage-gated sodium channels. METHOD: The effects of salicylate on voltage-gated sodium channels in freshly dissociated inferior colliculus neurons of rats were studied, using the whole-cell voltage clamp method. RESULT: Salicylate blocked sodium current (INa) in concentration-dependent manner (0.1-10 mmol/L). The IC50 value for the blocking action of salicylate was 1.43 mmol/L. Salicylate did not affect the conductance-voltage curve and the steady-state activation curve of INa. The steady-state INa inactivation curve of INa was shifted by about 9 mV in the hyperpolarizing direction. In addition, salicylate delayed the sodium channel recovery from INa inactivation by increasing the slow time constant. CONCLUSION: Our results suggest that salicylate causes a concentration-dependent blockade of INa and shifts the INa inactivation curve to more hyperpolarized potentials, which could be related to the mechanism of salicylate-induced tinnitus. PMID- 18159773 TI - Methadone maintenance therapy in residential therapeutic community settings: challenges and promise. AB - The therapeutic community (TC) and methadone maintenance treatment (MMT) have individually demonstrated consistent positive outcomes yet rarely have been combined. This article describes how a well-established residential therapeutic community integrated methadone maintenance treatment into its activities. Practical recommendations regarding how to incorporate MMT in a residential program are provided including topics for staff (training, coordination with the methadone service provider agency), patients (education, confronting stigma about methadone maintenance), and potential therapeutic activities (methadone group therapy). The implementation of these staff, patient, and therapeutic adaptations can assist residential substance abuse treatment programs in integrating these two addiction treatment modalities. PMID- 18159774 TI - Methamphetamine use in Dayton, Ohio: preliminary findings from the Ohio Substance Abuse Monitoring Network. AB - Since the 1990s, methamphetamine manufacturing and use have been spreading into midwestern and southern United States. However, most research on methamphetamine use has been conducted in the western United States. This exploratory study is based on qualitative interviews with 26 people recruited in Dayton, Ohio, for the Ohio Substance Abuse Monitoring Network, a statewide epidemiological surveillance system. Participants ranged in age from 21 to 57 years. The majority were White and male. The participants reported increases in methamphetamine availability and identified powder-type locally manufactured methamphetamine as the most commonly seen form of the drug. The participants reported extensive drug use histories, and were introduced to methamphetamine in the context of their involvement with pharmaceutical stimulants, crack, powder cocaine, and/or club drug scene. A large proportion of the participants continued to use crack or powder cocaine in addition to methamphetamine. Many primary methamphetamine users felt they were able to moderate methamphetamine use and fulfill their daily responsibilities related to work and family life. Ranking results and consensus analysis revealed that participants shared high agreement about drug-related risks, and perceived methamphetamine as a serious drug, although not as risky as heroin, crack or OxyContin. Implications for treatment and prevention are discussed. PMID- 18159775 TI - Examining the decay of HIV risk reduction outcomes following a community-friendly intervention targeting injection drug users in treatment. AB - It has become crucial for risk reduction interventions targeting injection drug users (IDUs) in treatment to be "community-friendly" and potent over time so that limited resources may be optimally utilized. This study examined (1) the extent to which observed post-intervention effects--including enhanced HIV-related knowledge, motivation, behavioral skills, and drug- and sex-risk reduction behavior--decayed over time and (2) whether repeating the intervention atfollow up provided additional benefit. Approximately 10 months after completing an adapted, substantially shortened, version of an evidence-based intervention, participants completed a follow-up assessment and then repeated the intervention. No evidence of decay was found. Even so, after repeating the intervention, a trend toward additional sex-risk reduction was observed for participants at higher risk for HIV. Findings point to the potential for an adapted evidence based intervention for IDUs to be both community-friendly and potent over time within community-based treatment settings. PMID- 18159776 TI - Implementing an evidence-based practice: Seeking Safety Group. AB - This article presents findings from a multisite study on adopting and implementing an evidence-based practice, Seeking Safety, for women with co occurring disorders and experiences of physical and sexual abuse. It focuses on what implementation decisions different sites made to optimize the compatibility of Seeking Safety with the site's needs and experiences and on issues posed by Rogers (1995) as relevant to successful diffusion of an innovative practice. A total of 157 clients and 32 clinicians reported on satisfaction with various aspects of the model. Cross-site differences are also examined. Results show that Seeking Safety appears to be an intervention that clinicians perceive as highly relevant to their practice, and one that adds value. Clients perceive the treatment as uniquely touching on their needs in a way that previous treatments had not. PMID- 18159777 TI - Understanding the working alliance between persons with substance abuse problems and strengths-based case managers. AB - A substantial body of literature has examined the importance of the working alliance in psychotherapy; few works have examined it in the context of case management Qualitative methods were used to examine how 26 persons with substance abuse problems perceived the working alliance with case managers who practice from the strengths perspective. Clients' narratives emphasized the personal qualities of the case manager and the nature of the client-case manager relationship. Their narratives also reflected two guiding principles of strengths based case management: personal control over goal-setting, and an emphasis on strengths. Most clients concluded that a positive working alliance helped them to build trust, self-worth, and self-esteem. PMID- 18159778 TI - Substance abuse treatment staff perceptions of intimate partner victimization among female clients. AB - Providing intimate partner violence (IPV)-related services to women enrolled in substance abuse treatment programs has the potential to reach a population disproportionately affected by IPV. Integrating basic IPV services into substance abuse treatment, however, poses challenges to organizations and staff. Using focus groups, the authors examined the experiences and attitudes of substance abuse treatment staff towards clients with IPV victimization experiences in order to elucidate factors that might affect the implementation of IPV services within substance abuse treatment. Seven focus groups were conducted with staff members from substance abuse treatment programs in New York City. Although participants believed that IPV is common and negatively affects client recovery, they felt competing time demands, complex confidentiality issues, insufficient training and lack of agency leadership would impede their provision of IPV-services. The study suggests that system-level assessment and change is needed to provide IPV-related services in substance abuse treatment settings. PMID- 18159779 TI - Adolescents' participation in Alcoholics Anonymous and Narcotics Anonymous: review, implications and future directions. AB - Youth treatment programs frequently employ 12-Step concepts and encourage participation in Alcoholics Anonymous (AA) and Narcotics Anonymous (NA). Since AA/NA groups are easily accessible at no charge and provide flexible support at times of high relapse risk they hold promise as a treatment adjunct in an increasingly cost-constricting economic climate. Yet, due to concerns related to adolescents' developmental status, skepticism exists regarding the utility of AA/NA for youth. This review evaluates the empirical evidence in this regard, identifies and discusses knowledge gaps, and recommends areas for future research. Searches were conducted in PsychINFO, Medline, relevant literature and by personal correspondence. Findings suggest youth may benefit from AA/NA participation following treatment, but conclusions are limited by four important factors: (1) a small number of studies; (2) no studies with outpatients; (3) existing evidence is solely observational; and (4) only partial measurement of the 12-Step construct. While surveys of adolescent SUD treatment programs indicate widespread clinical interest and application of adult-derived 12-Step approaches this level of enthusiasm has not been reflected in the research community. Qualitative research is needed to improve our understanding of youth specific AA/NA barriers, and efficacy, comparative effectiveness, and process studies are still needed to inform clinical practice guidelines for youth providers. PMID- 18159780 TI - An exploratory study of OxyContin use among individuals with substance use disorders. AB - This study surveyed 422 individuals being treated in a substance abuse treatment program that offers various levels of care in order to learn about OxyContin use among this population. Focus areas included exposure and use of OxyContin, how this medication was obtained, reasons for initial use, and whether users of OxyContin were drug naive or experienced users of opiate or non-opiate drugs. Whether OxyContin users who previously had never used opiates would report migrating to heroin was also explored. Findings revealed that 48% of the population had used OxyContin and of this population, only 1% had no history of prior substance use. Seventy percent of this sample obtained them from friends and 14% obtained them directly from physicians. Many reported that their friends obtained their OxyContin from physicians, bringing the percentage of people who directly or indirectly obtained OxyContin through a physician to 37%. While most users of OxyContin had a history of past opiate use, a small percentage had not, and of this later group, 73% migrated to using heroin. The majority of individuals who obtained OxyContin from a physician had a history of recreational or problematic opiate or non-opiate drug use. Finally, over time, 90% had stopped using this drug, although other drug use continued. PMID- 18159781 TI - Beliefs and social norms about codeine and promethazine hydrochloride cough syrup (CPHCS) use and addiction among multi-ethnic college students. AB - In this study a qualitative approach is used to investigate relevant beliefs and norms concerning the consumption, initiation, and perceived addiction of codeine and promethazine hydrochloride cough syrup (CPHCS) among 61 college-age students who identified themselves as current CPHCS users. In general, a majority of students stated that doctors and pharmacists were the greatest facilitators of CPHCS acquisition. A majority of students believed that their friends felt codeine promethazine use was "normal" and "cool" among college students their age, and that reinforcing factors, such as peer pressure and curiosity, contributed to initial CPHCS use. PMID- 18159782 TI - The creativity of Crumb: research on the effects of psychedelic drugs on the comic art of Robert Crumb. AB - This article investigates the influence of perception that is altered by psychedelic drugs on processes of creativity through a case study of the work of well-known comic artist Robert Crumb. Samples of Crumb's work before, during, and after the period of his use of psychedelic drugs are content analyzed and compared according to the categorization offered by Janiger and Dobkin de Rios (1989). The results of the comparison indicate that Robert Crumb's drug use significantly altered the stylistic approach of his artwork not only during the period of his drug use, but long after he had stopped using drugs. PMID- 18159783 TI - Gamma-hydroxybutyric acid (GHB) withdrawal: a case report. AB - GHB is an increasingly popular drug of abuse that can be associated in select cases with growing dependence and a severe withdrawal syndrome. While benzodiazepines are recommended for treatment of the withdrawal syndrome, some cases have been described as benzodiazepine-resistant. The authors describe treatment of such a case, which was unsuccessfully treated initially with benzodiazepines, then successfully treated with adjuvant atypical neuroleptics, and offer a possible neurochemical explanation for why such agents may be theoretically more effective than benzodiazepines in treating GHB withdrawal. PMID- 18159784 TI - Abuse of olanzapine by substance abusers. AB - Olanzapine has been used for over a decade for treatment of schizophrenia and bipolar disorder. The drug may have sedative properties for some patients, especially in large doses. The case reported here involves a 25-year-old male who abused olanzapine, both by itself and in combination with other drugs. Also described are the patient's reports of abuse of olanzapine by several of his acquaintances. The potential for abuse of olanzapine by substance abusers is discussed. PMID- 18159785 TI - Death by drug overdose: impact on families. AB - Death by overdose is loaded with social/moral stigmas, in addition to strong feelings of anger, helplessness, guilt and shame in the families. The objective of this study was to analyze the impact of these feelings on families facing death by overdose. Qualitative methodology was used to study six families with a history of death by overdose of one of their members. The interview was open, and guided by the question "What did you feel with the death of your family member by overdose and what was the impact of this death on your family as a whole?" The families were grouped into two categories: families who knew about the drug use of their family member, and families who were not aware of it. The reports show that secrecy regarding drug use followed by death by overdose arouses feelings of anger, guilt, helplessness, and deprives the family members of information that could allow them to take action. As regards families that were aware of the drug use, there seems to be a "veiled preparation" for a possible death by overdose, bringing about ambivalent situations of grief and relief. The report stresses how disturbing it is to lose a family member by overdose, and points to the need for psychological support for those families. PMID- 18159786 TI - Cognitive fitness. AB - Recent neuroscientific research shows that the health of your brain isn't, as experts once thought, just the product of childhood experiences and genetics; it reflects your adult choices and experiences as well. Professors Gilkey and Kilts of Emory University's medical and business schools explain how you can strengthen your brain's anatomy, neural networks, and cognitive abilities, and prevent functions such as memory from deteriorating as you age. The brain's alertness is the result of what the authors call cognitive fitness -a state of optimized ability to reason, remember, learn, plan, and adapt. Certain attitudes, lifestyle choices, and exercises enhance cognitive fitness. Mental workouts are the key. Brain-imaging studies indicate that acquiring expertise in areas as diverse as playing a cello, juggling, speaking a foreign language, and driving a taxicab expands your neural systems and makes them more communicative. In other words, you can alter the physical makeup of your brain by learning new skills. The more cognitively fit you are, the better equipped you are to make decisions, solve problems, and deal with stress and change. Cognitive fitness will help you be more open to new ideas and alternative perspectives. It will give you the capacity to change your behavior and realize your goals. You can delay senescence for years and even enjoy a second career. Drawing from the rapidly expanding body of neuroscience research as well as from well-established research in psychology and other mental health fields, the authors have identified four steps you can take to become cognitively fit: understand how experience makes the brain grow, work hard at play, search for patterns, and seek novelty and innovation. Together these steps capture some of the key opportunities for maintaining an engaged, creative brain. PMID- 18159787 TI - A leader's framework for decision making. A leader's framework for decision making. AB - Many executives are surprised when previously successful leadership approaches fail in new situations, but different contexts call for different kinds of responses. Before addressing a situation, leaders need to recognize which context governs it -and tailor their actions accordingly. Snowden and Boone have formed a new perspective on leadership and decision making that's based on complexity science. The result is the Cynefin framework, which helps executives sort issues into five contexts: Simple contexts are characterized by stability and cause-and effect relationships that are clear to everyone. Often, the right answer is self evident. In this realm of "known knowns," leaders must first assess the facts of a situation -that is, "sense" it -then categorize and respond to it. Complicated contexts may contain multiple right answers, and though there is a clear relationship between cause and effect, not everyone can see it. This is the realm of "known unknowns." Here, leaders must sense, analyze, and respond. In a complex context, right answers can't be ferreted out at all; rather, instructive patterns emerge if the leader conducts experiments that can safely fail. This is the realm of "unknown unknowns," where much of contemporary business operates. Leaders in this context need to probe first, then sense, and then respond. In a chaotic context, searching for right answers is pointless. The relationships between cause and effect are impossible to determine because they shift constantly and no manageable patterns exist. This is the realm of unknowables (the events of September 11, 2001, fall into this category). In this domain, a leader must first act to establish order, sense where stability is present, and then work to transform the situation from chaos to complexity. The fifth context, disorder, applies when it is unclear which of the other four contexts is predominant. The way out is to break the situation into its constituent parts and assign each to one of the other four realms. Leaders can then make decisions and intervene in contextually appropriate ways. PMID- 18159788 TI - If private equity sized up your business. AB - As the dust settles on the recent frenzy of private equity deals (including transactions topping $20 billion), what lessons can companies glean? Directors and executives of public companies may now be slightly less fearful of imminent takeover, yet the pressure remains: They face shareholders who wonder why they aren't getting private-equity-level returns. Rather than dismiss the value private equity has created as manipulated or aberrant, public company leaders should recognize the disciplined management that often underlies it. Pozen, a longtime leader in the financial services industry, finds that in the aftermath of buyouts, companies undergo five major thrusts of reform. These translate into five key questions that directors should pose to senior management: Have we left too much cash on our balance sheet instead of raising our cash dividends or buying back shares? Do we have the optimal capital structure, with the lowest weighted after-tax cost of total capital, including debt and equity? Do we have an operating plan that will significantly increase shareholder value, with specific metrics to monitor performance? Are the compensation rewards for our top executives tied closely enough to increases in shareholder value, with real penalties for nonperformance? Finally, does our board have enough industry experts who have made the time commitments and been given the financial incentives necessary to maximize shareholder value? The era of private equity is far from over - the top funds have become very large and are likely to play an influential role in future market cycles. Boards that ask these questions, and act on them, won't just beat the takeover artists to the punch. They will build stronger businesses. PMID- 18159789 TI - Solve the succession crisis by growing inside-outside leaders. AB - In his analysis of 1800 successions, Harvard Business School professor Bower found that companies performed significantly better when they appointed insiders to the job of CEO. Other researchers, including Jim Collins in Good to Great, have come to similar conclusions working from different data sets. Yet Bower finds far too many companies have no succession plans; as a result, when the time comes to name a new chief executive, more firms turn to outsiders. Both insider and outsider CEOs have strengths and weaknesses at the start. Insiders know the company and its people but are often blind to the need for radical change. Outsiders see the need for a new approach but can't make the necessary changes because they don't know the organization or industry sector well enough. What companies must do, then, is find a way to nurture what Bower calls inside outsiders--internal candidates who have outside perspective. Often such executives have spent much of their time away from the mainstream of the organization, and away from headquarters, living with new opportunities and threats. Before becoming CEO, Procter & Gamble's A.G. Lafley, for instance, worked for years building P&G's Chinese cosmetics operation rather than the core detergent business. IBM's Sam Palmisano was a champion of software and open systems at a time when Big Blue was essentially a closed-system, hardware oriented company. Nascent inside-outsiders should enter the CEO-training process by the time they are 30 and be given the opportunity to manage a whole business, so that they become good insiders. But they also need to be mentored with an eye toward preserving their outsider perspective, so they learn how to turn their new ideas into great businesses and are protected from old-timers who might be inclined to teach them a lesson. PMID- 18159790 TI - 8 ways to build collaborative teams. AB - Executing complex initiatives like acquisitions or an IT overhaul requires a breadth of knowledge that can be provided only by teams that are large, diverse, virtual, and composed of highly educated specialists. The irony is, those same characteristics have an alarming tendency to decrease collaboration on a team. What's a company to do? Gratton, a London Business School professor, and Erickson, president of the Concours Institute, studied 55 large teams and identified those with strong collaboration despite their complexity. Examining the team dynamics and environment at firms ranging from Royal Bank of Scotland to Nokia to Marriott, the authors isolated eight success factors: (1) "Signature" relationship practices that build bonds among the staff, in memorable ways that are particularly suited to a company's business. (2) Role models of collaboration among executives, which help cooperation trickle down to the staff. (3) The establishment of a "gift culture," in which managers support employees by mentoring them daily, instead of a transactional "tit-for-tat culture", (4) Training in relationship skills, such as communication and conflict resolution. (5) A sense of community, which corporate HR can foster by sponsoring group activities. (6) Ambidextrous leadership, or leaders who are both task-oriented and relationship-oriented. (7) Good use of heritage relationships, by populating teams with members who know and trust one another. (8) Role clarity and task ambiguity, achieved by defining individual roles sharply but giving teams latitude on approach. As teams have grown from a standard of 20 members to comprise 100 or more, team practices that once worked well no longer apply. The new complexity of teams requires companies to increase their capacity for collaboration, by making long-term investments that build relationships and trust, and smart near-term decisions about how teams are formed and run. PMID- 18159791 TI - Mapping your competitive position. AB - A price-benefit positioning map helps you see, through your customers' eyes, how your product compares with all its competitors in a market. You can draw such a map quickly and objectively, without having to resort to costly, time-consuming consumer surveys or subjective estimates of the excellence of your product and the shortcomings of all the others. Creating a positioning map involves three steps: First, define your market to include everything your customers might consider to be your product's competitors or substitutes. Second, track the price your customers actually pay (wholesale or retail? bundled or unbundled?) and identify what your customers see as your offering's primary benefit. This is done through regression analysis, determining which of the product's attributes (as described objectively by rating services, government agencies, R&D departments, and the like) explains most of the variance in its price. Third, draw the map by plotting on a graph the position of every product in the market you've selected according to its price and its level of primary benefit, and draw a line that runs through the middle of the points. What you get is a picture of the competitive landscape of your market, where all the products above the line command a price premium owing to some secondary benefit customers value, and all those below the line are positioned to earn market share through lower prices and reduced secondary benefits. Using examples as varied as Harley-Davidson motorcycles, Motorola cell phones, and the New York restaurant market, Tuck professor D'Aveni demonstrates some of the many ways the maps can be used: to locate unoccupied or less-crowded spaces in highly competitive markets, for instance, or to identify opportunities created through changes in the relationship between the primary benefit and prices. The maps even allow companies to anticipate--and counter-- rivals' strategies. R eprint RO711G PMID- 18159792 TI - Simple rules for making alliances work. AB - Corporate alliances are growing in number--by about 25% a year--and account for up to a third of revenues and value at many companies. Yet some 60% to 70% of them fail. What is going wrong? Because alliances involve interdependence between companies that may be competitors and may also have vastly different operating styles and cultures, they demand more care and handling than other business arrangements, say Hughes and Weiss, management consultants at Vantage Partners. The authors have developed five principles--based on their two decades of work with alliances -to complement the conventional advice on alliance management: (1) Focus less on defining the business plan and more on how you and your partner will work together. (2) Develop metrics pegged not only to alliance goals but also to performance in working toward them. (3) Instead of trying to eliminate differences, leverage them to create value. (4) Go beyond formal systems and structures to enable and encourage collaborative behavior. (5) Be as diligent in managing your internal stakeholders as you are in managing the relationship with your partner. Companies that have adopted these principles have radically improved their alliance success rate. Schering-Plough, for example, engages in a systematic "alliance relationship launch": four to six weeks of meetings at which the partners explore potential challenges, examine key differences and develop shared protocols for managing them, and establish mechanisms for day-to-day decision making. Blue Cross and Blue Shield of Florida measures the quality of alliance progress through regular surveys of both its own staff and its partners'. These companies have learned that the conventional advice is not so much wrong as incomplete. The five simple rules can help fill in the blanks. PMID- 18159793 TI - Are your engineers talking to one another when they should? AB - Communication may not be on managers' minds at companies that design complex, highly engineered products, but it should be. When mistakes take place, it's often because product-component teams fail to talk. The consequences can be huge: Ford and Bridgestone Firestone lost billions by not coordinating the design of the Explorer with the design of its tires. The major delays and cost overruns involved in the development of Airbus's A380 "superjumbo"--which most likely led to the CEO's exit--were a result of unforeseen design incompatibilities. To help managers mitigate such problems, the authors present a new application of the design structure matrix, a project management tool that maps the flow of information and its impact on product development. Drawing on research into how Pratt & Whitney handled the development of the PW4098 jet engine, they have developed an approach that uncovers (a) areas where communication should be occurring but is not (unattended interfaces, usually bad) and (b) areas where communication is occurring but has not been planned for (unidentified interfaces, usually good). After finding the unattended and unidentified interfaces, the next step is to figure out the causes of the critical ones. If a significant number of unattended interfaces cross organizational boundaries, executives may need to redraw organizational lines. Executives can then manage the remaining critical interfaces by extending the responsibilities of existing integration teams (those responsible for cross-system aspects, such as a jet engine's fuel economy) to include supervising the interaction, by dedicating teams to specific interfaces, or by formally charging teams already involved with the interfaces to oversee them. Finally, it's important to ensure that the teams are working with compatible design equipment; inconsistencies between CAD tools have cost Airbus dearly. PMID- 18159794 TI - A lifelong battle against disease. PMID- 18159796 TI - Shopping for a drug plan. PMID- 18159795 TI - Making science free to all. PMID- 18159797 TI - The right way to recover. PMID- 18159798 TI - Why science can't save the GOP. The stem-cell breakthrough doesn't make up for six years of hypocrisy and lost research. PMID- 18159799 TI - Is this disorder for real? PMID- 18159800 TI - Head games. Girls are getting more concussions than boys are. Why they are at risk. PMID- 18159802 TI - Reform bids draw dollars. In shift from 2000 and 2004, Democrats lead race to raise campaign funds from influence-seeking healthcare executives. PMID- 18159801 TI - Diet-pill dilemma. When willpower fails, are antiobesity drugs worth the risk of less-than-pleasant side effects? PMID- 18159803 TI - Quality - costs = access. A remade health system is key to universal coverage. PMID- 18159804 TI - One for all. My plan is the only true universal coverage initiative in this carpaign. PMID- 18159805 TI - Let the consumer rule. I seek excellence, affordability in a reformed system. PMID- 18159806 TI - Prevention in a free market. My plan: let consumers run their own care, with incentives to stay healthy. PMID- 18159807 TI - The solution is freedom. My plan for U.S. healthcare: let the informed patient make the decisions. PMID- 18159808 TI - I would solve the real problem. We have to rein in the cost of care by changing the way we pay for it. PMID- 18159809 TI - My cure for an ailing system. How I, as president, would achieve affordable, universal health coverage. PMID- 18159810 TI - Encourage all to keep costs down. We must take steps to return control of healthcare dollars, decisions to individuals. PMID- 18159811 TI - Coverage for all, at the right price. Individual choice and shared responsibility in a lower-cost healthcare system. PMID- 18159812 TI - What the other candidates are saying. Biden eyes SCHIP, Tancredo targets immigration, Thompson talks choice. PMID- 18159813 TI - The heart of the matter. A summary of what the top candidates are saying about healthcare policy. PMID- 18159814 TI - Modern healthcare's presidential straw poll. PMID- 18159815 TI - Ambitious agenda awaits. Rare opportunity coming to achieve change in system. PMID- 18159816 TI - Free the market. Government interference hampers healthcare reform. PMID- 18159817 TI - Use your political power wisely. Question to candidates: if elected, how will you improve healthcare in America? PMID- 18159818 TI - Some bacteria for brunch? Health-boosting probiotics are all the rage, but shop carefully. PMID- 18159819 TI - A birth control pill dilemma. PMID- 18159820 TI - [Human papilloma virus infection in pregnant women with normal pap-smears, HPV oncogenity and risk factors]. AB - The process of carcinogenesis in both types of cervical carcinoma is dependent on the infection of oncogenic types of HPV. HPV infection could be diagnosed on the basis of whether or not the DNA virus in present. In pregnant women the latent persistent infection easily changes into its active form. This process is related to changes in immunological response and concentration of the hormones. AIM: The main aim of the study was to evaluate the frequency of HPV infection in healthy pregnant women in second and third trimester of pregnancy and the presence of selected risk factors. MATERIAL AND METHODS: The study was conducted in 2005-2006 on hospitalized women in The Clinic of High Risk Pregnancy. The first group consisted of 180 pregnant women in 2nd trimester of pregnancy, the second comprised 220 pregnant women in the 3rd trimester. In all women the Pap-smears and diagnosis of DNA presence of high and low risk HPV from the border line of cervical epithelium were tested on the first day of the hospitalization. The PCR method using Human Papilloma Virus Typing Set was used. RESULTS: In 400 pregnant women the presence of HPV was found in 4.5%, type 16 was fund in 2.5%, type 18 in 1.7%. Combined infection 16 and 18 types was found in 0.2%. In 180 pregnant women in II trimester high risk HPV was found in 4.4%: 16 type was found in 2.8%, 18 type in 1.7%. In IlI trimester high risk HPV was found in 4.5%: 16 type was found in 2.3%, type 18 in 1.8%, combined 16 and 18 in 0.4%. Low risk HPV was found in 1.0%- type 6. In II trimester in 1.1%, In III trimester 0.9%. There were no differences between HPV infection rate in II and III trimester, pregnant women age, the gravity and HSV2 infection. The difference was found between high risk HPV infection and parity, cigarette smoking and oncological family history. CONCLUSION: Asymptomatic HPV infection has the same frequency in II and III trimester. High parity, cigarette smoking and oncological family history were connected with an increased rate of high risk HPV infection. All women in reproductive age should be checked for the HPV DNA. PMID- 18159821 TI - [Implementation of cytology images classification--the Bethesda 2001 System--in a group of screened women from Podlaskie region--effect evaluation]. AB - Increasing knowledge concerning carcinogenesis within cervical epithelium has forced us to make continues modifications of cytology classification of the cervical smears. Eventually, new descriptions of the submicroscopic cytomorphological abnormalities have enabled the implementation of Bethesda System which was meant to take place of the former Papanicolaou classification although temporarily both are sometimes used simultaneously. AIM: The aim of this study was to compare results of these two classification systems in the aspect of diagnostic accuracy verified by further tests of the diagnostic algorithm for the cervical lesion evaluation. MATERIALS AND METHODS: The study was conducted in the group of women selected from general population, the criteria being the place of living and cervical cancer age risk group, in the consecutive periods of mass screening in Podlaski region. The performed diagnostic tests have been based on the commonly used algorithm, as well as identical laboratory and methodological conditions. RESULTS: Performed assessment revealed comparable diagnostic accuracy of both analyzing classifications, verified by histological examination, although with marked higher specificity for dysplastic lesions with decreased number of HSIL results and increased diagnosis of LSILs. Higher number of performed colposcopies and biopsies were an additional consequence of TBS classification. Results based on Bethesda System made it possible to find the sources and reasons of abnormalities with much greater precision, which enabled causing agent treatment. CONCLUSION: Two evaluated cytology classification systems, although not much different, depicted higher potential of TBS and better, more effective communication between cytology laboratory and gynecologist, making reasonable implementation of The Bethesda System in the daily cytology screening work. PMID- 18159822 TI - [The value of the cytocins and the solubility of theirs receptors in the serum of the patients with the cervical cancer]. AB - The study encompassed 146 women with recognized invasive cervical cancer in 4 clinical stages. Patients underwent therapy in Oncologic Gynaecology Department of Wroclaw Medical University in 2001 and 2002 years. The correlation between pretreatment serum level of proangiogenic and inflammation factors--VEGF, sTNF R1, IL-6 and clinical stage or early effects of therapy were observed. The strong and statistically significant relationship between pretreatment serum level of IL 6 and sTNF-R1 and clinical stage with early effect of treatment were found. The statistically significant correlation between serum level of all investigated parameters and clinical stage of cervix cancer was noticed. VEGF wasn't an independent prognostic factor in the study, but the prognostic value of IL-6 was demonstrated. PMID- 18159823 TI - [Cytology and DNA HPV-testing in the era of HPV-vaccine]. AB - Human papillomavirus (HPV) persistent infection is the main factor leading to the cervical cancer carcinogenesis. Wide-spread public vaccination against HPV as primary prevention is expected to reduce cervical cancer incidence and mortality rates. It is essential to bear in mind that screening for precancerous lesions cannot be discontinued because vaccination will not protect the patients against HPV types which have not been included in the first and second generation of vaccines. PMID- 18159824 TI - [Mechanism of the cancerogenesis in cervix paraepidermal epithelium cells with chronic infection of oncogenic types of human papiloma virus]. AB - The human papillomavirus family is composed of a large number of different and variably related types, each of which is associated with a characteristic set of epithelial lesions. Each of the many identified types of human papillomavirus have shown considerable specificity of different anatomical sites and different characteristic lesions. HPV 6 and 11 are frequently associated with benign condylomas, while HPV 16 and 18 are associated with malignant progression and cervical cancer. The genome of papillomaviruses is composed of a circular double stranded DNA. Various open reading frames (ORFs) are located on only one DNA strand. The coding strand contains from 8 to 10 translational ORFs. Among them, only 6 to 8 were designated as early and 2 as late. The role of HPVs 16 and 18 in uterine cervix carcinoma has been well-documented, but their contriobution to carcinogenesis of other neoplasias is still questionable. PMID- 18159825 TI - [Clinical manifestations of HPV infections]. AB - Human papillomavirus (HPV) infections are the causal factors of benign diseases, precancerous lesions and malignancies of various locations. The degree of causality between HPV infections and the development of these disorders varies and reaches 100% in cervical cancer. Low risk HPVs stimulate epithelial cells to excessive proliferation, causing warty lesions, whereas high risk types also possess carcinogenic potential. In this article selected clinical manifestations of HPV infections and their effects have been discussed. PMID- 18159826 TI - [Tumor makrers in cervical cancer]. AB - IMeasurement of tumor markers level in human body fluids, mainly in serum may be useful for diagnosis, therapy monitoring and early recurrence detection. SCC-Ag, Cyfra 21-1, CEA, CA 125 and TPS are of a clinical value in uterine cervical cancer despite their diverse significance. Other biological markers that could be measured in serum as well as in tumor tissue are cytokines: VEGF, IL-6, IL-8, IL 4, IL-10, leptin, stromal drive factor (SDF- 1), although assessment of their importance needs further examination. Elevation of uterine cervical cancer hypoxia markers such as HIF 1a, CA 9, GLUT-1 is combined with poorer clinical outcome prognosis. PMID- 18159827 TI - [The role of colposcopy in Population Program of Cervical Cancer Early Detection]. AB - Cervical screening is the most effective method of controlling and fighting with cervical cancer. The article has presented the role and importance of cytology and colposcopy in the realization of The Population Programme of Cervical Cancer Early Detection. Furthermore, the significance of cytological examination during the first two stages of the programme, as well as the role of colposcopy in the realization of the third stage of the precise diagnosis among women with abnormal pap smears, have been presented as well. The goals and basic advice on performing colposcopy have been introduced. Methodology of colposcopy has been presented in detail. PMID- 18159828 TI - [Surgical treatment of cervical cancer]. AB - The article deals with modern views on the role of surgery in cervical cancer treatment. Objective indications for surgery in early and advance stage of the disease have been presented. The role of the operative technique, taking into consideration also the conservative procedures, has been defined. In the summary the authors emphasize the importance and necessity of cervical cancer patients being subjected to very careful preoperative diagnosis, and choice of the type of procedure made after very detailed analysis of all clinical and pathological features and patients' preference. PMID- 18159829 TI - [Carcinoma of the uterine cervix during pregnancy]. AB - Pregnancy offers a unique opportunity for the early diagnosis of cervical cancer, due to the fact that pregnant patients have many possibilities of gynecological and cytological examinations. There is much evidence that pregnant women have two to three-fold higher chance of having preneoplastic lesions and early, operable stages of disease diagnosed. The preneoplastic lesions do not require any intervention during pregnancy. However, precise, serial colposcopic examinations, completed by biopsy if necessary, must be seriously considered in order to exclude invasive cancer. The only indication for conization during pregnancy is to rule out or confirm microinvasive or invasive cancer, provided such diagnose can change the time and the way of delivery. Invasive cervical cancer diagnose is frequently associated with difficult medical and ethical decisions. The most proper approach should be considered, taking into account the benefit of the mother and the child. The decision is easier in the early stage of cancer, because it has been proven that six- to twelve-week delay of the beginning of the therapy does not deteriorate the cancer outcome but it enables the fetus to acquire sufficient lung maturity. Advanced carcinoma of the cervix forces us to take prompt therapeutic decisions. Both, the continuation of the pregnancy and the administration of neoadjuvant chemotherapy are still possible. PMID- 18159831 TI - Quality dentistry and profits--is there a relationship? PMID- 18159830 TI - [Agresive variants of uterine cervical cancers]. AB - Three different variants of biologically agressive cervical cancers, such as: basaloid carcinoma, glassy cell carcinoma and small cell endocrine carcinoma, have been reported. Hematoxylin and eosin, periodic acid-Schiff and immunohistochemical stains were used. The course of disease and aplication of treatment has been analyzed as well. Characteristic morphologic features of these cancers have been emphasized. Rather limited susceptibility for radiotherapy and chemiotherapy of these cancers has not gone unnoticed. The description of these types of cervical cancers might enable us to arrive at right conclusions, both clinical and diagnostic. PMID- 18159832 TI - Giving back to those in need. PMID- 18159833 TI - Doing some small part to provide relief. PMID- 18159834 TI - How does Pennsylvania stack up? An analysis of dental and medical visits for Pennsylvania's children. AB - The percent of Pennsylvania children with dental and medical visits is one of the highest in the country. In some rankings of the health system performance, the state fares quite well in comparison with the performance of other states and the District of Columbia. However, while the state ranks favorably in some areas, it fares not so favorably in other rankings, which include personal behavior, community environment, public health policies and health outcomes. This article highlights the major areas of both approval and concern. PMID- 18159835 TI - Observations. PMID- 18159836 TI - Testicular lymphoma: a case report. AB - This case report describes a 67-year-old man with testicular lymphoma. The typical symptom in patients when first seen with testicular lymphoma is a painless enlargement of the testicle. However, this patient had a long history of progressively worsening weakness in his right arm and leg, as well as a sudden onset of chest pain and dyspnea. Upon initial examination of this patient, a large mass was discovered in the scrotal and inguinal region on the right side. The patient subsequently underwent a right radical orchiectomy. When the diagnosis of lymphoma was confirmed, a proper chemotherapy regimen was initiated. This case report reviews the case, diagnosis, and management of testicular lymphoma. PMID- 18159837 TI - And the question is...Is the patient bleeding or clotting? A guide to appropriate testing and evaluation. AB - With recent advances in hemostasis, there has been an increased demand for clinical testing of patients with suspected bleeding and clotting disorders. There has also been an increase in the available testing that clinicians can order from the clinical laboratory. Unfortunately, to complicate matters further, some of the tests sound very much alike but test for very different conditions. Which test is indicated for what condition is not always clear, and can lead to erroneous testing with complicating results. A basic understanding of the hemostasis mechanism and the specific target of the test is imperative for correct test selection. PMID- 18159838 TI - Diagnosis and treatment of constipation in children: a survey of primary care physicians in West Virginia. AB - A questionnaire was mailed to all PCPs practicing in West Virginia. Demographics, clinical practice, treatment, referral practices, and familiarity with the constipation guideline were recorded. Results were compared between pediatricians (Ps) and family practitioners (FPs). Out of 718, 210 (78 Ps and 112 FPs) completed the survey. Compared to FPs, Ps reported a higher number of children diagnosed with constipation in their clinics. All PCPs prescribed various treatment modalities for constipation, but Ps used more lavage therapy (P< 0.001). Most of the PCPs referred patients to specialists for treatment failure (85%), and preferred to co-manage their patients (58%). The majority (67-86%) of physicians were not familiar with the published clinical guidelines for constipation in children. Most PCPs in West Virginia are not familiar with the clinical guideline for constipation in children. An educational campaign for constipation in children is clearly warranted. PMID- 18159839 TI - Internet report cards on quality: what exists and the evidence on impact. AB - Report cards based on publicly disclosed data abound. Consumers can use the internet to review grades on physicians, hospitals, nursing homes, home health agencies, and insurance plans. The reports differ in their combinations of mortality data, process measures, access scores, and satisfaction surveys. The internet sites also differ by access charge, data sources, and presentation of information. Public disclosure of quality data has had little impact on the behavior of consumers, larger purchasers of health care, and physicians. However, health care provider organizations have responded to the public reports of quality. Analysis of the impact of public report cards is lagging as web available reports rapidly grow and pay for performance programs emerge. PMID- 18159840 TI - Non-tropical pyomyositis in pediatric and adult patients. AB - Twelve cases of pyomyositis treated at a rural tertiary care referral center are reported and compared to other published cases of this disorder in the United States. About 350 cases have been reported nationally and over a quarter of them are in patients infected with human immunodeficiency virus (HIV). Pyomyositis, classically a disease of the healthy young male, starts insidiously and progresses to a pus filled abscess of a muscle or a group of muscles. It is a clinical diagnosis often misdiagnosed and therefore treatment is delayed. Imaging studies are conclusive in most cases by the time of presentation. Our series has a predominance of older females, no HIV positive patients and is unique in comparison to previously reported trends and cases. PMID- 18159841 TI - Acute pancreatitis: management update and future directions of pancreatic imaging. AB - Essential management points for AP are: 1. Intravenous fluids and adequate hydration are the mainstay of therapy. Antibiotics are not routinely indicated in most cases. 2. The most reliable marker for diagnosing biliary AP is a greater than threefold elevation of the serum alanine amniotrasferase (ALT), which has a positive predictive value of 95% for biliary acute pancreatitis. 3. Early ERCP with ES for stone extraction and biliary decompression has proved beneficial for patients with biliary pancreatitis and evidence of persistent or progressive biliary obstruction with elevated serum total bilirubin and ALT levels. 4. Most patients with severe necrotizing pancreatitis will need intensive care, imaging, possible fine needle aspiration (FNA) by interventional radiology (IR) if necrosis is present on CT and the patient's condition is worsening. 5. If sterile necrosis is found on the FNA aspirate then conservative therapy would be continued; but if infected necrosis is found, then consideration for surgical consultation is needed for possible surgical therapy with debridement and necreotectomy. PMID- 18159842 TI - Double ATV-related fatalities in West Virginia, 2000-2007. PMID- 18159843 TI - Efforts to reduce infant mortality. PMID- 18159844 TI - [Contribution of selected cellular adhesion molecules and proinflammatory cytokines in the pathogenesis of proliferative diabetic retinopathy]. AB - Proliferative diabetic retinopathy (PDR) is caused by structural and functional disorders of the retinal vessels. Complex pathobiological mechanisms of this process have not been clearly explained so far. PURPOSE: To prove the role of immunological-inflammatory process in the development of the proliferative diabetic retinopathy. MATERIAL AND METHODS: Forty six patients, aged 63.97+/-9 were enrolled in the study, treated for diabetes type 2 for 12.56+/-6,87 years. In 19 patients (12 women and 7 men aged 64.63+/-8.38, treated for diabetes for 12.47+/-6.75 years) a parallel analysis of given parameters was performed in both serum and vitreous body samples. Vitrectomy, in course of which the vitreous body samples were obtained, was performed in these patients because of vitreous haemorrhage, causing traction retinal detachment in some patients. The control group consisted of 15 patients (9 women and 6 men), aged 63.00+/-14.58, who underwent the vitrectomy procedure because of reasons other than diabetic retinopathy. The concentration values of the soluble forms of ICAM-1, VCAM-1, IL 6 and TNF-alpha were estimated both in the serum and vitreous body samples on the basis of the ELISA method. The biochemical tests were performed routinely in the clinical laboratory. RESULTS: Mean concentrations of the investigated parameters were significantly higher in the group of patients suffering from PDR, both in the serum and in the vitreous body samples, in comparison with the values observed in the control group. In the presence of HbAlc at the level of 9.21+/ 2.17% a directly proportional correlation between the TNF-alpha either in serum or in the vitreous body samples and VCAM-1 in the vitroeus body samples was found. It is worth mentioning that the increase of the TNF-alpha concentration in the vitreous body was significantly related to the rise of the VCAM-1. CONCLUSIONS: The above mention of findings prove the remarkable role of the immunological and inflammatory response in the pathogenesis of this specific vascular complication of diabetes. The research was a source of evidence which highlighted the necessity of optimal metabolic control of the disease in the prevention of vascular complications of diabetes. PMID- 18159845 TI - [The Pro12Ala polymorphism of the peroxisome proliferator-activated receptor gamma and immunological processes in patients with type 2 diabetes and insulin resistance]. AB - INTRODUCTION: The peroxisome proliferator-activated receptor gamma (PPARgamma), a transcriptor factor, regulates immunological and metabolic processes, which are important for carbohydrate and lipid metabolism. Various polymorphic forms of PPARgamma may promote diabetes mellitus and diabetic complications. AIM OF THE WORK: The assessment of TNFalpha gene expression in peripheral blood monocytes, serum TNFalpha concentration and anti-GAD and ICA antibodies in relation to the polymorphism Pro12Ala in patients with 2 diabetes. PATIENTS AND METHODS: 58 patients with type 2 diabetes (average age 59.0 +/- 11 years) and 18 healthy people were examined. The Pro12Ala polymorphism of PPARy gene were assessed using mini-sequence technic SnaPshot [ABIPRISM-310]. The TNFalpha gene expression were estimated using real-time PCR [Applied Bio-systems]. The TNFalpha concentration [Quantikin Immunoassay, R&D Systems] and ICA and GAD antibodies [immunofluorescence method, DRG] were evaluated in venous blood. RESULTS: A heterozygotous genotype Pro12Ala was estimated in 32 patients and a homozygotous genotype Pro12Pro in 21. Only 6 patients were positive for GAD antibodies and only 6 patients for ICA antibodies. The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes did not refer to the Pro12ala polymorphism of PPARy and neither to antibodies. CONCLUSION: 1) The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes do not refer to the Pro12ala polymorphism of PPARgamma in patients with type 2 diabetes. 2) The Pro12Ala genotype do not influence autoimmunologic processes of diabetes. PMID- 18159846 TI - [Glycemic index of meals and postprandial glycemia in patients with permanent neonatal diabetes due to Kir6.2 gene mutations]. AB - Activating mutations in the KCNJ11 gene encoding the ATP-sensitive potassium channel subunit of Kir6.2 result in the phenotype of permanent neonatal diabetes (PNDM). Patients with PNDM can be successfully transferred from insulin to sulphonylurea. It is not clear, however, whether the type of diet may play a role in the metabolic control in PNDM patients. This report describes two cases of patients with PNDM due to the R201H mutation coming from the Polish Nationwide Registry of PNDM treated with the same sulphonylurea (glipizide GITS). In one of them, diet was practically free (Pol1), the other one (Pol2) avoided high glycemic-index products. Both mutation carriers were submitted to a 72 h continuous glucose monitoring system (CGMS) (Medtronic, CA). Before the CGMS record, families were encouraged not to alter their usual pattern of food intake during recording periods and to use food diaries. The postprandial glycemia in Poll reached the maximal level of 9.5 mmo/l, 5 episodes of glycemia above 8.0 mmol/l lasting overall for about 6 hours followed the ingestion of high-glycemic index (>70) meals. Patient Pol2 did not use high-glycaemic-index-products and his postprandial blood glucose did not exceed 7.0 mmol/l. Following the CGMS record, an additional diet-oriented educational session with patient Poll and his parents was performed, Poll declared to avoid the intake of high-glycemic-index products. He remained on the same dose of Glipizide GITS. Results of home blood glucose monitoring performed 2 months later showed normoglycemia. We conclude that to achieve normoglycemia, patients with PNDM who are on sulphonylurea should refrain from eating high glycemic-index products. PMID- 18159847 TI - [Glucokinase gene mutations in gestational diabetes in Polish population. Prediction of diabetes mellitus development after delivery]. AB - Gestational Diabetes Mellitus (GDM) comprises different forms of glucose metabolism disturbances with first recognition during pregnancy. There are a number of publications that have suggested that diabetes with onset during pregnancy is not a monogeneous disease and apart from "classical" form of GDM, which precedes type 2 diabetes development, MODY 2 diabetes, caused by monogenic defect of glucokinase gene, is relatively frequent "subtype" of gestational diabetes. The aim of our study was to estimate the risk of diabetes mellitus development, including MODY 2 diabetes, between 6 months - 10 years after delivery in 225 women with gestational diabetes. Gucokinase gene mutations and polymorphisms were performed by direct sequencing of DNA. In the present study it was shown that the frequency of glucokinase gene mutation is 6.7% in the Polish population of gestational diabetic women and 17.8% of new onset or persistant diabetes recognised during 5 years after pregnancy could be a result of this mutation. We have also observed that risk of type 2 diabetes development is about 50% in the next 5 years after delivery in women with gestational diabetes and is associated with higher levels of BMI during or after delivery and with clinical and biochemical features of insulin resistance (high values of WHR abd HOMA-R). Moreover, our study suggests that c.1253+8 C-->T polymorphism in intron 9 of glucokinase gene could have a role in predisposition to type 2 diabetes in women with gestational diabetes. In summary, our results suggest that, because of high costs and time-consuming methods of genetic studies, the investigations of glucokinase gene mutations should be concentrated in women with gestational diabetes without clinical and biochemical features of insulin resistance, but with family history of diabetes in two generations. PMID- 18159848 TI - [Nail susceptibility to fungal infection in patients with type 1 and 2 diabetes under long term poor glycaemia control]. AB - Onychomycosis is a common disorder in adults. Its prevalence increases also in diabetics. The objective of the study was: 1) evaluation of finger and toe nail susceptibility to Candida albicans and Trichophyton mentagrophytes infection in patients with type 1 and type 2 diabetes with long term glycaemia under poor control as compared with healthy persons, and 2) checking whether or not various aetiology of type 1 and type 2 diabetes may influence the intensity of fungal nail infection. The materials comprised finger and toe nails sampled from 26 patients with type 1 diabetes (20 females and 6 males at average age 51 +/- 10 years), 25 patients with type 2 diabetes (17 females and 8 males at average age 58 +/- 4 years). Twenty two healthy volunteers (18 females and 4 males at average age 47 +/- 14 years) served as controls. All of the diabetics (except one with type 1 diabetes and four with type 2 diabetes) had increased fasting glycaemia; moreover, all of them had poor controlled long term glycaemia because the concentration of glycated haemoglobin HbAlc exceeded 7.5%. The patients with type 1 diabetes were treated with insulin while those with type 2 diabetes with diet only (one person), with gliclazide (sixteen persons), with glimepirid (five persons), and with metformin (four persons). Enhanced fingernail susceptibility to Candida albicans infection was detected in 38.5% of the patients with type 1 diabetes, in 28% of those with type 2 diabetes, and in 22.7% of the controls. Intensive toenail infection was found in 34.6%, 20%, and 22.7% respectively. Enhanced fingernail susceptibility to Trichophyton mentagrophytes infection was found in 30.8% of the patients with type 1 diabetes, in 48% of those with type 2 diabetes, and in 4.54% of the controls while intensive toenail infection in 15.4%, 20%, and 18,2% respectively. Statistical analysis gave evidence of statistically significant higher susceptibility to infection of finger and toe nails with T. mentagrophytes but not with C. albicans in both type 1 and type 2 diabetics, as compared with the controls. In the diabetics, no significant correlation was found between the susceptibility to C. albicans and T. mentagrophytes infection and the age of the patients, metabolic diabetes control (evaluated on the basis of serum glucose level and blood HbAlc concentration), as well as the duration of diabetes. PMID- 18159849 TI - [The influence of physical training on metabolic indices in men with myocardial infarction and impaired glucose tolerance]. AB - INTRODUCTION: Body mass reduction and regular physical training form part of a strategy to treat disorders of carbohydrate metabolism associated with obesity. Evidence shows that even a slight reduction in body mass may improve carbohydrate tolerance, lipid profile and insulin resistance, reduce insulin levels and finally delay or reduce risk of diabetes mellitus. Multiple studies, including prospective studies confirm the independent protective effects of physical training against future development of type 2 diabetes mellitus. Myocardial infarction is a severe complication of atherosclerosis. Patients with glucose intolerance have a 2-fold higher risk of dying. Impaired glucose tolerance is negatively associated with prognosis in patients after myocardial infarction. Glucose intolerance accompanies hyperinsulinemia, a major indicator of insulin resistance. AIM: The aim of the study was to analyze the effect of physical training on hyperinsulinemia/ insulin resistance in patients after myocardial infarction (MI) with impaired glucose tolerance (IGT). MATERIALS AND METHODS: 31 men aged 37-69 years (mean 51 +/- 7.4) with IGT 3.5 years after MI, in NYHA class I and II participated in the study. Group A (n=16 men) underwent supervised physical training and group B (n=15) received standard information on physical training. Tissue glucose disposal using normoglycemic glucose clamp technique, fasting insulinemia, glycemia during OGTT, lipid profile, BMI and body mass composition were obtained in all patients. RESULTS: The groups were matched for age. There were no differences in BMI, percent fat content, blood pressure, diet, smoking status and pharmacotherapy. Glycemia during baseline OGTT did not differentiate the groups, either. Analysis of insulinemia and glycemia during OGTT at baseline and at 12 weeks after regular physical training showed lower levels of insulinemia and glycemia compared with baseline levels in group A (fasting glycemia 6.41+/-0.46 vs. 4.8+/-0.32 mmol/l, p<0.001; fasting insulinemia 59.42+/-38.9 vs. 27.42+/-26.5 umol/l, p<0.001). A similar analysis in group B did not reveal any significant differences (fasting glycemia 6.17+/-0.49 vs. 6.18+/ 0.46 mmol/ l; fasting insulinemia 61.86+/-48.5 vs. 68.86+/-78.5 umol/l). Mean tissue glucose disposal in the whole study population was 2.96+/-1.03 mg/kg bm/min. CONCLUSIONS: The present findings indicate the beneficial effects of regular physical training on the reduction of fasting insulinemia in patients after myocardial infarction with insulin resistance and improve lipid profile. In patients after myocardial infarction standard recommendations concerning physical activity are not well realized. It seems that better compliance, more strict control and motivation of patients can result in better metabolic control. PMID- 18159850 TI - [Serological markers of chronic Chlamydia pneumoniae and cytomegalovirus (CMV) infection in patients with peripheral occlusive artery disease--an initial report]. AB - Involvement of infection agents in pathogenesis of atherosclerosis was described in several studies, particularly in patients with acute coronary syndrome or ischemic stroke. However, in very few studies an association of serological markers of chronic infection with peripheral occlusive artery disease was analysed. The prevalence and concentration of immunoglobulin G and A to Chlamydia pneumoniae and immunoglobulin G to CMV were measured in sera of 31 participants suffering from peripheral occlusive artery disease. Significant difference in the prevalance of immunoglobulin G to C. pneumoniae and CMV between study and control groups was documented. There was no such association in reference to immunoglobulin A to C. pneumoniae index. Serum concentration of all measured antibodies were significantly higher in the study group than in control. PMID- 18159851 TI - [Role of chronic infection and heat shock proteins in peripheral arterial disease]. AB - The activation of the immunologic system plays an immportant role in the initiation of atherogenesis, as shown in numerous studies. However the role of infectious agents in this process still remains controversial. The aim of this study was to investigate the involvement of heat shock protein as a link between infection and peripheral arterial disease. 31 patients suffering from lower limb ischemia were enrolled in the study. Patients were divided into 2 groups. Group I - patients with peripheral arterial disease, group II patients with diabetic macroangiopathy. The control group consisted of 11 healthy volunteers. Blood samples were taken from each participant in order to determine serum concentrations of anti Chlamydia pneumoniae, CMV and HSP 60/65 antibodies. Statistic analysis showed anti-C. pneumoniae IgG (p< 0.025) and anti-CMV IgG (p<0.0157) antibodies were significantly more frequent in both study groups in comparison with healthy controls. Antibodie levels were also found significantly higher than in controls. Mean concentration of anti-C. pneumoniae IgG in the study group was 69.67574 vs. 18.59722 [AU/ml] in the control group (p<0.01). Analogical anti CMV IgG levels in the study group were 337.6516 vs 121.3778 [AU/ml] in controls (p<0.025). Similar changes in antibody concentration were noticed for the C. pneumoniae IgA index. 0.835258 vs. 0.176333 (p< 0.005). Antibodies against HSP 60/65 were present in significantly higher titre (p<0.005). No significant differences in antibody levels were detected beteween groups I and II. The positive correlation between anti-C. pneumoniae Ig A (r=0.3910; p<0.03) and anti HSP 60/65 antibodies titre, as well as anti-C. pneumoniae Ig G (r= 0.7151; p<0.00009) and anti HSP 60/65 speaks for the heat shock protein involvement in atherosclerotic plaque development. PMID- 18159852 TI - [Evaluation of polyneuropathy severity in chronic renal failure patients on continuous ambulatory peritoneal dialysis or on maintenance hemodialysis]. AB - The aim of the study was to assess severity of injury of the peripheral nervous system of chronic renal failure patients on continuous ambulatory peritoneal dialysis (CAPD) or on maintenance hemodialysis (HD) diagnosed based on the electrophysiological testing as well as to establish the effect of adequate dialysis dose on peripheral nerve functioning. The study was performed in 53 patients (21 males, 32 females) on CAPD for mean 16.9 +/- 14.4 months and 68 patients (46 males, 22 females) on HD for mean 47.5 +/- 46.5 months. Both groups were age matched (48.9 +/- 12.4 vs. 50.6 +/- 11.9 yrs; p= NS). Patients with co existing diseases that could lead to disturbances in nerve conduction were excluded from the study group. METHODS: Electrophysiological testing was performed using Medelec Shaphire 2ME equipment. Nerve conductivity testing was performed in motor and sensory fibres of right ulnar nerve, motor fibres of right fibular nerve and left tibial nerve as well as in sensory fibres of right calf nerve. In the sensory nerves there were assessed: amplitude of the evoked sensory response, latency of the response and conduction velocity. In the motor nerves there were analyzed amplitude of the compound muscle action potential, latency of the evoked response, F-wave minimal latency and conduction velocity. Polyneuropathy was diagnosed if abnormal results obtained in electrophysiological examination were found in at least two out of four tested nerves. To assess dialysis adequacy in the CAPD group weekly Kt/V (wKt/ V) and weekly creatinine clearance (WCrCI) were used and in the HD patients the following indices were measured: Kt/V, PRU, TAC and TAD. RESULTS: Polyneuropathy diagnosed based on the above described criteria, was found in 59 (86.8%) HD patients and in 41 (77.4%) CAPD patients (p= NS). The results of motor conductivity testing showed significantly longer distal latency in HD patients in comparison to the CAPD patients in fibular nerve (4.9 +/- 0.9 vs. 4.5 +/- 0.8 ms; p< 0.05) and F-wave latency in ulnar nerve (29.3 +/- 3.1 vs. 27.3 +/- 2.5 ms; p< 0.05), in fibular nerve (52.4 +/- 6.4 vs. 48.5 +/- 7.94 ms; p< 0.05), in tibial nerve (54.8 +/- 7.8 vs. 50.6 +/- 7.12 ms; p< 0.05) and also significantly lower conduction velocity in ulnar nerve (51.9 +/- 5.9 vs. 55.6 +/- 6.99 m/s; p< 0.05), in fibular nerve (41.2 +/- 5.9 vs. 44.5 +/- 5.5 m/s; p< 0.05), in tibial nerve (40.1 +/- 5.81 vs. 42.7 +/- 4.6 m/s; p< 0.05). Mean value of sensory response amplitude evoked in ulnar nerve was significantly higher in the CAPD treated patients than in HD patients (21.2 +/- 14.8 vs. 15.1 +/- 11.4 microV; p< 0.05) and distal latency in calf nerve was significantly shorter (2.1 +/- 1.0 vs. 2.4 +/- 0.6 ms; p< 0.05). Analysing the effect of dialysis on peripheral nerve functioning in the group of CAPD patients a statistically significant relationship between the severity of peripheral nerve injury and WCrCl was demonstrated. Mean value of WCrCl in patients without features of neuropathy was significantly higher (83.3 +/- 28.98 l/week/ 1.73m2) in comparison with those with diagnosed polyneuropathy (59.9 +/- 15.9 l/week/1.73m2) (p< 0.01). Analyzing effect of WCrCl on conduction velocity in the tested nerves there was demonstrated that in patients with WCrCl value at least 60 l/week/1.73m2, mean velocity values were higher, and the statistically significant difference was found for sensory (p< 0.05) and motor (p< 0.01) fibers of ulnar nerve. Mean value of residual renal clearance in the CAPD treated patients without features of sensory and motor nerve injury was significantly higher (2.9 +/- 1.8 ml/min) when compared to the patients with diagnosed polyneuropathy (1.4 +/- 1.3 ml/min) (p= 0.006). In the hemodialyzed group no statistically significant relationship was found between conduction velocity in the tested nerves and indices of dialysis efficiency (Kt/V, PRU) as well as degree of exposure to uremic toxemia described by TAC and TAD. CONCLUSIONS: Polyneuropathy is a common complication in patients with chronic renal failure independently of a kind of the therapy. Well preserved residual renal function in CAPD patients plays an important role in improving effectiveness of the dialytic treatment, in consequence influencing preservation of proper peripheral nerve function. PMID- 18159853 TI - [Blood serum osteocalcin and beta-crosslaps concentrations in patients after renal transplantation]. AB - In 16 patients (7 males, 9 females), aged 47.2 +/- 15.9 years, blood serum concentrations of osteocalcin, beta-crosslaps, parathormone, calcium, phosphate, creatinine and urea were determined before renal transplantation and 3 and 6 months following the procedure. Three as well as six months following renal transplantation significant decrease in blood serum concentration of osteocalcin and beta-crosslaps are found. A significant positive correlation between osteocalcin and beta-crosslaps concentration was found in each investigated period. Six-months observation revealed only partial correction of bone metabolism following renal transplantation. PMID- 18159854 TI - [Microcirculatory disturbance in the course of acute pancreatitis]. AB - Although as many studies are sacrificed to acute pancreatitis (AP), the pathogenesis of the disease remains undiscovered. The microcirculatory disturbances during AP as a cause of necrotic changes in the parenchyma of the gland, described first time by Panum in 1886, have become again a very attractive theory. In the last decades, several studies were done to prove that microcirculatory impairment plays a crucial role in pathogenesis of severe AP. It had been proved that many mediators play important roles in this process including nitric oxide and endothelin balance. Vasoconstriction and vasodilatation named "vascular game" caused by cytokine accumulation and lower oxygenation of the tissue gives very interesting basis for theories about creating pancreatic necrosis. The severity of the course of AP also determinates damage of the microcirculation of the lungs, the liver and the bowels which leads to SIRS, MODS and MOF. The new experimental dates seems to be very promising and can became a basis for developing a new strategy of treatment in the most severe cases of AP. In this article we present up-to-date information about these theories and concepts. PMID- 18159855 TI - [The clinical picture of morphea]. AB - Skin scleroderma (LS) is characterised by stiffness of skin and/or deeper tissues. As opposed to systemic scleroderma, the involvement of internal organs and Raynaud phenomenon are predominately not observed in morphea. LS is quite rare disease, more frequent in women and young people. There are several useful classifications of skin scleroderma in literature, however the classification which concerns the shape and extension of LS lesion and depth of stiffness is concerned to be the most clear. LS is divided into: plaque morphoea, generalised morphoea, blistering morphoea, linear morphoea and deep morphoea. Different types of skin scleroderma lesions can be observed in one patient or can combine linear and deep fibrosis. Presented classification is clinically useful and it has prognostic and therapeutic implications. PMID- 18159856 TI - [Coexistence of Takayasu's disease and atherosclerosis in computed tomography angiography examination--case report]. AB - Takayasu disease is a syndrome of unknown etiology having an immunological basis and appearing mainly in young women. It leads to characteristic changes in arteries as confirmed by diagnostic imaging methods. The case of a 51-year-old woman with recognized Takayasu's disease and stenocardial complaints arising from atheroma of the coronary arteries is presented. Computed tomography angiography examination showed changes typical for both Takayasu's disease and atherosclerosis. The potential influence of the inflammation appearing in Takayasu's disease on atheroma is discussed. The clinical signs accompanying Takayasu's disease and diagnostic imaging methods for their detection are presented. PMID- 18159857 TI - How nurses will tackle the flu pandemic. PMID- 18159858 TI - 'We need to protect our patients from abuse and those who will inflict it'. PMID- 18159859 TI - Maidstone and Tunbridge Wells nurses speak out. PMID- 18159860 TI - Evidence on effectiveness of self-care support strategies. PMID- 18159861 TI - The sense of taste. PMID- 18159862 TI - Understanding invasive monitoring 1: indications. AB - This is the first part of a two-part unit on invasive monitoring, which aims to increase readers' understanding of the use of these methods and to reduce the incidence and impact of associated complications. This part explores the background and indications in relation to invasive line insertion. PMID- 18159864 TI - Primary care organisation of the year. Winner: Wirral Primary Care Trust. PMID- 18159863 TI - Acute healthcare organisation of the year. Winner: South Tees Hospitals Trust. PMID- 18159865 TI - Chronic disease management. Winner: Cornwall and Isles of Scilly PCT with Cornwall CC Department of Adult Social Care. PMID- 18159866 TI - Clinical service redesign. Winner: Salford Royal Foundation Trust and Salford PCT. PMID- 18159867 TI - Communications. Winner: Newham University Hospital Trust. PMID- 18159868 TI - Cost-effective partnership working. Winner: Birmingham East and North PCT. PMID- 18159869 TI - Good corporate citizenship. Good corporate citizenship. Winner: Nottingham University Hospitals Trust. PMID- 18159870 TI - Implementing NICE guidance. Winner: Oxleas Foundation Trust. PMID- 18159871 TI - Improving care with e-technology. Winner: NHS North West, Lancashire and South Cumbria Cardiac Network and Broomwell Healthwatch. PMID- 18159872 TI - Improving patient access. Winner: Bolton PCT. PMID- 18159873 TI - Information-based decision making. Winner: Leicestershire, Northamptonshire and Rutland Cancer Network. PMID- 18159874 TI - Mental health innovation. Winner: Oxfordshire PCT. PMID- 18159875 TI - Patient-centred care. Winner: Leeds PCT. PMID- 18159876 TI - Patient safety. Winner: Royal Wolverhampton Hospitals Trust. PMID- 18159877 TI - Primary care innovation. Winner: Bradford and Airedale Teaching PCT. PMID- 18159878 TI - Recruitment and retention. Winner: NHS Tayside. PMID- 18159879 TI - Reducing health inequalities. Winner: East Lancashire PCT. PMID- 18159880 TI - Skills development. Winner: The DESMOND Collaborative. PMID- 18159881 TI - Your liability for ED patients. PMID- 18159882 TI - Group practices pay better. PMID- 18159883 TI - Sinking reimbursement, harder work. PMID- 18159884 TI - The smarter way to faster billing collections. PMID- 18159885 TI - Coping with tough markets: private practice doctors fight for survival. PMID- 18159886 TI - District general hospitals face heavy specialist service losses. PMID- 18159887 TI - National director to back anti-privatisation charity. PMID- 18159888 TI - Social care green paper. How will government defuse the demographic time bomb? PMID- 18159889 TI - Noel Plumridge on finding the right GP. PMID- 18159890 TI - Mental health. The safety scandal. AB - *Sexual safety incidents are treated as part of mental health inpatient life. *Disbelief is built into the system. There is an attitude that patients cannot be believed because they are ill. *A lack of adequately trained and experienced staff can exacerbate poor levels of safety. PMID- 18159891 TI - Health inequalities. The great divide. PMID- 18159892 TI - Workforce planning. Dish of the day. PMID- 18159893 TI - Soft tissue augmentation. PMID- 18159894 TI - Tumescent anesthesia: evolution and current uses. AB - The tumescent technique of local anesthesia is one of the great techniques developed in the last 20 years. It has dramatically changed the way clinicians perform many cutaneous surgical procedures. Those that use the technique are forever grateful for the insight that Dr. Jeffrey Klein used to push the envelope and discover the safe doses of subcutaneous administration of lidocaine. PMID- 18159895 TI - Sentinel lymph node evaluations. PMID- 18159896 TI - Genetic mutations involved in melanoma: a summary of our current understanding. AB - The biomolecular understanding of melanoma is in flux. The importance of high penetrance genes involved in familial melanoma includes a significant number of mutations that directly lead to impairment of the checkpoints of the normal cell cycle. Furthermore, a greater understanding of the interaction between genetic factors and environmental factors, such as MC1R, CDKN2A, BRAF, and ultraviolet light, is emerging from landmark research. Although currently and with rare exception most clinicians still confine genetic testing to the realm of research, even in familial melanoma, continued and major advances in this arena may lead to development of new and revolutionary means of diagnosis and treatment, patterned on improved understanding of melanoma-related genetic mutations and resultant aberrations in cellular pathways. PMID- 18159897 TI - Management of cutaneous melanoma: a public health and individual patient care perspective. AB - The damaging impact of cutaneous melanoma on individuals and society is apparent. As dermatologists, we are in a unique position to help each patient on a personal level and make contributions with sweeping effects on a societal level. On an individual level, the successful care of each patient involves accurate and early diagnosis, proper education, sufficient biopsy, adequate surgical and medical managements, and long-term follow-up. It is also imperative to remember that each patient presents with his or her own set of unique challenges and needs. The general assessment of risk factors and predictions on survival outcome should only serve as a starting point for discussion during the consultation. A significant percentage of patients will defy these predictions. On a public health level, the concerted endeavor until now to reduce the incidence of the disease should be viewed as a partial success, considering the recent trends in the incidence and mortality rate of the disease. However, there are still many questions that need to be addressed before the implementation of more comprehensive and effective strategies to control MM mortality. It is only through this unrelenting pursuit by the medical and scientific community that may eventually lead to understanding, and perhaps a cure for, this deadly skin cancer that is easily visualized but has thus far proven difficult to control. PMID- 18159899 TI - Nephrogenic systemic fibrosis: a review and exploration of the role of gadolinium. AB - NSF is a new and emerging disease. Significant investigative work to date has led to an unexpected suspect-gadolinium-containing contrast agents. Considerable additional work is now underway to formulate specific recommendations about the use of these agents in the population of patients who have renal disease. Goals on the immediate research horizon include (1) the identification of risk factors and conditions that must be met to permit the development of NSF in patients who have renal disease, (2) the characteristics of contrast agents that make them more or less likely to induce NSF, and (3) the development of prophylactic or treatment strategies that can reduce the overall development and severity of NSF. The investigative process has already yielded new insight into the functions (and malfunctions) of the CF in the setting of NSF. As the CF is being increasingly implicated in other organ-specific and systemic fibrosing disorders, we can expect to see significant developments in the studies of allied disorders as well. PMID- 18159900 TI - The efficacy and safety of amoxicillin, trimethoprim-sulfamethoxazole, and spironolactone for treatment-resistant acne vulgaris. AB - In situations where acne has proved resistant to other systemic agents or they are not indicated and isotretinoin is not desired by patients, the authors find amoxicillin, TMP-SMX, and spironolactone, alone or in combination, useful alternatives. In women who have acne, spironolactone can be used, and in patients who have sulfa allergy, amoxicillin can be used. It is important to continue alternate topical therapy along with these interventions to augment the improvement and to assist in the eventual discontinuance of oral medication. All of the authors' patients were using topical therapy concomitantly and, although this may have contributed to improvement, the authors believe the addition of amoxicillin, TMP-SMX, or spironolactone contributes to the majority of improvement. Tetrospective chart analysis provides supportive data for amoxicillin, TMP-SMX, and spironolactone in the treatment of refractory acne vulgaris. PMID- 18159898 TI - Growth factors and oncogenes as targets in melanoma: lost in translation? PMID- 18159901 TI - The dermatopathology of new and emerging infectious diseases. PMID- 18159902 TI - Emerging cutaneous infections in the premature neonate. AB - As the age of viability of premature neonates continues to decline, early recognition of emerging cutaneous infections will become increasingly important. Fungal and bacterial infections should always be included in the differential diagnosis of unusual-appearing skin lesions in the premature neonate since prompt evaluation and aggressive therapy is paramount in this high-risk population. PMID- 18159903 TI - Genetic immunodeficiency diseases. AB - The approach to the patient with genetic immunodeficiency is multidisciplinary, and requires close interaction between the primary care physician, immunologist, and other specialists. Dermatologists may play a key role in both the diagnosis of immunodeficiency based on recurrent infection or specific cutaneous abnormalities and in the management of cutaneous complications. The availability of bone marrow and stem cell transplantation has been life-saving for many affected children. The underlying genetic basis is now known for most forms of immunodeficiency, which has facilitated confirmation of patient diagnosis and prenatal diagnosis. Gene therapy has already been initiated for severe combined immunodeficiency, and will certainly play a growing role in therapy of this group of disorders in the future. PMID- 18159905 TI - Bullous pemphigoid: physiopathology, clinical features and management. AB - There has been a considerable progress in the understanding of the physiopathology of BP during the past 2 decades. The insights into the humoral and cellular immune response against BP180 and BP230 have increased significantly. Nevertheless, the factors underlying the initiation of the disease leading to a disruption of self-tolerance remain unclear. Clinically, the disease shows protean presentations, and diagnostic delay is common. A practical, relevant, and unresolved question is how to identify patients suffering from BP at an early stage of the disease, when direct immunofluorescence microscopy findings still may be negative. The characterization of markers allowing the differentiation of BP from other pruritic eruptions occurring in the elderly population would be extremely helpful in daily practice. Finally, despite the knowledge that potent topical steroids are efficient in controlling the disease, management of BP sometimes remains difficult and requires systemic therapies. It is hoped that a better knowledge of the regulation of the autoimmune response in BP also will facilitate the design of novel immunomodulatory therapeutic approaches devoid of the severe side effects of current immunosuppressive treatments. PMID- 18159906 TI - Hidradenitis suppurativa: an update. AB - HS is a chronic, debilitating inflammatory dermatosis that is often refractory to treatment. Many medical therapies have been tried and some show efficacy. The introduction of new immunosuppressive medications has revolutionized the treatment of psoriasis in dermatology. These therapies have the potential to powerfully abort the molecular signals driving inflammation in HS and perhaps even induce a remission. This may offer patients an alternative to radical excisional surgery. However, more studies are needed to evaluate the long-term effects of these medications, particularly in light of their association with an increased risk of malignancy and infection. Evaluation of treatment efficacy has been hindered by a lack of uniform standards to track treatment response. The classic clinical classification system has limitations but is a useful scheme to guide selection of treatments. Further laboratory studies may be warranted, depending on the clinical presentation (Box 3). While there are many approaches to treating HS, the authors offer an algorithm based on disease severity (Fig. 9). It is important to address many patient-oriented concerns, such as the pain, dampness, and smell associated with draining sinuses. Communication with the patient about expectations is important to direct treatment. Often a multidisciplinary approach involving dermatology, plastic surgery, gynecology, and urology may be necessary to diagnose and treat the disease. Both medical and surgical options should be explored before embarking on therapy. PMID- 18159904 TI - Ichthyosis update: towards a function-driven model of pathogenesis of the disorders of cornification and the role of corneocyte proteins in these disorders. PMID- 18159907 TI - Dendritic cells and T cells in the regulation of cutaneous immunity. AB - DCs are a complex cell population in the skin consisting of epidermal LCs and dermal DCs, which differ in their anatomic location, antigen recognition, processing machinery, and migratory capacity. Cutaneous DCs (LCs as well as dermal DCs) function as sentinels that survey invading agents and transmit the information into immune responses by taking up exogenous antigens, migrating to draining LNs, and presenting the processed antigens to T cells resulting in T cell differentiation and activation. Indeed, further studies are needed to clarify the specific contribution of each cutaneous DC subpopulation to antigen presentation and induction of cutaneous immune responses. Recent results suggested to revisit the "paradigm" that exclusively immigrant LCs present skin acquired antigens to T cells upon reaching the draining LNs, because viral antigens, for instance, are presented by a DC subpopulation other than LCs after infection of mouse epidermis with herpes simplex virus [153]. Additionally, different DC subpopulations may sequentially present skin-acquired antigens, possibly serving as a regulatory mechanism of cell-mediated immunity and adding further complexity to established concepts. Nevertheless, cutaneous DCs are involved in several pathologies (including infections, inflammatory disorders, or skin cancers) and play a pivotal role in regulating the balance between immunity and peripheral tolerance. However, it is widely accepted that (cutaneous) DC in an immature state may have tolerogenic properties resulting in the induction or expansion of Tregs. CD4+CD25+ Tregs are essential for the control of immune responses in inflammatory, autoimmune, or cancer diseases, and it is well established that in particular the lineage-specific transcription factor Foxp3, as well as cytokines (including IL-2, IL-10, and TGF-beta), characteristic surface markers such as CTLA-4, and members of the TNF superfamily (e.g., RANKL) are critically involved in the thymic development, peripheral maintenance, and suppressive activity of CD4+CD25+ Tregs. Recently, new methods for generating and expanding Tregs in vitro have emerged and supported the use of CD4+CD25+ T cells as a novel strategy for the treatment of patients suffering from autoimmune diseases. In the future, a better understanding of Treg function in vivo and the interactions of Tregs and pathogenic effector T cells in autoimmune disorders may help to improve the design of Treg-based therapies. PMID- 18159908 TI - Emerging diseases in tropical dermatology. AB - All dermatologists worldwide should have at least passing familiarity with various tropical maladies that generate cutaneous manifestions. In addition to the standard infectious ailments associated with tropical environs, the authors have described herein five "emerging" illnesses that are gaining increasing attention for their capacity to cause human disease in those immigrating from, or traveling to, the tropical and subtropical world. PMID- 18159909 TI - [Paediatric orthopedics]. PMID- 18159910 TI - [Jumper's knee. Surgery and arthroscopic treatment with scraping and povidone collagen in high-performance athletes]. AB - The jumper's knee or patellar tendonitis is a common injury in the athlete with an incidence between 14% and 16% among high-performance athletes. In addition to an overuse injury, there are some intrinsic factors for its development. Conservative treatment is indicated for the initial form, but when it fails, surgical treatment should be performed with an appropriate rehabilitation program. MATERIAL AND METHODS: We retrospectively studied 18 high performance athletes in various disciplines, with an average age of 22 years, operated by arthroscopy and mini-arthrotomy scraping and application of povidone collagen sponge between March 2001 and December 2005. There after patients underwent a rehabilitation program specific for their return to their athletic activity. RESULTS: The patients returned to their sports activity in an average of 15 weeks, with functional knees without pain, with full range of motion. We did not find postoperative fibrosis. DISCUSSION: The results were similar and slightly better in time of return to sports activities to those reported in the world literature, with the difference in follow up. According to the clinical evaluation, treatment performed allows functional improvement with a return to athletic activity in a reasonable time. PMID- 18159911 TI - [Minimally invasive surgery in diaphyseal humeral fractures with helicoidal plate. One year result in seven patients]. AB - INTRODUCTION: The surgical treatment of choice in the first instance before a humeral diaphysis fracture surgery are intramedullary nails. So are simple plates or external fixators that in different studies reported a large percentage of complications. OBJECTIVE: To implement an alternative system of treatment that shows to be minimal invasive, painless and inexpensive. MATERIAL AND METHODS: Applied in 7 patients with a follow-up to a year. The type of study is prospective, cross-sectional observational and descriptive. RESULTS: The results were no cases of nonunion, no delayed unions or infections, no radial or circumflex nerve injury added postoperatively with a functional restoration of 90% between the 6th and the 8th weeks of the surgery, and a proper consolidation. CONCLUSIONS: This treatment method proved to be minimally invasive with no extra features in their application with a functional recovery between the 6th and 8th week after surgery. The total cost turned out to be minor in comparison to other surgical methods. PMID- 18159912 TI - [Prosthesis use in pediatric patients with fibular hipoplasia]. AB - OBJECTIVE: To assess the prosthesis adaptability at use of pedestal and transtibial prosthesis, recognize the average average age of amputation surgery, and the complications of the amputation in patients with fibular hemimelia. MATERIAL AND METHODS: 47 patients were evaluated, initially to adjust pedestal prosthesis and when this was not tolerated, we realize amputation and adaptation of transtibial prosthesis, we valued the average age of amputation surgery, the associate complications and the prosthesis adaptability we use the K system for functional ambulation. RESULTS: Of the 12 patients to actually use pedestal prosthesis 9 (75%) had a K.2 level, 3 (25%) had a K-3 level, of the 35 patients to use transtibial prosthesis 1 (3%) had a K-2 level, 19 (54%) K-3 level, and 15 (43%) in the K-4 level. The average age to amputation surgery was 3.9 years old, one case had a complication. DISCUSSION: The early prosthesis adaptability provides advantages in the functional ambulation, as demonstrated on the results. The goal of the transtibial amputation is to facilitate the prosthesis adaptability, is due to take advantage of the best functional adaptation the child in the first years of life, circumstance that improves the integral rehabilitation of the patient. PMID- 18159913 TI - [Etofenamate and the analgesic effect in the management of acute pain from spine in the emergency room]. AB - OBJECTIVE: To demonstrate that the application of etofenamate is effective in management of acute pain. MATERIAL AND METHODS: We conducted a prospective, longitudnal, 6 months trial, which included 22 women and 18 men with intense acute low back pain of mechanical or postural aetiology, patients who had pain of traumatic origin and needed radiographic studies by orthopaedic surgeon were excluded; the study subjects were treated with 1 g etofenamate intramuscularly and the analgesic effect was assessed by visual analog scale every 5 minutes for 30 minutes. RESULTS: Marked improvement in pain at 25 minutes in 35 subjects (87.5%). Pain did not improve in 5 subjects (four men, one woman, 12.5%) at 30 minutes. There were no adverse reactions to medication. DISCUSSION: The single dose of 1 g etofenamate is effective in the management of acute pain. Its use prevented 35 admissions with a cost savings of $70,000 pesos. Applying etofenamate caused satisfaction of the beneficiaries and emergency personnel, this drug could be an alternative treatment in medical services and first-level emergency. PMID- 18159914 TI - [Utility of platelet-rich plasma and growth factors bone in the bone defects. Regional Hospital Lic. Adolfo Lopez Mateos, ISSSTE]. AB - INTRODUCTION: The platelet-rich plasma is a concentrated of platelets with the presence of growth factors and proteins that serve as osteoconducer matrix for bone formation. We present the results obtained with the use of platelet-rich plasma and a hydroxyapatite and bovine collagen graft in the management of bone defects. MATERIAL AND METHODS: We studied eight patients with bone defects, treated surgically in whom platelet rich plasma and a hydroxyapatite and bovine collagen graft was used, with clinical and radiographic follow-up at 2, 4, 6, 10 and 18 weeks after surgery. RESULTS: Starting on week 7 since surgery, evidence of osteointegration and bone callus formation, during the weeks 10 and 14 most cases showed bone consolidation. A case without consolidation through week 18. DISCUSSION: The immediate response of the body tissue damaged is the accumulation of a large number of activated platelets, which release their granules and growth factors that promote the regeneration of tissues. It is possible to obtain platelet-rich plasma and accelerating the process of bone regeneration. Our study shows beneficial results with the use of platelet-rich plasma. PMID- 18159915 TI - [Lumbar osteochondroma and radicular compression. A case report]. AB - Most cartilage forming tumors are benign and rarely affect the spinal canal, their principal location is in humerus metaphysis, femur and tibia. They represent 2% of all tumors and 2.6% of the benign tumors of the spine, usually located at the cervical spine. It is speculated that the secondary ossification center may be aberrant cartilaginous tissue that causes the formation of osteochondroma. From 1 to 4% of osteochondromas involve the spine and are commonly included in the posterior elements of the vertebrae and if located near neurological structures causing irritation and spinal compression is rare but potentially catastrophic. The average age of clinical onset is at the 2nd and 3rd decade of life. CASE PRESENTATION: 50-year-old male who began his illness four years ago with pain in the lumbar region with right leg paresthesias, muscle strength 4/5 in the dermatomes L4 and L5, pain was increasing in intensity and frequently. The complete cure was to remove the exostosis of the lamina of right side of L4 with projection to the medullary canal and origin of the contact with the root of L4. We performed: clinical history, physical examination, X-rays, magnetic resonance imaging, electromyography, histology. It was performed: total removal of the tumor effectively and symptoms disappeared. As the spinal osteochondromas are rare and tend to occur in young adults. PMID- 18159916 TI - [Treatment of chondral lesions from knee injuries in the professional soccer players. Review of 34 cases]. AB - OBJECTIVE: Evaluate the results of Pridie chondroplasty as an efficient treatment to recover the complete activity in soccer players and compare this with others chondral repair techniques. MATERIAL AND METHODS: The patients were professional soccer players, all with a knee cartilage injury treated with the Pridie technique in an arthroscopic surgery between March 1999--December 2004. The age of the patients and the presence of a simple meniscal tear wasn't a fact to exclude a patient. Complex meniscal tear and a ACL rupture were an exclusion criteria. We deferred the support by 6 to 8 weeks. We initiated rehabilitation to the 4-5 post operating day. The follow up average was of 30 months. RESULTS: We included 34 patients, age rank 19-31 years (average 24.6), 11 of them with meniscal injury Degree I associated with chondral damage. In 26 patients (76.47%) the outcomes were good allowing them to take up again their high level sport activity. The rest (23.53%) had regular or bad results with decrement in the game level, 4 of which (11%) they retired of the professional practice in relation to the found injuries. CONCLUSIONS: The follow up time give us a good validation to establish that the used technique is a treatment of low cost, surgically simple with favorable outcomes and low morbidity comparable to the results obtained with other useful techniques of condral repair in the professional soccer player. PMID- 18159917 TI - [Comprehensive treatment in Achilles tendon rupture]. AB - Due to incapacity caused by calcaneal tendon injuries for the reintegration of patients back to their daily activities and/or sparts it is necessary to decrease the time of reinstatement of patients. At present these times have improved by a good surgical technique and an early rehabilitation, and the patient is returned quickly as he sees less disability. It is proposed in this paper a type of surgical treatment and an early rehabilitation program, which have shortened the time of disability and incorporation to their daily activities and sports to eight weeks in 10 patients with Achilles tendon plasty. PMID- 18159918 TI - [Instability from the distal radioulnar joint: reconstruction and evolution of dorsal triangular fibrocartilage complex ligament]. AB - Dorsal and palmar radio ulnar ligaments are part of the triangular fibrocartilage complex and are the principal stabilizer of this articulation. The distal radio ulnar joint provides not only pronation and supination to the forearm, but also it's basic for strength. The objective of this study is to evaluate the functional results of the dorsal radio ulnar joint ligament reconstruction in patient with distal radio ulnar instability. This is a prospective, longitudinal, experimental and auto-control assay. Functional preoperative data was analyzed, proper surgical technique and another findings. The patients were evaluated with a functional exploration, comparative dynamometry and DASH evaluation, they were also asked about their personal satisfaction. Between 1997 and 2004, this treatment was practiced in eleven patients (2 men and 9 women) with distal radio ulnar joint instability. With a mean age of 34.1 years. The dominant had was involved in 91% of patients. The mean complaint was pain for all patients, weakness in 9 patients and instability in 7. The mean follow up was 2 years 4 months, with a range from 6 months to 8 years. The patients refer totally satisfied, 8 of them, partially satisfied 2 and one patient insatisfied. PMID- 18159919 TI - [Distal tibial reconstruction and ankle arthrodesis in osteosarcoma (salvage technique). A case presentation]. AB - Treatment of bone sarcomas is aimed in healing and secondarily rescuing the limb. With the use of chemotherapy, radiation therapy, new prostheses and availability of graft, now most tumors can be resected, reducing functional deficit. OBJECTIVE: To present a surgical option in management of osteosarcoma of distal tibia. CASE REPORT: Fifteen year-old man, starting in April 2003 with a mass in the left ankle area. June 2003 pulsatile progressive pain started, increased with activity, decreased with rest. November 2003 the physical exam: limping by left lower limb, with a tumor in the posteromedial ankle surface (3 x 2 cm), hard, not movable, regular edges, painful on palpation, Radiographic observations (November 2003), showed blastic oval stone image (4 x 3.5 cm) with expanded posterior and medial cortices with periosteal reaction. The initial biopsy report (chondroblastoma) was not consistent with the diagnostical assumption. So resection was performed 10 cm wide, placing a methylmethacrylate spacer. The final histological report was positive to osteogenic sarcoma. The patient received 11 sessions of chemotherapy. The reconstruction of the distal tibia and arthrodesis with autologous graft from the fibula, heterologous graft and stabilization with transcalcaneal nail, was done in March 2005. RESULTS: Successful evolution without tumor activity with graft integration to arthrodesis and plantigrade gait. PMID- 18159920 TI - [Leonardo Zamudio Villanueva, MD]. PMID- 18159921 TI - In search of novel agents for therapy of tropical diseases and human immunodeficiency virus. AB - Malaria, sleeping sickness, Chagas' disease, Aleppo boil, and AIDS are among the tropical diseases causing millions of infections and cases of deaths per year because only inefficient chemotherapy is available. Since the targeting of the enzymes of the polyamine pathway may provide novel therapy options, we aimed to inhibit the deoxyhypusine hydroxylase, which is an important step in the biosynthesis of the eukaryotic initiation factor 5A. In order to identify new lead compounds, piperidines were produced and biologically evaluated. The 3,5 diethyl piperidone-3,5-dicarboxylates 11 and 13 substituted with 4-nitrophenyl rings in the 2 and 6 positions were found to be active against Trypanosoma brucei brucei and Plasmodium falciparum combined with low cytotoxicity against macrophages. The corresponding monocarboxylates are only highly active against the T. brucei brucei. The piperidine oximether 53 demonstrated the highest plasmodicidal activity. Moreover, compounds 11 and 53 were also able to inhibit replication of HIV-1. PMID- 18159922 TI - Structure-activity relationships of novel iron chelators for the treatment of iron overload disease: the methyl pyrazinylketone isonicotinoyl hydrazone series. AB - The design of novel Fe chelators with high Fe mobilization efficacy and low toxicity remains an important priority for the treatment of Fe overload disease. We have designed and synthesized the novel methyl pyrazinylketone isonicotinoyl hydrazone (HMPIH) analogs based on previously investigated aroylhydrazone chelators. The HMPIH series demonstrated high Fe mobilization efficacy from cells and showed limited to moderate antiproliferative activity. Importantly, this novel series demonstrated irreversible electrochemistry, which was attributed to the electron-withdrawing effects of the noncoordinating pyrazine N-atom. The latter functionality played a major role in forming redox-inactive complexes that prevent reactive oxygen species generation. In fact, the Fe complexes of the HMPIH series prevented the oxidation of ascorbate and hydroxylation of benzoate. We determined that the incorporation of electron-withdrawing groups is an important feature in the design of N, N, O-aroylhydrazones as candidate drugs for the treatment of Fe overload disease. PMID- 18159923 TI - 7-fluoroindazoles as potent and selective inhibitors of factor Xa. AB - We have developed a novel series of potent and selective factor Xa inhibitors that employ a key 7-fluoroindazolyl moiety. The 7-fluoro group on the indazole scaffold replaces the carbonyl group of an amide that is found in previously reported factor Xa inhibitors. The structure of a factor Xa cocrystal containing 7-fluoroindazole 51a showed the 7-fluoro atom hydrogen-bonding with the N-H of Gly216 (2.9 A) in the peptide backbone. Thus, the 7-fluoroindazolyl moiety not only occupied the same space as the carbonyl group of an amide found in prior factor Xa inhibitors but also maintained a hydrogen bond interaction with the protein's beta-sheet domain. The structure-activity relationship for this series was consistent with this finding, as the factor Xa inhibitory potencies were about 60-fold greater (DeltaDelta G approximately 2.4 kcal/mol) for the 7 fluoroindazoles 25a and 25c versus the corresponding indazoles 25b and 25d. Highly convergent synthesis of these factor Xa inhibitors is also described. PMID- 18159924 TI - Semisupervised model-based validation of peptide identifications in mass spectrometry-based proteomics. AB - Development of robust statistical methods for validation of peptide assignments to tandem mass (MS/MS) spectra obtained using database searching remains an important problem. PeptideProphet is one of the commonly used computational tools available for that purpose. An alternative simple approach for validation of peptide assignments is based on addition of decoy (reversed, randomized, or shuffled) sequences to the searched protein sequence database. The probabilistic modeling approach of PeptideProphet and the decoy strategy can be combined within a single semisupervised framework, leading to improved robustness and higher accuracy of computed probabilities even in the case of most challenging data sets. We present a semisupervised expectation-maximization (EM) algorithm for constructing a Bayes classifier for peptide identification using the probability mixture model, extending PeptideProphet to incorporate decoy peptide matches. Using several data sets of varying complexity, from control protein mixtures to a human plasma sample, and using three commonly used database search programs, SEQUEST, MASCOT, and TANDEM/k-score, we illustrate that more accurate mixture estimation leads to an improved control of the false discovery rate in the classification of peptide assignments. PMID- 18159926 TI - Effect of cholesterol on the release of amphotericin B from PEG-phospholipid micelles. AB - Micelles formed from PEG-DSPE solubilize high levels of the poorly water-soluble antifungal amphotericin B (AmB). AmB release from PEG-DSPE micelles is slow in buffer but remarkably rapid in the presence of bovine serum albumin (BSA). Sequential changes in the absorbance spectrum of AmB in PEG-DSPE micelles point to a rapid dissociation of incorporated drug in the presence of BSA. In this context, we have studied micelles formed from PEG-DSPE which coincorporate cholesterol (PEG-DSPE|cholesterol). (1)H NMR measurements point to a lower mobility of lipid in PEG-DSPE|cholesterol micelles compared to PEG-DSPE micelles. The absorbance spectrum of AmB incorporated in PEG-DSPE|cholesterol micelles is distinct from that in PEG-DSPE micelles, which may point to differences in the drug-micelle interaction. AmB release from PEG-DSPE|cholesterol micelles is slow in buffer and in the presence of BSA. The absorption spectrum of AmB in PEG DSPE|cholesterol micelles remained unchanged in BSA, further supporting stable incorporation and the slow release from the carrier. PMID- 18159927 TI - Structure-transfection activity studies of novel cationic cholesterol-based amphiphiles. AB - Inclusion of DOPE in lipoplex formulations has hampered the establishment of a correlation between cationic lipid structure, biological specificity, and transfection activity, simply because the presence of a helper lipid not only alters the physicochemical properties of the lipoplex but also modifies cell surface specific interactions during the process of transfection. To this end, four cationic cholesterol-based derivatives were synthesized by systematically varying the methylation of the polar headgroup, after which the physicochemical properties, in the absence of DOPE and serum, were correlated with their transfection activity and interaction with cell membranes. It was found that only the primary and secondary amine derivatives, AC-Chol and MC-Chol, respectively, are able to mediate in vitro cell transfection. These results were consistent with fusion experiments and cell internalization studies which illustrated that although cell surface binding occurs for all of the cationic lipids, only the active analogues were able to gain entry into the cytosol. Given the minute differences in the physical properties of these cationic derivatives, we speculate that the biological specificity of the active cationic derivatives either triggers endocytotic pathways leading to eventual endosomal fusion allowing cytoplasmic access to the packaged DNA or other endocytotic pathways that avoid lysosomal degradation. PMID- 18159925 TI - Cell signaling and nuclear receptors: new opportunities for molecular pharmaceuticals in liver disease. AB - Liver-enriched nuclear receptors (NRs) collectively function as metabolic and toxicological "sensors" that mediate liver-specific gene-activation in mammals. NR-mediated gene-environment interaction regulates important steps in the hepatic uptake, metabolism, and excretion of glucose, fatty acids, lipoproteins, cholesterol, bile acids, and xenobiotics. Hence, liver-enriched NRs play pivotal roles in the overall control of energy homeostasis in mammals. While it is well recognized that ligand-binding is the primary mechanism behind activation of NRs, recent research reveals that multiple signal transduction pathways modulate NR function in liver. The interface between specific signal transduction pathways and NRs helps to determine their overall responsiveness to various environmental and physiological stimuli. In general, phosphorylation of hepatic NRs regulates multiple biological parameters including their transactivation capacity, DNA binding, subcellular location, capacity to interact with protein-cofactors, and protein stability. Certain pathological conditions including inflammation, morbid obesity, hyperlipidemia, atherosclerosis, insulin resistance, and type-2 diabetes are known to modulate selected signal transduction pathways in liver. This review will focus upon recent insights regarding the molecular mechanisms that comprise the interface between disease-mediated activation of hepatic signal transduction pathways and liver-enriched NRs. This review will also highlight the exciting opportunities presented by this new knowledge to develop novel molecular and pharmaceutical strategies for combating these increasingly prevalent human diseases. PMID- 18159928 TI - Liposomal formulations of serratiopeptidase: in vitro studies using PAMPA and Caco-2 models. AB - The feasibility of using liposomes as a potential oral delivery system for the systemic delivery of other peptides and protein-based pharmaceuticals has been studied. Serratiopeptidase, a proteolytic enzyme, was used as a model drug. Liposomes were prepared by a thin film hydration method using various lipids, namely, soya lecithin, DMPC and DMPE. It was further investigated whether the liposomal formulations of serratiopeptidase altered the permeability/absorption of the drug using PAMPA, a non-cell-based assay, and Caco-2 assay, a cell monolayer system, mimicking in vivo GI epithelium cells. The entrapment efficiency of the formulations was found to be 62%, 84% and 86% for the liposomes of soya lecithin, DMPC and DMPE respectively. The effectiveness of the liposomal formulations against the pure drug in terms of permeability/absorption was compared. The effective permeability (log Pe) values from PAMPA study varied from -7.47 to -6.5 cm/s whereas for the serratiopeptidase it was -7.72 cm/s. The apparent permeability values calculated from Caco-2 assay varied from 1.25 x 10( 6) to 1.61 x 10(-6) cm/s whereas for the serratiopeptidase it was 1.25 x 10(-6) cm/s. The flux was found to be 3.88-4.96 microg/cm (2)/h for the formulations when compared to 3.208 microg/cm(2)/h for serratiopeptidase. The results obtained indicated that in comparison with the pure drug, incorporation of drug into liposomes improved the permeability. Thus it could be concluded that the liposomal formulations would improve the oral absorption of serratiopeptidase. PMID- 18159929 TI - Xenoreceptors CAR and PXR activation and consequences on lipid metabolism, glucose homeostasis, and inflammatory response. AB - Xenobiotic and drug metabolism and transport are managed by a large number of genes coordinately regulated by at least three nuclear receptors or xenosensors: aryl hydrocarbon receptor (AhR), constitutive androstane receptor (CAR, NR1I3), and pregnane X receptor (PXR, NR1I2). Initially characterized as xenosensors, it is now evident that CAR and PXR also trigger pleiotropic effects on liver function. Recent studies have shown the existence of crosstalk between xenosensors and other nuclear receptors or transcription factors controlling endogenous signaling pathways which regulate physiological functions. This review is focused on recent observations showing that activation of CAR and PXR alters lipid metabolism, glucose homeostasis, and inflammation by interfering with HNF4alpha, FoxO1, FoxA2, PGC1alpha, or NFkB p65. Such crosstalks explain clinical observations and provide molecular mechanisms allowing understanding how xenobiotics and drugs may affect physiological functions and provoke endocrine disruptions. PMID- 18159930 TI - Novel insulin thiomer nanoparticles: in vivo evaluation of an oral drug delivery system. AB - It was aim of the study to investigate the in vivo potential of a novel insulin thiomer complex nanoparticulate delivery system. Insulin loaded nanoparticles were obtained by the formation of hydrogen bonds between poly(vinyl pyrrolidone) (PVP) and poly(acrylic acid)-cysteine (PAA-Cys) or poly(acrylic acid) (PAA), respectively, in the presence of insulin. Dissolution behavior of insulin from tablets as well as nanoparticulate suspensions was evaluated in vitro. Serum insulin concentrations and reduction of blood sugar values were determined after oral administration of nanoparticles formulated as enteric coated tablets and suspensions. Results displayed a low serum insulin concentration and pharmacological efficacy in terms of blood sugar reduction after oral administration of enteric coated tablets. On the contrary, nanoparticulate suspensions led to significant serum insulin concentrations. Furthermore a 2.3 fold improvement of the AUC of insulin could be achieved due to the use of thiolated PAA instead of unmodified PAA. In addition, a blood sugar reduction of 22% was observed. Results demonstrate that this novel complex nanoparticulate formulation is an encouraging new attempt toward the noninvasive delivery of peptide drugs. PMID- 18159931 TI - Chitin nanocrystals prepared by TEMPO-mediated oxidation of alpha-chitin. AB - Chitin nanocrystals dispersed in water were successfully prepared by 2,2,6,6 tetramethylpiperidine-1-oxyl radical (TEMPO) mediated oxidation of alpha-chitin in water at pH 10 under specific conditions, followed by ultrasonic treatment. When the amount of NaClO added as co-oxidant in the oxidation was 5.0 mmol/g of chitin, the weight percentage of the water-insoluble fraction in the TEMPO oxidized chitin was 90%, and its carboxylate content reached 0.48 mmol/g. Since the TEMPO-oxidized chitin thus prepared had a crystallinity as high as that of the original alpha-chitin, the C6 carboxylate groups formed by TEMPO-mediated oxidation can be regarded as being present only on the chitin crystallite surfaces. No N-deacetylation occurred on the TEMPO-oxidized chitins. When the TEMPO-oxidized chitin was subjected to ultrasonic treatment in water, mostly individualized chitin nanocrystals were obtained, and the average nanocrystal length and width were 340 and 8 nm, respectively. PMID- 18159932 TI - Pathways of arachidonic acid peroxyl radical reactions and product formation with guanine radicals. AB - Peroxyl radicals were derived from the one-electron oxidation of polyunsaturated fatty acids by sulfate radicals that were generated by the photodissociation of peroxodisulfate anions in air-equilibrated aqueous solutions. Reactions of these peroxyl and neutral guanine radicals, also generated by oxidation with sulfate radicals, were investigated by laser kinetic spectroscopy, and the guanine oxidation products were identified by HPLC and mass spectrometry methods. Sulfate radicals rapidly oxidize arachidonic (ArAc), linoleic (LnAc), and palmitoleic (PmAc) acids with similar rate constants, (2-4) x 10 (9) M (-1) s (-1). The C centered radicals derived from the oxidation of ArAc and LnAc include nonconjugated Rn(.) ( approximately 80%) and conjugated bis-allylic Rba(.) ( approximately 20%) radicals. The latter were detectable in the absence of oxygen by their prominent, narrow absorption band at 280 nm. The Rn(.) radicals of ArAc (containing three bis-allylic sites) transform to the Rba(.) radicals via an intramolecular H-atom abstraction [rate constant (7.5 +/- 0.7) x 10 (4) s (-1)]. In contrast, the Rn(.) radicals of LnAc that contain only one bis-allylic site do not transform intramolecularly to the Rba(.) radicals. In the case of PmAc, which contains only one double bond, the Rba(.) radicals are not observed. The Rn(.) radicals of PmAc rapidly combine with oxygen with a rate constant of (3.8 +/- 0.4) x 10(9) M(-1) s(-1). The Rba(.) radicals of ArAc are less reactive and react with oxygen with a rate constant of (2.2 +/- 0.2) x 10 (8) M (-1) s (-1). The ArAc peroxyl radicals formed spontaneously eliminate superoxide radical anions [rate constant = (3.4 +/- 0.3) x 10 (4) M (-1) s (-1)]. The stable oxidative lesions derived from the 2',3',5'-tri- O-acetylguanosine or 2',3',5'-tri- O acetyl-8-oxo-7,8-dihydroguanosine radicals and their subsequent reactions with ArAc peroxyl radicals were also investigated. The major products found were the 2,5-diamino-4 H-imidazolone (Iz), dehydroguanidinohydantoin (Gh ox), and diastereomeric spiroiminodihydantoin (Sp) nucleosides from 2',3',5'-tri- O acetylguanosine and the Gh ox and Sp nucleosides from 2',3',5'-tri- O-acetyl-8 oxo-7,8-dihydroguanosine. In air-saturated aqueous solutions, covalent alkylated guanine adducts were not detected. PMID- 18159933 TI - An efficient and expeditious synthesis of di- and monosubstituted 2 aminoimidazoles. AB - A microwave-assisted protocol was developed for the construction of di- and monosubstituted 2-aminoimidazoles. The two-step reaction involves the synthesis of N-(1H-imidazol-2-yl)acetamides from readily available alpha-haloketones and N acetylguanidine, followed by deacetylation. Significant rate enhancement was observed for both steps of the protocol, and the overall reaction time was shortened to 20 min compared to 48 h of the conventional procedures. A representative set of di- and monosubstituted 2-aminoimidazoles was prepared using commercially available parallel reactors. PMID- 18159935 TI - Epothilones as lead structures for the synthesis-based discovery of new chemotypes for microtubule stabilization. AB - Epothilones are macrocyclic bacterial natural products with potent microtubule stabilizing and antiproliferative activity. They have served as successful lead structures for the development of several clinical candidates for anticancer therapy. However, the structural diversity of this group of clinical compounds is rather limited, as their structures show little divergence from the original natural product leads. Our own research has explored the question of whether epothilones can serve as a basis for the development of new structural scaffolds, or chemotypes, for microtubule stabilization that might serve as a basis for the discovery of new generations of anticancer drugs. We have elaborated a series of epothilone-derived macrolactones whose overall structural features significantly deviate from those of the natural epothilone scaffold and thus define new structural families of microtubule-stabilizing agents. Key elements of our hypermodification strategy are the change of the natural epoxide geometry from cis to trans, the incorporation of a conformationally constrained side chain, the removal of the C3-hydroxyl group, and the replacement of C12 with nitrogen. So far, this approach has yielded analogs 30 and 40 that are the most advanced, the most rigorously modified, structures, both of which are potent antiproliferative agents with low nanomolar activity against several human cancer cell lines in vitro. The synthesis was achieved through a macrolactone-based strategy or a high yielding RCM reaction. The 12-aza-epothilone ("azathilone" 40) may be considered a "non-natural" natural product that still retains most of the overall structural characteristics of a true natural product but is structurally unique, because it lies outside of the general scope of Nature's biosynthetic machinery for polyketide synthesis. Like natural epothilones, both 30 and 40 promote tubulin polymerization in vitro and at the cellular level induce cell cycle arrest in mitosis. These facts indicate that cancer cell growth inhibition by these compounds is based on the same mechanistic underpinnings as those for natural epothilones. Interestingly, the 9,10-dehydro analog of 40 is significantly less active than the saturated parent compound, which is contrary to observations for natural epothilones B or D. This may point to differences in the bioactive conformations of N-acyl-12-aza-epothilones like 40 and natural epothilones. In light of their distinct structural features, combined with an epothilone-like (and taxol-like) in vitro biological profile, 30 and 40 can be considered as representative examples of new chemotypes for microtubule stabilization. As such, they may offer the same potential for pharmacological differentiation from the original epothilone leads as various newly discovered microtubule-stabilizing natural products with macrolactone structures, such as laulimalide, peloruside, or dictyostatin. PMID- 18159936 TI - Function-oriented synthesis, step economy, and drug design. AB - This Account provides an overview and examples of function-oriented synthesis (FOS) and its increasingly important role in producing therapeutic leads that can be made in a step-economical fashion. Biologically active natural product leads often suffer from several deficiencies. Many are scarce or difficult to obtain from natural sources. Often, they are highly complex molecules and thus not amenable to a practical synthesis that would impact supply. Most are not optimally suitable for human therapeutic use. The central principle of FOS is that the function of a biologically active lead structure can be recapitulated, tuned, or greatly enhanced with simpler scaffolds designed for ease of synthesis and also synthetic innovation. This approach can provide practical access to new (designed) structures with novel activities while at the same time allowing for synthetic innovation by target design. This FOS approach has been applied to a number of therapeutically important natural product leads. For example, bryostatin is a unique natural product anticancer lead that restores apoptosis in cancer cells, reverses multidrug resistance, and bolsters the immune system. Remarkably, it also improves cognition and memory in animals. We have designed and synthesized simplified analogs of bryostatin that can be made in a practical fashion (pilot scale) and are superior to bryostatin in numerous assays including growth inhibition in a variety of human cancer cell lines and in animal models. Laulimalide is another exciting anticancer lead that stabilizes microtubules, like paclitaxel, but unlike paclitaxel, it is effective against multidrug resistant cell lines. Laulimalide suffers from availability and stability problems, issues that have been addressed using FOS through the design and synthesis of stable and efficacious laulimalide analogs. Another FOS program has been directed at the design and synthesis of drug delivery systems for enabling or enhancing the uptake of drugs or drug candidates into cells and tissue. We have generated improved transporters that can deliver agents in a superior fashion compared with naturally occurring cell-penetrating peptides and that can be synthesized in a practical and step-economical fashion. The use of FOS has allowed for the translation of exciting, biologically active natural product leads into simplified analogs with superior function. This approach enables the development of synthetically innovative strategies while targeting therapeutically novel structures. PMID- 18159938 TI - Increased levels of numerical chromosome aberrations after in vitro exposure of human peripheral blood lymphocytes to radiofrequency electromagnetic fields for 72 hours. AB - Mazor, R., Korenstein-Ilan, A., Barbul, A., Eshet, Y., Shahadi, A., Jerby, E. and Korenstein, R. Increased Levels of Numerical Chromosome Aberrations after In Vitro Exposure of Human Peripheral Blood Lymphocytes to Radiofrequency Electromagnetic Fields for 72 Hours. Radiat. Res. 169, 28-37 (2008). We investigated the effects of 72 h in vitro exposure of 10 human lymphocyte samples to radiofrequency electromagnetic fields (800 MHz, continuous wave) on genomic instability. The lymphyocytes were exposed in a specially designed waveguide resonator at specific absorption rates (SARs) of 2.9 and 4.1 W/kg in a temperature range of 36-37 degrees C. The induced aneuploidy of chromosomes 1, 10, 11 and 17 was determined by interphase FISH using semi-automated image analysis. We observed increased levels of aneuploidy depending on the chromosome studied as well as on the level of exposure. In chromosomes 1 and 10, there was increased aneuploidy at the higher SAR, while for chromosomes 11 and 17, the increases were observed only for the lower SAR. Multisomy (chromosomal gains) appeared to be the primary contributor to the increased aneuploidy. The effect of temperature on the level of aneuploidy was examined over the range of 33.5-40 degrees C for 72 h with no statistically significant difference in the level of aneuploidy compared to 37 degrees C. These findings suggest the possible existence of an athermal effect of RF radiation that causes increased levels of aneuploidy. These results contribute to the assessment of potential health risks after continuous chronic exposure to RF radiation at SARs close to the current levels set by ICNIRP guidelines. PMID- 18159939 TI - Operating regimes of signaling cycles: statics, dynamics, and noise filtering. AB - A ubiquitous building block of signaling pathways is a cycle of covalent modification (e.g., phosphorylation and dephosphorylation in MAPK cascades). Our paper explores the kind of information processing and filtering that can be accomplished by this simple biochemical circuit. Signaling cycles are particularly known for exhibiting a highly sigmoidal (ultrasensitive) input output characteristic in a certain steady-state regime. Here, we systematically study the cycle's steady-state behavior and its response to time-varying stimuli. We demonstrate that the cycle can actually operate in four different regimes, each with its specific input-output characteristics. These results are obtained using the total quasi-steady-state approximation, which is more generally valid than the typically used Michaelis-Menten approximation for enzymatic reactions. We invoke experimental data that suggest the possibility of signaling cycles operating in one of the new regimes. We then consider the cycle's dynamic behavior, which has so far been relatively neglected. We demonstrate that the intrinsic architecture of the cycles makes them act--in all four regimes--as tunable low-pass filters, filtering out high-frequency fluctuations or noise in signals and environmental cues. Moreover, the cutoff frequency can be adjusted by the cell. Numerical simulations show that our analytical results hold well even for noise of large amplitude. We suggest that noise filtering and tunability make signaling cycles versatile components of more elaborate cell-signaling pathways. PMID- 18159940 TI - Determinants of protein abundance and translation efficiency in S. cerevisiae. AB - The translation efficiency of most Saccharomyces cerevisiae genes remains fairly constant across poor and rich growth media. This observation has led us to revisit the available data and to examine the potential utility of a protein abundance predictor in reinterpreting existing mRNA expression data. Our predictor is based on large-scale data of mRNA levels, the tRNA adaptation index, and the evolutionary rate. It attains a correlation of 0.76 with experimentally determined protein abundance levels on unseen data and successfully cross predicts protein abundance levels in another yeast species (Schizosaccharomyces pombe). The predicted abundance levels of proteins in known S. cerevisiae complexes, and of interacting proteins, are significantly more coherent than their corresponding mRNA expression levels. Analysis of gene expression measurement experiments using the predicted protein abundance levels yields new insights that are not readily discernable when clustering the corresponding mRNA expression levels. Comparing protein abundance levels across poor and rich media, we find a general trend for homeostatic regulation where transcription and translation change in a reciprocal manner. This phenomenon is more prominent near origins of replications. Our analysis shows that in parallel to the adaptation occurring at the tRNA level via the codon bias, proteins do undergo a complementary adaptation at the amino acid level to further increase their abundance. PMID- 18159941 TI - Spike correlations in a songbird agree with a simple markov population model. AB - The relationships between neural activity at the single-cell and the population levels are of central importance for understanding neural codes. In many sensory systems, collective behaviors in large cell groups can be described by pairwise spike correlations. Here, we test whether in a highly specialized premotor system of songbirds, pairwise spike correlations themselves can be seen as a simple corollary of an underlying random process. We test hypotheses on connectivity and network dynamics in the motor pathway of zebra finches using a high-level population model that is independent of detailed single-neuron properties. We assume that neural population activity evolves along a finite set of states during singing, and that during sleep population activity randomly switches back and forth between song states and a single resting state. Individual spike trains are generated by associating with each of the population states a particular firing mode, such as bursting or tonic firing. With an overall modification of one or two simple control parameters, the Markov model is able to reproduce observed firing statistics and spike correlations in different neuron types and behavioral states. Our results suggest that song- and sleep-related firing patterns are identical on short time scales and result from random sampling of a unique underlying theme. The efficiency of our population model may apply also to other neural systems in which population hypotheses can be tested on recordings from small neuron groups. PMID- 18159942 TI - Hemolytic C-type lectin CEL-III from sea cucumber expressed in transgenic mosquitoes impairs malaria parasite development. AB - The midgut environment of anopheline mosquitoes plays an important role in the development of the malaria parasite. Using genetic manipulation of anopheline mosquitoes to change the environment in the mosquito midgut may inhibit development of the malaria parasite, thus blocking malaria transmission. Here we generate transgenic Anopheles stephensi mosquitoes that express the C-type lectin CEL-III from the sea cucumber, Cucumaria echinata, in a midgut-specific manner. CEL-III has strong and rapid hemolytic activity toward human and rat erythrocytes in the presence of serum. Importantly, CEL-III binds to ookinetes, leading to strong inhibition of ookinete formation in vitro with an IC(50) of 15 nM. Thus, CEL-III exhibits not only hemolytic activity but also cytotoxicity toward ookinetes. In these transgenic mosquitoes, sporogonic development of Plasmodium berghei is severely impaired. Moderate, but significant inhibition was found against Plasmodium falciparum. To our knowledge, this is the first demonstration of stably engineered anophelines that affect the Plasmodium transmission dynamics of human malaria. Although our laboratory-based research does not have immediate applications to block natural malaria transmission, these findings have significant implications for the generation of refractory mosquitoes to all species of human Plasmodium and elucidation of mosquito-parasite interactions. PMID- 18159943 TI - Analysis of cells targeted by Salmonella type III secretion in vivo. AB - The type III secretion systems (TTSS) encoded in Salmonella pathogenicity island 1 and -2 (SPI-1 and -2) are virulence factors required for specific phases of Salmonella infection in animal hosts. However, the host cell types targeted by the TTSS have not been determined. To investigate this, we have constructed translational fusions between the beta-lactamase reporter and a broad array of TTSS effectors secreted via SPI-1, SPI-2, or both. Secretion of the fusion protein to a host cell was determined by cleavage of a specific fluorescent substrate. In cultured cells, secretion of all six effectors could be observed. However, two to four days following i.p. infection of mice, only effectors secreted by SPI-2 were detected in spleen cells. The cells targeted were identified via staining with nine different cell surface markers followed by FACS analysis as well as by conventional cytological methods. The targeted cells include B and T lymphocytes, neutrophils, monocytes, and dendritic cells, but not mature macrophages. To further investigate replication in these various cell types, Salmonella derivatives were constructed that express a red fluorescent protein. Bacteria could be seen in each of the cell types above; however, most viable bacteria were present in neutrophils. We find that Salmonella is capable of targeting most phagocytic and non-phagocytic cells in the spleen but has a surprisingly high preference for neutrophils. These findings suggest that Salmonella specifically target splenic neutrophils presumably to attenuate their microbicidal functions, thereby promoting intracellular survival and replication in the mouse. PMID- 18159944 TI - Discordant evolution of the adjacent antiretroviral genes TRIM22 and TRIM5 in mammals. AB - TRIM5alpha provides a cytoplasmic block to retroviral infection, and orthologs encoded by some primates are active against HIV. Here, we present an evolutionary comparison of the TRIM5 gene to its closest human paralogs: TRIM22, TRIM34, and TRIM6. We show that TRIM5 and TRIM22 have a dynamic history of gene expansion and loss during the evolution of mammals. The cow genome contains an expanded cluster of TRIM5 genes and no TRIM22 gene, while the dog genome encodes TRIM22 but has lost TRIM5. In contrast, TRIM6 and TRIM34 have been strictly preserved as single gene orthologs in human, dog, and cow. A more focused analysis of primates reveals that, while TRIM6 and TRIM34 have evolved under purifying selection, TRIM22 has evolved under positive selection as was previously observed for TRIM5. Based on TRIM22 sequences obtained from 27 primate genomes, we find that the positive selection of TRIM22 has occurred episodically for approximately 23 million years, perhaps reflecting the changing pathogenic landscape. However, we find that the evolutionary episodes of positive selection that have acted on TRIM5 and TRIM22 are mutually exclusive, with generally only one of these genes being positively selected in any given primate lineage. We interpret this to mean that the positive selection of one gene has constrained the adaptive flexibility of its neighbor, probably due to genetic linkage. Finally, we find a striking congruence in the positions of amino acid residues found to be under positive selection in both TRIM5alpha and TRIM22, which in both proteins fall predominantly in the beta2-beta3 surface loop of the B30.2 domain. Astonishingly, this same loop is under positive selection in the multiple cow TRIM5 genes as well, indicating that this small structural loop may be a viral recognition motif spanning a hundred million years of mammalian evolution. PMID- 18159945 TI - Shp2 knockdown and Noonan/LEOPARD mutant Shp2-induced gastrulation defects. AB - Shp2 is a cytoplasmic protein-tyrosine phosphatase that is essential for normal development. Activating and inactivating mutations have been identified in humans to cause the related Noonan and LEOPARD syndromes, respectively. The cell biological cause of these syndromes remains to be determined. We have used the zebrafish to assess the role of Shp2 in early development. Here, we report that morpholino-mediated knockdown of Shp2 in zebrafish resulted in defects during gastrulation. Cell tracing experiments demonstrated that Shp2 knockdown induced defects in convergence and extension cell movements. In situ hybridization using a panel of markers indicated that cell fate was not affected by Shp2 knock down. The Shp2 knockdown-induced defects were rescued by active Fyn and Yes and by active RhoA. We generated mutants of Shp2 with mutations that were identified in human patients with Noonan or LEOPARD Syndrome and established that Noonan Shp2 was activated and LEOPARD Shp2 lacked catalytic protein-tyrosine phosphatase activity. Expression of Noonan or LEOPARD mutant Shp2 in zebrafish embryos induced convergence and extension cell movement defects without affecting cell fate. Moreover, these embryos displayed craniofacial and cardiac defects, reminiscent of human symptoms. Noonan and LEOPARD mutant Shp2s were not additive nor synergistic, consistent with the mutant Shp2s having activating and inactivating roles in the same signaling pathway. Our results demonstrate that Shp2 is required for normal convergence and extension cell movements during gastrulation and that Src family kinases and RhoA were downstream of Shp2. Expression of Noonan or LEOPARD Shp2 phenocopied the craniofacial and cardiac defects of human patients. The finding that defective Shp2 signaling induced cell movement defects as early as gastrulation may have implications for the monitoring and diagnosis of Noonan and LEOPARD syndrome. PMID- 18159946 TI - Experimental characterization of Cis-acting elements important for translation and transcription in halophilic archaea. AB - The basal transcription apparatus of archaea is well characterized. However, much less is known about the mechanisms of transcription termination and translation initation. Recently, experimental determination of the 5'-ends of ten transcripts from Pyrobaculum aerophilum revealed that these are devoid of a 5'-UTR. Bioinformatic analysis indicated that many transcripts of other archaeal species might also be leaderless. The 5'-ends and 3'-ends of 40 transcripts of two haloarchaeal species, Halobacterium salinarum and Haloferax volcanii, have been determined. They were used to characterize the lengths of 5'-UTRs and 3'-UTRs and to deduce consensus sequence-elements for transcription and translation. The experimental approach was complemented with a bioinformatics analysis of the H. salinarum genome sequence. Furthermore, the influence of selected 5'-UTRs and 3' UTRs on transcript stability and translational efficiency in vivo was characterized using a newly established reporter gene system, gene fusions, and real-time PCR. Consensus sequences for basal promoter elements could be refined and a novel element was discovered. A consensus motif probably important for transcriptional termination was established. All 40 haloarchaeal transcripts analyzed had a 3'-UTR (average size 57 nt), and their 3'-ends were not posttranscriptionally modified. Experimental data and genome analyses revealed that the majority of haloarchaeal transcripts are leaderless, indicating that this is the predominant mode for translation initiation in haloarchaea. Surprisingly, the 5'-UTRs of most leadered transcripts did not contain a Shine Dalgarno (SD) sequence. A genome analysis indicated that less than 10% of all genes are preceded by a SD sequence and even most proximal genes in operons lack a SD sequence. Seven different leadered transcripts devoid of a SD sequence were efficiently translated in vivo, including artificial 5'-UTRs of random sequences. Thus, an interaction of the 5'-UTRs of these leadered transcripts with the 16S rRNA could be excluded. Taken together, either a scanning mechanism similar to the mechanism of translation initiation operating in eukaryotes or a novel mechanism must operate on most leadered haloarchaeal transcripts. PMID- 18159947 TI - Patterns and implications of gene gain and loss in the evolution of Prochlorococcus. AB - Prochlorococcus is a marine cyanobacterium that numerically dominates the mid latitude oceans and is the smallest known oxygenic phototroph. Numerous isolates from diverse areas of the world's oceans have been studied and shown to be physiologically and genetically distinct. All isolates described thus far can be assigned to either a tightly clustered high-light (HL)-adapted clade, or a more divergent low-light (LL)-adapted group. The 16S rRNA sequences of the entire Prochlorococcus group differ by at most 3%, and the four initially published genomes revealed patterns of genetic differentiation that help explain physiological differences among the isolates. Here we describe the genomes of eight newly sequenced isolates and combine them with the first four genomes for a comprehensive analysis of the core (shared by all isolates) and flexible genes of the Prochlorococcus group, and the patterns of loss and gain of the flexible genes over the course of evolution. There are 1,273 genes that represent the core shared by all 12 genomes. They are apparently sufficient, according to metabolic reconstruction, to encode a functional cell. We describe a phylogeny for all 12 isolates by subjecting their complete proteomes to three different phylogenetic analyses. For each non-core gene, we used a maximum parsimony method to estimate which ancestor likely first acquired or lost each gene. Many of the genetic differences among isolates, especially for genes involved in outer membrane synthesis and nutrient transport, are found within the same clade. Nevertheless, we identified some genes defining HL and LL ecotypes, and clades within these broad ecotypes, helping to demonstrate the basis of HL and LL adaptations in Prochlorococcus. Furthermore, our estimates of gene gain events allow us to identify highly variable genomic islands that are not apparent through simple pairwise comparisons. These results emphasize the functional roles, especially those connected to outer membrane synthesis and transport that dominate the flexible genome and set it apart from the core. Besides identifying islands and demonstrating their role throughout the history of Prochlorococcus, reconstruction of past gene gains and losses shows that much of the variability exists at the "leaves of the tree," between the most closely related strains. Finally, the identification of core and flexible genes from this 12-genome comparison is largely consistent with the relative frequency of Prochlorococcus genes found in global ocean metagenomic databases, further closing the gap between our understanding of these organisms in the lab and the wild. PMID- 18159948 TI - Distinct roles and regulations for HoxD genes in metanephric kidney development. AB - Hox genes encode homeodomain-containing proteins that control embryonic development in multiple contexts. Up to 30 Hox genes, distributed among all four clusters, are expressed during mammalian kidney morphogenesis, but functional redundancy between them has made a detailed functional account difficult to achieve. We have investigated the role of the HoxD cluster through comparative molecular embryological analysis of a set of mouse strains carrying targeted genomic rearrangements such as deletions, duplications, and inversions. This analysis allowed us to uncover and genetically dissect the complex role of the HoxD cluster. Regulation of metanephric mesenchyme-ureteric bud interactions and maintenance of structural integrity of tubular epithelia are differentially controlled by some Hoxd genes during renal development, consistent with their specific expression profiles. We also provide evidence for a kidney-specific form of colinearity that underlies the differential expression of two distinct sets of genes located on both sides and overlapping at the Hoxd9 locus. These insights further our knowledge of the genetic control of kidney morphogenesis and may contribute to understanding certain congenital kidney malformations, including polycystic kidney disease and renal hypoplasia. PMID- 18159949 TI - Human subtelomeric WASH genes encode a new subclass of the WASP family. AB - Subtelomeres are duplication-rich, structurally variable regions of the human genome situated just proximal of telomeres. We report here that the most terminally located human subtelomeric genes encode a previously unrecognized third subclass of the Wiskott-Aldrich Syndrome Protein family, whose known members reorganize the actin cytoskeleton in response to extracellular stimuli. This new subclass, which we call WASH, is evolutionarily conserved in species as diverged as Entamoeba. We demonstrate that WASH is essential in Drosophila. WASH is widely expressed in human tissues, and human WASH protein colocalizes with actin in filopodia and lamellipodia. The VCA domain of human WASH promotes actin polymerization by the Arp2/3 complex in vitro. WASH duplicated to multiple chromosomal ends during primate evolution, with highest copy number reached in humans, whose WASH repertoires vary. Thus, human subtelomeres are not genetic junkyards, and WASH's location in these dynamic regions could have advantageous as well as pathologic consequences. PMID- 18159950 TI - Comparison of Helzel and OxyLite systems in the measurements of tumor partial oxygen pressure (pO2). AB - Wen, B., Urano, M., Humm, J. L., Seshan, V. E., Li, G. C. and Ling, C. C. Comparison of Helzel and OxyLite Systems in the Measurements of Tumor Partial Oxygen Pressure (pO(2)). Radiat. Res. 168, 67-75 (2008). It has been demonstrated in both experimental and human malignancies that hypoxic tumor cells are linked with aggressive disease phenotype. One of the methods to identify these cells is by direct physical measurement of tumor pO(2). This study compared pO(2) values measured with two systems, the Helzel Hypoximeter (successor of the polarographic Eppendorf electrode) and the Oxford-Optronix OxyLite (fiber-optic probe), in R3327-AT and R3327-AT/tkeGFP tumors. Partial oxygen pressure was measured in individual tumors with either system or in the same tumor with both systems. The similarities and discrepancies in pO(2) measurements between the two systems were also investigated when tumor-bearing animals were breathing pure oxygen. Our data showed a considerable heterogeneity in pO(2) values in each tumor using both the Helzel and OxyLite systems. Similar results were obtained with both systems for the mean and median pO(2) values, and the distributions of pO(2) values within the interval 0 < pO(2) < 40 mmHg (the range important for defining tumor hypoxia) were found to be statistically equivalent. However, the frequencies of high pO(2) values (>40 mmHg) and zero values measured by the two systems were statistically significantly different. PMID- 18159951 TI - Sustained metaphase arrest in response to ionizing radiation in a non-small cell lung cancer cell line. AB - Kodym, E., Kodym, R., Choy, H. and Saha, D. Sustained Metaphase Arrest in Response to Ionizing Radiation in a Non-small Cell Lung Cancer Cell Line. Radiat. Res. 169, 46-58 (2008). In solid tumors, non-apoptotic forms of tumor cell inactivation such as mitotic catastrophe appear to be predominant in the response to DNA-damaging agents. Despite its importance, the underlying molecular mechanisms of mitotic catastrophe have been only partially elucidated. We found that a large fraction of HCC2279 non-small cell lung cancer cells underwent mitotic catastrophe after irradiation. Cells were arrested in metaphase with chromosomal damage indicated by DNA fragments displaced from the metaphase plate and considerable numbers of residual gamma-H2AX foci. Although TP53 was nonfunctional, we detected a prompt radiation response on the level of checkpoint kinases. In contrast, CDC25A was the only checkpoint phosphatase that was responsive to radiation. CDC25B was not detectable, and CDC25C was constitutively phosphorylated at serine 216, leading to its cytoplasmic sequestration and functional inactivation. Therefore, radiation-induced mitotic catastrophe in HCC2279 cells appears to be induced by a combination of relative insufficiencies in the p53-mediated and checkpoint kinase-mediated pathways leading to premature entry into mitosis. Displaced chromosome fragments triggering an intra-M checkpoint in cells entering mitosis presumably result in a sustained metaphase arrest. The phenomenon found in these cells, which were derived directly from a human patient, might be responsible for therapy-induced genetic instability of tumors. PMID- 18159952 TI - Repeated 0.5-Gy gamma irradiation attenuates autoimmune disease in MRL-lpr/lpr mice with suppression of CD3+CD4-CD8-B220+ T-cell proliferation and with up regulation of CD4+CD25+Foxp3+ regulatory T cells. AB - Tago, F., Tsukimoto, M., Nakatsukasa, H. and Kojima. S. Repeated 0.5 Gy Gamma Irradiation Attenuates Autoimmune Disease in MRL-lpr/lpr Mice with Suppression of CD3(+)CD4(-)CD8(-)B220(+) T-Cell Proliferation and with Up-regulation of CD4(+)CD25(+)Foxp3(+) Regulatory T Cells. Radiat. Res. 169, 59-66 (2008). MRL lpr/lpr mice are used as a model of systemic lupus erythematosus. We previously reported attenuation of autoimmune disease in MRL-lpr/lpr mice by repeated gamma irradiation (0.5 Gy each time). In this study, we investigated the mechanisms of this attenuation by measuring the weight of the spleen and the population of highly activated CD3(+)CD4(-)CD8(-)B220(+) T cells, which are characteristically involved in autoimmune pathology in these mice. Splenomegaly and an increase in the percentage of CD3(+)CD4(-)CD8(-)B220(+) T cells, which occur with aging in nonirradiated mice, were suppressed in irradiated mice. The high proliferation rate of CD3(+)CD4(-)CD8(-)B220(+) T cells was suppressed in the irradiated animals. The production of autoantibodies and the level of IL6, which activates B cells, were also lowered by radiation exposure. These results indicate that progression of pathology is suppressed by repeated 0.5-Gy gamma irradiation. To uncover the mechanism of the immune suppression, we measured the regulatory T cells, which suppress activated T cells and excessive autoimmune responses. We found that regulatory T cells were significantly increased in irradiated mice. We therefore conclude that repeated 0.5-Gy gamma irradiation suppresses the proliferation rate of CD3(+)CD4(-)CD8(-)B220(+) T cells and the production of IL6 and autoantibodies and up-regulates regulatory T cells. PMID- 18159953 TI - Evidence for a Grotthuss-like mechanism in the formation of the rhamnose alkoxy radical based on periodic DFT calculations. AB - Pauwels, E., Declerck, R., Van Speybroeck, V. and Waroquier, M. Evidence for a Grotthuss-Like Mechanism in the Formation of the Rhamnose Alkoxy Radical Based on Periodic DFT Calculations. Radiat. Res. 169, 8-18 (2008). Molecular modeling adopting a periodic approach based on density functional theory (DFT) indicates that a Grotthuss-like mechanism is active in the formation of the radiation induced alkoxy radical in alpha-l-rhamnose. Starting from an oxidized crystal structure, a hydroxyl proton is transferred along an infinite hydrogen bond chain pervading the entire crystal. The result of this proton shuttling mechanism is a stable radical species dubbed RHop. Only after several reorientations of crystal waters and hydroxyl groups, the more stable radical form RO4 is obtained, which differs in structure from the former by the absence of only one hydrogen bond. Calculations of the energetics associated with the mechanism as well as simulated spectroscopic properties reveal that different variants of the rhamnose alkoxy radical can be observed depending on the temperature of irradiation and consecutive EPR measurement. Cluster calculations on both radical variants provide hyperfine coupling and g tensors that are in good agreement with two independent experimental measurements at different temperatures. PMID- 18159954 TI - Analysis of functional groups of differentially expressed genes in the peripheral blood of patients with cervical cancer undergoing concurrent chemoradiation treatment. AB - Chao, A., Wang, T. H., Lee, Y. S., Hong, J. H., Tsai, C. N., Chen, C. K., Tsai, C. S., Chao, A. S. and Lai, C. H. Analysis of Functional Groups Differentially Expressed Genes in the Peripheral Blood of Patients with Cervical Cancer Undergoing Concurrent Chemoradiation Treatment. Radiat. Res. 169, 76-86 (2008). We prospectively investigated the gene expression profiles of cervical cancer patients undergoing concurrent chemoradiation treatment. Up-regulated genes associated with anemia were analyzed. Peripheral blood of 20 patients (bulky stage IB-IVA cervical squamous cell carcinomas) undergoing concurrent chemoradiation treatment at four times was collected. Total RNA extracted by the PAXgene Blood RNA System was analyzed with microarrays and MetaCoretrade mark functional network analyses. Fifty-three genes were significantly differentially expressed during concurrent chemoradiation treatment. Fetal and embryonic hemoglobin genes were up-regulated when patients had been severely myelosuppressed. Twenty-eight genes correlated significantly with the hemoglobin genes are involved in responses to hypoxia and oxygenation, TGF-beta signaling, cell cycle suppression, G-protein signaling, and transcriptional regulation. c Myc has the highest rank in transcriptional co-regulation. In addition, IGKV1D-13 was significantly down-regulated in patients with severe hematological toxicity. These approaches identified biological processes in peripheral blood modulated by concurrent chemoradiation treatment and subsequent anemia. PMID- 18159955 TI - A systematic review of epidemiological associations between low and moderate doses of ionizing radiation and late cardiovascular effects, and their possible mechanisms. AB - Little, M. P., Tawn, E. J., Tzoulaki, I., Wakeford, R., Hildebrandt, G., Paris, F., Tapio, S. and Elliott, P. A Systematic Review of Epidemiological Associations Between Low and Moderate Doses of Ionizing Radiation and Late Cardiovascular Effects, and Their Possible Mechanisms. Radiat. Res. 169, 99-109 (2008). The link between high doses of ionizing radiation and damage to the heart and coronary arteries is established. In this paper, we systematically review the epidemiological evidence for associations between low and moderate doses (<5 Gy) of ionizing radiation and late-occurring cardiovascular disease. Risks per unit dose in epidemiological studies vary over at least two orders of magnitude, possibly a result of confounding factors. An examination of possible biological mechanisms indicates that the most likely causative effect of radiation exposure is damage to endothelial cells and subsequent induction of an inflammatory response, although it seems unlikely that this would extend to low-dose and low dose-rate exposure. However, a role for somatic mutation has been proposed that would indicate a stochastic effect. In the absence of a convincing mechanistic explanation of epidemiological evidence that is less than persuasive at present, a cause-and-effect interpretation of the reported statistical associations cannot be reliably inferred, although neither can it be reliably excluded. Further epidemiological and biological evidence will allow a firmer conclusion to be drawn. PMID- 18159956 TI - Apoptosis is induced by radiofrequency fields through the caspase-independent mitochondrial pathway in cortical neurons. AB - Joubert, V., Bourthoumieu, S., Leveque, P. and Yardin, C. Apoptosis is Induced by Radiofrequency Fields through the Caspase-Independent Mitochondrial Pathway in Cortical Neurons. Radiat. Res. 169, 38-45 (2008). In the present study, we investigated whether continuous-wave (CW) radiofrequency (RF) fields induce neuron apoptosis in vitro. Rat primary neuronal cultures were exposed to a CW 900 MHz RF field with a specific absorption rate (SAR) of 2 W/kg for 24 h. During exposure, an increase of 2 degrees C was measured in the medium; control experiments with neurons exposed to 39 degrees C were then performed. Apoptosis was assessed by condensation of nuclei with 4',6-diamino-2-phenylindole (DAPI) staining observed with an epifluorescence microscope and fragmentation of DNA with TdT-mediated dUTP nick-end labeling (TUNEL) analyzed by flow cytometry. A statistically significant difference in the rate of apoptosis was found in the RF field-exposed neurons compared to the sham-, 37 degrees C- and 39 degrees C exposed neurons either 0 or 24 h after exposure using both methods. To assess whether the observed apoptosis was caspase-dependent or -independent, assays measuring caspase 3 activity and apoptosis-inducing factor (AIF) labeling were performed. No increase in the caspase 3 activity was found, whereas the percentage of AIF-positive nuclei in RF-field-exposed neurons was increased by three- to sevenfold compared to other conditions. Our results show that, under the experimental conditions used, exposure of primary rat neurons to CW RF fields may induce a caspase-independent pathway to apoptosis that involves AIF. PMID- 18159957 TI - Radiosensitization of DNA by gold nanoparticles irradiated with high-energy electrons. AB - Zheng, Y., Hunting, D. J., Ayotte, P. and Sanche, L. Radiosensitization of DNA by Gold Nanoparticles Irradiated with High-Energy Electrons. Radiat. Res. 168, 19-27 (2008). Thin films of pGEM-3Zf(-) plasmid DNA were bombarded by 60 keV electrons with and without gold nanoparticles. DNA single- and double-strand breaks (SSBs and DSBs) were measured by agarose gel electrophoresis. From transmission electron micrographs, the gold nanoparticles were found to be closely linked to DNA scaffolds, probably as a result of electrostatic binding. The probabilities for formation of SSBs and DSBs from exposure of 1:1 and 2:1 gold nanoparticle:plasmid mixtures to fast electrons increase by a factor of about 2.5 compared to neat DNA samples. For monolayer DNA adsorbed on a thick gold substrate, the damage increases by an order of magnitude. The results suggest that the enhancement of radiosensitivity is due to the production of additional low-energy secondary electrons caused by the increased absorption of ionizing radiation energy by the metal, in the form of gold nanoparticles or of a thick gold substrate. Since short-range low-energy secondary electrons are produced in large amounts by any type of ionizing radiation, and since on average only one gold nanoparticle per DNA molecule is needed to increase damage considerably, targeting the DNA of cancer cells with gold nanoparticles may offer a novel approach that is generally applicable to radiotherapy treatments. PMID- 18159958 TI - Improved estimates of cancer site-specific risks for A-bomb survivors. AB - Pawel, D. J., Preston, D. L., Pierce, D. A. and Cologne, J. B. Improved Estimates of Cancer Site-Specific Risks for A-Bomb Survivors. Radiat. Res. 169, 87-98 (2008). Simple methods are investigated for improving summary site-specific radiogenic risk estimates. Estimates in this report are derived from cancer incidence data from the Life Span Study (LSS) cohort of A-bomb survivors that are followed up by the Radiation Effects Research Foundation (RERF). Estimates from the LSS of excess relative risk (ERR) for solid cancer sites have typically been derived separately for each site. Even though the data for this are extensive, the statistical imprecision in site-specific (organ-specific) risk estimates is substantial, and it is clear that a large portion of the site-specific variation in estimates is due to this imprecision. Empirical Bayes (EB) estimates offer a reasonable approach for moderating this variation. The simple version of EB estimates that we applied to the LSS data are weighted averages of a pooled overall estimate of ERR and separately derived site-specific estimates, with weights determined by the data. Results indicate that the EB estimates are most useful for sites such as esophageal or bladder cancer, for which the separately derived ERR estimates are less precise than for other sites. PMID- 18159959 TI - A modified alkaline comet assay for measuring DNA repair capacity in human populations. AB - Trzeciak, A. R., Barnes, J. and Evans, M. K. A Modified Alkaline Comet Assay for Measuring DNA Repair Capacity in Human Populations. Radiat. Res. 169, 110-121 (2008). Use of the alkaline comet assay to assess DNA repair capacity in human populations has been limited by several factors, including lack of methodology for use of unstimulated cryopreserved peripheral blood mononuclear cells (PBMCs), insufficient control of interexperimental variability, and limited analysis of DNA repair kinetics. We show that unstimulated cryopreserved PBMCs can be used in DNA repair studies performed using the comet assay. We have applied data standardization for the analysis of DNA repair capacity using negative and positive internal standards as controls for interexperimental variability. Our standardization procedure also uses negative controls, which provides a way to minimize the interference of interindividual variation in baseline DNA damage levels on DNA repair capacity measurements in populations. DNA repair capacity was assessed in a small human cohort using the parameters described in the literature including initial DNA damage, half-time of DNA repair, and residual DNA damage after 30 and 60 min. We have also introduced new DNA repair capacity parameter, initial rate of DNA repair. There was no difference in DNA repair capacity between fresh and cryopreserved PBMCs when measured by the Olive tail moment and tail DNA. The use of DNA repair capacity parameters in assessment of fast and slow single-strand break repair components is discussed. PMID- 18159960 TI - What can epidemiology tell us about risks at low doses? AB - Puskin, J. S. What Can Epidemiology Tell Us about Risks at Low Doses? Radiat. Res. 169, 122-124 (2008). Limitations on statistical power preclude direct detection and quantification of radiogenic cancer risks at very low (environmental) levels of low-LET radiation through epidemiological studies. Given this limitation and our incomplete understanding of cellular processes leading to radiation carcinogenesis, an "effective threshold" in the dose range of interest for radiation protection cannot yet be ruled out. Ongoing epidemiological studies of chronically exposed individuals receiving very low daily doses of radiation can be used, however, together with radiobiological data, to critically test whether such a threshold is plausible. PMID- 18159961 TI - Telomeres and double-strand breaks - all's well that "ends" well. ... AB - Bailey, S. Telomeres and Double-Strand Breaks - All's Well that "Ends" Well. ... Radiat. Res. 169, 1-7 (2008). Sometimes one's life (including one's science) makes a lot more sense when viewed from the perspective of time, reflected back on over a number of years. That has indeed been the case for me. Strangely enough, the story begins with chromosomes and "ends" with telomeres, both at Colorado State University. And, just as with chromosomes, a lot happened in between. Telomeres were first identified based on their function-they protected the physical ends of chromosomes from interaction with broken DNA ends created by ionizing radiation. While I was at Los Alamos National Laboratory, the sequence of human telomeres was discovered, making probes available that allowed us to re examine and provide direct support of these early observations; thus began my fascination with telomeres. Chromosome orientation in situ hybridization (CO FISH) also came onto the scene while I was in Los Alamos. This strand-specific modification of standard FISH, especially when combined with telomeric sequence probes, has proven to be a powerful approach that provides information not available by any other means. Applications have included pericentric inversion detection, distinction between leading- and lagging-strand telomeres, and identification of telomere-double-strand break (DSB) fusions. We also provided the first direct evidence that DSB repair proteins (DNA-PK in particular) are required for mammalian telomeric end capping, and we have been characterizing telomere dysfunction in NHEJ and HR repair-deficient backgrounds ever since. Cells must correctly distinguish between DNA ends represented by telomeres and DNA ends produced by DSBs if all is to end well. Just as these studies have provided new insight into the complex, often surprising, interactions at DNA ends, they also provoke new questions. Whereas it is now well established that DSB repair proteins associate with telomeres, most recently we've been asking whether the reverse scenario holds: Do telomere proteins interact with DSBs? We find that DSBs induced by ionizing radiations are not sufficient to recruit the essential telomere protein TRF2 as an early damage response, so perhaps this interplay is a one-way street. The rest of the story waits to unfold. PMID- 18159963 TI - Physical activity in the prevention of the most frequent chronic diseases: an analysis of the recent evidence. AB - BACKGROUND: Physical activity is widely recognized as a means for the primary prevention of chronic diseases as well as in patients' treatment and rehabilitation. Moreover, activity has beneficial effects on an individual's health and well-being. Despite the benefits of regular physical activity, the percentage of physically inactive adults in the world is high. Environmental and policy approaches aimed to increase physical activity require continual stress of the epidemiological evidence from studies investigating disease mechanisms as well as controlled clinical trials. PURPOSE: To update the evidence that physical activity/exercise is important for reducing the chronic diseases (cardiovascular and heart, diabetes, cancer, obesity, osteoporosis, and fall-related injuries, depression and emotional stress) and for mechanisms that may operate in the relation between physical activity and a disease risk. METHODS: Research studies published from 2004 through to March 2007 were identified through a review of the literature available on the NLM PubMed, Medline, Current Contents, and Elsevier Science Direct databases. RESULTS: Recent evidence on physical activity/exercise and reduction of chronic major diseases incidence and rehabilitation of patients replicates previous findings. The strongest evidence exists for colon cancer, breast cancer, and cardiovascular diseases. The maximal magnitudes of the risk reduction reported were: 75% for breast cancer, 49% for cardiovascular and heart diseases, 35% for diabetes, 22% for colorectal cancer. Increased physical activity also prevented the weight gain associated with aging at least 2-times greater in individuals who were more active compared with those who were inactive. Limited new findings has been reported for the beneficial role of physical activity in fall-related injures, depression and emotional distress. CONCLUSION: Recent evidence confirms previous findings that engaging in moderate physical activity is very important for the primary prevention of chronic diseases, decreasing all causes of mortality and that exercise is one of the determinants for physical and psychological well-being. The current evidence provides further support that physical activity can suppress concentrations of 17 alpha-estradiol in women. PMID- 18159964 TI - Female genital warts. AB - Genital warts are a clinical manifestation of HPV types 6 and 11, and are estimated to affect 1% of sexually active adults aged between 15 and 49. HPV leading to a broad spectrum of human diseases, ranging from benign warts to malignant neoplasms, depending on the location of the lesion, the immune status of the patient and the type of HPV. Current therapies for human papillomavirus associated disease are based on the excision or ablation of involved tissue and are associated with a high frequency of recurrent disease, discomfort and costs. PMID- 18159965 TI - Iran cancer incidence should be corrected for under-ascertainment in cancer cases in the elderly (aged 65+). AB - OBJECTIVE: This article is to calculate corrected Iran cancer incidence by a novel method to compensate under-ascertainment of cancer cases in the very elderly (aged 65+). STUDY DESIGN AND SETTING: Corrected age-specific rate for a certain cancer in age group 65+ was calculated from the age-specific rate of that cancer in age group 55-64 multiplied by the corresponding coefficient from reference cancer registry (sex- and age-specific coefficients from Finnish Cancer Registry, a nation-wide registry with high validity of data). All cancer data were obtained from GLOBOCAN 2002. RESULTS: The crude rate (and number of new cases) for "All sites excluding skin" was 13.6% (men 18.7%; women 8.1%) under estimated. The under-enumeration was 18.9% for the age-standardized rate (men 25.4%; women 11.8%). This means there were 58,000 new cancer cases (about 7,000 more than original) in 2002. Corrected incidence for the year 2050 was 26.1% higher (men 32.8%; women 17.3%) than the original estimate (49,000 more). Depending on cancer site and sex, percentage under-estimation varied remarkably. CONCLUSION: After correction, the estimates of number of new cases and incidence rates of Iran increase substantially. Without correction, cancer occurrence measures can be remarkably under-estimated which may lead to inadequate resource allocation for control measures. PMID- 18159966 TI - Can an appointment-letter intervention increase pap smear screening in Samliem, Khon Kaen, Thailand? AB - Our objective was to assess the efficiency of an appointment-letter intervention aimed to increase uptake of cervical cancer screening in women between 35 and 65 years of age. From January, 2007, we randomly recruited 320 women, not screened for at least 5 years, from the Samliem inner-city community, Khon Kaen, Northeast Thailand. A total of 150 women 35, 40, 45, 50, 55, 60 and 65 years of age were assigned to the intervention group according to Thai National Cancer Institute's ( TNCI) strategy. A further 170 women between 36-39, 41-44, 46-49, 51-54, 56-59 and 61-64 years of age were assigned to the control group. Baseline interviews were conducted for all women in both groups by one of the researchers in January, who also provided culturally-sensitive health education emphasizing the need for screening. Then appointment letters were sent only to women in the intervention group in February, with the last date for an appointment being March 31st. In April of 2007, immediately post-intervention, screening-coverage interviews were performed in both groups for comparison. There was a significant increase in the Pap smear screening-coverage rate in the intervention group compared with the control group (44.67% vs. 25.88%, p=0.001). Therefore, the appointment-letter intervention produced a significant effect on increasing Pap smear coverage in this group of women. PMID- 18159967 TI - Patho-epidemiology of Cancer Cervix in Karachi South. AB - INTRODUCTION: The present study was conducted with the objective of examining descriptive epidemiological and pathological characteristics of cancer cervix in Karachi South, an all urban district population of Karachi, Pakistan. METHODOLOGY: A total of 74 cases of cancer cervix, ICD-10 (International Classification of Diseases 10th Revision) category C53 were registered at the Karachi Cancer Registry, for Karachi South, during a 3 year period, 1st January, 1995 to 31st December 1997. RESULTS: The age standardized incidence rate (ASR) world and crude incidence rate (CIR) per 100,000 were 6.81 (5.2, 8.43) and 3.22 (2.49 to 3.96). Cancer cervix accounted for approximately 3.6% of all cancers in females and was the sixth malignancy in hierarchy. The mean age of the cancer cases was 53.27 years [standard deviation (SD) 11.6; 95% confidence interval (CI) 50.58, 55.96; range (R) 32-85 years)]. The distribution by religion was Muslims (90.5%), Christians (8.1%) and Hindus (1.4%). There were no cases reported in Parsees. The frequency distribution by ethnicity was Urdu speaking Mohajirs (20.3%), Punjabis (17.6%), Gujrati speaking Mohajirs (4.1%), memon Mohajirs (8.1%), Sindhis (10.8%), Baluchs (8.1%), Pathans (5.4%) and Afghan migrants (2.7%). The ethnicity was not known in approximately a fourth (23.0%) of the cases. The socio-economic distribution was 27.0% financially deprived class, 24.4% lower middle class and 48.7% upper middle and affluent classes. The majority of the women were married (86.5%); a smaller number were unmarried (2.7%) or widows (10.8%). The age-specific curves showed a gradual increase in risk from the fourth up till the seventh decade, followed by an actual apparent decrease in risk after 64 years of age. The peak incidence was observed in the 60 64 year age group. The morphological categorization was squamous cell carcinoma (86.5%), adenocarcinoma (10.9%) and adenosquamous carcinoma (2.6%). The majority of cases presented with moderately differentiated or grade 2 lesions (45.9%). There were no in-situ cases. Approximately half the cancers (58.1%) had spread regionally and 8.1% to a distant site at the time of diagnosis. Odds ratios (OR) were calculated for socioeconomic residential categories, religion, ethnicity, age groups and education. The OR for socioeconomic residential categories ranged between 0.69 and 2.9 with a marginally higher risk in the lower [OR 2.09 (95% CI .97; 4.49)] and lower middle class [OR 2.08 (95%CI 0.95; 4.58)]. Hindus [OR 1.2 (95% CI 0.18; 2.2)] had a slightly higher risk then the Muslims [OR 0.14 (95% CI 0.17; 1.2)]. A higher risk was also observed for Christians [OR 7.76 (95% CI 1.74; 34.5)]. CONCLUSION: The incidence of cervical cancer in Karachi South (1995 97) reflects a low risk population with a late presentation and a high stage disease at presentation. It is suggested that cervical screening if implemented should focus on once a life time methodology involving 36-45 year old women. This should be combined with HPV vaccination for the young and public health education for all. A regular cervical screening program would require mobilization of considerable financial, structural and human resources along with training for personnel. This may burden the already stretched health resources of a developing country. PMID- 18159968 TI - Lymphovascular space invasion as a prognostic determinant in uterine cancer. AB - The objective of this study was to evaluate the clinical significance of lymphovascular space invasion (LVSI) in patients with uterine cancer in terms of lymph node metastasis, recurrence and survival rate. A total of 190 patients with newly diagnosed uterine cancer who underwent total abdominal hysterectomy (TAH), bilateral salpingo-oophorectomy (BSO), omentectomy, peritoneal washing or ascitic fluid collection, and pelvic/paraaortic lymph node sampling at Chiang Mai University Hospital between January 1999 and December 2004 were evaluated. All medical records and histopathologic slides were retrospectively reviewed to determine the relationship between LVSI and clinicopathological characteristics. LVSI was present in 79 patients (42%) and significantly correlated with lymph node metastasis (p0.001), BMI 25 kg/m2 (p0.001), advanced FIGO stage (p 0.001), poor histologic grade (p0.001), and deep uterine invasion (p0.001). Patients with LVSI, when stratified by FIGO stage, also had a significant lower 5-year survival rate. For those who had disease recurrence, LVSI and histologic grade were found to be independent prognostic factors in a multivariate analysis. LVSI was one of the prognostic determinants for disease recurrence and associated with poor survival in patients with uterine cancer. PMID- 18159969 TI - Chronic lymphocytic leukemia in the recent 10 years and treatment effects of Fludarabin. AB - OBJECTIVE: CLL (Chronic Lymphocytic Leukemia) is the most common form of leukemia in the western world and because of prolonged survival of patients, the prevalence is high. Chemotherapy is usually not indicated in early and stable disease and using Chlorambucil with or without steroids has been the drug of choice in the treatment of CLL for many years . Clinical studies have shown that using Fludarabin can cause a complete response in significant number of untreated and/or previously treated CLL patients. The aim of this study is evaluating of CLL patients and determining the effects of treatment with Fludarabin. METHODS: A retrospective (descriptive/cross sectional) study of CLL patients who admitted to Hematology and Oncology Research Center of Tabriz university of Medical Sciences, between 1995-2005 was made and 126 patients enrolled. Collection of data was carried out according to special questionnaire and response to Fludarabin was analyzed by SPSS 11 software. RESULTS: The patients mean age of diagnosis was 63.7 years (SD=8.9), 69.8% were males. Illness and fatigue were the commonest presenting symptoms in 54% and lymphadenopathy was the most common clinical sign in 88.9%. Most of the patients were in stage C in Binet system (52.4%) and/or stage IV in Rai system (44.4%). Chemotherapy with chlorambucil and Prednisolone was the most common regimen used (60.3%) and 49.2% of patients were in partial remission with this treatment. Forty two patients treated with Fludarabin and 50% were in partial remission, 35% in static disease, 10% in progressive disease and 5% in complete remission (P=0.053). CONCLUSION: The median survival with Fludarabin was 43.9 months (SD=27.2) and in the case of Chlorambucil+Prednisolone and CVP or Chop it was 45 months (SD=26.5) and 50 months (SD=32.2), respectively (P>0.05). P value in the relationship with survival and response to Fludarabin was more than 0.05. Above all, Fludarabin is the choice treatment as first and second line therapy, as well as for patients who have failed therapy with standard regimens. PMID- 18159970 TI - Lack of initiation activity of 4-oxo-2-hexenal, a peroxidation product generated from omega-3 polyunsaturated fatty acids, in an in vivo five-week liver assay. AB - Peroxidation products formed from polyunsaturated lipids have DNA damaging potential. 4-oxo-2-hexenal (4-OHE), generated by the oxidation of omega-3 fatty acids, has been demonstrated to be mutagenic in vitro as assessed in the Ames test. To examine the carcinogenic risk of 4-OHE in vivo, initiation activity was investigated in a five-week liver assay, established to be effective for screening of carcinogenic potential of mutagens. Seven-week-old male F344 rats underwent two-thirds partial hepatectomy (PH) and were administered 4-OHE intragastrically at doses of 128, 80, 64, 40, 32, 20, or 0 mg/kg body weight (b.w.) at 18 hours thereafter, then being fed on diet containing 0.015% 2 acetylaminofluorene from weeks 2 to 4. All rats were given with 0.8 ml/kg b.w. CCl4 at week 3. At week 5, all survivors were sacrificed and initiation activity was assessed with reference to induction of glutathione S-transferase placental form (GST-P) positive foci in the liver. Mortality was significantly increased to 72.7% in the 128 mg/kg b.w. dose group compared with 0.9% in the control group. However, the average number of GST-P positive foci in the "128" dose group was 3.26-/+1.66 foci/cm2, not significantly different from the control value (2.78?1.33). Areas of GST-P positive foci were also similar (1.11-/+0.5 and 1.53 /+1.33 mm2/cm2 in "128" and the control groups, respectively). These results showed 4-OHE to have no significant initiation activity in. PMID- 18159971 TI - Prostate-specific antigen levels among Chinese, Malays and Indians in Singapore from a community-based study. AB - The purpose of this study was to examine the distribution of prostate-specific antigen levels among Chinese, Malays and Indians in Singapore, taking the effect of age into consideration. The study was carried out as part of the Singapore Prostate Awareness Week from 23-26th February 2004. Men above 50 years old went to four government-restructured hospitals to participate in the study. Participants filled up a questionnaire and provided 5 ml of blood for measurement of PSA levels using the Abbott IMx Total PSA assay (Abbott Laboratories). 3,486 men responded to the study, comprising 92.8% Chinese, 3.0% Malays, 2.5% Indians and 1.8% Others. 92.7% of them had PSA levels of 4 microg/L or less. There were no significant differences (p<0.05) between the mean PSA levels of Chinese (1.60 microg/L), Malays (1.39 microg/L), Indians (1.23 microg/L) and Others (1.70 microg/L). PSA levels were significantly associated with age (Spearman's r= 0.27, p<0.01). PSA levels increased with each 10-year age group and these trends were significant (p<0.0001) across both PSA group levels and age groupings. In the 50-60 years age groups, the prevalence of PSA levels >4 mug/L were 1.1% and 3.7% respectively. This rose rapidly to 11.3% and 23.5% for age groups >60-70 and >80 years respectively. Our study shows that the median PSA levels in the Caucasian population in the USA are higher than those of Chinese, Malays and Indians in Singapore. PSA levels were positively associated with age. It may be more appropriate to offer PSA testing to men who are >60 years old rather than the current >50 years. PMID- 18159972 TI - Appropriate interval for repeat excision in women undergoing prior loop electrosurgical excision procedure for cervical neoplasia. AB - The objective of the present study was to evaluate the impact of intervals on complications and pathological examination in women undergoing a repeat loop electrosurgical excision procedure (LEEP) for cervical neoplasia. During October 2004 and January 2007, 78 women who had undergone repeat LEEP at Chiang Mai University Hospital, were prospectively evaluated. The mean age was 47.5 years (range; 27-69 years). The mean duration of uncomplicated vaginal bleeding was 4.4 days (range; 1-20 days). The occurrence of persistent vaginal bleeding was noted in 9 women. Among 78 women, 2 (2.56%) and 7 (8.97%) experienced intraoperative and postoperative hemorrhage, respectively. Six (7.69%) had postoperative infection. These complications were not significantly different from those observed in women undergoing first LEEP in the same period (P=0.56). There was no significant difference in the incidence of perioperative complications and the incidence of non-evaluable cone margins among women who undergoing repeat LEEP within 4-6 weeks, between 6-8 weeks, and more than 8 weeks after first LEEP. In conclusion, repeat LEEP could be safely performed 4-12 weeks after the first procedure without any impact on pathological specimen examination. PMID- 18159973 TI - Knowledge and attitudes of physicians in Iran with regard to chronic cancer pain. AB - BACKGROUND AND AIM: The knowledge and attitudes of health care professionals with regard to pain and its impact on the patient are among identified barriers that prevent health care professionals from providing effective treatment for pain. The purpose of the present study was to evaluate knowledge about and attitudes towards cancer pain and its management in Iranian physicians with patient care responsibilities. METHODS: We surveyed 122 physicians in six university hospitals in Tehran. Fifty-five (45.1%) questionnaires were completed. RESULTS: The majority of physicians (76%) recognized the importance of pain management priority and about one half of the physicians acknowledged the problem of inadequate pain management in their settings. Most cited inability to access professionals who practice specialized methods in this field, and inadequate staff knowledge of pain management as barriers to good pain management. A large majority of them expressed dissatisfaction with their training for pain management in medical school and in residency. Furthermore a considerable widespread knowledge deficit among all medical subspecialties and all levels of experience was noted which was significantly more profound in the non-oncologists group and only correlated poorly with number of total treated patients in past 6 months. CONCLUSION: The most significant barrier to the effective management of pain in cancer patients in Iran is deficit in knowledge as identified in this survey. A combination of an active continuing education program on both the international guidelines with routine professional education and dissemination of guidelines is needed to bring about significant improvement in cancer pain control. PMID- 18159974 TI - Prevalence and characteristics of late postoperative voiding dysfunction in early stage cervical cancer patients treated with radical hysterectomy. AB - BACKGROUND: Although effective as a primary treatment for early-stage cervical cancer, radical hysterectomy is associated with significant long-term morbidities, most commonly, voiding dysfunction. OBJECTIVE: To examine prevalence and characteristics of voiding dysfunction following radical hysterectomy for early-stage cervical cancer. METHODS: One hundred-eighty seven patients with FIGO stage IA2-IIA cervical cancer who underwent class II-III radical hysterectomy with systematic pelvic lymphadenectomy between January 1, 2002 and June 31, 2005 were interviewed with questionnaire on voiding function. Medical records were also reviewed for operative and pathologic data. RESULTS: The prevalence of symptomatic bladder dysfunction was 25.1%. There was no statistically significant association between rates of bladder dysfunction and all examined clinical/operative factors. The most common pattern of bladder dysfunction were incomplete emptying in 25 (13.4%) and urgency and nocturia in 21 (11.2%) each. CONCLUSION: Voiding dysfunction is a common and clinically significant long-term complication following radical hysterectomy. The pattern of dysfunction reflects combined surgical disruption of both parasympathetic and sympathetic innervations of the pelvis. PMID- 18159975 TI - Effect of homeopathic medicines on transplanted tumors in mice. AB - Ultra low doses used in homeopathic medicines are reported to have healing potential for various diseases but their action remains controversial. In this study we have investigated the antitumour and antimetastatic activity of selected homeopathic medicines against transplanted tumours in mice. It was found that Ruta graveolens 200c and Hydrastis canadensis 200c significantly increased the lifespan of Ehrlich Ascites Carcinoma and Dalton's Lymphoma Ascites induced tumour-bearing animals by 49.7%, and 69.4% respectively. Moreover there was 95.6% and 95.8% reduction of solid tumour volume in Ruta 200c and Hydrastis 200c treated animals on the 31st day after tumour inoculation. Hydrastis 1M given orally significantly inhibited the growth of developed solid tumours produced by DLA cells and increased the lifespan of tumour bearing animals. Some 9 out of 15 animals with developed tumors were completely tumour free after treatment with Hydrastis 1M. Significant anti-metastatic activity was also found in B16F-10 melanoma-bearing animals treated with Thuja1M, Hydrastis 1M and Lycopodium1M. This was evident from the inhibition of lung tumour nodule formation, morphological and histopathological analysis of lung and decreased levels of gamma-GT in serum, a cellular marker of proliferation. These findings support that homeopathic preparations of Ruta and Hydrastis have significant antitumour activity. The mechanism of action of these medicines is not known at present. PMID- 18159976 TI - Risk factors of breast cancer in north of Iran: a case-control in Mazandaran Province. AB - INTRODUCTION: Breast cancer is the most common cancer among Iranian women. This study aimed to determine risk factors for breast cancer in the north of Iran. METHOD: A matched case-control study was conducted in Mazandaran province of Iran in 2004 of 250 biopsy proven cases of breast cancer and 500 neighbor controls that were matched by age within a 3 year period. Statistical analysis was carried out using conditional logistic regression with the backward elimination method and crude and adjusted odds ratios with related 95% CIs were estimated with Stata 8.0 software RESULTS: Multivariate analysis showed that higher education (OR=4.70, 95%CI: 1.71-12.88), late menopause (OR=4.18, 95%CI: 2.54-6.88), history of induced abortion (OR=1.62, 95%CI: 1.13-2.31), positive first-degree family history of breast cancer (OR=3.14, 95%CI: 1.37-7.20), and BMI (OR=1.02, 95%CI: 1.01-1.03) were risk factors for breast cancer. Furthermore, having more episodes of full term pregnancy (OR=0.87, 95%CI: 0.80-0.95), longer duration of breast feeding (OR=0.993, 95%CI: 0.989-0.997) and parity more than 2 were shown to be protective factors. CONCLUSIONS: Our study revealed the role of some modifiable determinants of breast cancer that can be focused by public health intervention in the northern community of Iran. Accordingly, the women who have one or more of the following risk factors should take the special attention to risk of breast cancer: obesity, being menopause, positive family history of breast cancer and history of induced abortion. The protective effect of longer duration of breast feeding should be encouraged too. PMID- 18159977 TI - Pediatric brain tumours at a tertiary care hospital in Karachi. AB - The objectives of this study were to determine the epidemiology of brain tumors during infancy and childhood and to define and segregate childhood brain tumors vis-a-vis their morphological characteristics. The present study includes pediatric brain tumors, ICD-10 category C71 encountered during 10 years (January 1989 through December 1998) at a tertiary care hospital in Karachi. Eighty one cases were included, 58 (71.6%) in males and 23 (28.4%) in females with a male to female ratio of 2.5:1. The cases were divided into 3 age groups each covering five years of life (0-4, 5-9, 10-14 years), with the greatest number in the second age group i.e. 5-9 years followed by the third age group and the 0-4 year age group. The mean age for all cases, both genders was 8.8 years (95% CI 7.9; 9.6) with a marginal variation for cases occurring in the cerebrum and cerebellum. The malignancies occurred at a younger age in the males for each subcategory by site and morphology. The morphological distribution of cases was astrocytoma (28 cases, 34.6%), primitive neuroectodermal tumor or PNET (40 cases; 49.4%), ependymoma (8 cases, 10%), mixed glioma (4 cases; 5%) and a case of oligodendroglioma. The 81 malignancies included in this study were further categorized by site into two groups, supratentorial (27 cases; 33.3%) and infratentorial (54 cases; 66.7%). The morphological categorization of supratentorial tumors was astrocytoma (17 cases; 63%), ependymoma (5 cases; 18.5%), mixed glioma (2 cases; 7.4%). PNET with rhabdoid differentiation, oligodendroglioma and pinealoblastoma comprised 1 case (3.7%) each. The 17 supratentorial astrocytoma were sub-categorized as follows - pilocytic astrocytoma (5 cases; 29.4%), grade II astrocytoma (6 cases; 35.3%); grade III astrocytoma (2 cases; 11.8%), anaplastic astrocytoma (1 case; 5.9%) and glioblastoma multiforme (3 cases; 17.7%). The morphological categorization of infratentorial tumors was astrocytoma (11 cases; 20.4%), medulloblastoma (38 cases; 70.4%), ependymoma (3 cases; 5.6%) and mixed glioma - astroependymoma (2 cases, 3.7%). The morphological sub-categorization of infratentorial astrocytoma was pilocytic astrocytoma (7 cases, 63.6%), with gemistocytic astrocytoma, grade II, grade III and anaplastic astrocytoma comprising 1 (9.1%) case each. The morphological categorization of medulloblastoma was classical medulloblastoma (15 cases; 39.5%), desmoplastic medulloblastoma (8 cases; 21.1%), medulloblastoma with astrocytic differentiation (12 cases; 31.5%), medulloblastoma with neural differentiation (2 cases; 5.3%), and neuroblastic medulloblastoma (1 case; 2.6%). The pediatric brain tumors in Karachi reflect a developing country scenario, with a strong male predisposition and a late presentation with a peak in the 5-9 year age group. There is a predominance of medulloblastoma and a paucity of astrocytomas. The current study is a single institution study and needs cautious interpretation. Population-based studies are required to determine the cancer burden due to pediatric malignancies of the brain in this population and for the morphological categorization of brain tumors in Karachi. PMID- 18159978 TI - Cancer incidence in the Middle Eastern population of California, 1988-2004. AB - International statistics suggest lower cancer incidence in the Middle East and Middle Eastern (ME) immigrants in Europe, Australia, and Canada, but little is known from the United States. This study compares cancer rates in ME population with other race/ethnic groups in California from 1988 through 2004. ME cases in California cancer registry were identified by surname and ME population was estimated from U.S. Census data. Cancer rates for ME countries was obtained from Globocan. The ME incidence rate ratios for all sites combined in male and female were 0.77 and 0.82, respectively and were statistically significant. ME rates were significantly lower for cancers of the colon, lung, skin melanoma, female breast and prostate, and were significantly higher for cancers of the stomach, liver, thyroid, leukemia, and male breast. Cancer incidence in ME population in California was 2.4 times higher than rates in home countries. Incidence trends in ME males remained fairly stable but in females shows a slight decline in recent years. Cancer incidence in ME population is lower than non-Hispanic white and non Hispanic Black, but is higher than rates for Hispanics and Asians, and ME countries. Improved data quality, chronic infections, acculturation, and access to screening services are some of the factors responsible for the observed pattern. PMID- 18159979 TI - Comparing Cox regression and parametric models for survival of patients with gastric carcinoma. AB - BACKGROUND: Researchers in medical sciences often tend to prefer Cox semi parametric instead of parametric models for survival analysis because of fewer assumptions but under certain circumstances, parametric models give more precise estimates. The objective of this study was to compare two survival regression methods - Cox regression and parametric models - in patients with gastric adenocarcinomas who registered at Taleghani hospital, Tehran. METHODS: We retrospectively studied 746 cases from February 2003 through January 2007. Gender, age at diagnosis, family history of cancer, tumor size and pathologic distant of metastasis were selected as potential prognostic factors and entered into the parametric and semi parametric models. Weibull, exponential and lognormal regression were performed as parametric models with the Akaike Information Criterion (AIC) and standardized of parameter estimates to compare the efficiency of models. RESULTS: The survival results from both Cox and Parametric models showed that patients who were older than 45 years at diagnosis had an increased risk for death, followed by greater tumor size and presence of pathologic distant metastasis. CONCLUSION: In multivariate analysis Cox and Exponential are similar. Although it seems that there may not be a single model that is substantially better than others, in univariate analysis the data strongly supported the log normal regression among parametric models and it can be lead to more precise results as an alternative to Cox. PMID- 18159980 TI - Tobacco use among students and school personnel in India. AB - BACKGROUND: Tobacco usage is addictive and causative for several diseases and premature death. Concerted efforts by the individual and society are needed for control and for surveillance. The habit is initiated during early youth and this age group requires constant monitoring and timely appropriate action to curtail usage. The WHO FCTC has recommended actions to monitor and limit the tobacco use in young age groups. One of the actions is to examine the prevalence of tobacco habits in school children 13-15 years of age and of personnel employed in schools. METHODS: WHO & CDC designed the study systems for Global Youth Tobacco Survey (GYTS) and Global School Personnel Survey (GSPS). In 2006 we conducted GYTS and GSPS in several parts of the country. The schools were chosen by strict sampling procedure and a well structured, self-administered questionnaire was used to obtain information on tobacco usage from 13 to 15 year old students of chosen schools and personnel of these schools. RESULTS: Current use of any tobacco product was 14.1% among students (17.3% boys, 9.8% girls) and among school personnel it was 29.2%(35.0% males and 13.7% females). The prevalence was highest among male students in North East (34%) and the lowest was 4.9% among female students of western states. Cigarettes and Bidi smoking were more prevalent among boys. Smokeless tobacco use prevalence rate varied between 20% and 4.5% among boys and between 21.5% and 1.6% among girl students. Among male school personnel, the prevalence varied from 57.9% in NE to 25.7% in South. Among females 26.5% were tobacco users in the NE and in Western region it was 6.6%. CONCLUSION: It is essential to adopt forceful strategies, which are area specific, in order to undo the harm inflicted by tobacco use upon the individuals &society. Periodic surveys for surveillance of trends are essential to evaluate the outcome of programmes among students and school personnels. PMID- 18159981 TI - The risk factors of prostate cancer: a multicentric case-control study in Iran. AB - Prostate cancer (PC), in Iran, is the third most frequently diagnosed visceral cancer among men and the seventh most common underlying cause of cancer mortality. We evaluated the relation between speculated factors and PC risk using data from a multicentric case-control study conducted in Iran from 2005 to 2007 on 130 cases of incident, clinicopathologically confirmed PC, and 75 controls admitted to the same network of hospitals without any malignant disease. Odds ratios (OR) and corresponding 95% confidence intervals (CIs) were estimated using conditional logistic regression models. The risk of PC was increased with aging (OR: 5.35, 95% CI: 2.17-13.19; P<0.0001), and with the number of sexual intercourse >or=2 times/week (OR: 3.14, 95% CI: 1.2-8.2; P=0.02). One unit elevation in serum estradiol and testosterone concentration was related to increase (OR: 1.04, 95% CI: 1.01-1.06; P=0.006) and decrease (OR: 0.79; 95% CI: 0.64-0.96; P=0.02) of PC risk, respectively. Cases were less likely to have a history of diabetes (OR: 0.34, 95% CI: 0.12-0.98; P=0.04). Increasing in dietary consumption of lycopene and fat was associated with declined (OR: 0.45, 95% CI: 0.09-2.12) and increased (OR: 2.38, 95% CI: 0.29-19.4) PC development, respectively. Other factors including educational level, marriage status, dietary meat consumption, vasectomy and smoking have not been shown to affect PC risk in the Iranian population. Our study adds further information on the potential risk factors of PC and is the first epidemiologic report from Iran. However, justification of these results requires more well-designed studies with a larger number of participants. PMID- 18159982 TI - Chinese and Vietnamese adult male smokers' perspectives regarding facilitators of tobacco cessation behavior. AB - INTRODUCTION: National surveys show a low prevalence of tobacco cigarette smoking within the Asian American/Pacific Islander population. However, smoking rates loom higher when data is disaggregated by ethnicity and gender. Nevertheless, few data are available on how smokers in this population quit smoking. The aim of this study was to collect first-hand perspectives from adult male Chinese and Vietnamese current and former smokers who were patients at a community clinic in Seattle, Washington, in order to understand the facilitators toward smoking cessation and the methods that they might use to quit smoking. METHODS: A telephone survey was administered to age-eligible male Chinese and Vietnamese clinic patients who were current or former smokers. A total of 196 Chinese and 198 Vietnamese (N=394) adult male current and former smokers were contacted from a pool culled from the clinic database. RESULTS: Descriptive analysis using SPSS software revealed ethnicity-specific differences between current and former smokers regarding influences on smoking cessation behavior as well as uptake and endorsement of cessation methods. Family encouragement and physician recommendations were significant facilitators on the cessation process. Will power and self-determination were frequently mentioned by both Vietnamese and Chinese smokers as helpful methods to quit smoking. Vietnamese smokers were more resourceful than Chinese smokers in their use of smoking cessation methods. CONCLUSION: Even with access to cessation classes at a health clinic, half of current smokers indicated that they had no intention to quit. Such attitudes underscore the need for promotion of effective smoking cessation programs as well as successful strategies for reaching smokers. These conclusions are particularly important for Chinese smokers, who were comparatively less resourceful in their use of smoking cessation methods. Future studies should explore integrating the concept of will power with current mainstream state-of-the-art smoking cessation programs. PMID- 18159983 TI - Study of the interaction surface for the c-Src-imatinib complex by a molecular dicing technique. AB - With the beginning of the new millennium, a new and exciting era for cancer therapy has begun with the appearance of molecular targeted drugs. Imatinib is a clinically well-tolerated small molecule that exerts selective, dual inhibition of the transforming growth factor beta (TGFbeta) and platelet-derived growth factor (PDGF) pathways. Imatinib is also suggested as a chemopreventive for recurrent and metastatic malignancies. An interesting point to be clarified regarding the mechanism of imatinib is its interaction with c-Src. Fortunately, complexing of c-Src and imatinib has recently reported, which provides a basis for further study of the interactions between the two molecules. In the present study, the author used the technique named molecular dicing to study the interaction surface between the two molecules. Accordingly, the interaction surface in c-Scr and imatinib could be identified. PMID- 18159984 TI - Association of CYP1A1, CYP1A2, GSTM1 and NAT2 gene polymorphisms with colorectal cancer and smoking. AB - We investigated CYP1A1*2A, CYP1A1*2C, CYP1A2*1C, CYP1A2*1F, GSTM1 and NAT2 gene polymorphisms, involving enzymes which metabolize many carcinogens, with reference to colorectal cancer risk. The distribution of these genotypes was not associated with risk overall. However, the CYP1A1*2A T/C genotype showed a significant association with colorectal cancer risk in never-smokers (odds ratio [OR], 3.06; 95% confidence interval [95% CI], 1.11-8.40; p = 0.030). The risk of the NAT2 rapid genotype in never-smokers was also statistically significantly increased (OR, 5.38; 95%CI, 1.80-16.1; p = 0.003). Furthermore, the joint effects of NAT2 rapid plus other genotypes were associated with colorectal cancer overall (OR, 3.12; 95%CI, 1.15-8.51; p = 0.026, for NAT2 rapid plus combined CYP1A1*2C Ile/Val and Val/Val, OR, 3.25; 95%CI, 1.09-9.74; p = 0.035, for NAT2 rapid plus CYP1A2*1C G/G, and OR, 4.20; 95%CI, 1.09-16.1; p = 0.037, for NAT2 rapid plus GSTM1 null, respectively). In never-smokers, the joint effects of NAT2 rapid plus other genotypes were remarkable (OR, 15.9; 95%CI, 1.87-135.8; p = 0.011, for NAT2 rapid plus combined CYP1A1*2A T/C and C/C, OR, 5.71; 95%CI, 1.49-21.9; p = 0.011, for NAT2 rapid plus combined CYP1A1*2C Ile/Val and Val/Val, and OR, 9.14; 95%CI, 2.05-40.7; p = 0.004, for NAT2 rapid plus CYP1A2*1F A/A, respectively). The joint effect of CYP1A2*1F A/A plus CYP1A2*1C G/G genotypes was also increased in never smokers (OR, 6.16; 95%CI, 1.26-30.1; p = 0.025). Our findings suggest that the CYP1A1*2A T/C and NAT2 rapid genotypes is associated with colorectal cancer susceptibility without smoking exposure. These results also indicate that the NAT2 in combination with CYP1A1*2C, CYP1A2*1C, or GSTM1 genotypes may strongly confer susceptibility to colorectal cancer. In particular, the combination of NAT2 plus CYP1A1*2A, CYP1A1*2C, or CYP1A2*1F genotypes, and that of CYP1A2*1F plus CYP1A2*1C genotype may define a group of persons who are genetically susceptible to colorectal cancer in never smokers. PMID- 18159985 TI - Cancer cachexia: pathophysiologic aspects and treatment options. AB - Cancer cachexia is a syndrome characterized with progressive weight loss and abnormal wasting of fat and muscle tissue, and affects 40 to 85% of all terminally ill patients, accounting more than 20% of all cancer deaths. Current treatment for cancer cachexia principally depends on its prevention rather than reversing the present disease state, and the clinical results are far from being satisfactory. Although the exact mechanism and predisposing factors have yet to be clarified in detail, our growing knowledge about the pathophysiology and biochemical changes considering this life threatening condition should help in development of future therapeutic strategies. In the present paper, the current preclinical and clinical features considering the pathophysiology and treatment of cancer related cachexia are reviewed. PMID- 18159986 TI - Cancer registration in Pakistan: contemporary state of affairs. AB - An estimated 7.6 million mortalities were attributed to cancers in 2005 across the globe, and the figures keep mounting. The existing prevalence rate for all reportable cancers is at 25 million, projected to ascend to 30 million around year 2020. Some 70% of these will burden the health care services in under resourced countries. However, reliable statistics on the incidence, prevalence, mortality and survival rates (five and ten year) are limited. A National Cancer Control Program (NCCP) in Pakistan has existed since 1994, but little is known to the public and negligible information has been made available to medical services providers. We have a poverty of local bio-medical literature related to the specialties like oncology and cancer epidemiology. Only 175 papers are indexed by Pakmedinet, seven being relevant to the cancer registration in Pakistan, an impetus for programmed documentation, and to serve as a foundation for approach to Cyber-medicine. Infrastructure and ample requisite manpower exists locally and a number of local institutions as well as outside sources are teaming up. However, there are still quite a few issues to be looked into, including the question of who will own the data arising from this pooling of assets? A community-based approach is mandatory to allay public concerns over confidentiality and possible use of the registry data. Already, there are hurdles in attaining public approval after the intentions to found a National Cancer Registry were ventilated by the Pakistan Medical Research Council. PMID- 18159987 TI - Use of lay health workers in a community-based chronic disease control program. AB - The increasing burden of non-communicable diseases in the developing world, and in particular diabetes, cancer and circulatory diseases, is an unfortunate fact of life. At the same time infection-related diseases, including sexually transmitted HIV-AIDS and HPV-dependent cervical cancer, remain important. One approach to alleviating the resultant stress on national health provision is to expand the knowledge base at the community level with contributions by lay health workers (LHWs). Here we take a brief look at the available literature and propose a model for intervention incorporating two way dialogue with the general populace to find effective means to package expertise in the medical/research community for lay consumption. Our argument is that particular attention should be paid to socioeconomic and behavioural aspects and to disease surveillance at the local level in order to be able to accurately assess the impact of interventions. For this purpose, we need to marshal volunteers from within communities taking account of their problems and motivations. Included are provision of assistance in setting up physical exercise programs, quit tobacco campaigns, alcohol awareness programs, running disease screening exercises and general help by providing advice as to risk and protective factors and clinical treatments, with an special focus on palliative care. PMID- 18159988 TI - Are there any real Helicobacter pylori infection-negative gastric cancers in Asia? AB - The great variability in gastric cancer rates across Asia, with very high incidences in Japan and Korea, and exceedingly low incidences in ethnic Malays, whether in Malaysia or Indonesia, appears largely due to variation in Helicobacter pylori infection rates. While between 2% and 10.6% of gastric cancers in a recent Japanese survey were considered to be negative for bacterial infection on the basis of seropositivity and H. pylori-dependent mucosal atrophy, it is notoriously difficult to preclude past infection. The situation is greatly complicated by reported differences in the etiology of gastric cardia and non cardia cancers. In the Western world there do appear to be tumours arising close to the esophageal-gastric junction which are not related to H. pylori and associated inflammation, but in most Asian populations these appear to be very rare. Therefore preventive efforts, and particularly screening, should be focused on markers of bacterial infection, with avoidance of unnecessary exposure to X ray radiation. PMID- 18159989 TI - Cooperation partners in information sharing within the context of an Asian cancer network. AB - It would be a great mistake to analyze the health situation in Asia relying on the focus on individualism inherent in the sense of values of Europeans and Americans. Cooperation across fields is indispensable for effective control of the epidemic of disease we are facing in the 21st century. We need to concentrate efforts on bringing together specialists, not only within the various areas of medical practice, but also across such fields as economics, politics and information technology (IT). Asia differs from Europe and America in that it does not have any group political structure and therefore we must rely on voluntary integration of our efforts if we are to achieve the most effective application of our combined resources. Non-intervention in internal affairs is naturally a very important condition for success. Sharing of information while abiding by national regulations regarding medical data confidentiality does pose difficulties, but gentle persuasion to standardize processes with a shared commitment to overcoming problems should reduce opposition. Our common purpose in maintaining healthy societies, whether we be scientists, medical staff, economists, computer specialists or politicians, provides the bond. Ways and means by which this bond can be strengthened deserve our attention. PMID- 18159990 TI - Serum biomarker concentrations and outcome after pediatric traumatic brain injury. AB - Predicting outcome after pediatric traumatic brain injury (TBI) is important for providing information to families and prescribing rehabilitation services. The study objective was to assess whether biomarkers concentrations obtained at the time of injury are associated with outcome. Serial serum concentrations of neuron specific enolase (NSE), S100B and myelin basic protein (MBP) were measured in 152 children with acute TBI. Outcome was assessed with the Glasgow Outcome Scale (GOS) score and/or GOS-Extended Pediatric (GOS-E Peds). Spearman's rank correlation and binary logistic regression assessed the relationship between biomarker concentrations and outcome. For all biomarkers and time points, higher biomarker concentrations were associated with worse outcome. Initial and peak NSE concentrations and initial MBP concentrations were more strongly correlated with outcome in children < or =4 years compared with those >4 years of age. Using binary logistic regression to evaluate the simultaneous affect of all biomarkers on outcome, there was significant overall model fit predicting a dichotomous GOS from biomarker concentrations with a 77% correct classification rate and a negative and positive predictive value of 97% and 75%, respectively. We conclude that NSE, S100B, and MBP concentrations obtained at the time of TBI may be useful in predicting outcome. Future studies should focus on assessing the differential benefit of biomarkers compared with clinical variables and in assessing a continuous rather than categorical outcome variable. PMID- 18159991 TI - Correlation between APOE -491AA promoter in epsilon4 carriers and clinical deterioration in early stage of traumatic brain injury. AB - The objective of this work was to investigate the relationship between apolipoprotein E (APOE) promoters (G-219T, C-427T, A-491T) polymorphisms and the clinical deterioration in early stage of traumatic brain injury (TBI) in a cohort of Chinese patients. In this study, we used the cohort of patients which has been reported previously. A total of 110 subjects with TBI (80 males and 30 females, with mean age of 43.87 years) were admitted from December 2003 to May 2004, and demographic and clinical data were collected. The clinical deterioration of patient's condition in acute stage (<7 days after TBI) was judged by either of the following criteria: decrease of Glasgow Coma Scale (GCS) score (compared with initial admission GCS), increase in hematoma volume or delayed hematoma both detected by repeated computed tomography (CT) scanning compared to that on admission. Venous blood was collected from patients with TBI on admission to determine the APOE promoter polymorphisms. The APOE genotyping was performed by means of polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP). chi(2) test and logistic regression analyses were done by SPSS. In 110 Chinese patients, the distributions of APOE genotypes and alleles matched Hardy Weinberg Law, and 19 subjects presented with deteriorated clinical condition in acute stage after hospitalization. chi(2) test showed insignificant differences in association of APOE promoter polymorphisms with clinical deterioration (p>0.05). But logistic regression analyses, after adjusting patients' age, injury severity and injury mechanism etc, showed that -491AA (OR=11.681, p=0.009, 95%, CI 1.824-74.790) and APOE epsilon4 were all risk factors, with injury severity and alcohol-drinking as other risk factors. In Chinese population, as a significant but not independent risk factor, only APOE -491AA promoter in epsilon4 carriers is apt to the clinical deterioration and may contribute to the poor outcome after TBI. PMID- 18159992 TI - Comparative magnetic resonance imaging at 1.5 and 3 Tesla for the evaluation of traumatic microbleeds. AB - Traumatic microbleeds (TMBs) can be regarded as a radiological marker of diffuse axonal injury (DAI). We sought to investigate the impact of the field strengths on the depiction of TMBs by T2*-weighted gradient echo magnetic resonance imaging (MRI). By the use of comparative MRI of 14 patients (age range, 22-62 years) on 1.5- and a 3 T (Tesla) systems at a median time interval of 61 months after traumatic brain injury (TBI), we found 239 (range 0.5-48.5, median 7.5) TMBs at 1.5 T, and 470 (range 2-118, median 18.5) TMBs at 3 T, respectively (p=0.001). However, in all but one patients MRI at 1.5 T also clearly showed TMBs. A significant negative correlation between the number of TMBs and the time interval TBI-MRI was observed, which was weaker for the imaging at 3 T (r(s)=-0.798; p=0.001; and r(s)=-0.649; p=0.012, respectively). In conclusion, T2*-weighted gradient-echo MRI at 3 T is superior as compared to MRI at 1.5 T for the detection of TMBs. Nevertheless, in clinical practice, MRI at 1.5 T seems to be sufficient for this purpose. MRI at 3 T may be appropriate if there is a strong clinical suspicion of DAI, despite unremarkable routine MRI, and possibly also if evidence of DAI is sought after a long interval from trauma. PMID- 18159993 TI - Effect of neuregulin-1 on histopathological and functional outcome after controlled cortical impact in mice. AB - Neuregulin-1 is a pleiotropic endogenous growth factor that is neuroprotective in experimental models of cerebral ischemia. We tested the hypothesis that pretreatment with neuregulin-1 would be similarly protective after traumatic brain injury in mice. Mice were administered neuregulin-1 or equal amounts of vehicle intravenously immediately before controlled cortical impact. Injured mice were subjected to motor and cognitive testing, and brain tissue loss was quantitated at 4 weeks. Compared to vehicle, pretreatment with neuregulin-1 had no effect on brain tissue loss, motor function, or acquisition of a spatial learning task. However, neuregulin-1 treated mice showed improved retention of spatial memory versus vehicle-treated mice in subsequent probe trials (p<0.05). These proof-of-principle data suggest that neuregulin-1 may improve some functional outcomes after brain trauma. Further studies are therefore warranted to more carefully explore molecular mechanisms, dose-responses, and relationships between morphological outcome and long-term recovery. PMID- 18159994 TI - Closed head injury causes hyperexcitability in rat hippocampal CA1 but not in CA3 pyramidal cells. AB - Traumatic brain injury frequently elicits epileptic seizures hours or days after the impact. The mechanisms on cellular level are poorly understood. Because posttraumatic epilepsy appears in many cases as a temporal-lobe epilepsy which originated the hippocampus, we studied trauma-induced hyperexcitability on the cellular level in this brain area. We used the model of closed head injury to analyse the electrophysiological changes in CA1 and CA3 pyramidal cells and in interneurones of the CA1 field, which is extremely sensitive to ischemia. We found that morphologically closed head injury (CHI) led to a gradual progressive, cell type specific time course in neuronal degeneration. To analyse electrophysiological impairment we measured resting membrane potential, recorded spontaneous action potentials and induced action potentials by current pulses at different times after CHI. We found a dramatic increase in the frequency of spontaneous action potentials of CA1 but not of CA3 pyramidal cells after CHI. This hyperexcitability was maximal at 2 h (4.5-fold higher than sham), was also observed at 24 h after CHI and disappeared after 3 days. We found that CA1 interneurones responded by a much weaker increase of AP frequency after CHI. We conclude that the strong hyperexcitability after CHI is cell-type specific and transient. The understanding of the complex neuronal interactions probably offers a promising possibility for pharmacological intervention to prevent posttraumatic epilepsy. PMID- 18159995 TI - Simultaneous detections of 27 cytokines during cerebral wound healing by multiplexed bead-based immunoassay for wound age estimation. AB - Quantification of 27 cytokines following cerebral wounding was performed for wound age estimation. The cytokines evaluated included interleukin (IL)-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12 p40, IL-12 p70, IL-15, IL 17, IL-18, basic fibroblast growth factor (bFGF), granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), Interferon-gamma (IFN-gamma), keratinocyte derived cytokine (KC), leukemia inhibitory factor (LIF), macrophage-colony stimulating factor (M-CSF), monokine inducible by interferon gamma (MIG), macrophage inflammatory protein (MIP)-1 alpha, MIP 2, platelet-derived growth factor BB (PDGF BB), regulated upon activation, normal T-cell expressed, and secreted (Rantes), tumor necrosis factor alpha (TNF-alpha), and vascular endothelial growth factor (VEGF). The proliferation of glial cells as well as the infiltration of inflammatory cells were also evaluated. Although astroglia proliferated from 72 hours post-injury, inflammatory cell dynamics were generally steady. Among cytokines analyzed in the present study, IL-1beta, IL-5, IL-6, IL-12 p40, G-CSF, IFN-gamma, KC, LIF, MIP2, and PDGF BB increased during the early phase of cerebral wound healing, and M-CSF increased during the middle phase, while IL-15, IL-18, and MIG increased during the late phase. In contrast, IL-1alpha, IL-10, IL-12 p70, and TNF-alpha were suppressed throughout the cerebral wound healing process. Based on our findings, quantitative cytokine analyses at the cerebral wound site may be a useful tool for wound age estimation. Further, this study suggests that multiplex data gained from the same sample using a single methodology demonstrates highly accurate cytokine interactions during the process of cerebral wound healing. PMID- 18159996 TI - Nitric oxide spin-trapping and NADPH-diaphorase activity in mature rat brain after injury. AB - Traumatic brain injury (TBI) induces inflammatory reactions, and one of the essential mediators of this reaction is nitric oxide (NO). The action of this compound is still under study because no clear consensus has been reached about its exact action in the central nervous system. Further, it is unknown if, in the damaged brain, its neuroprotective activity outweighs its putative neurodegenerative properties. Using ferrous-diethyldithiocarbamate chelate, a lipophilic spin trap for NO detection by electron paramagnetic resonance (EPR) spectroscopy, we followed NO production in injured brain of mature Wistar rats. To relate changes in the amount of NO in the lesioned brain to the activity of NO synthase (NOS), this study also used NADPH-diaphorase staining. Our data show a rapid drop of NO concentration in the damaged brain below control values. This phenomenon persisted over several hours postinjury and varied with brain region. This decrease in NO concentration was accompanied by a simultaneous increase in the number of NADPH-diaphorase-positive cells, perhaps indicative of increased NOS activity. It is therefore assumed that, in the lesioned brain, a very rapid removal of NO occurs via its transformation to other reactive species such as peroxynitrite, which further adversely influence the damaged tissue. PMID- 18159997 TI - L-arginine reverses alterations in drug disposition induced by spinal cord injury by increasing hepatic blood flow. AB - High hepatic extraction drugs--such as phenacetin, methylprednisolone, and cyclosporine--exhibit an increased bioavailability after acute spinal cord injury (SCI) due to an impaired clearance. For these drugs, metabolic clearance depends on hepatic blood flow. Thus, it is possible that pharmacokinetic alterations can be reversed by increasing liver perfusion. Therefore, we evaluated the effect of L-arginine, a nitric oxide precursor, on the pharmacokinetics of a prototype drug with high hepatic extraction, and on hepatic microvascular blood flow (MVBF) after acute SCI. Pharmacokinetics of i.v. phenacetin was studied in rats 24 h after a severe T-5 spinal cord contusion; animals being pretreated with L arginine 100 mg/kg i.v. or vehicle. MVBF was assessed under similar experimental conditions using laser Doppler flowmetry. SCI significantly altered phenacetin pharmacokinetics. Clearance was significantly reduced, resulting in a prolonged half-life and an increase in bioavailability, while volume of distribution was decreased. Pharmacokinetic alterations were reversed when injured rats were pretreated with L -arginine. It was also observed that L-arginine significantly increased hepatic MVBF in injured rats, notwithstanding it exhibited a limited effect on sham-injured animals. Our data hence suggest that L-arginine is able to reverse SCI-induced alterations in phenacetin pharmacokinetics due to an impaired hepatic MVBF, likely by increased nitric oxide synthesis leading to vasodilation. Further studies are warranted to examine the potential usefulness of nitric oxide supplementation in a clinical setting. PMID- 18159998 TI - Co-transplantation of neural stem cells and NT-3-overexpressing Schwann cells in transected spinal cord. AB - Spinal cord transection results in severe neurological sequelae, and to date, there is no effective treatment. Because of the limited capacity for axonal regeneration in the spinal cord, recovery is minimal. Recently, efforts have been made to establish, by grafting neural tissue, a functional relay-station between the severed stumps of the injured cord. Previously, we used co-transplantation of neural stem cells (NSCs) and Schwann cells (SCs) to improve functional recovery of transected spinal cord. However, this effort has been partially impeded by limited neuronal differentiation of transplanted NSCs. To circumvent this problem, we have pre-differentiated NSCs toward neurons in vitro with the application of retinoic acid (RA) prior to cell grafting. Further, we genetically modified SCs to overexpress human neurotrophin-3 (hNT-3). When these cells were co-transplanted into the transected spinal cord of rats, injured animals had partial improvement (both functionally and structurally), including improved Basso, Beattie, and Bresnahan (BBB) scores, increased axonal regeneration/remyelination, and reduced neuronal loss. However, this pre differentiation of NSCs in vitro only mildly improved neuronal differentiation of NSCs in vivo. PMID- 18159999 TI - Characterization of the induced neuropeptide Y-like immunoreactivity in primary sensory neurons following complete median nerve transection. AB - In this study, we examined characteristics of the neuropeptide Y-like immunoreactive (NPY-LI) dorsal root ganglion (DRG) neurons after complete median nerve transection (CMNT). With fluorogold (FG) injection into normal median nerves, numerous FG-labeled DRG neurons were localized predominantly in the C6 and C7 DRGs, where the focal regions were examined after CMNT. With NPY immunohistochemistry, a few NPY-LI neurons were detected in the ipsilateral but not contralateral DRGs after FG injection into the nerve. As early as 3 days after CMNT a few NPY-LI neurons could be detected, reaching a maximum in the DRGs at 4 weeks, subsiding thereafter over 20 weeks. The NPY-LI DRG neurons were primarily medium-sized and large neurons. With FG injection into the transected median nerve, we found that approximately 99% of NPY-LI neurons were labeled for FG, suggesting that they were derived from the injured but not intact DRG neurons. Using double fluorescent dyes tracing, we detected that some of the injured DRG neurons were NPY-LI neurons that projected to the cuneate nucleus (CN). Following dorsal rhizotomy, our data indicated that after CMNT the induced NPY-LI fibers in the ipsilateral CN originated exclusively from the injured DRG neurons. Taken together, these findings suggest that injury-induced NPY-LI fibers in the CN may originate from the injured DRG neurons via the median primary afferent fibers, affect the excitability of cuneothalamic projection neurons (CTNs), and involve neuropathic sensation following CMNT. PMID- 18160001 TI - HIV in the workplace in Botswana: incidence, prevalence, and disease severity. AB - Few detailed epidemiologic data exist regarding the impact of HIV infection on the workplace in the developing world. In addition, most HIV surveys examine only prevalence, without data on incidence or disease severity. In June 2003, we conducted a voluntary anonymous HIV serosurvey among employees of the Debswana Mining Company, the largest nongovernmental employer in Botswana. Among the 3558 participants, annual HIV incidence was estimated to be 3.4%, and HIV prevalence was 23.8%. HIV-infected participants had a median CD4(+) lymphocyte count of 427 cells/mm(3) (interquartile range 269-642), with 13.3% of samples <200 cells/mm(3). The high incidence and prevalence of HIV, and the proportion of workers with <200 CD4(+) lymphocytes/mm(3), demonstrate the impact of HIV infection in this workplace and the potential negative impact on productivity. These results highlight the need for routine HIV testing and access to antiretro viral treatment for workers in countries such as Botswana. PMID- 18160002 TI - An efficient tool for surveying CRF01_AE HIV type 1 resistance in Thailand to combined stavudine-lamivudine-nevirapine treatment: mutagenically separated PCR targeting M184I/V. AB - Under programs organized by the government of Thailand, HIV-1-infected patients have been treated since 2002 with several regimens, including a tablet known as GPOvir, which contains lamivudine, stavudine, and nevirapine. The aim of this study was to establish an effective assay, based on mutagenically separated PCR (MS-PCR), with the goal of surveying GPOvir-resistant HIV-1 cases. To determine the target mutation point for the assay, we analyzed the patterns of acquired drug resistance in plasma samples from GPOvir-failed cases. Of 428 HIV-1-infected individuals treated with GPOvir at Lampang Hospital in northern Thailand from 2002 to 2004, 66 had detectable viral loads after 3 months of treatment. The HIV 1 sequences of these 66 GPOvir-failed cases and 55 pre-GPOvir baseline samples were analyzed. The most prevalent drug resistance mutation among the samples was the lamivudine resistance M184I/V mutation. Based on this finding, we developed a new MS-PCR assay to detect the M184I/V mutation, and evaluated the assay performance for detecting GPOvir-resistant CRF01_AE cases. Comparing the results of M184I/V MS-PCR and sequence analyses, we found a concordance rate of 95% and an overall sensitivity of the M184I/V MS-PCR for detecting GPOvir-resistant cases of 79%. Considering the relatively low price of the assay, approximately $12.50 per sample, M184I/V MS-PCR may be a candidate for monitoring a large number of GPOvir-treated patients, particularly in developing nations. PMID- 18160003 TI - Variation of CD4 count and percentage during pregnancy and after delivery: implications for HAART initiation in resource-limited settings. AB - We studied whether the use of T-lymphocyte CD4 (CD4) absolute count instead of CD4 percentage could affect the decision process regarding HAART initiation in African HIV-infected pregnant women. A prospective cohort in Abidjan, Cote d'Ivoire before HAART was available. Participating women received a perinatal antiretroviral prophylaxis (zidovudine + single-dose of nevirapine). CD4 count and percentage were measured by flow cytometry at baseline (32 weeks of amenorrhea) and at 1 month after delivery. A signed-rank test was used to compare the distributions of the CD4 absolute count and percentage values. A total of 325 HIV-1-infected pregnant women were included. At baseline, their median CD4 count was 355 cells/mm(3) and the median CD4 percentage was 24.8%; 17.8% of women had a CD4 count <200 cells/mm(3) and 14.8 % had a CD4 percentage <15%. One month after delivery, the median CD4 count was 489 cells/mm(3) (vs. baseline: p < 0.001), the median CD4 percentage was 25.6% (vs. baseline: p = 0.107), 9.5% of women had CD4 count <200 cells/mm(3) (vs. baseline: p < 0.001), and 15.1% of women had a CD4 percentage <15% (vs. baseline: p = 0.823). When combining the CD4 count and the WHO clinical stage, the proportion of women who met the WHO 2006 criteria for initiating HAART was 28.3% at baseline but 17.2% only at 1 month after delivery (p < 0.001). Between the prepregnancy and the postdelivery periods, the CD4 count experienced a significant increase, whereas the CD4 percentage remained unchanged. To accurately target the most appropriate time to start HAART, the CD4 percentage could be more reliable than the absolute count in sub-Saharan African pregnant women. PMID- 18160004 TI - Comparison of methods to detect recent HIV type 1 infection in cross-sectionally collected specimens from a cohort of female sex workers in the Dominican Republic. AB - Interest in estimating HIV-1 incidence using specimens obtained as part of cross sectional surveys has led to the development of new methods to detect recent HIV 1 infection through the testing of a single anti-HIV-positive specimen. These assays are based on quantitative and qualitative differences in anti-HIV-1 antibodies between recent and long-standing infections. An ongoing vaccine preparedness study enrolled female sex workers in the Dominican Republic. Specimens from women found to be HIV positive at baseline were tested for recent HIV-1 infection using the detuned assay, avidity index, and BED-CEIA assay. An unweighted kappa statistic in pairwise comparisons was used to estimate the correlation of recent HIV-1 infection detection by the three methods. Nineteen (3.9%) of 482 women were positive for HIV-1 infection. The incidence of HIV infection was 1.4% [95% confidence interval (CI): 0.2, 5.3], 0.9%(95% CI: 0.1, 4.4), and 1.0%(95% CI: 0.1, 4.4) using detuned assay, avidity index, and BED-CEIA techniques, respectively. The overall agreement between both detuned assay and avidity index and detuned assay and BED-CEIA was 94%(kappa = 0.8, 95% CI; 0.3, 1.0). The correlation was highest between BED-CEIA and avidity index methods (100%; kappa = 1.0). All three methods performed similarly in detecting recent HIV-1 infection in this region dominated by clade B HIV-1 infection. Although incidence estimates were slightly higher using the detuned assay method, they were not significantly different. These assays may be of value in both clinical research and practice. The utility of individual assays for recent infection detection will depend upon operating characteristics, HIV-1 subtype limitations, and selection of appropriate assay cutoff values. PMID- 18160005 TI - HIV type 1 genetic variability in central Brazil. AB - This study analyzed the genes pol and env to determine the genetic variability of HIV-1 in Central Brazil. Forty-one isolates of HIV-1-infected individuals had protease, reverse transcriptase, and C2C3/ env amplified by nested PCR and sequenced. The subtype was determined by the program REGA and phylogenetic analyses. The samples identified as putative recombinant forms were analyzed by SimPlot. A high prevalence of subtype B (95.1%) was observed, followed by mosaic viruses B/F (4.9%). The amino acid sequences from 30 HIV-1 isolates were analyzed for the antigenic intrasubtype diversity. The most prevalent gp120 V3 loop motif was the GPGR (United States/Europe) (43.3%), described in B and F subtypes, followed by the GPGK tetrapeptide (10%). The Brazilian variant B" (GWGR), GFGR, and GLGR tetrapeptides were found in 6.7%. Other V3 variants were found in eight isolates (26.7%). Phylogenetic tree analysis was also performed in order to verify the relationship of the HIV-1 samples from Central Brazil with other HIV-1 sequences that circulate in Brazil. The subtype B sequences from Central Brazil formed a polyphyletic cluster in the tree, indicating that these strains are similar to those from other geographic regions. These results contribute to the understanding of HIV in Brazil, and may prove useful for the development of vaccine candidates. PMID- 18160006 TI - Evaluation and implementation of rapid HIV tests: the experience in 11 African countries. AB - Rapid HIV testing enables HIV prevention and care services to expand to settings with limited laboratory infrastructure. The World Health Organization recommends that prior to their implementation, rapid HIV tests first be evaluated at a reference laboratory and then at points-of-service, and that measures to ensure quality of testing be in place. We describe the experience of 11 African countries that implemented rapid HIV testing using this process. A questionnaire regarding rapid test evaluations and quality assurance measures was administered to personnel in ministries of health, reference laboratories, and Centers for Disease Control and Prevention programs in 11 African countries. In reference laboratory evaluations, median sensitivity was above 99% for 10 of 15 rapid tests and median specificity was above 99% for 13 of 15 rapid tests. Similar results were observed in evaluations at point-of-service sites. Rapid testing algorithms have been implemented in over 600 sites in nine of the countries. Concordance between point-of-service rapid testing and reference laboratory retesting of samples ranged from 95.7% to 99.5% (median: 98.7%). A systematic approach to the evaluation and implementation of rapid HIV tests was useful in ensuring accurate, decentralized testing even when conducted by persons with limited laboratory experience. PMID- 18160007 TI - Brain magnetic resonance imaging white matter lesions are frequent in HTLV-I carriers and do not discriminate from HAM/TSP. AB - Human T lymphotropic virus (HTLV)-I is known to cause HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and other pronounced disease in less than 4% of those infected. However, evidence is accumulating that a proportion of HTLV-I carriers have neurological and urological symptoms without fulfilling criteria for HAM/TSP. Brain white matter (WM) lesions on magnetic resonance imaging (MRI) are frequently seen in HAM/TSP. HTLV-I carriers with MRI scans for other neurological diagnoses have WM lesions more frequently than expected. We studied 10 patients with HAM/TSP and 20 HTLV-I carriers without overt neurological disease and evaluated clinical characteristics, viral load, total, small, large, confluent WM lesion number, and lesion volume on MRI. Cerebral WM lesions were found in of 85% of HTLV-I carriers and 80% of HAM/TSP patients. Lesion number, size or location was no different between carriers and HAM/TSP. Cognitive function was lower in HAM/TSP (p = 0.045) but did not correlate with WM lesion number. Viral load and peripheral blood mononuclear cell interferon production correlated positively (p = 0.001) but did not correlate with lesion number or volume. Conventional brain MRI frequently shows WM lesions in HTLV-I-infected individuals suggesting potential early central nervous system inflammation with rare development of progressive disease. PMID- 18160008 TI - A lopinavir/ritonavir-based once-daily regimen results in better compliance and is non-inferior to a twice-daily regimen through 96 weeks. AB - We assessed the safety and efficacy and evaluated the adherence to lopinavir/ritonavir (LPV/r) dosed QD or BID in antiretroviral-naive, HIV-1 infected subjects through 96 weeks of treatment. A randomized, open-label, multicenter comparative study was conducted. A total of 190 antiretroviral-naive subjects with plasma HIV-1 RNA above 1000 copies/ml and any CD4(+) T cell count were enrolled. Subjects were randomized (3:2) to LPV/r 800/200 mg QD (n = 115) or 400/100 mg BID (n = 75). Subjects received TDF 300 mg and FTC 200 mg QD. Adherence to LPV/r through 96 weeks was measured using MEMS((R)) monitors. Median baseline VL and CD4(+) T cell count were 4.8 log(10) copies/ml and 216 cells/mm(3), respectively. Prior to week 96, 37% (QD) and 39% (BID) of subjects discontinued, primarily due either to adverse events (17% QD, 9% BID) or to loss to follow-up or nonadherence (12% QD, 17% BID). The proportion of subjects with VL <50 copies/ml [57% QD, 53% BID; p = 0.582 (ITT NC = F)], change in CD4 count (244 cells/mm(3) QD, 264 cells/mm(3) BID; p = 0.513), and evolution of resistance did not differ between groups through 96 weeks. Diarrhea (17% QD, 5% BID, p = 0.014) was the most common moderate or severe, study drug-related adverse event. Adherence to LPV/r was higher for the QD group than the BID group and declined over time in both groups. Time to loss of virologic response was significantly associated with adherence to LPV/r in both groups. LPV/r QD resulted in virologic response similar to LPV/r BID through 96 weeks in antiretroviral-naive subjects. Adherence was significantly higher in the QD group. PMID- 18160009 TI - Persistence of genotypic resistance to nelfinavir among women exposed to prophylactic antiretroviral therapy during pregnancy. AB - We assessed the development of drug resistance in women exposed to antiretroviral therapy (ART) for prevention of mother-to-child transmission (PMTCT) after 24 weeks postpartum in a prospective cohort of HIV-1-infected women. HIV-1-infected women, who received prophylactic ART during pregnancy, had genotypic resistance testing performed at the start (T1) of and 24 weeks after ART interruption (T2). The women had CD4 counts >250 cells/ml and no AIDS defining conditions. Of the 30 eligible women, the median age was 27 years [25-75% interquartile range (IQR): 21 32] and the median gestational age of ART initiation was 22 weeks (IQR: 19-27): 19 (63.3%) received zidovudine (ZDV) plus lamivudine (3TC) plus nelfinavir (NFV). At entry, most women (96.7%) were asymptomatic (CDC93 A1/A2), with a median CD4 count of 446 (IQR: 353-686) and median viral load (VL) of 8560 copies/ml (IQR: 3,252-19,515). No HIV-1 vertical transmission was observed. HIV subtype B was the most prevalent (70%). The development of new mutations associated with ART resistance was analyzed at T2. NFV resistance was observed in 4 out of 17 (23.5%) patients exposed to this drug: two major mutations D30N (1/17) and L90M (1/17) and minor mutations (N88S, 2/17). Mutations on positions 44, 69, and 118 (1/28) were present on reverse transcriptase (RT) analysis. No new nonnucleoside reverse transcriptase inhibitor (NNRTI)-associated mutation was observed. In this cohort, ART regimens were very efficient at blocking HIV vertical transmission. However, the high rate of NFV-resistant mutations observed in the postpartum period indicates the need for discussion of ART choices during pregnancy and the potential impact on future therapeutic options for these women. Women previously exposed to ART for PMTCT who will start HIV treatment should have genotypic resistance testing performed. PMID- 18160010 TI - HIV proteins regulate bone marker secretion and transcription factor activity in cultured human osteoblasts with consequent potential implications for osteoblast function and development. AB - A high incidence of decreased bone mineral density (BMD) has increasingly been associated with HIV infection. In this study mesenchymal stem cell (MSC) and human osteoblast (hOB) cell lines were treated with HIV tat, HIV rev, HIV p55 gag, HIV gp120 and HTLV env (100 ng/ml, 24 h). Cells were then analyzed for calcium deposition, alkaline phosphatase (ALP) activity, and lipid levels using established methods. Real-time PCR with gene-specific primers was used to quantify the mRNA levels of the transcription factors RUNX-2 and PPARgamma, transcription factors known to be pro-osteogenic and pro-adipogenic, respectively. The levels of secreted bone markers and transcription factor activity were determined using commercial assays. In OBs, HIV p55-gag and gp120 were seen to reduce calcium deposition, ALP activity, levels of secreted BMP-2, 7, and RANK-L, and the expression and activity of RUNX-2. The levels of osteocalcin were also significantly reduced by p55-gag treatment, while gp120 also increased PPARgamma activity. Lipid levels were also increased by gp120 treatment. The ability of MSCs to develop into functioning OBs was also affected by the presence of HIV proteins, with p55-gag inducing a decrease in osteogenesis, while rev induced an increase. HIV proteins can potentially modulate OB development and function in vitro via modulation of bone maker secretion and RUNX-2 and PPARgamma transcription factor activity. PMID- 18160011 TI - Biological and genetic evolution of HIV type 1 in two siblings with different patterns of disease progression. AB - To investigate the immunological and virological factors that may lead to different patterns of disease progression characteristic of HIV-1-infected children, two HIV-1-infected siblings, a slow and a fast progressor, were followed prospectively before the onset of highly active antiretroviral therapy. Viral coreceptor usage, including the use of CCR5/CXCR4 chimeric receptors, macrophage tropism, and sensitivity to the CC-chemokine RANTES, has been studied. An autologous and heterologous neutralizing antibody response has been documented using peripheral blood mononuclear cells- and GHOST(3) cell line-based assays. Viral evolution was investigated by env C2-V3 region sequence analysis. Although both siblings were infected with HIV-1 of the R5 phenotype, their viruses showed important biological differences. In the fast progressor there was a higher RANTES sensitivity of the early virus, an increased trend to change the mode of CCR5 receptor use, and a larger genetic evolution. Both children developed an autologous neutralizing antibody response starting from the second year with evidence of the continuous emergence of resistant variants. A marked viral genetic and phenotypic evolution was documented in the fast progressor sibling, which is accompanied by a high viral RANTES sensitivity and persistent neutralizing antibodies. PMID- 18160012 TI - Characterization of gp120 hydrolysis by IgA antibodies from humans without HIV infection. AB - Antibody hydrolysis of the superantigenic gp120 site and HIV-1 neutralization was studied as a potential anti-HIV mechanism in uninfected humans. gp120 hydrolysis by purified serum and salivary antibodies was determined by electrophoresis and peptide sequencing, the proteolytic mechanism was analyzed using electrophilic peptide analogs, and viral neutralization was studied using peripheral blood mononuclear cells as hosts. Polyclonal and monoclonal IgA but not IgG preparations selectively catalyzed the cleavage of HIV gp120 at rates sufficient to predict biologically relevant protection against the virus. The IgA hydrolytic reaction proceeded by noncovalent recognition of gp120 residues 421-433, a component of the superantigenic site of gp120, coordinated with peptide bond cleavage via a serine protease-like mechanism. The Lys-432-Ala-433 bond was one of the cleavage sites. Infection of peripheral blood mononuclear cells by a primary isolate of HIV was neutralized by the IgA but not IgG fractions. The neutralizing activity was specifically inhibited by an electrophilic inhibitor of the catalytic activity. The existence of catalytic IgAs to gp120 in uninfected humans suggests their role in resistance to HIV. PMID- 18160013 TI - Immunogenicity in macaques of the clinical product for a clade B DNA/MVA HIV vaccine: elicitation of IFN-gamma, IL-2, and TNF-alpha coproducing CD4 and CD8 T cells. AB - The clinical product for a clade B HIV DNA/MVA vaccine expressing Gag, Pol, and Env has been tested for immunogenicity in macaques. Responding T cells were at the threshold for detection following DNA priming at weeks 0 and 8 but underwent sharp expansions and contractions following MVA boosting at weeks 16 and 24. Both CD4 and CD8 T cell responses had high frequencies of cytokine coproducing cells with >50% of the memory cells coproducing multiple cytokines including IL-2. The highest responses were elicited to Gag, followed by Env and then Pol. In two of six macaques, the vaccine also elicited low levels of neutralizing Ab for easy to neutralize clade B isolates. PMID- 18160014 TI - Reverse transcriptase mutations in Cambodian CRF01_AE isolates after antiretroviral prophylaxis against HIV Type 1 perinatal transmission. AB - This study explores amino acid changes of the reverse transcriptase (rt) of CRF01_AE isolates from pregnant women naive to antiretroviral drugs before and 2, 6, and 52 weeks after exposure to single dose nevirapine (sdNVP). Results based on 51 observations showed that the proportion of isolates with nonnucleoside reverse transcriptase inhibitor (NNRTI) RMs in the group treated with sdNVP (n = 35) increased from 0% pre-NVP to 22.9% at week 2 postpartum (pp) and 22.9% at week 6 pp. In the group treated with zidovudine + sdNVP (n = 16), the proportion with RM was 31.3% and 18.8% at weeks 2 and 6 pp, respectively. Only a few RMs were still detected at week 52 pp. No apparent subtype-specific treatment-related mutations were detected. NNRTI RM occurrence in CRF01_AE strains is similar to subtype A, D, and CRF02_AG strains after exposure to antiretroviral drugs for PMTCT. PMID- 18160015 TI - HIV type 1 subtype A epidemic in injecting drug user (IDU) communities in Iran. AB - HIV-1 C2-V3 subgenomic regions of the env gene from Iranian seropositive injecting drug users (IDUs) living in Mashhad have been analyzed to evaluate molecular and phylogenetic relationships with IDUs living in Tehran and identify possible common founder virus isolates. The results show that the viral sequences of the Iranian IDUs are strongly related and form a single cluster within the A subtype related to African Ugandan/Kenyan sub-Saharan isolates. Pairwise nucleotide alignment shows higher average divergence values within the Mashhad group than the Tehran group. Furthermore, the Mashhad sequences show much less conserved amino acid residues in the V3 loop than the Tehran sequences. These data represent the first macro-analysis of the HIV-1 molecular evolution in the Iran and Middle East epidemics and may be extremely relevant to guide the development and implementation of diagnostic as well as preventive/therapeutic approaches in this region. PMID- 18160016 TI - Complete genome sequencing of a non-syncytium-inducing HIV type 1 subtype D strain from Cape Town, South Africa. AB - The fast growing HIV-1 epidemic in South Africa is mainly caused by HIV-1 group M subtype C, spreading via heterosexual transmission. In South Africa HIV-1 subtype B and D viruses were responsible for the initial epidemic during the 1980s, primarily in the homosexual population. This study describes the full-length PCR amplification and sequencing of an HIV-1 subtype D strain recovered from plasma from a sample taken during 1990. This is only the second full-length non syncytium-inducing (NSI) subtype D strain described. Although restricted, the subtype D strain is still being detected in the South African population. PMID- 18160017 TI - HIV type 1 subtype C and CB Pol recombinants prevail at the cities with the highest AIDS prevalence rate in Brazil. AB - HIV-1 B is predominant in Brazil, but HIV-1 C has increasingly been reported in the south of the country. However, many samples clustering with clade C are actually a recombinant, with a small B segment at RT (CRF31). Samples (209) from the three cities with the highest aids prevalence rate are analyzed. Partial polymerase sequences from HIV RNA made it possible to determine HIV clades and recombination patterns and to identify primary drug resistance mutations (DRMs). The incidence was estimated with a BED assay. HIV-1 C and CRF31 patterns were twice as frequent as clade B at all sites, but the proportion of C and CRF31 patterns was significantly different among sites. The incidence estimate for SC was 2.6 persons-years. Infection in recent or younger cases showed no association with clade C. Surveillance DRM was observed in 8.3% (95% CI 5-13), mostly to NNRTIs. Clade F pol genomes had significantly more primary DRM. PMID- 18160018 TI - Characterization of HIV type 1 reverse transcriptase mutations in infants infected by mothers who received peripartum nevirapine prophylaxis in Jos, Nigeria. AB - This study was carried out to characterize HIV-1 reverse transcriptase (RT) mutations in vertically infected infants in Jos, Nigeria. DNA was extracted from peripheral blood mononuclear cells of 102 infants, aged 0 to 6 months, born to HIV-1-infected mothers who had received peripartum single-dose nevirapine prophylaxis. PCR-based diagnosis revealed that 14 infants (13.7%) were infected with HIV-1. Phylogenetic analyses of RT revealed wide viral diversity, with CRF02_AG, subtype G, subsubtype A3, CRF06_cpx, and a subtype D recombinant present in the population. Four of 13 (31%) infants had NNRTI resistance mutations--V179I (2 infants), Y181C, and V179E. Intriguingly, subtype G sequences did not have NNRTI mutations but rather carried a Q207N mutation, which may undergo negative selection under drug pressure. Our data suggest wide diversity for vertically transmitted HIV-1 viruses in Nigeria and highlight the potential significance of transmitting rare mutations in subtype G. PMID- 18160019 TI - Analysis of HIV type 1 gp41 sequences in diverse HIV type 1 strains. AB - The HIV fusion inhibitor enfuvirtide (ENF/Fuzeon) targets the env gp41 transmembrane domain. Mutations in gp41 are associated with ENF resistance. We developed a prototype assay to genotype a 676-bp region spanning the heptad repeat domains (HR1 and HR2) of HIV-1 gp41. Plasma samples were collected from 126 HIV-1-infected blood donors in Cameroon, Brazil, Uganda, South Africa, Thailand, and Argentina. Based on analysis of gag p24, pol integrase, and env gp41 genes, the panel was composed of subtypes A/A2 (18), B (11), C (14), D (10), F/F2 (9), G (7), CRF01_AE (9), CRF02_AG (33), and recombinant strains (15). Genotyping was successful for 119 of the 126 samples (94.4%). Although numerous amino acid polymorphisms were detected in some samples, none had primary mutations associated with ENF resistance. The gp41 HIV-1 research reagents developed by Celera are useful tools for genotyping analysis of the gp41 region in diverse HIV-1 strains. PMID- 18160020 TI - New insights into the origin of the HIV type 1 subtype A epidemic in former Soviet Union's countries derived from sequence analyses of preepidemically transmitted viruses. AB - The HIV-1 subtype A epidemic affecting injecting drug users (IDU) in former Soviet Union (FSU) countries started dramatically in Odessa, southern Ukraine, in 1995, and is caused by a variant of monophyletic origin, often designated IDU-A. We phylogenetically analyzed one near full-length genome and two partial sequences of three HIV-1 subtype A viruses collected in St. Petersburg, Russia, heterosexually transmitted in 1992-1994. The sequences branched basally to the IDU-A clade, together with eight viruses from Odessa collected in 1993, all presumably acquired heterosexually, and two viruses from the Democratic Republic of Congo. Of all other FSU sequences in databases, only those from three recently collected viruses, one from Ukraine and two from northwestern Russia, at least one of them acquired heterosexually, branched basally to the IDU-A cluster. The results indicate that the FSU IDU-A variant derives from a strain that initially propagated heterosexually in Ukraine and originated in central Africa. PMID- 18160021 TI - Host genetic analysis of HIV type 1 subtype CRF01_AE (E)-infected Thai patients with different rates of disease progression. PMID- 18160022 TI - Long-term treatment with a Yang-invigorating Chinese herbal formula produces generalized tissue protection against oxidative damage in rats. AB - Previous work in our laboratory has shown that long-term treatment with Vigconic 28 (VI-28), a Yang-invigorating Chinese herbal formula used for the promotion of overall wellness in Chinese medicine, can enhance the mitochondrial functional ability and antioxidant capacity in various tissues of both male and female rats. To investigate whether the VI-28 treatment regimen could afford tissue protection against oxidative injury, the effects of long-term VI-28 treatment (80 or 240 mg/kg/d x 30) on oxidative stress-induced tissue damage in various organs (brain, heart, liver, and kidney) were examined in female rats. The results indicated that long-term VI-28 treatment invariably protected against oxidative tissue damage in the rat models of cerebral/myocardial ischemia-reperfusion injury, CCl4 hepatotoxicity, and gentamicin nephrotoxicity. The tissue protection was associated with increases in the levels and activities of mitochondrial antioxidant components as well as with the preservation of mitochondrial structural integrity. This was evidenced by decreases in the sensitivity of mitochondria to Ca2+-induced permeability transition, and in the levels of mitochondrial malondialdehyde production, Ca2+ loading, and cytochrome c release in the tissues examined. Interestingly, the VI-28 treatment increased red cell CuZn-superoxide dismutase (CuZn-SOD) levels, and these levels correlated positively with the degree of tissue protection afforded by long-term VI-28 treatment in rats. The generalized tissue protection afforded by long-term VI-28 treatment may have clinical implications in the prevention of age-related diseases, and VI-28 treatment may possibly delay the aging process. PMID- 18160023 TI - Age-specific modulation of genes involved in lipid and cholesterol homeostasis by dietary zinc. AB - The association between zinc status and lipid-related metabolic diseases, such as coronary heart disease, has produced conflicting results. Since heterogeneity of the adult population-or inter-individual variation-may impact on response to dietary zinc, nutritional genomic approaches were used to investigate the impact of age on zinc-regulated gene expression in lymphocytes. Many genes involved in lipid and cholesterol homeostasis were found to be differentially regulated by zinc in an age-dependent manner. Additionally, the individual genes affected and the extent of differential regulation varied with respect to donor age, suggesting that the age-related phenotype may alter zinc action. PMID- 18160024 TI - Can lipofuscin accumulation be prevented? AB - During normal autophagic degradation of mitochondria and iron-containing proteins in lysosomes, iron is released intralysosomally where it may react with hydrogen peroxide forming hydroxyl radicals (Fenton reaction). Depending on their rate of formation, these highly reactive radicals can cross-link intralysosomal material, leading to lipofuscin formation, or destabilize the lysosomal membrane, which induces apoptosis/necrosis. Since the sensitivity of lysosomes to oxidative stress can be manipulated by altering the intralysosomal level of redox-active iron, it follows that lipofuscin formation might also be influenced. It is suggested that pulse doses of iron chelators that easily penetrate membranes could be used to diminish lipofuscinogenesis. PMID- 18160025 TI - Fatty acid profile of erythrocyte membranes as possible biomarker of longevity. AB - Offspring of long-lived individuals are a useful model to discover biomarkers of longevity. The lipid composition of erythrocyte membranes from 41 nonagenarian offspring was compared with 30 matched controls. Genetic loci were also tested in 280 centenarians and 280 controls to verify a potential genetic predisposition in determining unique lipid profile. Gas chromatography was employed to determine fatty acid composition, and genotyping was performed using Taqman assays. Outcomes were measured for erythrocyte membrane percentage content of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids (omega-6 and omega-3), geometrical isomers of arachidonic and oleic acids, and total trans fatty acids. Also, allele and genotyping frequencies at endothelial-nitric oxide synthase and delta-5/delta-6 and delta-9 desaturase loci were considered. Erythrocyte membranes from nonagenarian offspring had significantly higher content of C16:1 n-7, trans C18:1 n-9, and total trans-fatty acids, and reduced content of C18:2 n-6 and C20:4 n-6. No association was detected at endothelial nitric oxide synthase and delta-5/delta-6 and delta-9 desaturase loci that could justify genetic predisposition for the increased trans C18:1 n-9, monounsaturated fatty acids and decreased omega-6 synthesis. We concluded that erythrocyte membranes derived from nonagenarian offspring have a different lipid composition (reduced lipid peroxidation and increased membrane integrity) to that of the general population. PMID- 18160026 TI - Extracts of Valeriana officinalis L. s.l. show anxiolytic and antidepressant effects but neither sedative nor myorelaxant properties. AB - Extracts of Valeriana officinalis L. s.l. are used for treating mild sleep disorders and nervous tension. Despite intensive research efforts, the pharmacological actions accounting for the clinical efficacy of valerian remain unclear. Thus, it was the aim of this study to evaluate CNS-related effects of different valerian extracts using behavioral paradigms (mice and rats). Following oral administration two commercially available preparations (extraction solvents: 45% methanol m/m and 70% ethanol v/v), a 35% ethanolic v/v extract and a refined extract derived from it (patented special extract phytofin Valerian 368) were tested for sedative (locomotor activity, ether-induced anaesthesia) and anxiolytic (elevated plus maze) activity. Using the forced swimming and the horizontal wire test the latter two extracts were additionally tested for antidepressant and myorelaxant properties. Up to maximum dosages of 500 or 1000 mg/kg bw none of the valerian extracts displayed sedative effects. Neither spontaneous activity was reduced nor the duration of ether-induced narcosis was prolonged. In contrast, results obtained in the elevated plus maze test revealed a pronounced anxiolytic effect of the 45% methanolic and 35% ethanolic extract as well as of phyotofin Valerian 368 in a dose range of 100-500 mg/kg bw. Additionally and different from its primary extract (35% ethanolic extract) phytofin Valerian 368 showed antidepressant activity in the forced swimming test after subacute treatment. Myorelaxant effects were not observed in dosages up to 1000 mg/kg bw. Due to these findings it is proposed that not sedative but anxiolytic and antidepressant activity, which was elaborated particularly in the special extract phytofin Valerian 368, considerably contribute to the sleep enhancing properties of valerian. PMID- 18160027 TI - LC-MS/MS methods for the detection of isoprostanes (iPF2alpha-III and 8,12-iso iPF2alpha-VI) as biomarkers of CCl4-induced oxidative damage to hepatic tissue. AB - Isoprostanes are formed after peroxidation of arachidonic acid and are promising biomarkers for reactive oxygen species. A LC-MS/MS based method was developed for the quantitation of two isoprostanes (iPF2alpha-III and 8,12-iso-iPF2alpha-VI) in hepatocytes, tissue and urine samples of rats. A column switching method was used to reduce sample preparation to a minimum. Precision was 9.4% and accuracy was between 96 and 114% for free iPF2alpha-III in tissue at concentrations from 1.9 to 6.1 ng/g. Treatment of rats with CCl4 to induce oxidative stress resulted in a dose-dependent increase (two- to three-fold) of iPF2alpha-III and 8,12-iso iPF2alpha-VI in liver and kidney. For both isoprostanes an increase of four- to five-fold was observed in CCl4 treated hepatocytes and six- to eight-fold in CCl4 treated and glutathione depleted hepatocytes. In conclusion, the presented method is sensitive, specific and precise to be applied for the quantitation of iPF2alpha-III and 8,12-iso-iPF2alpha-VI which are shown to increase by CCl4 treatment in vitro and in vivo. PMID- 18160028 TI - Latin American science moves into the spotlight. AB - With new incentives, some governments in Latin America are starting to increase investment in basic and applied research. Although the news is encouraging, scientists in this region still face many challenges ahead. PMID- 18160029 TI - Somatic mutation and autoimmunity. PMID- 18160030 TI - Fruit fly family fun. AB - A recent comparative analysis of the sequenced genomes of 12 Drosophila species (Drosophila 12 Genomes Consortium, 2007; Stark et al., 2007) reveals a comprehensive picture of the evolution of small animal genomes and greatly improves computational predictions of functional elements in the D. melanogaster reference sequence. PMID- 18160031 TI - RNA polymerase I: a multifunctional molecular machine. AB - In this issue, Kuhn et al. (2007) report the complete structure of the 14-subunit yeast RNA polymerase (Pol) I enzyme at 12 A resolution using cryo-electron microscopy (cryo-EM). Their study reveals that three subunits of Pol I perform functions in transcription elongation that are outsourced to the transcription factors TFIIF and TFIIS in the analogous Pol II transcription system. PMID- 18160032 TI - A dead end for microRNAs. AB - Dead end (Dnd1) is an RNA-binding protein that mediates germ-cell viability and suppresses the formation of germ-cell tumors. Kedde et al. (2007) now provide evidence that Dnd1 mediates these effects by counteracting microRNA-mediated silencing of mRNAs. PMID- 18160033 TI - Template switching: from replication fork repair to genome rearrangements. AB - Genome rearrangements are a hallmark of human genomic disorders and occur largely through recombination mechanisms. In this issue, Lee et al. (2007) show that the complex nonrecurrent rearrangements observed in the dysmyelinating disorder Pelizaeus-Merzbacher disease (PMD) are likely to be caused by a replication mechanism involving template switching. PMID- 18160034 TI - Modulation of longevity by environmental sensing. AB - Organisms from bacteria to humans have the capacity to gauge the quality of their respective environments. Recent advances in understanding how various types of environmental conditions are sensed and interpreted by cells and by organisms have established a critical role for these systems in the modulation of physiology, health, and aging. PMID- 18160035 TI - A DNA replication mechanism for generating nonrecurrent rearrangements associated with genomic disorders. AB - The prevailing mechanism for recurrent and some nonrecurrent rearrangements causing genomic disorders is nonallelic homologous recombination (NAHR) between region-specific low-copy repeats (LCRs). For other nonrecurrent rearrangements, nonhomologous end joining (NHEJ) is implicated. Pelizaeus-Merzbacher disease (PMD) is an X-linked dysmyelinating disorder caused most frequently (60%-70%) by nonrecurrent duplication of the dosage-sensitive proteolipid protein 1 (PLP1) gene but also by nonrecurrent deletion or point mutations. Many PLP1 duplication junctions are refractory to breakpoint sequence analysis, an observation inconsistent with a simple recombination mechanism. Our current analysis of junction sequences in PMD patients confirms the occurrence of simple tandem PLP1 duplications but also uncovers evidence for sequence complexity at some junctions. These data are consistent with a replication-based mechanism that we term FoSTeS, for replication Fork Stalling and Template Switching. We propose that complex duplication and deletion rearrangements associated with PMD, and potentially other nonrecurrent rearrangements, may be explained by this replication-based mechanism. PMID- 18160036 TI - Tel2 regulates the stability of PI3K-related protein kinases. AB - We report an unexpected role for Tel2 in the expression of all mammalian phosphatidylinositol 3-kinase-related protein kinases (PIKKs). Although Tel2 was identified as a budding yeast gene required for the telomere length maintenance, we found no obvious telomeric function for mammalian Tel2. Targeted gene deletion showed that mouse Tel2 is essential in embryonic development, embryonic stem (ES) cells, and embryonic fibroblasts. Conditional deletion of Tel2 from embryonic fibroblasts compromised their response to IR and UV, diminishing the activation of checkpoint kinases and their downstream effectors. The effects of Tel2 deletion correlated with significantly reduced protein levels for the PI3K related kinases ataxia telangiectasia mutated (ATM), ATM and Rad3 related (ATR), DNA-dependent protein kinase catalytic subunit ataxia (DNA-PKcs). Tel2 deletion also elicited specific depletion of the mammalian target of rapamycin (mTOR), suppressor with morphological effect on genitalia 1 (SMG1), and transformation/transcription domain-associated protein (TRRAP), and curbed mTOR signaling, indicating that Tel2 affects all six mammalian PIKKs. While Tel2 deletion did not alter PIKK mRNA levels, in vivo pulse labeling experiments showed that Tel2 controls the stability of ATM and mTOR. Each of the PIKK family members associated with Tel2 in vivo and in vitro experiments indicated that Tel2 binds to part of the HEAT repeat segments of ATM and mTOR. These data identify Tel2 as a highly conserved regulator of PIKK stability. PMID- 18160037 TI - Functional architecture of RNA polymerase I. AB - Synthesis of ribosomal RNA (rRNA) by RNA polymerase (Pol) I is the first step in ribosome biogenesis and a regulatory switch in eukaryotic cell growth. Here we report the 12 A cryo-electron microscopic structure for the complete 14-subunit yeast Pol I, a homology model for the core enzyme, and the crystal structure of the subcomplex A14/43. In the resulting hybrid structure of Pol I, A14/43, the clamp, and the dock domain contribute to a unique surface interacting with promoter-specific initiation factors. The Pol I-specific subunits A49 and A34.5 form a heterodimer near the enzyme funnel that acts as a built-in elongation factor and is related to the Pol II-associated factor TFIIF. In contrast to Pol II, Pol I has a strong intrinsic 3'-RNA cleavage activity, which requires the C terminal domain of subunit A12.2 and, apparently, enables ribosomal RNA proofreading and 3'-end trimming. PMID- 18160038 TI - CENP-B controls centromere formation depending on the chromatin context. AB - The centromere is a chromatin region that serves as the spindle attachment point and directs accurate inheritance of eukaryotic chromosomes during cell divisions. However, the mechanism by which the centromere assembles and stabilizes at a specific genomic region is not clear. The de novo formation of a human/mammalian artificial chromosome (HAC/MAC) with a functional centromere assembly requires the presence of alpha-satellite DNA containing binding motifs for the centromeric CENP-B protein. We demonstrate here that de novo centromere assembly on HAC/MAC is dependent on CENP-B. In contrast, centromere formation is suppressed in cells expressing CENP-B when alpha-satellite DNA was integrated into a chromosomal site. Remarkably, on those integration sites CENP-B enhances histone H3-K9 trimethylation and DNA methylation, thereby stimulating heterochromatin formation. Thus, we propose that CENP-B plays a dual role in centromere formation, ensuring de novo formation on DNA lacking a functional centromere but preventing the formation of excess centromeres on chromosomes. PMID- 18160039 TI - The ATPase SpoIIIE transports DNA across fused septal membranes during sporulation in Bacillus subtilis. AB - The FtsK/SpoIIIE family of ATP-dependent DNA transporters mediates proper chromosome segregation in dividing bacteria. In sporulating Bacillus subtilis cells, SpoIIIE translocates much of the circular chromosome from the mother cell into the forespore, but the molecular mechanism remains unclear. Using a new assay to monitor DNA transport, we demonstrate that the two arms of the chromosome are simultaneously pumped into the forespore. Up to 70 molecules of SpoIIIE are recruited to the site of DNA translocation and assemble into complexes that could contain 12 subunits. The fusion of the septal membranes during cytokinesis precedes DNA translocation and does not require SpoIIIE, as suggested by analysis of lipid dynamics, serial thin-section electron microscopy, and cell separation by protoplasting. These data support a model for DNA transport in which the transmembrane segments of FtsK/SpoIIIE form linked DNA conducting channels across the two lipid bilayers of the septum. PMID- 18160040 TI - Architecture of a coat for the nuclear pore membrane. AB - The symmetric core of the nuclear pore complex can be considered schematically as a series of concentric cylinders. A peripheral cylinder coating the pore membrane contains the previously characterized, elongated heptamer that harbors Sec13 Nup145C in its middle section. Strikingly, Sec13-Nup145C crystallizes as a hetero octamer in two space groups. Oligomerization of Sec13-Nup145C was confirmed biochemically. Importantly, the numerous interacting surfaces in the hetero octamer are evolutionarily highly conserved, further underlining the physiological relevance of the oligomerization. The hetero-octamer forms a slightly curved, yet rigid rod of sufficient length to span the entire height of the proposed membrane-adjacent cylinder. In concordance with the dimensions and symmetry of the nuclear pore complex core, we suggest that the cylinder is constructed of four antiparallel rings, each ring being composed of eight heptamers arranged in a head-to-tail fashion. Our model proposes that the hetero octamer would vertically traverse and connect the four stacked rings. PMID- 18160041 TI - STIM2 is a feedback regulator that stabilizes basal cytosolic and endoplasmic reticulum Ca2+ levels. AB - Deviations in basal Ca2+ levels interfere with receptor-mediated Ca2+ signaling as well as endoplasmic reticulum (ER) and mitochondrial function. While defective basal Ca2+ regulation has been linked to various diseases, the regulatory mechanism that controls basal Ca2+ is poorly understood. Here we performed an siRNA screen of the human signaling proteome to identify regulators of basal Ca2+ concentration and found STIM2 as the strongest positive regulator. In contrast to STIM1, a recently discovered signal transducer that triggers Ca2+ influx in response to receptor-mediated depletion of ER Ca2+ stores, STIM2 activated Ca2+ influx upon smaller decreases in ER Ca2+. STIM2, like STIM1, caused Ca2+ influx via activation of the plasma membrane Ca2+ channel Orai1. Our study places STIM2 at the center of a feedback module that keeps basal cytosolic and ER Ca2+ concentrations within tight limits. PMID- 18160042 TI - Genome-wide high-resolution mapping of exosome substrates reveals hidden features in the Arabidopsis transcriptome. AB - The exosome complex plays a central and essential role in RNA metabolism. However, comprehensive studies of exosome substrates and functional analyses of its subunits are lacking. Here, we demonstrate that as opposed to yeast and metazoans the plant exosome core possesses an unanticipated functional plasticity and present a genome-wide atlas of Arabidopsis exosome targets. Additionally, our study provides evidence for widespread polyadenylation- and exosome-mediated RNA quality control in plants, reveals unexpected aspects of stable structural RNA metabolism, and uncovers numerous novel exosome substrates. These include a select subset of mRNAs, miRNA processing intermediates, and hundreds of noncoding RNAs, the vast majority of which have not been previously described and belong to a layer of the transcriptome that can only be visualized upon inhibition of exosome activity. These first genome-wide maps of exosome substrates will aid in illuminating new fundamental components and regulatory mechanisms of eukaryotic transcriptomes. PMID- 18160043 TI - A predictive model for transcriptional control of physiology in a free living cell. AB - The environment significantly influences the dynamic expression and assembly of all components encoded in the genome of an organism into functional biological networks. We have constructed a model for this process in Halobacterium salinarum NRC-1 through the data-driven discovery of regulatory and functional interrelationships among approximately 80% of its genes and key abiotic factors in its hypersaline environment. Using relative changes in 72 transcription factors and 9 environmental factors (EFs) this model accurately predicts dynamic transcriptional responses of all these genes in 147 newly collected experiments representing completely novel genetic backgrounds and environments-suggesting a remarkable degree of network completeness. Using this model we have constructed and tested hypotheses critical to this organism's interaction with its changing hypersaline environment. This study supports the claim that the high degree of connectivity within biological and EF networks will enable the construction of similar models for any organism from relatively modest numbers of experiments. PMID- 18160045 TI - SnapShot: Noncanonical Wnt Signaling Pathways. PMID- 18160044 TI - Atypical AAA+ subunit packing creates an expanded cavity for disaggregation by the protein-remodeling factor Hsp104. AB - Hsp104, a yeast protein-remodeling factor of the AAA+ (ATPases associated with various cellular activities) superfamily, and its homologs in bacteria and plants mediate cell recovery after severe stress by disaggregating denatured proteins through a poorly understood mechanism. Here, we present cryo-electron microscopy maps and domain fitting of Hsp104 hexamers, revealing an unusual arrangement of AAA+ modules with the prominent coiled-coil domain intercalated between the AAA+ domains. This packing results in a greatly expanded cavity, which is capped at either end by N- and C-terminal domains. The fitted structures as well as mutation of conserved coiled-coil arginines suggest that the coiled-coil domain plays a major role in the extraction of proteins from aggregates, providing conserved residues for key functions in ATP hydrolysis and potentially for substrate interaction. The large cavity could enable the uptake of polypeptide loops without a requirement for exposed N or C termini. PMID- 18160046 TI - Rare variant of hypoxia-inducible factor-1alpha (HIF-1A) and breast cancer risk in Korean women. AB - BACKGROUND: Hypoxia inducible factor 1 alpha (HIF-1A) is activated by low oxygen condition tension, a key regulator of the gene involved in the cellular response to hypoxia. Tumors exhibiting extensive hypoxia are more aggressive than tumors oxygenized better. METHODS: To evaluate the potential role of the polymorphisms of HIF-1A in the etiology of breast cancer, histologically confirmed incident breast cancer cases (n=1599) and control subjects (n=1536) were recruited. RESULTS: Two selected SNPs (Ex15+197C>T and P582S) were not associated with overall breast cancer risk (TT vs. CC: OR=0.9, 95% CI=0.6-1.5, Ser/Ser vs. Pro/Pro: OR=5.5, 95% CI=0.7-45.4, respectively). However, when stratified analyses were performed, significant associations were observed between Ser/Ser genotype at codon 582 and breast cancer risk among women with larger tumor size (>2 cm) (OR=10.1, 95% CI=1.1-91.1) or without lymph node involvement (OR=9.3, 95% CI=1.1-79.4), although confidence intervals were wide. CONCLUSIONS: Our findings support the hypothesis that the HIF-1Alpha P582S variant may confer susceptibility to subgroups of breast cancer in Korean women. However, further study is warranted due to low statistical power caused by very low minor allele frequencies. PMID- 18160047 TI - Differences in circulating insulin levels following glargine administration. AB - OBJECTIVES: To evaluate two-site non competitive insulin immunoassays [BI-INSULIN assay (IRMA) and an electro-chemiluminescent immunoassay (ECLIA)] in routine practice. DESIGN AND METHODS: We studied 145 consecutive patients attending our Endocrine Department for general endocrine explorations and metabolic status investigations. RESULTS: The ECLIA yielded higher results than IRMA in all but six patients treated by glargine. CONCLUSIONS: Insulin levels measured by Elecsys in patients treated by glargine are directly related to glargine biotransformation into detectable metabolite (M1). PMID- 18160048 TI - Lyl1 interacts with CREB1 and alters expression of CREB1 target genes. AB - The basic helix-loop-helix (bHLH) transcription factor family contains key regulators of cellular proliferation and differentiation as well as the suspected oncoproteins Tal1 and Lyl1. Tal1 and Lyl1 are aberrantly over-expressed in leukemia as a result of chromosomal translocations, or other genetic or epigenetic events. Protein-protein and protein-DNA interactions described so far are mediated by their highly homologous bHLH domains, while little is known about the function of other protein domains. Hetero-dimers of Tal1 and Lyl1 with E2A or HEB, decrease the rate of E2A or HEB homo-dimer formation and are poor activators of transcription. In vitro, these hetero-dimers also recognize different binding sites from homo-dimer complexes, which may also lead to inappropriate activation or repression of promoters in vivo. Both mechanisms are thought to contribute to the oncogenic potential of Tal1 and Lyl1. Despite their bHLH structural similarity, accumulating evidence suggests that Tal1 and Lyl1 target different genes. This raises the possibility that domains flanking the bHLH region, which are distinct in the two proteins, may participate in target recognition. Here we report that CREB1, a widely-expressed transcription factor and a suspected oncogene in acute myelogenous leukemia (AML) was identified as a binding partner for Lyl1 but not for Tal1. The interaction between Lyl1 and CREB1 involves the N terminal domain of Lyl1 and the Q2 and KID domains of CREB1. The histone acetyl transferases p300 and CBP are recruited to these complexes in the absence of CREB1 Ser 133 phosphorylation. In the Id1 promoter, Lyl1 complexes direct transcriptional activation. We also found that in addition to Id1, over-expressed Lyl1 can activate other CREB1 target promoters such as Id3, cyclin D3, Brca1, Btg2 and Egr1. Moreover, approximately 50% of all gene promoters identified by ChIP-chip experiments were jointly occupied by CREB1 and Lyl1, further strengthening the association of Lyl1 with Cre binding sites. Given the newly recognized importance of CREB1 in AML, the ability of Lyl1 to modulate promoter responses to CREB1 suggests that it plays a role in the malignant phenotype by occupying different promoters than Tal1. PMID- 18160049 TI - Tocopherols and tocotrienols in membranes: a critical review. AB - The familiar role of tocols (tocopherols and tocotrienols) as lipid-soluble chain terminating inhibitors of lipid peroxidation is currently in the midst of a reinterpretation. New biological activities have been described for tocols that apparently are not dependent on their well-established antioxidant behaviour. These activities could well be real, but there remain large gaps in our understanding of the behaviour of tocols in membranes, especially when it comes to the alpha-, beta-, gamma-, delta-chroman methylation patterns and the seemingly special nature of tocotrienols. It is inappropriate to make conclusions and develop models based on in vivo (or cell culture) results with reference to in vitro measurements of antioxidant activity. When present in biological membranes, tocols will experience a large variation in the local composition of phospholipids and the presence of neutral lipids such as cholesterol, both of which would be expected to change the efficiency of antioxidant action. It is likely that tocols are not homogeneously dispersed in a membrane, but it is still not known whether any specific combination of lipid head group and acyl chains are conferred special protection from peroxidation, nor do we currently appreciate the structural role that tocols play in membranes. Tocols may enhance curvature stress or counteract similar stresses generated by other lipids such as lysolipids. This review will outline what is known about the location and behaviour of tocols in phospholipid bilayers. We will draw mainly from the biophysical literature, but will attempt to extend the discussion to biologically relevant phenomena when appropriate. We hope that it will assist researchers when designing new experiments and when critically assessing the results, in turn providing a more thorough understanding of the biochemistry of tocols. PMID- 18160050 TI - Identifying the site of spin trapping in proteins by a combination of liquid chromatography, ELISA, and off-line tandem mass spectrometry. AB - An off-line mass spectrometry method that combines immuno-spin trapping and chromatographic procedures has been developed for selective detection of the nitrone spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) covalently attached to proteins, an attachment which occurs only subsequent to DMPO trapping of free radicals. In this technique, the protein-DMPO nitrone adducts are digested to peptides with proteolytic agents, peptides from the enzymatic digest are separated by HPLC, and enzyme-linked immunosorbent assays (ELISA) using polyclonal anti-DMPO nitrone antiserum are used to detect the eluted HPLC fractions that contain DMPO nitrone adducts. The fractions showing positive ELISA signals are then concentrated and characterized by tandem mass spectrometry (MS/MS). This method, which constitutes the first liquid chromatography-ELISA mass spectrometry (LC-ELISA-MS)-based strategy for selective identification of DMPO-trapped protein residues in complex peptide mixtures, facilitates location and preparative fractionation of DMPO nitrone adducts for further structural characterization. The strategy is demonstrated for human hemoglobin, horse heart myoglobin, and sperm whale myoglobin, three globin proteins known to form DMPO trappable protein radicals on treatment with H(2)O(2). The results demonstrate the power of the new experimental strategy to select DMPO-labeled peptides and identify sites of DMPO covalent attachments. PMID- 18160051 TI - Protein, lipid, and DNA radicals to measure skin UVA damage and modulation by melanin. AB - Afro-Caribbeans have a lower incidence of skin cancer than Caucasians, but the effectiveness of melanin as a photoprotective pigment is debated. We investigated the UVA and solar irradiation of ex vivo human skin and DMPO using electron spin resonance spectroscopy, to determine whether pigmented skin is protected by melanin against free radical damage. Initial ascorbate radicals in Caucasian skin were superseded by lipid and/or protein radical adducts with isotropic (a(H)=1.8 mT) and anisotropic spectra comparable to spectra in irradiated pig fat (a(H)=1.9 mT) and BSA. DNA carbon-centered radical adducts (a(H)=2.3 mT) and a broad singlet were detected in genomic DNA/melanin but were not distinguishable in irradiated Caucasian skin. Protein and lipid radicals (n=6 in Caucasian skin) were minimal in Afro-Caribbean skin (n=4) and intermediate skin pigmentations were variable (n=3). In irradiated Afro-Caribbean skin a shoulder to the melanin radical (also in UVA-irradiated pigmented melanoma cells and genomic DNA/melanin and intrinsic to pheomelanin) was detected. In this sample group, protein (but not lipid) radical adducts decreased directly with pigmentation. ESR/spin trapping methodology has potential for screening skin susceptibility to aging and cancer-related radical damage and for measuring protection afforded by melanin, sunscreens, and antiaging creams. PMID- 18160052 TI - Novel redox-dependent regulation of NOX5 by the tyrosine kinase c-Abl. AB - We investigated the mechanism of H(2)O(2) activation of the Ca(2+)-regulated NADPH oxidase NOX5. H(2)O(2) induced a transient, dose-dependent increase in superoxide production in K562 cells expressing NOX5. Confocal studies demonstrated that the initial calcium influx generated by H(2)O(2) is amplified by a feedback mechanism involving NOX5-dependent superoxide production and H(2)O(2). H(2)O(2) NOX5 activation was inhibited by extracellular Ca(2+) chelators, a pharmacological inhibitor of c-Abl, and overexpression of kinase dead c-Abl. Transfected kinase-active GFP-c-Abl colocalized with vesicular sites of superoxide production in a Ca(2+)-dependent manner. In contrast to H(2)O(2), the Ca(2+) ionophore ionomycin induced NOX5 activity independent of c-Abl. Immunoprecipitation of cell lysates revealed that active GFP-c-Abl formed oligomers with endogenous c-Abl and that phosphorylation of both proteins was increased by H(2)O(2) treatment. Furthermore, H(2)O(2)-induced NOX5 activity correlated with increased localization of c-Abl to the membrane fraction, and NOX5 proteins could be coimmunoprecipitated with GFP-Abl proteins. Our data demonstrate for the first time that NOX5 is activated by c-Abl through a Ca(2+) mediated, redox-dependent signaling pathway and suggest a functional association between NOX5 NADPH oxidase and c-Abl. PMID- 18160053 TI - The effects of frataxin silencing in HeLa cells are rescued by the expression of human mitochondrial ferritin. AB - Frataxin is a ubiquitous mitochondrial iron-binding protein involved in the biosynthesis of Fe/S clusters and heme. Its deficiency causes Friedreich's ataxia, a severe neurodegenerative disease. Mitochondrial ferritin is another major iron-binding protein, abundant in the testis and in sideroblasts from patients with sideroblastic anemia. We previously showed that its expression rescued the defects caused by frataxin deficiency in the yeast. To verify if this occurs also in mammals, we silenced frataxin in HeLa cells. This caused a reduction of growth, inhibition of the activity of aconitase and superoxide dismutase-2 and reduction of cytosolic ferritins without alteration of mitochondrial iron content. None of these effects were evident when silencing was done in cells expressing mitochondrial ferritin. These data indicate that frataxin has some roles in controlling the balance between different mitochondrial iron pools that are partially in common with those of mitochondrial ferritin. PMID- 18160054 TI - Quest for the cardiovascular holy grail: mammalian myocardial regeneration. PMID- 18160056 TI - The MRL mouse repairs both cryogenic and ischemic myocardial infarcts with scar. AB - OBJECTIVE: The MRL mouse strain shows extraordinary wound healing capacities. Some years ago, Leferovich et al. (Proc Natl Acad Sci U S A 2001;98:9830-35) have reported the absence of scar formation after cryogenically-induced right ventricular myocardial infarcts in adult MRL mice. An independent group (Oh et al., Cardiovasc Pathol 2004;13:203-6) found that MRL mice repair left ventricular ischemic infarcts after coronary artery ligation with regular scar formation. Given the divergent outcomes in infarct healing in MRL mice reported by those two studies, we have investigated whether MRL mice heal myocardial infarcts without scar both in the cryoinjury and in the coronary ligation model. METHODS AND RESULTS: Four different protocols of cryogenically induced right and left ventricular injury, as well as permanent ligation of the left anterior descending coronary artery, were tested in adult MRL and control C57Bl/6 mice. At 60 days after experimental infarction, MRL mice showed pronounced scarring of the affected right and left ventricular areas, with no significant differences in infarct size and thickness between MRL and C57Bl/6 mice using any of the five experimental protocols. Analysis of cell proliferation by 5-bromo-2'-deoxyuridine (BrdU) incorporation into the DNA did not show any difference between the two strains of mice after infarction. Histological analysis of infarct areas using picrosirius red staining did not show differences in extent of collagen and distribution between the two mouse strains. CONCLUSIONS: MRL mice heal myocardial infarcts with scar formation in response to ischemic as well as to cryogenic injuries. PMID- 18160055 TI - Absence of regeneration in the MRL/MpJ mouse heart following infarction or cryoinjury. AB - BACKGROUND: Myocardial infarcts in mammals heal by scar formation rather than formation of new muscle tissue. The MRL/MpJ [Murphy Roths large (MRL) derived by the Murphy group of the Jackson Laboratory (MpJ)] mouse, however, has been reported to exhibit minimal scarring and subsequent cardiac regeneration after cryoinjury of the right ventricle. Other groups have reported that permanent and temporary ligation of the coronary artery resulted in scarring without regeneration. METHODS: To clarify these contradictory results, we studied the temporal evolution of infarcts in MRL/MpJ and C57BL/6 control mice from 1 to 90 days post injury and the effects of intrathoracic cryoinjury to 28 days. RESULTS: After infarction, the conversion from necrotic myocardium to granulation tissue and then to scar proceeded identically in the two groups. Infarct DNA synthesis, measured by incorporation of a 5-bromo-2-deoxyuridine pulse, peaked at 4 days in both strains and did not differ between strains at any time point. Endothelial cell and total vascular density in the both the infarcted and noninfarcted cardiac tissue did not differ between groups at any time. Histological analysis of directly cryoinjured right and left ventricular myocardium showed indistinguishable wound healing in both strains, and final scar size was identical in each group. CONCLUSIONS: These studies demonstrate that both myocardial infarcts and cryoinjuries in MRL/MpJ mice heal by typical scar formation rather than muscle regeneration, in a manner very similar to C57BL/6 controls. We conclude that the MRL mouse is not a model for myocardial regeneration. PMID- 18160057 TI - Cryoinjury: a model of myocardial regeneration. AB - INTRODUCTION: Although traditionally adult cardiomyocytes are thought to be unable to divide, recent observations provide evidence for cardiomyocyte proliferation after myocardial injury. Myocardial cryoinjury has been shown to be followed by neovascularization. We hypothesize that, in addition to neovascularization, cardiomyocyte proliferation after myocardial cryoinjury contributes to regeneration. METHOD: Cryolesions were applied to the left ventricle of mouse hearts. Inflammatory cell infiltration (F4/80, neutrophils), neovascularization (CD31), and cardiomyocyte proliferation (5-bromo-2 deoxyuridine, Ki-67, mitotic spindle) were determined at different time points (2 70 days) after cryoinjury. RESULTS: Between Days 7 and 14 after injury, a 150- and 280-fold increase in number of proliferating cardiomyocytes was observed, as compared to controls. At the same time, numerous proliferating capillaries were found in between the proliferating cardiomyocytes. Presence of high numbers of macrophages in the cryolesion preceded and coincided with this proliferation. The area of cryolesion decreased significantly between Days 7 (23+/-5%) and 14 (8+/ 2%) after cryoinjury. Moreover, regeneration of viable, nonhypertrophied myocardium was observed. After 14 days, cardiomyocyte proliferation decreased to numbers observed in controls, and concomitantly, the number of macrophages strongly decreased. CONCLUSION: Our data show that adult cardiomyocytes proliferate in sufficiently high numbers to effectuate myocardial regeneration after left ventricular cryoinjury in mice. PMID- 18160058 TI - The MRL mouse heart does not recover ventricular function after a myocardial infarction. AB - INTRODUCTION: Murphy Roth Large (MRL) mice have a remarkable regenerative capacity. A recent report demonstrated rapid cardiac healing in these mice following cryogenically induced right ventricular injury, suggesting the potential for new regenerative therapies to restore cardiac function in mammals. We therefore evaluated the cardiac regenerative wound-healing response and functional recovery of MRL mice in response to a clinically relevant left ventricular coronary ligation. METHODS: Female MRL/MpJ+/+ and C57BL/6 mice underwent left coronary artery ligation. Cardiac function was evaluated by echocardiography at Days 0, 5, 15, and 60. At Day 96, invasive hemodynamics were assessed by pressure-volume loops using a Millar catheter. Hearts were perfusion fixed for histomorphometric analysis at Days 5, 15, and 96. Some hearts were fresh frozen at Days 5 and 15 for immunohistochemical analysis and digital quantitation of blood vessel density (CD31) and cellular proliferation (Ki67). RESULTS: MRL mice healed ear punch wounds (89% reduction in area) more extensively than C57BL/6 mice (28% reduction in area) but did not differ functionally from C57BL/6 animals before or after coronary ligation. In addition, blood vessel density and cell proliferation were similar between the two strains. CONCLUSIONS: Although MRL mice rapidly healed ear injury, they did not exhibit regeneration of the left ventricle or enhanced functional improvement in response to coronary ligation. The prospect of cardiac regeneration after myocardial infarction will require further studies designed to elucidate the possible mechanisms of functional restoration. PMID- 18160059 TI - A comparison of genetic chromosomal loci for intracranial, thoracic aortic, and abdominal aortic aneurysms in search of common genetic risk factors. AB - BACKGROUND: Genetic factors are likely to be involved in the pathogenesis of intracranial, ascending thoracic aorta, and infrarenal aortic abdominal aneurysms. Common genetic risk factors for these three types of aneurysms have been suggested. This review describes the results of whole-genome linkage studies on intracranial, thoracic aorta, and aortic abdominal aneurysms, and compares the genomic loci identified in these studies in search of possible common genetic risk factors for the three aneurysmal types. METHODS: A literature search of all whole-genome linkage studies performed on intracranial, thoracic aorta, and aortic abdominal aneurysms was performed. The genomic loci identified in these studies were described and compared in search of similarities between them. RESULTS: Five chromosomal regions on 3p24-25, 4q32-34, 5q, 11q24, and 19q that may play a role in the pathogenesis of two or more aneurysmal types were identified: 3p24-25 for thoracic aorta and intracranial aneurysms; 4q32-34 for aortic abdominal and intracranial aneurysms; 5q for thoracic aorta and intracranial aneurysms; 11q24 for thoracic aorta, aortic abdominal, and intracranial aneurysms; and 19q for aortic abdominal and intracranial aneurysms. CONCLUSIONS: Five chromosomal regions that may include common genetic factors for intracranial, thoracic aorta, and aortic abdominal aneurysms were identified. Further studies are needed to explore these chromosomal regions in different aneurysm patient groups and may further help to unravel the disease pathogenesis of aneurysms in general. PMID- 18160060 TI - Genetic insights into normal and abnormal heart development. AB - Congenital heart defects (CHDs) affect 1-2% of newborn children and are the leading cause of death in infants under 1 year of age. CHDs represent the single largest class of birth defects and account for 25% of all human congenital abnormalities. Numerous epidemiologic studies have established the heritable nature of CHDs. However, despite the remarkable progress of the past decade, very few CHD-causing genes have been identified so far. Molecular and genetic analysis of heart development--which requires the execution of specific genetic programs- has led to the identification of essential cardiac regulators and mutations that are linked to human CHD. Elucidation of the mechanisms of action of these transcription factors has also provided a molecular framework that will continue to help furthering our understanding of the molecular basis of normal and abnormal heart growth. This review will summarize present knowledge of cardiac development and illustrate how analysis of heart development has helped understand the genetic basis of some CHDs and how these advances could translate into better prevention, diagnosis, and care of congenital heart disease. PMID- 18160061 TI - Low-grade myofibroblastic sarcoma of the heart causing sudden death. AB - A 5-year-old Asian girl collapsed at school and was dead on arrival to the hospital. At autopsy, she had a tumor arising from her aortic valve. Histology revealed a low-grade myofibroblastic sarcoma, which we believe occluded a coronary artery ostium, leading to sudden death. Primary cardiac neoplasms are rare. The most common primary cardiac neoplasm is the cardiac myxoma. Although uncommon, the most prevalent primary malignant tumors of the heart are sarcomas, and regardless of the subtype, the prognosis is typically poor. We present a case of a primary cardiac sarcoma presenting as sudden death in a 5-year-old Asian girl. Pathologic examination of the lesion confirmed that this was an atypical myofibroblastic neoplasm, consistent with a low-grade sarcoma. Pediatric cardiac sarcomas are rare in general, and this is the first reported case of a valvular low-grade myofibroblastic sarcoma presenting in a child. PMID- 18160062 TI - Inhibition of the proliferation of human periodontal ligament fibroblasts by hyaluronidase. AB - Hyaluronan (HA) exists in various living tissues as one of the major matrix macromolecules, and is well known to play an integral role in cell differentiation and proliferation. The present study was conducted to elucidate whether or not the proliferation of periodontal ligament (PDL) cells are affected specifically by the degradation of HA by hyaluronidasze (HAase). Human PDL fibroblasts were isolated and cultured with and without 15-150U/ml bovine testicular HAase from 1 to 11 days after seeding. The cells were also cultured with anti-CD44 antibody of 2 microg/ml. For the control against the anti-CD44 antibody treatment, 2 microg/ml IgG was used. The HA-dependent pericellular matrix was visualized by particle-exclusion assay. The number of cells was counted by MTT assay during the proliferation. The mRNA levels of HA synthases (HASs), HAases (HYALs) and CD44s were examined by a quantitative real-time PCR analysis. The cell proliferation was inhibited by the treatment with HAase and anti-CD44 antibody in cultured PDL fibroblasts. HASs mRNAs were down-regulated, whereas HYALs mRNAs were up-regulated significantly by the treatment with HAase and anti-CD44 antibody. The CD44s mRNA level exhibited no significant changes. These results suggest that HA may contribute to modulate the proliferation of cultured human PDL cells through a CD44-mediated mechanism. PMID- 18160063 TI - Structural studies of the O-polysaccharide chain from the lipopolysaccharide of symbiotically enhanced mutant Mlo-13 of Mesorhizobium loti NZP2213. AB - The O-specific polysaccharide (OPS) of Mesorhizobium loti mutant Mlo-13 was obtained by mild-acid degradation of the lipopolysaccharide and studied by sugar and ethylation analyses, Smith degradation, as well as (1)H and (13)C NMR spectroscopy. The O-polysaccharide has a trisaccharide repeating unit composed of 6-deoxy-L-talose (L-6dTal), 4-O-acetyl-6-deoxy-L-talose (L-6dTal-4Ac) and L rhamnose residues. An analysis of NMR spectra revealed that the rhamnose residue was substituted nonstoichiometrically with O-methyl group at C-3. The following structure of the repeating unit of the Mlo-13 OPS was established: [structure: see text]. PMID- 18160064 TI - Activation of apoptosis, but not necrosis, during Mycobacterium tuberculosis infection correlated with decreased bacterial growth: role of TNF-alpha, IL-10, caspases and phospholipase A2. AB - Monocyte/macrophage cell death is an important event during mycobacterial infection. To get insights about the influence of mononuclear phagocyte maturation in this event we compared the response to Mycobacterium tuberculosis (Mtb) infection of fresh isolated monocytes and monocyte-derived macrophages (MDM) from healthy tuberculin positive individuals. Both monocytes and MDM underwent apoptosis, however, there was a higher numbers of apoptotic macrophages with active Caspases 8 and 9. We also compared Mtb-induced cell death in U937 pro monocytes and PMA-differentiated cells (U937D). In response to Mtb infection, U937D cells underwent apoptosis and promonocytes both apoptosis and necrosis. There were high number of U937D cells producing TNF-alpha and high number of IL 10+ promonocytes. These evidences suggest that U937 could be a valid model to study the mechanisms that rule Mtb-induced cell death. Experiments with the cell line and fresh isolated mononuclear cells with pharmacological inhibitors showed that induction of necrosis involved calcium and cAMP signals resulting in IL-10 production. Necrosis also correlated with Caspase 3, PLA2 activity and bacterial growth. In U937D cells and monocytes from healthy donors there was activation of calcium, TNF-alpha and Caspase 8 activation and decreased bacterial load. Understanding the mechanisms that control the dichotomy events between apoptosis and necrosis/oncosis associated with cell maturity might open new strategies to better control the course of mycobacterial infections. PMID- 18160065 TI - Reduction of cryoprotectant toxicity in cells in suspension by use of a sodium free vehicle solution. AB - Propane-1,2-diol (PD) possesses physico-chemical properties that make it a useful biological vitrifying agent but, although of relatively low toxicity, it still has substantial damaging effects on cells. This study aimed to identify possible toxic mechanisms using primary cell cultures from vascular tissue: these were exposed to the cryoprotectant at room temperature to avoid any possibility of hypothermic injury. Toxicity was evaluated by measuring the ability of the cells to divide in culture after exposure to the cryoprotectant. A variety of interventions, which addressed either possible consequences of PD exposure, or known mediators of other types of cell injury, were utilized in an attempt to inhibit PD toxicity. Some comparative studies with dimethyl sulphoxide (Me2SO) exposure were also made. Replacing sodium in the vehicle solution with choline was the only intervention that reduced PD toxicity. It did so both in smooth muscle cells, where the loss of functional capacity was reduced from 56% to 13%, and in endothelial cells. where the reduction was from 40% to 18%. Similar observations were also made in smooth muscle cells exposed to Me2SO. We failed to find evidence for a role of pH regulation, for oxidative injury and/or an involvement of redox-active iron as a mediator of the injury. The results strongly suggest that the influx of sodium into the cell provides one mechanism whereby both PD and Me2SO exert their toxic effects. We suggest that the use of choline-based vehicle solutions in cryopreservation would be beneficial. PMID- 18160066 TI - Heritability of refractive value and ocular biometrics. AB - The aim of this work was to analyse genetic influences on ocular refractive value and axial length using the hypothesis of a polygenic control. The genealogical records of 55 families were used in the analyses. The cohort included 723 individuals and clinical data were collected for 445 individuals with a mean age of 37.86 years. Ocular refraction was determined by standard autorefractometry. Axial length was evaluated by scan ultrasonography. Gender, age and ethnic origin were included as covariates in the statistical analyses. Using variance component analysis via a Markov Chain Monte Carlo (MCMC) method, we estimated the heritability of refractive value and axial length in the pedigree. We then performed a segregation analysis, using Loki, a (MCMC) linkage analysis program for multilocus inheritance models, examining different inheritance models with polygenic components. Polygenic control was modelled under an additive infinitesimal model (which assumes infinite loci with small effects, with additive actions) and under a finite locus model (i.e. several causal loci). The estimates of heritability were 0.20 (95% confidence interval (CI) 0.04-0.36) for refractive value and 0.20 (95% CI 0.03-0.43) for axial length. Segregation analyses suggested that ocular refraction and axial length are under a polygenic control. A finite number of genes were identified with or without a polygenic, infinitesimal component. Ocular refraction is mildly-moderately heritable in the studied population. PMID- 18160067 TI - Flavonoids protect retinal ganglion cells from ischemia in vitro. AB - Retinal ischemia is a common cause of visual impairment and blindness. However, despite the significant advances that have been made in understanding the pathophysiology of retinal ischemia, effective treatments are still lacking. The goal of these studies was to use an in vitro model to identify molecules that could be neuroprotective for retinal ganglion cells exposed to ischemia. Ischemia was induced in the rat retinal ganglion cell line, RGC-5, using iodoacetic acid (IAA). Brief treatment with IAA resulted in RGC-5 cell death within 24 h by a non apoptotic mechanism. Similar to ischemia in vivo, IAA treatment caused a rapid loss of ATP to approximately 50% of control levels. In contrast, changes in markers of oxidative stress occurred more slowly and included an increase in reactive oxygen species and a decrease in glutathione. Specific flavonoids were able to prevent the cell death caused by IAA treatment. Some of the flavonoids also prevented the loss of ATP as well as the changes in markers of oxidative stress. In contrast, classical antioxidants had only a very modest effect on IAA induced cell death. These results suggest that specific flavonoids may be useful in preventing ischemia-induced retinal ganglion cell death in vivo. PMID- 18160068 TI - Calcium homeostasis: reassessment of the actions of parathyroid hormone. AB - The purpose of this report is to adjust our interpretation of the actions of parathyroid hormone (PTH) to include its action on the bone mineral noncollagenous protein interactions at all bone surfaces. The three primary areas that respond to PTH are: (1) all bone surface areas in contact with the extracellular fluid (ECF), (2) the kidney, and indirectly the intestinal tract, and (3) the bone remodeling sequence. The primary rapid action of the hormone is to set and maintain the free calcium concentration of the ECF. This it does by raising the equilibrium level at bone mineral surfaces. It affects the noncollagenous protein bone mineral process to raise the free calcium level in the ECF from the base level of 3.5mg/100ml to the physiological level of 5.0mg/100ml. Maintaining the higher level requires continuous secretion of parathyroid hormone. The action of PTH at bone surfaces tends to be catabolic in nature in regard to bone loss. The hormone also acts on the kidney to raise the threshold for calcium reabsorption and by stimulation of renal hydroxylation of vitamin D to increase intestinal absorption of calcium. Its action here is to supply the ECF with calcium derived from food intake. This is the extent of PTH action on renal processes. PTH acts on all three steps in the bone remodeling process. While its total function here is not clear, the result is a net increase in the synthesis of collagen. The report concludes by comparing the actions of PTH as proposed here to the functions of PTH that have been proposed in the past. PMID- 18160069 TI - Vaccination with recombinant oncosphere antigens reduces the susceptibility of sheep to infection with Taenia multiceps. AB - Taenia multiceps is a cestode parasite, the larval stage of which encysts in the brain of sheep, goats and cattle causing an often fatal condition. The parasite also causes zoonotic infections in humans. Homologues of the recombinant oncosphere vaccine antigens from Taenia ovis and other Taenia species were identified in T. multiceps. Sequencing of the associated T. multiceps genes and cloning of the encoding mRNA has revealed conserved features in the genes and proteins. The T. multiceps oncosphere proteins, designated Tm16 and Tm18, contain a predicted secretory signal and fibronectin type III domain. The recombinant Tm16 and Tm18 proteins were successfully expressed in Escherichia coli as fusion proteins with GST. The antigens, formulated with Quil A adjuvant, were tested in a vaccine trial in sheep. The antigens stimulated immunity in sheep against challenge infection with T. multiceps eggs. Five of nine control sheep died due to a challenge infection with T. multiceps whereas none of 20 vaccinated animals died as a result of the parasite challenge (P=0.001). In addition, vaccination with the Tm16 protein, or Tm16 plus Tm18, induced significant protection against the number of parasites encysting in the brain as a result of the challenge infection (P=0.023, P=0.015, respectively). No clear relationship was apparent between the level of specific serum antibody in vaccinated animals and either the presence or absence of parasites or the number of parasites that occurred in some of the vaccinated animals. We believe this study is the first description of recombinant vaccine-related investigations for T. multiceps. The recombinant oncosphere antigens identified may allow development of effective vaccination strategies against T. multiceps infection in sheep. They raise the potential for the development of a combined vaccine with the Echinococcus granulosus EG95 antigen for prevention of T. multiceps as well as preventing the transmission of cystic hydatid disease. PMID- 18160070 TI - The relation of leptin and insulin with obesity-related cardiovascular risk factors in US adults. AB - Previous studies of leptin with cardiovascular disease (CVD) risk factors have been limited by clinical samples or lack of representation of the general population. This cross-sectional study, designed to examine whether leptin or insulin may mediate the endogenous relation of obesity with metabolic, inflammatory, and thrombogenic cardiovascular risk factors, included 522 men and 514 women aged >or=40 years who completed a physical examination during the third National Health and Nutrition Examination Survey. Participants were free of existing CVD, cancer (except non-melanoma skin cancer), diabetes, or respiratory disease. In multivariable analyses adjusted for race/ethnicity and lifestyle factors, waist circumference (WC) was positively associated with blood pressure, triglyceride, LDL cholesterol, total cholesterol:HDL ratio, apolipoprotein B, C reactive protein (CRP), and fibrinogen concentrations, and negatively associated with HDL cholesterol and apolipoprotein A1 levels. The associations of WC with the metabolic CVD risk factors were largely attenuated after adjustment for insulin levels, while the associations of WC with the inflammatory and thrombogenic factors (CRP and fibrinogen, respectively) were largely explained by adjustment for leptin concentrations. However, leptin levels were not independently associated with CRP and fibrinogen in men and CRP in women when adjusted for WC. Positive associations of leptin and insulin with fibrinogen in women, independent of WC, were noted. These results suggest that insulin may be an important mediator of the association of obesity with metabolic but not inflammatory or thrombogenic CVD risk factors, while leptin does not appear to influence cardiovascular risk through a shared association with these risk factors. However, we cannot rule out the possibility that leptin and insulin influence cardiovascular risk in women through independent effects on fibrinogen concentrations. PMID- 18160071 TI - Omega-3 fatty acids and coronary heart disease risk: clinical and mechanistic perspectives. AB - The most common omega-3 fatty acids contain 18-22 carbons and a signature double bond at the third position from the methyl (or n, or omega) end of the molecule. These fatty acids must be obtained in the diet as they cannot be synthesized by vertebrates. They include the plant-derived alpha-linolenic acid (ALA, 18:3n-3), and the fish-oil-derived eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3). Normally, very little ALA is converted to EPA, and even less to DHA, and therefore direct intake of the latter two is optimal. EPA and DHA and their metabolites have important biologic functions, including effects on membranes, eicosanoid metabolism, and gene transcription. Studies indicate that the use of fish oil is associated with coronary heart disease risk reduction. A number of mechanisms may be responsible for such effects. These include prevention of arrhythmias as well as lowering heart rate and blood pressure, decreasing platelet aggregation, and lowering triglyceride levels. The latter is accomplished by decreasing the production of hepatic triglycerides and increasing the clearance of plasma triglycerides. Our focus is to review the potential mechanisms by which these fatty acids reduce cardiovascular disease risk. PMID- 18160072 TI - Comprehensive two-dimensional gas chromatography in the screening of persistent organohalogenated pollutants in environmental samples. AB - Separations of eight persistent organohalogenated classes of pollutants, organochlorinated pesticides (OCPs), polychlorinated biphenyls (PCBs), polychlorinated diphenyl ethers (PBDEs), polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated naphthalenes (PCNs), polychlorinated terphenyls (PCTs) and toxaphene (CTT) by comprehensive two-dimensional gas chromatography (GC x GC) were evaluated. Columns with different polarity and selectivity, including ZB-5, HT-8, DB-17 and BP-10, were selected as first dimension and combined with columns of increasing polarity in the second dimension, i.e. HT-8, BPX-50 and Carbowax (or Supelcowax-10). In total nine column combinations were tested. Because the main interest of the study was fast screening of the test xenobiotic families in complex matrices, in all cases, attention was primarily focussed on group-type separation. Nevertheless, within group separation was also considered, especially for those classes containing particularly toxic congeners, such as PCBs and PCDD/Fs. Although none of the assayed column sets allowed the simultaneous and complete separation of all pollutants classes, some column combinations provided satisfactory separations among selected families and the rest of pollutants investigated. That was, for instance, the case of HT-8 x BPX-50 for PBDEs and PCDD/Fs, DB-17 x HT-8 for PCNs and OCPs and BP-10 x BPX-50 for CTT, PCDD/Fs and PBDEs. The feasibility of the proposed approach for the fast screening of the target classes of pollutants in complex samples was illustrated by the analysis of food and marine fat samples prepared using simplified miniaturised sample treatment methods. PMID- 18160073 TI - pH-gradient counter-current chromatography isolation of natural antioxidant chlorogenic acid from Lonicera japonica Thumb. using an upright coil planet centrifuge with three multi-layer coils connected in series. AB - A new pH-gradient counter-current chromatography method for the isolation of chlorogenic acid from flowers and buds of Lonicera japonica Thumb. has been successfully established using a novel upright coil planet centrifuge with three multi-layer coils connected in series with 600 mL capacity. The crude extracts were first prepared by direct extraction with hot water and following concentration to remove the solution. Then the two-phase solvent system composed of ethyl acetate-n-butanol-water (2:1:3, v/v) was applied to the separation. Its neutral upper phase was used as stationary phase, whereas both its neutral lower phase and base lower phase with 10mM NH(3) were employed as mobile phase with gradient elution in the head to tail mode. As a result, 330 mg quantity of crude extract was purified in one-step separation for 180 min, yielding 20.5mg chlorogenic acid with over 98% purity. Structure of the compound is further identified by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and nuclear magnetic resonance (NMR). PMID- 18160074 TI - Separation of rare earth elements by anion-exchange chromatography using ethylenediaminetetraacetic acid as mobile phase. AB - Rare earth elements (REEs) form anionic complexes which can be separated isocratically by anion-exchange chromatography using ethylenediaminetetraacetic acid (EDTA) as the mobile phase. For easy detection and identification, inductively coupled plasma (ICP) MS was used as detection method. From pH 3.5 to 7.5, retention increases from La to Sm and then decreases again up to Lu. Above pH 8.5, the retention of the light lanthanides increased drastically. It seems that the stoichiometry and the charge of the REE-EDTA complexes change with the elution pH. This strange elution behaviour can be easily tuned for particular applications by selecting the elution pH. For example, at pH values between 5.5 and 7.5 most isobaric and polyatomic interferences which occur in ICP-MS detection of the lanthanides are eliminated. A mechanism for the stepwise formation of the REE-EDTA complexes as a function of pH is proposed. PMID- 18160075 TI - Editorial on "Extending the molecular application range of gas chromatography" by E. Kaal and H.-G. Janssen. PMID- 18160076 TI - Dynamical types of bacteria and bacteriophages interaction: shielding by debris. AB - The dynamics of the bacteria and bacteriophages interaction process was explored in depth, and laid on a firm basis through simulation and analysis. A modified Campbell model of phage-bacteria interaction was used to simulate three interacting species: bacteria, phages and bacterial debris, and their time behavior in terms of three parameters, in selected range values: the phages burst size (beta), the dissolution rate of the bacterial debris (q), and the lysing time delay (tau). Six types of dynamical behavior were identified, occupying various zones in the two-dimensional (2D) plane (q, beta) for several values of tau, allowing the determination of the region where shielding by bacterial debris is in effect. The problem of the occurrence of stability transitions between stable and unstable dynamical regimes was also addressed analytically, and compared with simulation results. PMID- 18160077 TI - From prebiotic chemistry to cellular metabolism--the chemical evolution of metabolism before Darwinian natural selection. AB - It is generally assumed that the complex map of metabolism is a result of natural selection working at the molecular level. However, natural selection can only work on entities that have three basic features: information, metabolism and membrane. Metabolism must include the capability of producing all cellular structures, as well as energy (ATP), from external sources; information must be established on a material that allows its perpetuity, in order to safeguard the goals achieved; and membranes must be able to preserve the internal material, determining a selective exchange with external material in order to ensure that both metabolism and information can be individualized. It is not difficult to understand that protocellular entities that boast these three qualities can evolve through natural selection. The problem is rather to explain the origin of such features under conditions where natural selection could not work. In the present work we propose that these protocells could be built by chemical evolution, starting from the prebiotic primordial soup, by means of chemical selection. This consists of selective increases of the rates of certain specific reactions because of the kinetic or thermodynamic features of the process, such as stoichiometric catalysis or autocatalysis, cooperativity and others, thereby promoting their prevalence among the whole set of chemical possibilities. Our results show that all chemical processes necessary for yielding the basic materials that natural selection needs to work may be achieved through chemical selection, thus suggesting a way for life to begin. PMID- 18160078 TI - Spatio-temporal modelling explains the effect of reduced plasma membrane Ca2+ efflux on intracellular Ca2+ oscillations in hepatocytes. AB - In many non-excitable eukaryotic cells, including hepatocytes, Ca(2+) oscillations play a key role in intra- and intercellular signalling, thus regulating many cellular processes from fertilisation to death. Therefore, understanding the mechanisms underlying these oscillations, and consequently understanding how they may be regulated, is of great interest. In this paper, we study the influence of reduced Ca(2+) plasma membrane efflux on Ca(2+) oscillations in hepatocytes. Our previous experiments with carboxyeosin show that a reduced plasma membrane Ca(2+) efflux increases the frequency of Ca(2+) oscillations, but does not affect the duration of individual transients. This phenomenon can be best explained by taking into account not only the temporal, but also the spatial dynamics underlying the generation of Ca(2+) oscillations in the cell. Here we divide the cell into a grid of elements and treat the Ca(2+) dynamics as a spatio-temporal phenomenon. By converting an existing temporal model into a spatio-temporal one, we obtain theoretical predictions that are in much better agreement with the experimental observations. PMID- 18160079 TI - Industrial pollution at Bagnoli (Naples, Italy): benthic foraminifera as a tool in integrated programs of environmental characterisation. AB - In the 20th century an important industrial plant operated on the coastal area of Bagnoli. After its closing, an integrated study of environmental characterisation aimed at restoration started. The survey conducted was based on chemical and sedimentological analyses integrated with benthic foraminifera analyses. Statistical analysis of the data shows sectors with a distinct type and degree of pollution. Particularly, pollution linked to the silty sediment fraction, mainly due to Pb and Zn, was recognised in front of the southern sector of the plant. The study of benthic foraminifera provides evidence for a pollution-tolerant character in some species like Haynesina germanica and Quinqueloculina parvula. In addition, two species among the 113 recognised show high percentages of abnormal specimens. These percentages show a statistical correlation with some pollutants (PAHs, Mn, Pb and Zn). In addition, Energy Dispersive Spectrometry shows small amounts of Fe ions included in deformed tests of Miliolinella subrotunda. Because the number of these deformations is positively correlated to the concentration of PAHs, Mn and Zn, the inability of some specimens to exclude the foreign elements from the crystalline reticulum of the test could be attributed to the potential toxic effect of these pollutants. PMID- 18160080 TI - Role of orexin receptors in the nucleus accumbens in dopamine-dependent turning behaviour of rats. AB - The role of orexin receptors in the nucleus accumbens shell in rat turning behaviour of rats was studied. Unilateral injection of neither the orexin 1 and 2 receptor agonist orexin A (2 microg) nor the orexin 1 receptor antagonist SB 334867 (20 ng) into the nucleus accumbens shell elicited turning behaviour. Unilateral injection of a mixture of dopamine D(1) (SKF 38393) and D2 (quinpirole) receptor agonists into the nucleus accumbens shell has been found to elicit contraversive pivoting. Orexin A (1 and 2 microg) dose-dependently potentiated the contraversive pivoting induced by a mixture of SKF 38393 (1 microg) and quinpirole (10 microg) injected into the nucleus accumbens shell whereas SB 334867 (10 and 20 ng) did not significantly affect the pivoting. The potentiating effect of orexin A (2 microg) on the dopaminergic pivoting was not significantly inhibited by SB 334867 (10 and 20 ng) injected into the nucleus accumbens shell. The contraversive pivoting induced by a mixture of SKF 38393 (1 microg) and quinpirole (10 microg) injected into the nucleus accumbens shell was also potentiated by the orexin 2 receptor agonist orexin B (0.5, 1 and 2 microg), which alone did not elicit turning behaviour. These results suggest that orexin 2 receptors in the nucleus accumbens shell play a modulatory role in rat turning behaviour. PMID- 18160081 TI - Affective blindsight in the intact brain: neural interhemispheric summation for unseen fearful expressions. AB - The emotional valence of facial expressions can be reliably discriminated even in the absence of conscious visual experience by patients with lesions to the primary visual cortex (affective blindsight). Prior studies in one such patient (GY) also showed that this non-conscious perception can influence conscious recognition of normally seen emotional faces. Here we report a similar online interaction across hemispheres between conscious and non-conscious perception of emotions in normal observers. Fearful and happy facial expressions were presented either unilaterally (to the left or right visual field) or simultaneously to both visual fields. In bilateral displays, conscious perception of one face in a pair was prevented by backward masking after 20 ms, while the opposite expression remained normally visible. The results showed a bidirectional influence of non conscious fear processing over conscious recognition of happy as well as fearful expressions. Consciously perceived fearful faces were more readily recognized when they were paired with invisible emotionally congruent fearful expressions in the opposite field, as compared to the single presentation of the same unmasked faces. On the other hand, recognition of unmasked happy faces was delayed by the simultaneous presence of a masked fearful face. No such effect was reported for masked happy expressions. These findings show that non-conscious processing of fear may modulate ongoing conscious evaluation of facial expressions via neural interhemispheric summation even in the intact brain. PMID- 18160082 TI - Limited distribution of natural cyanamide in higher plants: occurrence in Vicia villosa subsp. varia, V. cracca, and Robinia pseudo-acacia. AB - Cyanamide (NH2CN) has recently been proven to be a natural product, although it has been synthesized for over 100 years for agricultural and industrial purposes. The distribution of natural cyanamide appears to be limited, as indicated by our previous investigation of 101 weed species. In the present study, to investigate the distribution of natural cyanamide in Vicia species, we monitored the cyanamide contents in V. villosa subsp. varia, V. cracca, and V. amoena during their pre-flowering and flowering seasons. It was confirmed that V. cracca was superior to V. villosa subsp. varia in accumulating natural cyanamide, and that V. amoena was unable to biosynthesize this compound under laboratory condition examined. The localization of cyanamide in the leaves of V. villosa subsp. varia seedlings was also clarified. In a screening study to find cyanamide biosynthesizing plants, only Robinia pseudo-acacia was found to contain cyanamide among 452 species of higher plants. We have investigated 553 species to date, but have so far found the ability to biosynthesize cyanamide in only three species, V. villosa subsp. varia, V. cracca and R. pseudo-acacia. PMID- 18160083 TI - Glycosylation of hesperetin by plant cell cultures. AB - The biotransformation of hesperetin by cultured cells of Ipomoea batatas and Eucalyptus perriniana was investigated. Three glycosides, hesperetin 3'-O-beta-D glucopyranoside (33 microg/g fr. wt of cells), hesperetin 3',7-O-beta-D diglucopyranoside (217 microg/g fr. wt of cells), and hesperetin 7-O-[6-O-(beta-D glucopyranosyl)]-beta-d-glucopyranoside (beta-gentiobioside, 22 microg/g fr. wt of cells), together with three hitherto known glycosides, hesperetin 5-O-beta-d glucopyranoside (23 microg/g fr. wt of cells), hesperetin 7-O-beta-D glucopyranoside (57 microg/g fr. wt of cells), and hesperetin 7-O-[6-O-(alpha-L rhamnopyranosyl)]-beta-D-glucopyranoside (beta-rutinoside, hesperidin, 13 microg/g fr. wt of cells), were isolated from cultured suspension cells of E. perriniana that had been treated with hesperetin. Oligosaccharide chains were regioselectively formed at the C-7 position of hesperetin to afford beta gentiobioside and beta-rutinoside. On the other hand, cultured I. batatas cells converted hesperetin into hesperetin 3'-O-beta-D-glucopyranoside (60 microg/g fr. wt of cells), hesperetin 5-O-beta-D-glucopyranoside (23 microg/g fr. wt of cells), and hesperetin 7-O-beta-D-glucopyranoside (110 microg/g fr. wt of cells). PMID- 18160084 TI - Relationship between health-related fitness and educational and income levels in Spanish women. AB - OBJECTIVES: The aim of this study was to determine whether a relationship exists between health-related fitness, taken as an indicator of regular physical activity, and educational and income levels in adult Spanish women. STUDY DESIGN: Descriptive, correlation, cross-sectional study. METHODS: A stepwise stratification procedure according to population size, age and level of physical activity according to a previous epidemiological survey was used for sampling. Two thousand and thirty-eight women gave their written consent to participate (62.8% of those invited). The final sample consisted of 1709 healthy women (aged 18-88 years). Subjects were categorized into high, medium and low level groups for education and income. All participants were assessed for morphological and physical health-related fitness. Three-way MANCOVA (age as covariate) and Bonferroni's post hoc test were used to determine the differences between groups. RESULTS: No significant relationships were found between age-adjusted educational and income levels. The lowest values for health-related fitness were found in the lowest educational and income groups (P<0.001). The higher the level of education and income, the better the values for all fitness variables (P<0.001), except anterior trunk flexibility. CONCLUSIONS: A positive relationship was found between health-related fitness and educational and income levels, which appeared to be most evident in the lowest educational and income groups. This implies that health-related promotion policies in Spain should stress the importance of regular physical activity in social classes with low levels of education and income. PMID- 18160085 TI - [Another aspect of the face."How to validate what we do?"]. PMID- 18160086 TI - [Osteosynthesis of the temporal articular tubercle]. AB - This technical note stresses the interest of the temporal articular tubercle osteosynthesis in zygomatic bone fractures. An impaction of the zygoma can lead to valgus of the temporal bone's zygomatic process with diastasis in the temporal articular tubercle. If the frontal and maxillary processes are comminuted, osteosynthesis of the temporal articular tubercle allows recovering the initial projection of the zygomatic bone. The adequate alignment of the sphenozygomatic suture is also reliable for an anatomical reduction of the fracture. PMID- 18160087 TI - Comparative evaluation of direct agglutination test, rK39 and soluble antigen ELISA and IFAT for the diagnosis of visceral leishmaniasis. AB - Five serological tests for the diagnosis of visceral leishmaniasis (VL) were compared: a direct agglutination test (DAT) based on freeze-dried antigen (DAT fd); a locally produced DAT (DAT-LPC); an IgG ELISA against rK39 (ELISA-rK39); an IgG ELISA for Leishmania chagasi (ELISA-L. chagasi); and an IgG IFAT against L. chagasi. Serum samples from 88 patients with VL, 20 non-infected individuals and 85 patients with others infectious diseases were evaluated. The sensitivity rates were: DAT-fd, 96.6%; DAT-LPC, 95.5%; ELISA-rK39, 88.6%; ELISA-L. chagasi, 89.8%; and IFAT, 92.0% (P>0.05). The specificity for the control groups varied from 53.3% to 100%. DAT-fd had the highest efficiency (97.4%), followed by DAT-LPC (91.7%) and ELISA-rK39 (90.7%). Our data suggest that DAT-fd, DAT-LPC and ELISA rK39 are useful tests for the diagnosis of VL and could replace IFAT as the routine diagnostic test in Brazil. PMID- 18160088 TI - Ultrastructure and morphology of midgut visceral muscle in early pupal Aedes aegypti mosquitoes. AB - These studies focus on the pupal Aedes aegypti midgut muscularis for the first 26 h following larval-pupal transition. The midgut muscularis of Ae. aegypti pupae during this first half of the pupal stadium is a grid of both circularly and longitudinally oriented muscle bands, arranged in a manner resembling that of the larvae. While many muscle bands exhibit signs of degeneration during the time period studied, not all bands degrade, nor is this degradation simultaneous. Band deterioration involves destruction of internal elements while the muscle fiber plasma membrane remains intact. Deterioration of contractile elements may involve proteosome-like structures and associated enzymes. Many features of the larval muscularis including cruciform cells, bifurcating circular bands, and bifurcating longitudinal bands of muscle are retained during the time period investigated. Neuromuscular junctions along some muscle bands are retained through at least 16 h into the pupal stadium. The selective nature of muscle fiber degradation, coupled with the retention of larval features and neural input, may allow for limited functionality of the muscularis during metamorphosis. Evidence of sexual dimorphism in the midgut muscularis of male and female Ae. aegypti pupae was not observed during the time period studied. PMID- 18160089 TI - Tissue distribution in mice of BPP 10c, a potent proline-rich anti-hypertensive peptide of Bothrops jararaca. AB - The snake venom proline-rich peptide BPP 10c is an active somatic angiotensin converting enzyme (sACE) inhibitors. Recently we demonstrated that the anti hypertensive effect of BPP 10c is not related to the inhibition of sACE alone, thus suggesting that this enzyme is not its only target for blood pressure reduction. In the present work, a biodistribution study in Swiss mice of [(125)I] BPP 10c in the absence or in the presence of a saturating concentration of captopril, a selective active-site inhibitor of sACE, demonstrated that: (1) [(125)I]-BPP 10c was present in several organs and the renal absorption was significantly high; (2) [(125)I]-BPP 10c showed a clear preference for the kidney, maintaining a high concentration in this organ in the presence of captopril for at least 3h; (3) The residual amount of [(125)I]-BPP 10c in the kidney of animals simultaneously treated with captopril suggest that the peptide can interact with other targets different from sACE in this organ. We also showed that Cy3-labeled BPP 10c was internalized by human embryonic kidney cells (HEK 293T). Taken together, these results suggest that sACE inhibition by captopril affects the tissue distribution of [(125)I]-BPP 10c and that the anti hypertensive effects of BPP 10c are not only dependent on sACE inhibition. PMID- 18160090 TI - Permeabilization of E. coli K12 inner and outer membranes by bothropstoxin-I, A LYS49 phospholipase A2 from Bothrops jararacussu. AB - Although lacking catalytic activity, the Lys49-PLA(2)s damage artificial membranes by a Ca(2+)-independent mechanism, and demonstrate a potent bactericidal effect. The relationship between the membrane-damaging activity and bactericidal effect of bothropstoxin-I (BthTx-I), a Lys49-PLA(2) from the venom of Bothrops jararacussu, was evaluated for the wild-type protein and a series of site-directed mutants in the active site and C-terminal regions of the protein. The membrane permeabilization effect against the inner and outer membranes of Escherichia coli K12 was evaluated by fluorescence changes of Sytox Green and N phenyl-N-naphthylamine, respectively. With the exception of H48Q, all mutants reduced the bactericidal activity, which correlated with a reduction of the permeabilization effect both against the inner bacterial membrane. No significant differences in the permeabilization of the bacterial outer membrane were observed between the native, wild-type recombinant and mutant proteins. These results suggest different permeabilization mechanisms against the inner and outer bacterial membranes. Furthermore, the structural determinants of bacterial inner membrane damage identified in this study correlate with those previously observed for artificial membrane permeabilization, suggesting that a common mechanism of membrane damage underlies the two effects. PMID- 18160091 TI - Optokinetic nystagmus in harbor seals (Phoca vitulina). AB - Harbor seals experience motion due to self-motion and to movement in the external world. However, motion vision has not been studied yet in marine mammals moving in the underwater world. To open up this research, optokinetic nystagmus (OKN) as a basic motion sensing and retinal image stabilizing reflex was studied in four harbor seals during stimulation with moving black-and-white stripe patterns. All seals responded with optokinetic eye movements. Detailed measurements obtained with one animal revealed a moderate gain for horizontal binocular OKN. Monocularly stimulated, the seal displayed a symmetrical OKN with slightly stronger responses to leftward moving stimuli, and, surprisingly, a symmetrical OKN was found in the vertical domain. PMID- 18160092 TI - Survival of Campylobacter jejuni in mineral bottled water according to difference in mineral content: application of the Weibull model. AB - The aim of the study was to examine the hypothesis proposed by Evans et al. [2003. Hazards of healthy living: bottled water and salad vegetables as risk factors for Campylobacter infection. Emerg. Infect. Dis. 9(10), 1219-1225] that mineral bottled water accidentally contaminated by Campylobacter jejuni would represent a risk factor for Campylobacter infection. Culturability of C. jejuni cells inoculated in low- and high-mineral bottled water during storage at 4 degrees C in the dark was performed by surface plating and modelled using the Weibull model. The loss of C. jejuni culturability observed in all conditions tested was shown to be dependent on strain, preculture condition and water composition. Following inoculation of C. jejuni, the rapid loss of culturability was not correlated to complete cell death as the passage into embryonated eggs enabled recovery of cells from the viable but non-culturable state. In conclusion, the sanitary risk associated with contaminated bottled water cannot be excluded although it is presumably low. Culture conditions, strain and water type must be taken into account in the evaluation of the risk factors as they influence significantly Campylobacter survival in water. PMID- 18160093 TI - Environmental quality assessment in estuarine ecosystems: use of biometric measurements and fecundity of the ragworm Nereis diversicolor (Polychaeta, Nereididae). AB - The ability to cope with environmental stress may be expensive in terms of energy and this cost of tolerance is suspected to have negative counterparts such as reduced growth and fecundity. To date, condition indices based on biometric measurements are currently used in bivalves or fish but do not exist in endobenthic worms, despite their interest as bioindicators for the sedimentary compartment in which the major part of pollutants is stored in aquatic environments. In the present work, several biometric variables (jaw and total body length, number of segments, the length of the first three segments L3, wet or dry weight) were measured in the ragworm Nereis diversicolor originating from clean (Authie) and polluted (Seine) estuaries (France) to study size-weight relationships. The production of oocytes by females (which represent 80-90% of the population) was quantified in the same specimens. In females from the polluted site, the condition was shown to be altered and, in parallel, the production of oocytes was lower than in females from the comparatively clean site. From an operational point of view, we recommend the use of the length L3 and the wet weight. Because fecundity is mainly dependent on size and weight in N. diversicolor, biometric measurements would be useful tools for assessing the biological quality of estuarine sediments. PMID- 18160094 TI - Residues of 18 organochlorine pesticides in 30 traditional Chinese medicines. AB - We analyzed 30 different traditional Chinese medicines (TCMs) to determine levels of contamination with organochlorine pesticides (OCPs). We tested for 18 pesticides: benzene hexachloride (BHC, including alpha-BHC, beta-BHC, gamma-BHC, delta-BHC), heptachlor, heptachlor epoxide, tecnazene, pentachloronitrobenzene (PCNB), hexachlorobenzene, aldrin, methyl pentachlorophenyl sulphide (MPCPS), alpha-endosulfan, trans-chlorodane, cis-chlorodane, p,p'-DDE, o,p'-DDT, p,p'-DDD, and p,p'-DDT. The sample extracts were analyzed by GC-ECD. A total of 280 samples of 30 different TCMs were analyzed. Our results showed that 75.8% of samples contained at least one of the above pesticides. The pesticides detected were tecnazene, hexachlorobenzene, PCNB, heptachlor, aldrin, alpha-BHC, beta-BHC, gamma-BHC, delta-BHC, p,p'-DDE, o,p'-DDT, p,p'-DDD p,p'-DDT. More than 50% of samples contained alpha-BHC (55.8%) and PCNB (55.8%); hexachlorobenzene was detected in 40.9% of samples, tecnazene in 19.5%, gamma-BHC in 16.7% and p,p'-DDE in 16.0%. Less than 10% of samples contained beta-BHC, delta-BHC, heptachlor, aldrin, o,p'-DDT, p,p'-DDT and p,p'-DDD. None of the 280 samples contained heptachlor epoxide, MPCPS, alpha-endosulfan, trans-chlorodane, or cis-chlorodane. Concentrations of OCPs in four samples exceeded the maximum allowable residue limits (MRLs) specified in the PRC Pharmacopoeia 2005. The results indicate that the most common contaminants among the 280 samples were alpha-BHC, PCNB, hexachlorobenzene, and tecnazene. Our results suggest that contamination of TCMs with OCPs is widespread. The MRLs for other OCPs commonly found in TCMs (e.g. hexachlorobenzene and tecnazene) should be set as quickly as possible, and the MRLs of OCPs in other TCMs should be established. PMID- 18160095 TI - Calibration and field application of a solvent-based cellulose membrane passive sampling device for the monitoring of polar herbicides. AB - A passive sampler device selective for hydrophilic analytes was constructed from cellulose membrane (40microm thickness) pre-stained with ruthenium red for 96 168h to impede degradation of the cellulose. The sampling device consisted of pre stained cellulose membrane tubing containing a binary mixture of the solvents 1 dodecanol and 2,2,4-trimethylpentane as the sequestering medium. A laboratory flow-through system was used to investigate the rates of uptake of herbicides into the solvent mixture of the device and their release. The target herbicides were diuron, atrazine, metolachlor and molinate. Uptake of the herbicides into the solvent mixture of the cellulose membrane device was linear for up to 22 days, and daily sampling rates were determined. Release half-lives from the solvent mixture of the sampling device varied from 14 days for diuron, 15 days for atrazine, 84 days for metolachlor and 28 days for molinate. A field study was undertaken to determine if herbicide concentrations in agricultural drainage water derived from the passive sampler devices deployed for periods from 7 to 22 days, using the laboratory-derived sampling rates, would compare closely with time-weighted average herbicide concentrations determined from extractions of daily composite water samples. The concentrations of diuron, atrazine, metolachlor and molinate determined using the cellulose membrane devices were within twofold of the cumulated mean of the daily drainage water extractions. PMID- 18160096 TI - Intra-specific variation in resting metabolic rate in MF1 mice is not associated with membrane lipid desaturation in the liver. AB - The 'membrane pacemaker' hypothesis provides a putative mechanistic linkage between variations in energy metabolism, rates of ageing and lifespan across different species. Within species we have found positive associations between longevity and metabolism, which contrast the inter-specific trends. It is of interest to know therefore how levels of lipid desaturation in membranes are linked to variation in metabolism between individuals within species. We explored this problem by extracting membrane fatty acids from the livers of mice that varied in their metabolic rate, in a strain (MF1) where we have previously demonstrated a positive association between metabolism and lifespan. We measured resting metabolic rate (RMR) in 60 mice, each measured on three occasions, and measured their body compositions using dual energy X-ray absorptiometry (DXA). We selected 28 individuals that exhibited a wide variation in their mean resting metabolic rates (RMR) and extracted membrane lipids from the livers of these mice post mortem and analysed them for the patterns of contribution of different fatty acids. We then sought associations between the levels of membrane desaturation and the individual variability in RMR, using the proportional contributions of each fatty acid as predictors in a stepwise regression or by re-describing the variation in fatty acyl lipids using a PCA analysis and then seeking associations between scores on the derived components and RMR. We used whole animal RMRs and also RMR with the effects of body composition (fat free mass) removed. The level of individual variation in RMR was consistent with our previous observations. There was a significant positive association (p=0.019) between the proportion of palmitic acid (16:0) in the membranes and RMR, which was strengthened (p=0.014) when we adjusted RMR for differences in fat free mass. The proportion of palmitic acid (16:0) explained 20.9% of the individual variation in residual RMR. There was no association between RMR or mass adjusted RMR and the proportional representation of any other fatty acid, including 22:6 (DHA) predicted by the membrane pacemaker hypothesis to be of particular significance. High levels of saturated fatty acids in the membranes of mice with high rates of metabolism may contribute to their greater longevity, but the mechanism tying together increased membrane saturation with elevated RMR remains unclear. PMID- 18160097 TI - Platelet as a physiological model to investigate apoptotic mechanisms in Alzheimer beta-amyloid peptide production. AB - Although neuronal apoptosis in Alzheimer's disease is generally interpreted as the consequence of the toxicity of extracellular beta-amyloid (Abeta) peptide aggregates, some experimental results provide evidence that the Abeta overproduction can be the result of a primary neuronal degeneration. As platelets are considered a good model where to study proteolytic processing of the amyloid precursor protein (APP), we exposed platelets to the proapoptotic agent ionomycin and analyzed Abeta40 and Abeta42 levels in the intracellular and extracellular compartments. The activation of apoptotic pathways in platelets has been verified by mitochondrial membrane depolarization, exposure of phosphatidylserine, protease activation and morphological changes. A significative increase in intraplatelet Abeta40, but not Abeta42, was observed after 10 min treatment with ionomycin. Thus, the activation of apoptotic pathways in platelets determines an altered processing of APP leading to elevated levels of intracellular Abeta40. The specific intracellular production of Abeta40 represents a potential threat to the cells since very high local Abeta40 concentration increases the risk of its aggregation and toxicity. As a result, Abeta40 might be dangerous even before it becomes secreted rendering neurons highly vulnerable. PMID- 18160099 TI - Multi-parametric flow cytometric cell cycle analysis using TO-PRO-3 iodide (TP3): detailed protocols. AB - TO-PRO-3 iodide (TP3), a monomeric cyanine nucleic acid stain with a peak absorbance at 642 nm and emission at 661 nm, is best excited by a helium-neon (HeNe) laser (633nm). It was tested in monocytes and different cell lines under conditions of different fixatives, dye concentrations, labeling kinetics and RNAse concentrations for mono-, bi- and tri-parametric flow cytometric cell cycle analysis to establish the best protocol for DNA analysis in terms of G1 peak CV, G2/G1 ratio and minimal amount of debris. A linear increase in G1 peak position was found from 0.1 to 2 microM TP3 concentrations. Fixatives 70% ethanol or 1% methanol-free formaldehyde, followed by 70% ethanol, resulted in the best DNA histograms. Although different protocols were found to be cell-type specific, in general, excellent results were obtained with 30 min incubation with 0.5 microM TP3 plus RNAse in almost all cell lines tested. These data show that TP3 is an alternative method to propidium iodide (PI), the most commonly used DNA-specific probe in flow cytometry. The most important advantage of using TP3 in combination with other fluorochromes, such as fluorescein isothiocyanate (FITC) or phycoerythrin (PE) in bi- or tri-parametric flow cytometric analysis, is that there is no need for fluorescence compensation for the TP3 signals. PMID- 18160098 TI - The role of telomere biology in bone marrow failure and other disorders. AB - Telomeres, consisting of nucleotide repeats and a protein complex at chromosome ends, are essential in maintaining chromosomal integrity. Dyskeratosis congenita (DC) is the inherited bone marrow failure syndrome (IBMFS) that epitomizes the effects of abnormal telomere biology. Patients with DC have extremely short telomere lengths (<1st percentile) and many have mutations in telomere biology genes. Interpretation of telomere length in other IBMFSs is less straightforward. Abnormal telomere shortening has been reported in patients with apparently acquired hematologic disorders, including aplastic anemia, myeolodysplasia, paroxysmal nocturnal hemoglobinuria, and leukemia. In these disorders, the shortest-lived cells have the shortest telomeres, suggestive of increased hematopoietic stress. Telomeres are also markers of replicative and/or oxidative stress in other complex disease pathways, such as inflammation, stress, and carcinogenesis. The spectrum of related disorders caused by mutations in telomere biology genes extends beyond classical DC to include marrow failure that does not respond to immunosuppression, idiopathic pulmonary fibrosis, and possibly other syndromes. We suggest that such patients be categorized as having an inherited disorder of telomere biology. Longitudinal studies of patients with very short telomeres but without classical DC are necessary to further understand the long term sequelae, such as malignancy, osteonecrosis/osteoporosis, and pulmonary and liver disease. PMID- 18160100 TI - Image analysis of the AgNOR response in ras-transformed human breast epithelial cells. AB - The argyrophylic staining of the nucleolar organizer regions (AgNOR positive response) in interphase nuclei is often related directly to the cellular demand for ribosome biogenesis and is considered of relevance in studies of tumor pathology. Transformation of human breast epithelial MCF-10A cells by the c-Ha ras oncogene results in altered growth, invasiveness and tumorigenicity in nude mice. Since ras transformation may be associated with a more intense nucleolar activity, we examined the influence of transfection by the Ha-ras oncogene on AgNOR staining response in MCF-10A cells. Following assessment of the AgNOR response with video image analysis, the AgNOR-positive areas and the AgNOR area/nuclear area ratio, but not the number of AgNOR aggregates or dots per nucleus, were found to be much higher after ras transformation. A role of the Ha ras transformation on the nucleolar activity of the MCF-10A is thus suggested as assessed by the AgNOR staining. Based on data in the literature, it is also hypothesized that a decreased wild-type p53 level, possibly promoted by the ras transformation, may be associated with the increased AgNOR response. PMID- 18160101 TI - A cytoarchitectonic and myeloarchitectonic study of the insular cortex of the bull, Bos taurus. AB - The mammalian Insula is characterised by considerable morphologic variability, however, it shows a cytoarchitectonic homogeneity within the species so far studied. Three cytoarchitectonic areas are generally recognisable in the Insula: an "agranular", a "dysgranular" and a "granular" area. The numerous functions attributed to the Insula (visceral sensory, visceral motor, somatosensorial, supplementary motor area, area speech and/or language related, etc.) have recently attracted renewed interest. Considering the involvement of the Insula in the control of gastroenteric motility and the structural and functional complexity of this region in ruminants, it seemed interesting to analyse the cyto and myelo-architectonic arrangement of the Insula of the bull, Bos taurus. Unlike that of the other species described, all the insular Neocortex of the bull, is of the "agranular" type. The latter includes at least four variants that differ according to the organisation of the cortical layer pattern. Considering the available evidence in laboratory animals and primates of a role played by the agranular insular cortex in gastroenteric motility control, the trademark presence of a very much extended insular agranular cortex in the Insula of the bull may be related to the necessity of motor activity control of a gastroenteric complex that is extensively more developed than in the monogastric species. PMID- 18160102 TI - In vivo activity of ABT-869, a multi-target kinase inhibitor, against acute myeloid leukemia with wild-type FLT3 receptor. AB - Neoangiogenesis plays an important role in leukemogenesis. We investigated the in vivo anti-leukemic effect of ABT-869 against AML with wild-type FLT3 using RFP transfected HL60 cells with in vivo imaging technology on both the subcutaneous and systemic leukemia xenograft models. ABT-869 showed a five-fold inhibition of tumor growth in comparison with vehicle control. IHC analysis revealed that ABT 869 decreased p-VEGFR1, Ki-67 labeling index, VEGF and remarkably increased apoptotic cells in the xenograft models. ABT-869 also reduced the leukemia burden and prolonged survival. Our study supports the rationale for clinically testing an anti-angiogenesis agent in AML with wild-type FLT3. PMID- 18160103 TI - The biological and behavioral basis of upper limb asymmetries in sensorimotor performance. AB - Asymmetries in upper limb performance are a fundamental aspect of human behavior. This phenomenon, commonly known as handedness, has inspired a great deal of research over the course of the past century garnering interest across a multitude of scientific domains. In the present paper, a thorough review of this literature is provided focusing on the current state of knowledge regarding neuro anatomical and behavior-based arm asymmetries. It is hoped that this information will provide a basis for new insights regarding the design and implementation of future studies regarding arm laterality. PMID- 18160104 TI - Expression of SH2D2A in T-cells is regulated both at the transcriptional and translational level. AB - The T-cell specific adapter protein (TSAd) encoded by the SH2D2A gene is up regulated in activated human CD4+ T-cells in a cAMP-dependent manner. Expression of SH2D2A is important for proper activation of T-cells. Here, we show that SH2D2A expression is regulated both at the transcriptional and translational level. cAMP signaling alone induces TSAd-mRNA expression but fails to induce increased TSAd protein levels. By contrast, TCR engagement provides signals for both TSAd transcription and translation. We further show that cAMP signaling can prime T-cells for a more prompt expression of TSAd protein upon TCR stimulation. Our study thus points to a novel mechanism for how cAMP signaling may modulate T cell activation through transcriptional priming of resting cells. PMID- 18160105 TI - Solid-phase extraction enhances detection of beta-amyloid peptides in plasma and enables Abeta quantification following passive immunization with Abeta antibodies. AB - We have previously developed a solid-phase extraction (SPE) procedure to enable the detection of beta-amyloid (Abeta) peptides in brain tissue from non transgenic animals. We have now adapted these methods to enrich the Abeta fraction in cerebrospinal fluid (CSF) and plasma. Human CSF and plasma and Tg2576 mouse plasma were subjected to guanidine denaturation followed by SPE in 96-well cassettes. The resulting eluates could be concentrated significantly to enhance detection of low-abundance Abeta peptides by immunoassay. The concentrated eluates diluted in a linear fashion with consistent recovery between SPE columns. This technique was therefore used to facilitate quantification of Abeta1-X, 1-40, 1-42, and 1-38 peptides in normal human CSF and plasma samples. SPE sample preparation was also applied to the plasma of mice dosed peripherally with a monoclonal antibody raised against Abeta. When such samples were assayed directly, the presence of the systemically administered antibody interfered with the subsequent immunoassay, by preventing detection of antibody-bound Abeta. After subjecting plasma from antibody-treated animals to denaturation and SPE, the antibody-antigen complex was disrupted, and the Abeta fraction could be isolated from the antibody-containing fraction. Application of this method allowed for detection of a 100-fold increase in plasma Abeta1-40 following treatment of Tg2576 mice or wild type littermate control mice with Abeta40 specific monoclonal antibody 9TL. Given the availability of a variety of SPE matrices, we hypothesize that these methods could facilitate plasma antigen retrieval using multiple therapeutic antibody approaches. PMID- 18160106 TI - Protective and cross-protective mucosal immunization of mice by influenza virus type A with bacterial adjuvant. AB - Mucosal immunization by inactivated viruses often fails to evoke a sufficient immune response. Intensive efforts have been made to enhance the response by suitable adjuvants. We used the G+ nonpathogenic delipidated bacterium Bacillus firmus with pronounced immunostimulatory properties as an adjuvant for immunizing mice with inactivated influenza virus type A. BALB/c mice were immunized intratracheally with inactivated influenza A H1N1 and H3N2 viruses. The production of antibodies in sera and secretions was determined by the ELISA. The local situation in the lungs was assessed histologically and by testing the cytokine expression. The protective and cross-protective effect against infection was tested in in vivo experiments after infection with influenza virus A H1N1. B. firmus as adjuvant increased both systemic and mucosal antibody responses, improved protection against homologous virus and induced cross-protection against virus H1N1 after immunization with virus H3N2. PMID- 18160107 TI - Human phagocytes lack the ability to kill Mycobacterium gordonae, a non pathogenic mycobacteria. AB - Non-pathogenic mycobacteria, like Mycobacterium gordonae, are rarely associated to disease. The analysis of the mechanisms which are successful against them in the human host may provide useful information to understand why they fail against the pathogenic M. tuberculosis. We have developed an infection model to test the ability of human phagocytes to kill two strains of M. gordonae, HL184G and an attenuated variety, HL184Gat. As controls we included a strain of M. tuberculosis (HL186T) and another one of L. pneumophila (ATCC13151). We observed that human phagocytes lack the intrinsic ability to eliminate either M. gordonae or M. tuberculosis, but they can kill the attenuated strain. We found a relationship between pathogenicity and the pattern of cytokine production. Thus, both the pathogenic M. tuberculosis and Legionella pneumophila, but not the non-pathogenic M. gordonae, induced the production of significantly different levels of IL 1beta, IL-6 and TNF-alpha in monocytes and IL-8 in neutrophils. Although both monocytes and neutrophils killed HL184Gat, but not HL184G, the patterns of cytokine production induced by either strain were identical. Addition of INF gamma and/or TNF-alpha did not enhance the antimycobacterial activity of phagocytes. PMID- 18160109 TI - The reduction in electric footshocks perception interferes with the amnesic effect of scopolamine. AB - In a passive avoidance paradigm, administration of scopolamine in mice seems to provoke forgetting of electric shocks they received when entering into the black compartment. We observed that, in fact, scopolamine reduced the shocks perception. Since, on the hot plate test, scopolamine did not affect latencies of avoidance reactions, this effect did not correspond to analgesia. Thus, in our experimental conditions, scopolamine effect did not exclusively result from a deficit of passive avoidance learning but likely resulted from its anti-sweating properties, thus reducing the shocks perception. PMID- 18160108 TI - Neurotoxic effects of DSP-4 on the central noradrenergic system in male zebra finches. AB - When administered systemically, the noradrenergic neurotoxin N-(2-chloroethyl)-N ethyl-2-bromobenzylamine (DSP-4) appears to target the noradrenergic innervation originating in the locus coeruleus causing long-term decrements in noradrenergic function. In songbirds, DSP-4-treatment decreased female-directed singing by males and copulation solicitation responses of females to male songs. However, DSP-4 treatment in songbirds did not lower measures of NE function in the brain to the same extent as it does in mammals. The current study had two goals: determining if two DSP-4 treatments 10 days apart would cause significant decrements in noradrenergic function in male zebra finches and determining if, as in other species, the noradrenergic innervation of midbrain and cortical areas would be profoundly affected while hypothalamic areas were spared. Dopamine-beta hydroxylase immunoreactivity (DBH-ir) was quantified in thirteen brain regions (five vocal control nuclei, one auditory nucleus, two hypothalamic nuclei, and five additional areas that demonstrated high DBH labeling in controls). Within 20 days, DSP-4 treatment profoundly reduced the number of DBH-ir cells in both the locus coeruleus and ventral subcoeruleus. Unlike a previous study, DBH labeling delineated four out of five vocal control nuclei and an auditory nucleus. As expected, DSP-4 treatment significantly decreased DBH labeling in all areas examined in the mesencephalon and telencephalon without significantly affecting DBH-ir in hypothalamic areas. This double treatment regime appears to be much more effective in decreasing noradrenergic function in songbirds than the single treatment typically used. PMID- 18160110 TI - Ecotoxicity of CdTe quantum dots to freshwater mussels: impacts on immune system, oxidative stress and genotoxicity. AB - The purpose of this study was to examine the toxic effects of cadmium-telluride (CdTe) quantum dots on freshwater mussels. Elliption complanata mussels were exposed to increasing concentrations of CdTe (0, 1.6, 4 and 8 mg/L) and cadmium sulfate (CdSO(4), 0.5mg/L) for 24h at 15 degrees C. After the exposure period, they were removed for assessments of immunocompetence, oxidative stress (lipid peroxidation) and genotoxicity (DNA strand breaks). Preliminary experiments revealed that CdTe dissolved in aquarium water tended to aggregate in the particulate phase (85%) while 15% of CdTe was found in the dissolved phase. Immunotoxicity was characterized by a significant decrease in the number of hemocytes capable of ingesting fluorescent beads, and hemocyte viability. The cytotoxic capacity of hemocytes to lyse mammalian K-562 cells was significantly increased, but the number of circulating hemocytes remained unchanged. Lipid peroxidation was significantly increased at a threshold concentration of 5.6 mg/L in gills and significantly reduced in digestive glands at a threshold concentration <1.6 mg/L CdTe. The levels of DNA strand breaks were significantly reduced in gills at <1.6 mg/L CdTe. In digestive glands, a transient but marginal increase in DNA strand breaks occurred at the lowest concentration and dropped significantly at the higher concentrations. A multivariate analysis revealed that the various response patterns differed based on the concentration of CdTe, thus permitting the identification of biomarkers associated with the form (colloidal vs. molecular) of cadmium. PMID- 18160111 TI - Distinctive accumulation patterns of Cd(II), Cu(II), and Cr(VI) in tissue of the South American teleost, pejerrey (Odontesthes bonariensis). AB - The time- and concentration-dependent accumulation patterns of three environmentally relevant metals, with different chemical and biological properties, were comparatively evaluated in tissue of South American fish, pejerrey (Odontesthes bonariensis). Six-month-old juvenile pejerrey were exposed to 1, 5, or 10 microgl(-1) of Cd(2+); 10, 50, or 100 microgl(-1) of Cu(2+); or 100, 500, or 1000 microgl(-1) of Cr(6+) during 16 days. Tested concentrations ranged from those that caused no significant accumulation to those that induced growth reduction or even mortality (Cd and Cu). Concentration-dependent accumulation of Cd and Cr presented similar patterns, characterized by a linear and exponential relationship in the gill and liver, respectively, with a greater slope at longer exposure times. Differently, essential Cu showed a linear relationship in both tissues, and no slope increase was observed in the gill. The gill time-dependent accumulation pattern of Cd exhibited relatively long times to half-saturation (t(1/2(1))=36.6 days) and high-saturation values (S((1))=205.4 nmolg(-1)). The opposite pattern was observed for Cu (t(1/2(100))=0.6 days; S((100))=24.4 nmolg(-1)), while Cr showed an intermediate position (t(1/2(1000))=2.8 days; S((1000))=50.2 nmolg(-1)). In the liver, times to half saturation for the three metals were as follows: Cd=4.7 days; Cu=2.5 days; Cr=1.6 days. Conversely, different saturation concentrations were observed among metals, Cu (S((100))=102) presenting the highest values, Cd (S((10))=4.4) the lowest, and Cr presenting an intermediate position (S((1000))=24.7). Bioconcentration of non essential Cd was high in the gill (2000-fold) and low in the liver (50-fold). On the other hand, the essential element Cu was poorly retained by the gill (15 fold) and accumulated mainly in the liver (50-fold). Differently, Cr was quickly and evenly accumulated by both organs, but barely bioconcentrating (2.5- and 1.5 fold levels as compared to the original in the gill and liver, respectively). The ratio of gill/liver concentration in exposed fish clearly differed from that of non-exposed fish, and was characteristic for each metal (Cd>1; Cu<1; Cr<9), representing a potential tool to assess exposure. The accumulation patterns of studied metals in the tissue of pejerrey corresponded more with those reported for sensitive than for tolerant fish species. Distinctive tissue accumulation patterns in relation to growth reduction and non-acute lethality suggests different target tissues for sub-chronic effects, and would partially explain differences in the relative toxicity of these metals. PMID- 18160112 TI - Assessing effects of a mining and municipal sewage effluent mixture on fathead minnow (Pimephales promelas) reproduction using a novel, field-based trophic transfer artificial stream. AB - The Junction Creek watershed, located in Sudbury, ON, Canada receives effluent from three metal mine wastewater treatment plants, as well as a municipal wastewater (MWW) discharge. Effects on fish have been documented within the creek (decreased egg size and increased metal body burdens). It has been difficult to identify the cause of the effects observed due to the confounded nature of the creek. The objectives of this investigation were to assess the: (1) effects of a mine effluent and municipal wastewater (CCMWW) mixture on fathead minnow (FHM; Pimephales promelas) reproduction in an on-site artificial stream and (2) importance of food (Chironomus tentans) as a source of exposure using a trophic transfer system. Exposures to CCMWW through the water significantly decreased egg production and spawning events. Exposure through food and water using the trophic transfer system significantly increased egg production and spawning events. Embryos produced in the trophic-transfer system showed similar hatching success but increased incidence and severity of deformities after CCMWW exposure. We concluded that effects of CCMWW on FHM were more apparent when exposed through the water. Exposure through food and water may have reduced effluent toxicity, possibly due to increased nutrients and organic matter, which may have reduced metal bioavailability. More detailed examination of metal concentrations in the sediment, water column, prey (C. tentans) and FHM tissues is recommended to better understand the toxicokinetics of potential causative compounds within the different aquatic compartments when conducting exposures through different pathways. PMID- 18160113 TI - The endoplasmic reticulum chaperone calreticulin is recruited to the uropod during capping of surface receptors in Entamoeba histolytica. AB - Calreticulin (CRT), an intracellular chaperone protein, is crucial for proper folding and transport of proteins through the endoplasmic reticulum (ER). It has recently been identified as a critical regulator of some several different cellular functions such as migration, phagocytosis of apoptotic cells and cytotoxic T lymphocyte- or natural killer cell-mediated lysis. Characterization of CRT isolated from parasites may thus help to decipher the contribution of this protein in the parasites' biology and host-parasite interactions. Here, we report descriptive data on the localization of Entamoeba histolytica's CRT at rest and following cap formation by Concanavalin A. As expected, CRT from E. histolytica localizes in the ER. However, the protein was surprisingly found to localize to the parasite surface and, furthermore, to concentrate in the uropod following activation of surface receptors by capping with Concanavalin A. PMID- 18160114 TI - A Presenilin-like protease associated with Plasmodium falciparum micronemes is involved in erythrocyte invasion. AB - We describe identification of a Plasmodium falciparum microneme protease involved in RBC invasion. From the yeast two-hybrid screening of a P. falciparum cDNA library, we have identified a 47 kDa membrane protein that interacted with the 5ABC domain of human RBC band 3. This protein shared homology with a Presenilin type aspartyl protease, the signal peptide peptidase (SPP). An antibody raised against a predicted exposed region of this protein reacted specifically to a single band of approximately 47 kDa in the P. falciparum protein extract. Immunofluorescence microscopy suggested that this protein co-localized with the microneme protein EBA-175 in schizonts, and immunoelectron microscopy established that it is primarily localized to micronemes in merozoites. Functional characterization of Plasmodium falciparum signal peptide peptidase (PfSPP), demonstrates that an antibody to PfSPP blocks RBC invasion by P. falciparumin vitro. Native and recombinant PfSPP bound directly to the 5ABC domain of band 3 in solution and the binding of PfSPP to RBCs was chymotrypsin-sensitive, but trypsin and neuraminidase-resistant. Together, these results suggest that host band 3 interacts with PfSPP during RBC invasion presumably following parasite microneme discharge. PfSPP is the first microneme-associated intramembrane aspartyl protease identified in the apicomplexan parasites that interacts with a major transmembrane receptor on host erythrocytes. PMID- 18160115 TI - Recognition and perception of elder abuse by prehospital and hospital-based care providers. AB - The purposes of this study were to evaluate the extent of exposure, knowledge and attitudes of prehospital care providers (PCPs) and hospital care providers (HCPs) to elder abuse and neglect. A 20-question survey was designed to determine the providers' perception, knowledge and ability to identify patients that were potential victims of elder abuse and/or neglect. The surveys were distributed at four Maryland statewide conferences during 2006. A total of 645 surveys were distributed at the start of the individual conferences and 400 completed surveys were returned. Of the respondents, 272 (68.2%) were PCP (emergency medical services=EMSs) and 127 (31.8%) were HCP. During the past 12 months, 51.3% of those surveyed did not have reason to suspect any patients were exposed to abuse or neglect, although 60.5% admitted little or no contact with the elderly. In an attempt to determine respondent's ability to recognize potential abuse and neglect patients, scenario-type questions were used. Respondents believed a decubital ulcer (bedsore) was a positive indicator (83.5%) of abuse/neglect and 92.8% indicated that the elderly could suffer from injuries similar to "shaken baby syndrome". When questioned about skin bruises as a possible indicator of abuse, only 69.3% of the respondents identified it as a possible sign of abuse. Seventy-one percent of respondents indicated that burns are not common in the elderly and could be another sign of elder abuse. One-in-three providers indicated they would suspect other reasons (dementia, depression, etc.) for the report of a sexual assault in an elderly patient. Eighty-nine percent of providers were aware that healthcare providers in the State of Maryland are required to report suspected elder and vulnerable patient abuse and/or neglect to law enforcement or social services' agencies. When asked to define elder abuse as a medical or social problem, 25.0% of providers stated that it was a social problem. Over 95% of the providers suspected the existence of abuse, neglect and domestic violence among the elderly were not rare events. In Maryland, there are a limited number of specific educational programs dealing with abuse and neglect of the elderly. A statewide training program is needed to ensure PCP and HCP can recognize the signs and symptoms of elder abuse and neglect, and to ensure that the providers are aware of their legal requirements for reporting the abuse to the proper state or local agencies. PMID- 18160116 TI - Simulating the BSE epidemic and multiplication factor in dairy herds in Japan. AB - With the objective of evaluating the effectiveness of an administrative guidance on the use of ruminant meat-and-bone meal in ruminant feed, effective from April 1996 to September 2001, we developed a model to simulate the evolution of the BSE epidemic and to estimate the BSE multiplication factor (K) in the Japanese dairy population. The output that provided the best fit to the number of BSE cases both observed and predicted to date suggest that the probability that bovine MBM was fed back to cattle was 14.2-75.2% and 0.129-0.570% during the periods from 1992 to April 1996 and from April 1996 to October 2001, respectively. Given these estimates, the value of K would have peaked in 1995 at 40-48 and then declined to 0.32-0.67 between 1997 and 2001. These results suggest that the administrative guidance was effective in reducing the amount of MBM fed to cattle by a factor of 104-141 and was perhaps enough to drive the epidemic towards extinction. PMID- 18160117 TI - Direct detection of RNA transcription by FRET imaging using fluorescent protein probe. AB - We have constructed a reporter system for intracellular direct detection of RNA transcription that consists of two biomolecular components. The first part is a GFP-based recombinant protein probe (YRG0C-11ad) containing the RNA-binding Rev peptide between ECFP and EYFP. The second component is RRE-RNA, which specifically binds to the Rev-peptide. Cells stably expressing YRG0C-11ad were identified by an increased FRET signal after direct transfection or intracellular transcription of RRE-RNA. In addition, the signal increase is more noticeable if tandemly repeated RRE-RNA is used as the reporter. Untranslatable non-coding RNAs are regarded as regulators of cellular gene expression, but they are difficult to study using indirect reporter systems that are dependent on translational products. Direct detection of reporter RNA would be a useful method for the detection of intracellular promoter activity during transcription of untranslatable RNAs. PMID- 18160118 TI - Species-specific deletion of the viral attachment glycoprotein of avian metapneumovirus. AB - The avian metapneumovirus (AMPV) genome encodes the fusion (F), small hydrophobic (SH), and attachment glycoprotein (G) as envelope glycoproteins. The F and G proteins mainly function to allow viral entry into host cells during the early steps of the virus life cycle. The highly variable AMPV G protein is a major determinant for distinguishing virus subtypes. Sequence analysis was used to determine if any differences between avian or mammalian cell propagated subtype C AMPV could be detected for the 1.8kb G gene. As a result, the complete 1.8kb G gene was found to be present when AMPV was propagated in our immortal turkey turbinate (TT-1) cell line regardless of passage number. Surprisingly, AMPV propagated for 15 or more passages in mammalian Vero cells revealed an essentially deleted G gene in the viral genome, resulting in no G gene mRNA expression. Although the Vero cell propagated AMPV genome contained a small 122 nucleotide fragment of the G gene, no other mRNA variants were detected from either mammalian or avian propagated AMPV. The G gene truncation might be caused by cellular molecular mechanisms that are species-specific. The lack of viral gene deletions suggests that avian cell propagated AMPV will provide a better alternative host for live recombinant vaccine development based on a reverse genetics system. PMID- 18160119 TI - Low discomfort and pain associated with intensified insulin therapy in children and adolescents. AB - OBJECTIVE: Discomfort during insulin injection and self-monitoring of blood glucose (SMBG) is a potential obstacle in diabetes therapy, but its prevalence and extent in relation to clinical variables is uncertain. RESEARCH DESIGN AND METHODS: We prospectively assessed treatment-associated discomfort and pain in an unselected cohort of patients (60 boys and 52 girls; mean age 14.6+/-3.0 years, mean A1C 8.0+/-1.4%) with type 1 diabetes and multiple daily self-injections of insulin, using visual analogue/verbal rating scales (range, 0-10) and a six-item questionnaire. RESULTS: Pain during insulin injection was absent to very low in 91.9% of patients, and its intensity was independent of age, gender, diabetes duration, current A1C, injection volume, or type of insulin. Injection was more unpleasant than SMBG in 64.2% of patients (mean difference of pain score, 1.0+/ 1.7, p<0.0001). Injection into the upper arm was less painful than into the thigh and abdomen. Surprisingly, painlessness of injection and SMBG was not judged an important treatment goal by 22.0 and 32.9% of patients, respectively. Logistic difficulties (41.2% of responses) and time requirements (23.8%), but not pain (10.1%), were considered most relevant problems. CONCLUSIONS: In young patients with access to optimized diabetes care, pain during insulin injection and SMBG is infrequent or mild, and not widely perceived as problematic, thus encouraging the use of multiple daily injection treatment. PMID- 18160120 TI - 2-year effects of pioglitazone add-on to sulfonylurea or metformin on oral glucose tolerance in patients with type 2 diabetes. AB - AIM: We report the effectiveness of the thiazolidinedione, pioglitazone, as add on medication to metformin or sulfonylurea in reducing post-load serum glucose levels, as assessed by 3-h oral glucose tolerance testing (OGTT). METHODS: Adult patients with Type 2 diabetes took part in one of two large-scale, 2-year clinical trials. One study compared pioglitazone treatment as add-on to failing metformin therapy (N=317) with add-on gliclazide treatment to metformin (N=313). The other study compared combination therapy with pioglitazone added to existing failing sulfonylurea therapy (N=319) with metformin treatment in addition to sulfonylurea (N=320). HbA(1c) and fasting plasma glucose concentrations were measured at baseline and throughout the study and at the final visit at week 104. At selected centers (N=299 patients), a 3-h OGTT was performed at baseline and at week 104. RESULTS: At week 104, mean HbA(1c) reduction from baseline was 0.89% for pioglitazone and 0.77% for gliclazide addition to metformin (p=0.200) and 1.03% with pioglitazone and 1.16% with metformin addition to sulfonylurea (p=0.173) in the total patient cohort. In the 299 patients who underwent OGTT, 2 years of treatment with pioglitazone, whether added to existing metformin or sulfonylurea medication, resulted in decreases in glucose excursions after an oral glucose load without increasing post-load serum insulin concentrations. In contrast, gliclazide in combination with metformin therapy caused increases in both post-load serum glucose and insulin excursions after 2 years, whereas metformin add-on to sulfonylurea did not have a significant effect on post-load serum glucose concentrations and resulted in an increase in insulin levels. CONCLUSIONS: There were no significant differences in HbA(1c) levels between groups. However, 2-year treatment with pioglitazone as an add-on to either failing metformin or sulfonylurea therapy improved post-load glucose excursions without affecting insulin secretion. In contrast, glucose excursions were not improved by gliclazide or metformin add-on therapy, despite increases in post load insulin levels. These data suggest that pioglitazone reduces peripheral insulin resistance via mechanisms different from those of metformin. PMID- 18160121 TI - Identification of a ferrochelatase mutation in a Chinese family with erythropoietic protoporphyria. AB - BACKGROUND/AIMS: Erythropoietic protoporphyria (EPP) is a rare autosomal dominant disorder of heme biosynthesis characterized by a partial decrease in ferrochelatase (FECH) activity leading to excessive accumulation of protoporphyrin. While a majority of EPP patients only exhibit photosensitivity, a small percentage of patients also develop liver complications and need liver transplantation. METHODS: In this study, we have sequenced the ferrochelatase gene of a Chinese EPP patient who suffered from EPP-related liver complications. RESULTS: A nonsense mutation in exon 4, 343C>T, introducing a premature stop codon at position arginine 115, was identified in the proband as well as her symptomatic mother and brother, but was absent in her father. All the family members with overt photosensitivity also carried the low-expressed allele IVS3 48c, whose prevalence in the Chinese Han population was determined to be 41.35% and which was also functional in producing an aberrant 63 bp insertion. CONCLUSION: We describe the first FECH mutation identified in the Chinese Han population and report a high frequency of the hypomorphic IVS3-48c allele in China. PMID- 18160122 TI - Cost-effective practices in the blood service sector. AB - OBJECTIVES: The objective of this study is to recommend alternative policies, which are tested on a computer simulation model, towards a more cost-effective management of the blood supply chain in the UK. METHODS: With the use of primary and secondary data from the National Blood Service (NBS) and the supplied hospitals, statistical analysis is conducted and a detailed discrete event simulation model of a vertical part of the UK supply chain of blood products is developed to test and identify good ordering, inventory and distribution practices. RESULTS: Fewer outdates, group substitutions, shortages and deliveries could be achieved by blood banks: holding stock of rare blood groups of red blood cells (RBC), having a second routine delivery per weekday, exercising a more insensitive ordering point for RBC, reducing the total crossmatch release period to less than 1.5 days, increasing the transfusion-to-crossmatch ratio to 70%, adhering to an age-based issuing of orders, holding RBC stock of a weighted average of approximately 4 days. CONCLUSIONS: The blood supply simulation model can offer useful pieces of advice to the stakeholders of the examined system which leads to cost reductions and increased safety. Moreover, it provides a great range of experimental capabilities in a risk-free environment. PMID- 18160123 TI - Hybrid (intravenous and oral) administration of vinorelbine plus cisplatinum followed by oral vinorelbine as first-line therapy of advanced non-small cell lung cancer: a phase II study. AB - BACKGROUND: The combination of alternate i.v./oral (hybrid) administration of vinorelbine (VNR) plus cisplatin (CDDP), followed by oral VNR, could result in a more suitable first-line regimen for patients (pts) with advanced non-small cell lung cancer (aNSCLC) in the outpatient setting. METHODS: The induction treatment consisted of CDDP 80 mg/m(2) i.v. and VNR 25 mg/m(2) i.v. day 1 and VNR 60 mg/m(2) oral day 8, every 3 weeks for 4 courses. A dose escalation of VNR to 80 mg/m(2) oral from day 8 of the second course and to 30 mg/m(2) i.v. from day 1 of the third course was planned in the absence of G3-4 toxicity. Pts with disease control after 4 courses underwent consolidation treatment with oral VNR 80 mg/m(2) days 1 and 8 every 3 weeks up to intolerance or progression. RESULTS: Fifty-three pts entered the study: 80% males; median age 63 years (range 43-71); median ECOG PS 0 (range 0-1); histotype: adenocarcinoma 59%, epidermoid 31%, undifferentiated 10%; disease stage: IIIB 22%, IV 70%, recurrent disease 8%. The objective response was as follows: 1 (2%) CR, 20 (38%) PR, 16 (30%) SD, 11 (21%) PD and 5 (9%) pts were not assessable. Median TTP and OS were 6 and 10 months, respectively. G3-4 neutropenia was observed in 23 and 24% of pts in the induction and in the consolidation phases, respectively, with febrile neutropenia in 6 pts (11%) and 2 (8%), respectively. G3-4 non-haematological toxicity was rare, being represented by nausea-vomiting and neurotoxicity in 3 pts (6%) in the induction phase. CONCLUSIONS: This combination regimen including hybrid administration of VNR plus CDDP is feasible, tolerable and effective as a first-line treatment in pts with aNSCLC. PMID- 18160124 TI - Pectin methylesterases induce an abrupt increase of acidic pectin during strawberry fruit ripening. AB - The decrease of strawberry (Fragariaxananassa Duch.) fruit firmness observed during ripening is partly attributed to pectolytic enzymes: polygalacturonases, pectate lyases and pectin methylesterases (PMEs). In this study, PME activity and pectin content and esterification degree were measured in cell walls from ripening fruits. Small green, large green, white, turning, red and over-ripe fruits from the Elsanta cultivar were analyzed. Using the 2F4 antibody directed against the calcium-induced egg box conformation of pectin, we show that calcium bound acidic pectin was nearly absent from green and white fruits, but increased abruptly at the turning stage, while the total pectin content decreased only slightly as maturation proceeded. Isoelectrofocalisation performed on wall protein extracts revealed the expression of at least six different basic PME isoforms. Maximum PME activity was detected in green fruits and steadily decreased to reach a minimum in senescent fruits. The preliminary role of PMEs and subsequent pectin degradation by pectolytic enzymes is discussed. PMID- 18160125 TI - Pre-exposure of calli to ozone promotes tolerance of regenerated Lycopersicon esculentum cv. PKM1 plantlets against acute ozone stress. AB - Studies were performed to evaluate the effects of pre-exposure of calli to ozone in promoting tolerance of the regenerated Lycopersicon esculentum cv. PKM1 (tomato) plantlets against acute ozone stress (AOS). Calli induced from tomato leaf explants were subjected to pre-treatment with ozone: T(1)=100 ppb, T(2)=200 ppb and T(3)=300 ppb. For the control (C) calli, charcoal-filtered air was supplied to test differential sensitivity of regenerated plantlets to acute ozone stress. All treated calli were subsequently transferred to shooting, rooting medium and acclimatized. The plantlets regenerated from the respective ozone (T(1), T(2), T(3))-treated calli are referred to here as T(1), T(2), T(3) plantlets and the plantlets regenerated from control calli are referred to as control plantlets. The frequencies of regeneration of tomato plantlets from the calli were T(1)=86%, T(2)=82% and T(3)=67%, and 92% regeneration was obtained from control calli. In order to evaluate the ozone tolerance, all the regenerated plantlets were exposed to the acute ozone exposure (AOE). After AOE, the T(2) plantlets endured remarkably well by experiencing reduced ozone stress, which was evident from the lower level of hydrogen peroxide and oxidative stress-related enzymes such as ascorbate peroxidase (EC 1.11.1.11) and superoxide dismutase (EC 1.15.1.1) activities relative to T(3), T(1) and C plantlets. All T(2) plantlets showed enhanced tolerance against AOE by upholding enhanced soluble phenol content, a higher level of foliar and apoplastic ascorbic acid, elevated dehydroascorbate reductase (EC 1.8.5.1) and glutathione content. The present study reveals that the calli pre-exposed to T(2) ozone treatment resulted in an increase in the level of antioxidants and provided the plants greater protection against acute ozone stress. PMID- 18160126 TI - Effect of mycorrhization on the isoflavone content and the phytoestrogen activity of red clover. AB - Red clover, known for its estrogenic activity due to its isoflavones content (biochanin A, genistein, daidzein and formononetin), was inoculated with the arbuscular mycorrhizal fungus Glomus mosseae. Once the symbiotic fungus was well established, plants were harvested and we determined the root and shoot dry weight as well as the P-content. In roots and leaves the levels of biochanin A, genistein, daidzein and formononetin were quantified by reversed-phase HPLC and the estrogenic activity of the leaves was measured by a transactivation assay using a yeast two-plasmid system. Mycorrhization increased the levels of biochanin A in the root and the shoot and reduced the levels of genistein in the shoot of red clover. The levels of the other isoflavones were not affected. The shoot biomass of mycorrhizal plants more than doubled compared with non mycorrhizal control plants, and this growth-stimulating effect of arbuscular mycorrhiza did not affect the estrogenic activity of red clover. In a control P treatment, the biomass of red clover was greatly enhanced. However, the estrogenic activity was reduced. These results suggest that, in contrast to an enhanced shoot biomass production after P application with a reduced estrogenic activity, with arbuscular mycorrhiza the shoot biomass of red clover can be enhanced without a negative effect on estrogenic activity. PMID- 18160127 TI - Abscisic acid-induced hydrogen peroxide is required for anthocyanin accumulation in leaves of rice seedlings. AB - The role of hydrogen peroxide (H(2)O(2)) in abscisic acid (ABA)-induced anthocyanin accumulation in detached and intact leaves of rice seedlings was investigated. Treatment with ABA resulted in an accumulation of anthocyanins in detached rice leaves. Dimethylthiourea, a chemical trap for H(2)O(2), was observed to be effective in inhibiting ABA-induced accumulation of anthocyanins. Inhibitors of NADPH oxidase (diphenyleneiodonium chloride and imidazole), phosphatidylinositol 3-kinase (wortmannin and LY 294002), and a donor of nitric oxide (N-tert-butyl-alpha-phenylnitrone), which have previously been shown to prevent ABA-induced H(2)O(2) accumulation in detached rice leaves, inhibited ABA induced anthocyanin increase. Exogenous application of H(2)O(2), however, was found to increase the anthocyanin content of detached rice leaves. In terms of H(2)O(2) accumulation, intact (attached) leaves of rice seedlings of cultivar Taichung Native 1 (TN1) are ABA sensitive and those of cultivar Tainung 67 (TNG67) are ABA insensitive. Upon treatment with ABA, H(2)O(2) and anthocyanins accumulated in leaves of TN1 seedlings but not in leaves of TNG67. Our results, obtained from detached and intact leaves of rice seedlings, suggest that H(2)O(2) is involved in ABA-induced anthocyanin accumulation in this species. PMID- 18160128 TI - Weight and weddings: women's weight ideals and weight management behaviors for their wedding day. AB - We conducted a descriptive cross-sectional survey to examine wedding-specific body weight ideals and weight management behaviors among women preparing for their wedding (n=272). The average bride-to-be was overweight according to clinical body mass index standards and idealized a significantly lower wedding weight. Most were attempting to lose weight and reported a small but significant weight loss using a combination of weight loss behaviors. Many women used one or more extreme weight loss behaviors. Additionally, some women reported purchasing a smaller-sized wedding dress, potentially to constrain behavior. Prior to weddings, interventions promoting a healthy, sustainable lifestyle may be useful to discourage extreme weight loss behaviors and emphasize healthy long-term weight management. PMID- 18160129 TI - Effects of estradiol on regulation of corticotropin-releasing factor gene and interleukin-6 production via estrogen receptor type beta in hypothalamic 4B cells. AB - Corticotropin-releasing factor (CRF) is produced in the hypothalamic paraventricular nucleus (PVN) in response to stress and stimulates the release of adrenocorticotropic hormone in the corticotrophs. Estrogens acting centrally are able to modulate the stress responses. In fact, direct estrogenic regulation of CRF gene expression has been demonstrated in various tissues. However, the mechanisms responsible for the actions of estrogens on CRF regulation in the PVN remain undetermined. We investigated whether estradiol (E2) contributes to the regulation of CRF gene and promoter activity in hypothalamic 4B cells. Furthermore, the involvement of E2 in the regulation of interleukin (IL)-6 and its role in hypothalamic 4B cells was explored. We demonstrated the dominant expression of estrogen receptor type beta (ERbeta) and found that a physiologically relevant dose of E2 and an ERbeta agonist stimulated CRF gene transcription in hypothalamic 4B cells. E2 stimulated IL-6 transcriptional activity via ERbeta, and subsequently the levels of IL-6 mRNA and protein. We also found that treatment with IL-6 significantly reduced cell viability. Thus, these data suggest the important effects of E2 on the regulation of CRF gene and IL-6 production via ERbeta in hypothalamic 4B cells. PMID- 18160130 TI - Comments on the through-space singlet energy transfers and energy migration (exciton) in the light harvesting systems. AB - Recent findings on the photophysical investigations of several cofacial bisporphyrin dyads for through space singlet and triplet energy transfers raised several serious questions about the mechanism of the energy transfers and energy migration in the light harvesting devices, notably LH II, in the heavily studied purple photosynthetic bacteria. The key issue is that for simple cofacial or slipped dyads with controlled geometry using rigid spacers or spacers with limited flexibilities, the fastest possible rates for singlet energy transfer for three examples are in the 10 x 10(9)s(-1) (i.e. just in the 100 ps time scale) for donor-acceptor distances approaching 3.5-3.6 A. The reported time scale for energy transfers between different bacteriochlorophylls, notably B800*-->B850, is in the picosecond time scale despite the long Mg...Mg separation of approximately 18 A. Such a short rate drastically contrasts with the well accepted Forster theory. This article reviews the modern knowledge of the structure, bacteriochlorophyll a transition moments, and photophysical processes and dynamics in LH II, and compares these parameters with the recently investigated model bisporphyrin dyads build upon octa-etio-porphyrin chromophores and rigid and semi-rigid spacers. The recently discovered role of the rhodopin glucoside residue called carotenoid will be commented as the possible relay for energy transfer, including the possibility of uphill processes at room temperature. In this context, the concept of energy migration, called exciton, may also be affected by relays and uphill processes. Also, it is becoming more and more apparent that the presence of an irreversible electron transfer reaction at the reaction center, i.e. electron transfer from the special pair to the phyophytin macrocycle and so on, renders the rates for energy transfer and migration more rapid precluding all possibility of back transfers. PMID- 18160131 TI - Simultaneous measurements of fast optical and proton current kinetics in the bacteriorhodopsin photocycle using an enhanced spectrophotometer. AB - A one-of-a-kind high speed optical multichannel spectrometer was designed and built at NIH and described in this journal in 1997 [J.W. Cole, R.W. Hendler, P.D. Smith, H.A. Fredrickson, T.J. Pohida, W.S. Friauf. A high speed optical multichannel analyzer. J Biochem Biophys Methods 1997;35:16-174.]. The most unique aspect of this instrument was the ability to follow an entire time course from a single activation using a single sample. The instrument has been used to study rapid kinetic processes in the photon-driven bacteriorhodopsin photocycle and electron transport from cytochrome c to cytochrome aa3 and from cytochrome aa3 to oxygen. The present paper describes a second generation instrument with a number of important enhancements which significantly improve its capabilities for multichannel kinetic studies. An example application is presented in which the kinetics of photon-induced proton flow across the biological membrane is measured simultaneously with the individual steps of the photocycle determined optically. Matching the time constants for the two processes indicates which molecular transformations are associated with major proton movements. PMID- 18160132 TI - A fast and gentle method for the isolation of myrosinase complexes from Brassicaceous seeds. AB - Myrosinase is a beta-thioglucosidase glucohydrolase that catalyses the hydrolysis of the thioglucoside bond in glucosinolates, allelochemicals present in Brassicaceous plants. These isoenzymes have been found to form complexes with other proteins; however, traditional isolation procedures involving ammonium sulphate precipitation and/or ion exchange chromatography do not allow for the isolation of these complexes. The present paper reports a fast and gentle procedure for the isolation of myrosinases in the complex form. Partial purification by Con A affinity chromatography followed by Sephadex G-200 gel filtration allowed for the isolation of myrosinase complexes from seeds of Brassica carinata, B. oleracea var. capitata, B. napus and Sinapis alba. Myrosinases in the Brassicas formed complexes of different molecular weight (500 600 kDa, 270-350 kDa and 140-200 kDa) whereas in seeds of S. alba it was only possible to isolate and detect 140-200 kDa complexes. In all species the complexes were formed by isoenzymes with isoelectric points between 4.8 and 5.6 and in some cases up to 6.8. SDS-PAGE confirmed that the myrosinase isoenzymes were composed by several protein subunits of molecular weights ranging between 10 and 110 kDa. The relative amount and enzymatic activity of the myrosinase complexes varied amongst the species studied. The isolation of myrosinase complexes in their native form is of great importance for the study of the hydrolysis of glucosinolates under autolysis conditions. PMID- 18160133 TI - A new detection method for arginine-specific ADP-ribosylation of protein -- a combinational use of anti-ADP-ribosylarginine antibody and ADP-ribosylarginine hydrolase. AB - Arginine-specific ADP-ribosylation is one of the posttranslational modifications of proteins by transferring one ADP-ribose moiety of NAD to arginine residues of target proteins. This modification, catalyzed by ADP-ribosyltransferase (Art), is reversed by ADP-ribosylarginine hydrolase (AAH). In this study, we describe a new method combining an anti-ADP-ribosylarginine antibody (alphaADP-R-Arg Ab) and AAH for detection of the target protein of ADP-ribosylation. We have raised alphaADP R-Arg Ab with ADP-ribosylated histone and examined the reactivity of the antibody with proteins treated by Art and/or AAH, as well as in situ ADP-ribosylation system with mouse T cells. Our results indicate that the detection of ADP ribosylated protein with alphaADP-R-Arg Ab and AAH is a useful tool to explore the target proteins of ADP-ribosylation. We applied the method to search endogenously ADP-ribosylated protein in the rat, and detected possible target proteins in the skeletal muscle, which has high Art activity. PMID- 18160135 TI - A benchmark test for a quantitative assessment of simple neuron models. AB - Several methods and algorithms have recently been proposed that allow for the systematic evaluation of simple neuron models from intracellular or extracellular recordings. Models built in this way generate good quantitative predictions of the future activity of neurons under temporally structured current injection. It is, however, difficult to compare the advantages of various models and algorithms since each model is designed for a different set of data. Here, we report about one of the first attempts to establish a benchmark test that permits a systematic comparison of methods and performances in predicting the activity of rat cortical pyramidal neurons. We present early submissions to the benchmark test and discuss implications for the design of future tests and simple neurons models. PMID- 18160134 TI - Imaging of glutamate in brain slices using FRET sensors. AB - The neurotransmitter glutamate is the mediator of excitatory neurotransmission in the brain. Release of this signaling molecule is carefully controlled by multiple mechanisms, yet the methods available to measure released glutamate have been limited in spatial and/or temporal domains. We have developed a novel technique to visualize glutamate release in brain slices using three purified fluorescence (Forster) energy resonance transfer (FRET)-based glutamate sensor proteins. Using a simple loading protocol, the FRET sensor proteins diffuse deeply into the extracellular space and remain functional for many tens of minutes. This allows imaging of glutamate release in brain slices with simultaneous electrophysiological recordings and provides temporal and spatial resolution not previously possible. Using this glutamate FRET sensor loading and imaging protocol, we show that changes in network excitability and glutamate re-uptake alter evoked glutamate transients and produce correlated changes in evoked cortical field potentials. Given the sophisticated advantages of brain slices for electrophysiological and imaging protocols, the ability to perform real-time imaging of glutamate in slices should lead to key insights in brain function relevant to plasticity, development and pathology. This technique also provides a unique assay of network activity that compliments alternative techniques such as voltage-sensitive dyes and multi-electrode arrays. PMID- 18160136 TI - Non-right-handedness is associated with migraine and soft bipolarity in patients with mood disorders. AB - BACKGROUND: There is a substantial body of data showing differences in the functioning of the two hemispheres in unipolar depressive and bipolar disorders. Migraine is a frequent co-morbid disorder in these patients, and it has been proposed that migraine may be associated with left-handedness. It would therefore be interesting to study migraine and handedness in a population of patients with mood disorders. METHODS: A total of 201 patients with an index episode of either major depression or mania were interviewed with a semi-structured interview based partly on DSM-IV criteria and partly on TEMPS-I for affective temperaments. The criteria of the Headache Classification Committee of the International Headache Society were used to establish the diagnosis of migraine. Hand preference was assessed using the Edinburgh inventory, and the patients were classified as having right-, left-, or mixed-handedness. RESULTS: In the whole group 117 patients had migraine (58%) and 59 (29%) were classified as having non-right hand preference. There was a significant increased prevalence of non-right-handedness in the migraine group (37% vs. 19%, p=0.021, Chi-square test; OR 2.5; 95% CI 1.3 4.8, p=0.007). In patients with cyclothymic, hyperthymic or irritable temperaments the prevalence of non-right-handedness (42%) was significantly higher (p=0.013, Chi-square test; OR 2.2, 95% CI 1.2-4.3) compared to patients with a depressive or no affective temperament (24%). The prevalence of non-right handedness was also significantly higher both in patients with co-morbid eating disorders (48% vs. 26%, p=0.008 Chi-square test; OR 2.7, 95% CI 1.3-5.9, p=0.01) and asthma (45% vs. 26%, p=0.026 Chi-square test; OR 2.3, 95% CI 1.1-5.1, p=0.029). LIMITATIONS: Non-blind evaluation of affective diagnosis, migraine and handedness. CONCLUSIONS: Our main finding supports the hypothesis that non-right handedness is associated with migraine and bipolar affective temperaments ("soft bipolarity") in a sample of patients with major affective disorders. PMID- 18160137 TI - Subjective and objective quality of life, levels of life skills, and their clinical determinants in outpatients with schizophrenia. AB - The purpose of the present study is to investigate the relationships among subjective and objective quality of life (QOL), and levels of life skills, and their clinical determinants in outpatients with schizophrenia by using schizophrenia disease-specific QOL measures. Data collected from 64 outpatients were analyzed. Subjective QOL was measured with the Schizophrenia Quality of Life Scale (SQLS) and objective QOL with the Quality of Life Scale (QLS). Patients' family members completed the Life Skills Profile (LSP). Clinical symptoms were also assessed with several scales including the Brief Psychiatric Rating Scale (BPRS) and the Calgary Depression Scale for Schizophrenia (CDSS). Only the motivation/energy scale, but not the other scales of the SQLS, correlated with the QLS. The LSP rated by the family showed significant correlations with both the SQLS and the QLS. The CDSS score predicted each scale of the SQLS, and the BPRS negative symptoms score predicted the QLS. The LSP was predicted by the BPRS negative symptoms score and the CDSS score independently. These results indicate that the patient's QOL could be predicted by the life skills measured by a family member and suggest that active treatment for depressive and negative symptoms might be recommended to improve the patient's QOL and life skills. PMID- 18160138 TI - LC MALDI-TOF MS/MS and LC ESI FTMS analyses of HLA-B27 associated peptides isolated from peripheral blood cells. AB - Peptides eluted from peripheral blood cells of HLA-B*2705 healthy donor were analyzed by LC MALDI MS/MS and LC ESI FTMS techniques. The sequences of 92 peptide ligands identified from one healthy blood donor by LC MALDI-TOF MS/MS were compared with those previously published from in vitro long-term cell cultures available in SYFPEITHI database and splenocytes. It was found that 18 sequences confirmed within 1ppm mass error by LC ESI FTMS were already described and 3 of them matched with those previously reported from HLA-B*2705 splenocytes. Another 38 sequences validated within the same mass error were not found in SYFPEITHI database and are identified here for the first time. Finally, 36 sequences (5 sequences already published in SYFPEITHI database) were evaluated by LC MALDI-TOF MS/MS but no matches in the list of monoisotopic masses obtained from LC ESI FTMS were found. PMID- 18160139 TI - Extra-esophageal acid reflux induced adenotonsillar hyperplasia: case report and literature review. AB - The etiology of adenotonsillar hyperplasia is not well understood. A 3-year-old child presented with obstructive sleep apnea believed to be secondary to enlarged tonsils and adenoids. Subglottic stenosis was encountered during intubation; the adenotonsillectomy was cancelled. Severe extra-esophageal reflux was identified and treated. At follow-up endoscopy 3 weeks later, the tonsils and adenoids were no longer enlarged or obstructing the airway. The role of extra-esophageal reflux in the pathogenesis adenotonsillar hyperplasia is discussed. PMID- 18160140 TI - Comparison between serum and saliva for the detection of hepatitis A virus RNA. AB - Due to the ease of collection, oral fluid is being investigated as an alternative to serum for diagnostic and epidemiological purposes. However, for prospective studies involving hepatitis A virus (HAV) RNA detection, a standard methodology must be developed. In the present study, nested RT-PCR and real-time PCR were optimized and evaluated for HAV detection and quantification, using oral fluid from healthy volunteers (n=20) and paired serum/oral fluid samples from individuals involved in a hepatitis A outbreak (n=78). Using nested RT-PCR, HAV RNA was detected in 50% of oral fluid and in 42% of serum samples from acute cases, as well as in 12% of all samples from cases without IgM and total anti HAV. Using real-time PCR, HAV RNA was detected in 61% of oral fluid and in 71% of serum samples from acute cases, as well as in 17 and 12%, respectively, from patients without HAV markers. Mean viral loads were 1.7+/-3.24 x 10(3)copies/ml in oral fluid and 2.8+/-6.46 x 10(3)copies/ml in serum. Although nested RT-PCR and real-time PCR both detected HAV RNA in oral fluid, real-time PCR was more sensitive. Oral fluid sample testing could be used as a noninvasive method of detecting HAV RNA during HAV outbreaks. PMID- 18160141 TI - Controlled inactivation of recombinant viruses with vitamin B2. AB - Inactivated viruses are important tools for vaccine development and gene transfer. 8-Methoxypsoralen (8-MOP) and long-wavelength ultraviolet irradiation (LWUVI) inactivates many viruses. Toxicity limits its use in animals and humans. Toxicological and photosensitizing properties of riboflavin make it suitable for virus inactivation in preparations for biological use. Viruses expressing beta galactosidase were mixed with either 8-MOP (1.5mM) or riboflavin (50 microM) and exposed to LWUVI (365 nm) for 2 h. Virus activity was determined by limiting dilution. The half-life of the adenovirus preparation treated with 8-MOP was 8.28 ns(-1) and 36.5 ns(-1) after treatment with riboflavin. Despite the difference in half-life, both preparations were completely inactivated within 45 min. In contrast, the half-lives for adeno-associated virus (AAV) preparations were similar (63 ns(-1) 8-MOP vs. 67 ns(-1) riboflavin). Each AAV preparation was fully inactivated within 90 min. The half-life of lentivirus was 193.4 ns(-1) after treatment with 8-MOP and 208 ns(-1) after exposure to riboflavin. Virus treated with riboflavin was inactivated within 20 min. Virus exposed to 8-MOP was inactivated in 90 min. DNA and RNA viruses can be inactivated by riboflavin and LWUVI and used in physiological systems sensitive to other photochemicals. PMID- 18160142 TI - Altered sensitivity of an R5X4 HIV-1 strain 89.6 to coreceptor inhibitors by a single amino acid substitution in the V3 region of gp120. AB - The replication of several R5X4 strains is blocked by single CXCR4 inhibitors such as AMD3100 or T140 although the target cells express both CXCR4 and CCR5 in vitro. To identify which region(s) of the Env are involved in the increased sensitivity to CXCR4 inhibitors, we isolated a T140-escape mutant using R5X4 HIV 1 strain 89.6. An isolated mutant harbored a single amino acid substitution in the V3 region of the Env (arginine 308 to serine R308S). Luciferase-reporter HIV 1 pseudotyped with the mutant Env showed that the substitution conferred total resistance to CXCR4 antagonists but increased sensitivity to a CCR5 antagonist TAK-779 in the infection of the cells expressing both CCR5 and CXCR4. Analyses using the cells expressing a single coreceptor showed that the mutant Env predominantly and efficiently utilized CCR5 rather than CXCR4 while retaining R5X4 phenotype. These results indicated that the sensitivities of the R5X4 strain to coreceptor inhibitors were altered by a single amino acid substitution in the V3 region of gp120. PMID- 18160143 TI - Effects of alcohol exposure during development on play behavior and c-Fos expression in response to play behavior. AB - Developmental exposure to alcohol can produce characteristic physiological and cognitive deficits, often termed Fetal Alcohol Spectrum Disorder (FASD). More recently, social deficits have been shown to occur both in FASD and animal models of FASD; the behavioral and neural bases of these deficits remain to be determined. It was hypothesized that changes in sensory processing may in part underlie the social deficits seen in FASD. This study used a rat model of FASD and social play, a behavior critical to adult social functioning, to begin to examine this hypothesis. Somatosensory cues from dorsal contact to the nape of the neck, critical to the initiation of pinning, were systematically degraded by administration of different doses of xylocaine, a topical anesthetic. Neuronal activity after 1h of play was assessed by measurement of c-Fos immunoreactivity (IR) in different brain regions. Ethanol-exposed rats showed an increased frequency of pinning during social play and were more sensitive to the degradation of somatosensory cues compared to the control groups, suggesting difficulties in processing somatosensory cues. Neuronal activity in the somatosensory cortex induced by play was significantly decreased in the ethanol exposed group compared to the non-treated group. The c-Fos IR in the nucleus accumbens was altered in a sexually dimorphic manner in the ethanol-exposed group. Thus, the behavioral and brain measures are consistent with the hypothesis that ethanol exposure during development induces alterations in social play via deficits in processing somatosensory cues that are important to social play. PMID- 18160144 TI - Risks to human health and estuarine ecology posed by pulling out creosote-treated timber on oyster farms. AB - Five oyster farms in Port Stephens, Australia were studied to identify consequences of using creosote-treated posts and the risks posed by removing the posts. Gas chromatography coupled with mass spectrometry (GC/MS) was used to measure polycyclic aromatic hydrocarbons (PAHs) and phenols in sediments, timber, water and oyster tissue. Before posts were removed, the total PAHs in surface sediment on farms was 24.1 mgkg(-1) dry weight. This increased to 45.5 mgkg(-1) dry weight after the posts were pulled out and remained significantly higher 6 months later at 59.7 mgkg(-1) dry weight. A similar increase was found in deeper sediments. The sediment attached to creosote-treated posts had a total concentration of PAHs of 484-2642 mgkg(-1) dry weight, while the corresponding value for the sediment on tar-treated posts was only 30.7 mgkg(-1) dry weight. The surface timber of creosote-treated posts had high levels of PAHs and an average post contained 43 g of PAHs. The total PAHs dispersed to the environment when a creosote-treated post was pulled out was at least 0.67 g. The main species were PAHs with low-molecular weights: fluoranthene, phenanthrene, pyrene, acenaphthylene and chrysene. Benzo(a)pyrene represented 1-10% of PAHs in most samples. Bioassays with creosote-contaminated sediment revealed that Sydney rock oysters (Saccostrea glomerate) and Pacific oysters (Crassostrea gigas) accumulated PAHs at (mgkg(-1) wet tissue weight): 11.3-15.3 and 35.5-47.9, respectively, when exposed for 5 days to water with < 1 microgl(-1) PAHs. Wild oysters growing on creosote-treated posts had high levels of phenols (0.09-6.92 mgkg(-1) wet weight) and PAHs (0.59-1.01 mgkg(-1) wet weight). The dilemma posed by removing creosote-treated posts and dispersing carcinogenic, bioavailable contaminants needs to be managed in light of risks to human health and estuarine ecology. PMID- 18160145 TI - Nutritional risk and cognitive impairment in the elderly. AB - The purpose of this study was to determine the relationship between nutritional risk and cognitive impairment in the elderly living in the community. Data obtained from 2934 subjects (912 men and 2022 women) aged above 60 years was analyzed from the Gwangju Dementia and Mild Cognitive Impairment Study (GDEMCIS). The study questionnaire comprised demographic characteristics, history of current and past illnesses, drug history, Korean version-Mini Mental State Examination (K MMSE), and Nutritional Screening Initiative (NSI) checklist. Additionally, we examined the blood pressure, fasting serum glucose level, lipid profile, body mass index, and ApoE genotype. Of the total, 1942 (66.2%) demonstrated good nutritional state (NSI checklist score< or =2) and 992 (33.8%) were at moderate or high nutritional risk (NSI checklist score>2). Multiple logistic regression analysis revealed that moderate or high nutritional risk subjects were associated with an increased risk of cognitive impairment (K-MMSE score< or =17) after adjustment for age, sex, educational level, and Korean version of Short form Geriatric Depression Scale (K-SGDS) score (Odds ratio=OR=1.71, 95%; confidence interval=CI=1.17-2.50). These results suggest that nutritional risk may be associated with cognitive impairment in the elderly. PMID- 18160146 TI - Green tea does not inhibit iron absorption. PMID- 18160147 TI - Pathophysiologic mechanisms of left ventricular apical ballooning in takotsubo cardiomyopathy. PMID- 18160149 TI - Prevalence of prehypertension, hypertension and, associated risk factors in Mongolian and Han Chinese populations in Northeast China. AB - BACKGROUND: The present study aimed to determine the prevalence of prehypertension, hypertension, and associated risk factors between Mongolian and Han populations in Northeast China. METHODS: The study was conducted in 2004 2006, and used a multistage cluster sampling method to select a representative sample. A total of 9236 Mongolian people and 36,154 Han people in the same area, age 35 years or older, were examined. The survey on blood pressure and associated risk factors was carried out. RESULTS: Overall, the prevalence of prehypertension for Mongolian people was 43.6%, for Han people was 44.3%. The prevalence of hypertension in Mongolian was higher than in Han (42% vs 36.7%, p<0.05). The prevalence of hypertension was positively correlated with age, smoking, body mass index (BMI), waist circumference (WC), lipid disorder, diabetes, salt intake and family history of hypertension in Mongolian, whereas it was positively correlated with age, female, smoking, drinking, BMI, WC, lipid disorder, diabetes, salt intake and family history of hypertension in Han. The rates of awareness, treatment and control in Mongolian and Han were very low (29.7% vs 29.2%, 23.6% vs 23.5%, 0.7% vs 1.2%). CONCLUSIONS: Hypertension and prehypertension were common in Mongolian and Han populations in Northeast China, and it were associated with many risk factors. The percentages of hypertensives who were aware, treated, and controlled were unacceptably low. These results place great emphasis on the urgent need for a public health program to improve the detection, prevention and treatment of hypertension in the rural area of Northeast of China. PMID- 18160150 TI - Asymptomatic double site of left ventricular outflow tract obstruction: subvalvular aortic stenosis associated with valvular aortic stenosis. AB - Left ventricular outflow tract obstruction can occur at the supravalvular, valvular, or subvalvular level. Each level of obstruction is associated with distinct symptomatology, natural history, and operative approach. We describe a case of asymptomatic double site of left ventricular outflow tract obstruction: subvalvular aortic stenosis associated with valvular aortic stenosis. PMID- 18160151 TI - Cerebral transient ischemic attack due to left ventricular thrombus formation in a young patient with silent myocardial infarction and normal coronary arteries. AB - Among many cardiac diseases related to cerebral stroke, left ventricular thrombus formation due to silent myocardial infarction with normal coronary arteries represents a rare cause of cerebral ischemia. We describe an unusual case of cerebral ischemia due to cardiac thrombus formation in a young patient with silent myocardial infarction and normal coronary arteries in which echocardiography and cardiac MR imaging clearly showed the embolic source. PMID- 18160152 TI - Correlation between glomerular filtration and markers of the atherosclerotic process in young patients with myocardial infarction. AB - In 41 young post myocardial infarction (MI) patients we investigated the relations between glomerular filtration rate (GFR) and markers of atherosclerosis: flow-mediated dilation of the brachial artery (FMD), intima media thickness (IMT) of the common carotid arteries and high sensitive C reactive protein (hsCRP). GFR was within normal values in all patients. Importantly, we found that GFR was significantly related to all three markers. Thus, our study supports the idea of the generalized nature of the atherosclerotic process and early involvement of the kidney. PMID- 18160153 TI - Gender impact on in-hospital outcomes after percutaneous coronary intervention. AB - BACKGROUND: Percutaneous coronary intervention (PCI) is a widely performed technique for coronary revascularization. Men and women seem to have different in hospital outcomes, although results remain unclear. We assessed the gender impact on in-hospital outcomes in men and women undergoing PCI. METHODS: In a single center, from 1998 to 2002, a total of 413 patients who underwent PCI were included in this analysis. From a dedicated database, the presence of risk factors, angiographic characteristics, left ventricular function and in-hospital outcomes were recorded and compared between men (n=244) and women (n=169). For comparison, unpaired t test and chi-square were used for continuous and dichotomous variables respectively. An alpha <0.05 was considered significant. RESULTS: Women were older (65.3+/-10.39 x 60+/-10.68 years, p=0.001), presented better ejection fraction (67.01+/-12.28% x 64.26+/-14.31%, p=0.028) and higher prevalence of stable angina (63.90% x 45.90%, p<0.001) than men. There were no differences related to family history for cardiovascular disease, hypertension, diabetes or hyperlipidemia; but there was an observed reduction in tobacco use (17.71% x 32.31%, p<0.001) by women. Similar rates of in-hospital successful procedures (94.3% x 94.90%, p=0.93) and death (0.41%x1.18%, p=0.40) were observed in men and women, respectively. Neither vascular complications nor stroke occurred. There was a higher incidence of urgent surgical myocardial revascularization (1.77%x0%, p=0.036) and a trend in the combined outcome of death/surgical myocardial revascularization (2.69% x 0.41%, p=0.06) in the women's group. CONCLUSION: Women present a higher incidence of urgent surgical myocardial revascularization following percutaneous coronary intervention when compared to men. PMID- 18160154 TI - Diastolic heart failure in the elderly: underlying mechanisms and clinical relevance. AB - Diastolic heart failure (DHF) is now firmly established as a significant contributor to the heart failure syndrome. However, compared to the better studied systolic dysfunction heart failure relatively little is known about this form of the syndrome. Epidemiological data have demonstrated that it is particularly important in the elderly likely reflecting the combination of several changes occurring in the myocardium occurring with advancing years, including progressive fibrosis and stiffening of the myocardium, the impact of hypertension over the years and the increased likelihood of ischaemic heart disease. This review will focus on the relevant aetiological factors in DHF, possible pathophysiological mechanisms and outline new and evolving therapeutic strategies for this problem. PMID- 18160156 TI - Quantifying fungal viability in air and water samples using quantitative PCR after treatment with propidium monoazide (PMA). AB - A method is described to discriminate between live and dead cells of the infectious fungi Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Mucor racemosus, Rhizopus stolonifer and Paecilomyces variotii. To test the method, conidial suspensions were heat inactivated at 85 degrees C or held at 5 degrees C (controls) for 1 h. Polycarbonate filters (25 mm diameter, 0.8 microm pore size) were placed on "welled" slides (14 mm diameter) and the filters treated with either PBS or PMA. Propidium monoazide (PMA), which enters dead cells but not live cells, was incubated with cell suspensions, exposed to blue wavelength light-emitting diodes (LED) to inactivate remaining PMA and secure intercalation of PMA with DNA of dead cells. Treated cells were extracted and the live and dead cells evaluated with quantitative PCR (QPCR). After heat treatment and DNA modification with PMA, all fungal species tested showed an approximate 100- to 1000-fold difference in cell viability estimated by QPCR analysis which was consistent with estimates of viability based on culturing. PMID- 18160157 TI - The impact of adjuvant radiation therapy on survival in women with uterine carcinosarcoma. AB - BACKGROUND AND PURPOSE: Uterine carcinosarcoma is an aggressive neoplasm and the benefit of adjuvant radiation therapy (RT) is unclear. This retrospective study analyzes the influence of RT on survival using a large population database. MATERIALS AND METHODS: Data were obtained from the Surveillance, Epidemiology, and End Results (SEER) program of the US National Cancer Institute. Women with uterine carcinosarcoma who underwent primary surgery were eligible. Survival rates and multivariate analyses were performed by standard methods. RESULTS: Of the 2461 women in the analysis, 890 received adjuvant RT. Five-year rates of overall survival were 41.5% and 33.2% (P<0.001) and uterine-specific survival were 56.0% and 50.8% (P=0.005), for women receiving RT compared to those who did not. Women with stages I-III disease experienced a benefit in overall survival (HR 0.87, P=0.03) while women with stage IV disease experienced benefits in overall (HR 0.63, P<0.001) and uterine-specific survival (HR 0.63, P=0.004) with RT. CONCLUSIONS: RT predicted for improved overall and disease specific survival in women with uterine carcinosarcoma within the SEER database. The benefit in disease specific survival was restricted to stage IV disease. These benefits may indicate a role for adjuvant RT in future prospective trials in the treatment of uterine carcinosarcoma. PMID- 18160158 TI - The relationship between waiting time for radiotherapy and clinical outcomes: a systematic review of the literature. AB - PURPOSE: To synthesize the direct clinical evidence relating waiting times (WTs) for radiotherapy (RT) to the outcomes of RT. METHODS AND MATERIALS: We did a systematic review of the literature between 1975 and 2005 to identify clinical studies describing the relationship between WTs and outcomes of RT. Only high quality (HQ) studies that had adequately controlled for confounding factors were included in the primary analysis. WTs that had originally been reported as a categorical variable were converted to a continuous variable based on the distribution of WTs in each category. Meta-analyses were done using a fixed effect model. RESULTS: The systematic review identified 44 relevant studies. Meta analyses of 20 HQ studies of local control demonstrated a significant increase in the risk of local failure with increasing WT, RRlocal recurrence/month =1.14, 95% Confidence Intervals (CI): 1.09-1.21. For post-operative RT for breast cancer; RRlocal recurrence/month =1.11, 95%CI: 1.04-1.19. For post-operative RT for head and neck cancer, RRlocal recurrenc/month =1.28, 95%CI: 1.08-1.52. For definitive RT for head and neck cancer, RRlocal recurrence/month =1.15, 95%CI: 1.02-1.29. There was little evidence of any association between WTs and the risk of distant metastasis. Meta-analyses of the 6 HQ studies of breast cancer showed RRmetastasis/month =1.04, 95%CI: 0.98-1.09. Meta-analyses of 4 HQ studies of breast cancer showed no significant decrease in survival with increasing WT, RRdeath/month =1.06, 95%CI: 0.97-1.16, but there was a marginally significant decrease in survival in 4 HQ studies of head and neck cancer, RRdeath/month =1.16, 95%CI: 1.02-1.32. CONCLUSIONS: The risk of local recurrence increases with increasing WTs for RT. The increase in local recurrence rate may translate into decreased survival in some clinical situations. WTs for RT should be as short as reasonably achievable. PMID- 18160159 TI - Obtaining the intrinsic electron spectrum of linear accelerators using the relation between the current of the bending magnet and the absorbed dose in water. AB - PURPOSE: To present a novel methodology to model the intrinsic electron spectra of a linear accelerator and its situation with respect to the energy window. METHODS: The spectra are obtained by fitting the variation of R(50) and the maximum dose rate measured in a water phantom with the bending magnet current. The obtained spectra are verified with a realistic Monte Carlo simulation of the accelerator. RESULTS: The intrinsic spectra and their relative position with respect to the energy window of the bending magnet have been obtained for a Siemens Mevatron KDS and an ELEKTA SL20 accelerators. CONCLUSIONS: Using this method in the commissioning and scheduled revisions of the accelerator, the tuning of the current of the bending magnet could be done in such a way that both the quality of the beam and the dose rate would reach a better long-term stability. PMID- 18160160 TI - Modulation of the mismatch negativity (MMN) to vowel duration changes in native speakers of Finnish and German as a result of language experience. AB - While crucial for phoneme distinctions in the Finnish language, mere vowel duration is of lower relevance as a phonetically distinctive cue in the German language. To investigate the pre-attentive processing of vowel duration between these two languages, the mismatch negativity (MMN), a component of the auditory event-related potential (ERP) that is an index of automatic auditory change detection, was measured in Finnish and German native speakers for vowel duration changes embedded in the pseudoword sasa. Vowel duration changes thereby were presented as a shortening or a lengthening of either the first- or second syllable vowel. An additional non-speech condition measured the MMN to duration and frequency changes in tones. In both language groups, diminished MMN amplitudes for the shortening of vowel duration in the word-final syllable suggested a generally more difficult discrimination of vowel duration in a word final position. Further, shorter MMN latencies for the Finns than the Germans for vowel duration as well as tone duration deviants suggested a generally higher sensitivity to duration contrasts in the Finnish language group. No latency difference between the groups was found for tone frequency processing. Moreover, the Finns, but not the Germans, showed a leftward shift of the MMN scalp distribution for changes in vowel duration, whereas the MMN topography was highly similar between both groups in the tone condition. An enhanced phonetic processing of vowel duration changes and possibly an enhanced processing of sound duration in general is thus indicated for the Finns as a result of their extensive linguistic experience with phonetically distinctive vowel duration contrasts in the native language. PMID- 18160161 TI - Affective visual event-related potentials: arousal, valence, and repetition effects for normal and distorted pictures. AB - Event-related potentials (ERPs) were used to assess arousal (low, high), valence (negative, positive), and stimulus repetition effects for normal and distorted images from the International Affective Pictures System (IAPS). Distorted stimuli were constructed by dividing each image into small squares and rearranging the segments randomly to produce a "scrambled" picture. Event-related potentials (ERPs) were elicited by presenting the normal and scrambled images as target stimuli, with a repeated visual pattern used as the standard stimulus. Participants (N=32) were instructed to press a button to the targets and ignore the standard. Stimulus repetition effects were assessed by presenting each stimulus twice in the normal and scrambled condition. High-arousal stimuli yielded larger late positive components for both the normal and scrambled pictures. No overall valence effects were obtained, but arousal and valence influenced component amplitudes for middle-latency ERPs from the scrambled stimuli. For the normal pictures, stimulus repetition was associated with increased component amplitudes for all potentials and decreased RTs of all affective categories. For the scrambled pictures, no repetition changes were obtained. The findings suggest that stimulus arousal level contributes more than valence to affective ERP measures for normal as well as perceptually distorted pictures. Stimulus repetition engages memory for previous normal picture items but is not influenced by affective category. Theoretical implications are discussed. PMID- 18160162 TI - Mycoflora and natural occurrence of aflatoxins and fumonisin B1 in cassava and yam chips from Benin, West Africa. AB - The presence of fungi, aflatoxins and fumonisin B1 in cassava and yam chips (during 28 processing and storage) were evaluated during two consecutive seasons in two agroecological zones of Benin (Northern Guinea Savannah, NGS and Sudan Savannah, SS). The Benin samples were assessed for moisture content, fungal infestation and total aflatoxin and fumonisin B1 contamination. During the two seasons, samples collected from the NGS, had moisture contents ranging from 10.0 to 14.7% in cassava chips and from 11.4 to 15.3% in yam chips. In samples from the SS, moisture content ranged from 10.1 to 14.5% and 11.1 to 14.5% in cassava and yam chips, respectively. A. flavus was the predominant fungal species. The maximum cfu/g in cassava and yam chips was 8950 and 6030, respectively. Other fungal species isolated included P. chrysogenum, M. piriformis, Phoma sorghina, F. verticillioides, R. oryzae and Nigrospora oryzae. High performance liquid chromatography analysis of both cassava and yam chips showed no contamination by either aflatoxins or fumonisin B1. PMID- 18160163 TI - Real-time PCR based procedures for detection and quantification of Aspergillus carbonarius in wine grapes. AB - Aspergillus carbonarius is the main species responsible for ochratoxin A accumulation in wine grapes and consequently, its rapid and sensitive detection is increasingly investigated. A new real-time PCR (RTi-PCR) based procedure was developed for the rapid and specific detection and quantification of A. carbonarius in wine grapes. The procedure includes the use of the pulsifier equipment to remove conidia from grapes which prevents releasing of PCR inhibitors, and DNA extraction with the EZNA Fungal DNA kit. It reduced the time for A. carbonarius DNA extraction from grapes to 30 min. Two specific primers (AcKS10L/AcKS10R) delimiting a 161 bp fragment, and a probe were designed and directed to the beta-ketosynthase domain of a polyketide synthase from A. carbonarius. Specificity was confirmed by testing primers towards purified DNA from 52 fungal strains, including reference and food isolates. Quantification was linear over at least 5 log units using both serial dilutions of purified DNA and calibrated conidial suspensions from A. carbonarius. The SYBR-Green I and TaqMan RTi-PCR approaches established were able to detect at least 2.4 and 24 genomic equivalents, respectively, using purified DNA. Results obtained from conidial suspensions, after DNA extraction, showed that at least 5 conidia per reaction should be present for a positive result with SYBR-Green I and 50 in the case of TaqMan. The quantification of fungal genomic DNA in artificially inoculated wine grapes performed successfully, with a minimum threshold of 10(3) conidia mL(-1) for accurate quantification. The developed RTi-PCR assay is a promising tool in the prediction of potential ochratoxigenic risk, even in the case of low-level infections, and suitable for a rapid, automated and high throughput analysis. PMID- 18160164 TI - Inactivation of conidia of Penicillium chrysogenum, P. digitatum and P. italicum by ethanol solutions and vapours. AB - A fractional factorial design, 2(5-1) experiments, was used for assessing the influence of 5 factors: water activity, aw [0.7, 0.9], temperature, T ( degrees C) [10, 30], mode of application, A [liquid, vapour], ethanol concentration, E (% w/w) [5, 10] and time, t (d) [1, 4] on the inactivation of spores of Penicillium chrysogenum, P. digitatum and P. italicum. Survival was determined by germination at optimal conditions within 3d. The experimental response was log (N 0/Nt), where N 0 and Nt (spore ml(-1)) the concentrations of viable spores at t=0 and t respectively. By a decreasing order of sensitivity to ethanol, moulds were ranked as followed: P. digitatum, P. italicum and P. chrysogenum. A greater inactivation for P. digitatum, P. italicum, that were the most sensitive moulds to ethanol, was obtained by applying vapour rather than ethanol solution. The order of significance of the main factors depended upon the mould. The key factor for explaining inactivation of P. chrysogenum was water activity. But, temperature was the main factor for explaining inactivation of P. digitatum and P. italicum. In the more drastic conditions, (i.e., 0.7 aw, 30 degrees C, 10% w/w ethanol), all spores were inactivated by applying liquid solution for 4d. PMID- 18160165 TI - Involvement of cell fatty acid composition and lipid metabolism in adhesion mechanism of Listeria monocytogenes. AB - The cell fatty acid (FA) composition of adhered and floating cells of two Listeria monocytogenes strains, selected on the basis of the biofilm forming ability, was taken into consideration. Anteiso-15:0, iso-C15:0 and anteiso-17:0 resulted the principal FAs in floating cells. On the other hand the adhered cells of the two strains presented a FAs profile characterized by a prevalence of straight FAs such as hexadecanoic acid (16:0) and octadecanoic acid (18:0). When adhered and free floating cells of the two strains were separately exposed to acid stress, an increase of the individual and total branched FAs was observed particularly in the floating cells of the two strains. The acid stress gave rise in the adhered cells of strain endowed with the lower biofilm forming ability to a relevant intracellular accumulation of straight medium chain FAs. Straight long chain and medium chain free fatty acids (FFAs) were released in the culture supernatants particularly by the strain CH4 endowed with a high biofilm forming ability. The exposure of the latter strain to its own supernatants (conditioned media-CMs) enhanced the C16:0 and C18:0 accumulation in the medium. An unusual quantity of straight medium chain hydrocarbons like n-decane and n-tridecane has been detected particularly in the supernatants of strain. Moreover, long chain hydrocarbons up to nonadecane were released by both strains when exposed to acid stress. Since adhesion is connected with the hydrophobic link of the cells with glass and ionic interactions, these hydrophobic molecules could exercise a key role in the adhesion and de-adhesion mechanisms. PMID- 18160166 TI - DNA oligonucleotide-assisted genetic manipulation increases transformation and homologous recombination efficiencies: Evidence from gene targeting of Dictyostelium discoideum. AB - Artificial gene alteration by homologous recombination in living cells, termed gene targeting, presents fundamental and considerable knowledge of in vivo gene function. In principle, this method can possibly be applied to any type of genes and transformable cells. However, its success is limited due to a low frequency of homologous recombination between endogenous targeted gene and exogenous transgene. Here, we describe a general gene-targeting method in which co transformation of DNA oligonucleotides (oligomers) could significantly increase the homologous recombination frequency and transformation efficiency. The oligomers were simply designed such that they were identical to both the ends of the homologous flanking regions of the targeting construct. Using this strategy, both targeted alleles of diploid cells were simultaneously replaced in a single transformation procedure. Thus, the simplicity and versatility of this method applicable to any type of cell may increase the application of gene targeting. PMID- 18160167 TI - Lipase-catalyzed synthesis of glucose fatty acid ester using ionic liquids mixtures. AB - Novozym 435-catalyzed synthesis of 6-O-lauroyl-d-glucose in ionic liquids (ILs) was investigated. The highest lipase activity was obtained in water-miscible [Bmim][TfO] which can dissolve high concentration of glucose, while the highest stability of lipase was shown in hydrophobic [Bmim][Tf(2)N]. The optimal activity and stability of lipase could be obtained in [Bmim][TfO] and [Bmim][Tf(2)N] mixture (1:1, v/v). Specifically, the activity of lipase was increased from 1.1 to 2.9 micromolmin(-1)g(-1) by using supersaturated glucose solution in this mixture, compared with reaction using saturated solution. After 5 times reuse of lipase, 86% of initial activity was remained in this mixture, while the residual activity in pure [Bmim][TfO] was 36%. Therefore, the productivity obtained by using ILs mixtures was higher than those in pure ILs. PMID- 18160168 TI - Molecular characterization and phylogenetic analysis of H1N1 and H3N2 human influenza A viruses among infants and children in Thailand. AB - The annual influenza outbreaks can cause a high mortality rate among infants and children. In the tropics, influenza shows no clear dependence on seasons. In the present study, we performed molecular and phylogenetic analysis of H1N1 and H3N2 influenza virus isolated from infants and children diagnosed with respiratory tract illness between February 2006 and February 2007. A total of 33 samples (10.92%) were found positive for human influenza virus infection. Characterization of the hemagglutinin gene revealed conserved sequences at the receptor-binding site as well as variations due to amino acid substitutions at the antigenic site, potentially resulting in an N-linked glycosylation site. As for the neuraminidase gene, amino acid substitutions were found in N1 and N2 but not directly at the catalytic or framework sites of this enzyme. Based on the phylogenetic tree, the hemagglutinin 1 (HA1) region and the neuraminidase (NA) gene of both H1N1 and H3N2 isolated subtypes clustered with the current vaccine strain for the Northern Hemisphere 2007-2008. This finding contributes to understanding the evolution of influenza A viruses in humans and is useful for surveillance and vaccine strain selection. PMID- 18160169 TI - In vitro toxicity and permeation of cyclodextrins in Calu-3 cells. AB - Cyclodextrins (CDs) can improve the pulmonary drug delivery by increasing aqueous solubility, absorption and bioavailability of drugs. Although the systemic absorption of CDs from gastrointestinal tract is very limited, their systemic absorption after pulmonary administration cannot be excluded. The aims of this study were 1) to evaluate the in vitro toxicity of various CDs (alpha-, beta-, gamma-, hydroxypropyl-alpha-, hydroxypropyl-beta- and randomly methylated-beta CD) in pulmonary Calu-3 cells and Calu-3 cell layers using MTT and LDH cytotoxicity tests, and 2) to study the permeation of natural CDs (alpha-, beta- and gamma-CD) at non-toxic concentrations across Calu-3 cell layers. Randomly methylated-beta-CD evoked cell death and membrane damage in Calu-3 cells at lower concentrations compared to the other CDs tested. In terms of their toxicity, gamma-CD, hydroxypropyl-beta-CD and hydroxypropyl-alpha-CD were the safest to the Calu-3 cells. Based on the cumulative penetrated amount at 4 h, the apparent permeability coefficients for alpha-, beta- and gamma-CD were 6.77+/-2.23 x 10( 8), 6.68+/-0.84 x 10(-8) and 6.71+/-0.74 x 10(-8) cm/s, respectively. In conclusion, this study indicates that 1) in terms of their local safety, hydroxypropylated CDs and natural gamma-CD seem to be the safest of the tested CDs in pulmonary drug delivery, and 2) cyclodextrins may be absorbed into the systemic circulation from the lungs. PMID- 18160170 TI - The contribution of phagocytic activity of liver macrophages to the accelerated blood clearance (ABC) phenomenon of PEGylated liposomes in rats. AB - We earlier reported that PEGylated liposomes lose their long-circulating property when they are administered twice in the same animal within certain intervals. We recently proposed that anti-PEG IgM elicited by the first dose PEGylated liposomes selectively binds to the surface of a second dose, subsequently leading to substantial complement activation and complement-receptor mediated uptake of the second dose by hepatic Kupffer cells. In this study we found, by using a single-pass liver perfusion technique, that the first dose does not increase the intrinsic phagocytic activity of the Kupffer cells. It was also found that only serum obtained from rats that had received a first dose is able to enhance the hepatic uptake of test dose. The conditioned-serum-dependent hepatic uptake was completely abolished by pre-treatment of the serum at 56 degrees C for 30 min, which inhibits the complement activity. Conclusively, our results strongly support our earlier proposal that complement activation caused by anti-PEG IgM elicited by the first dose is a major cause of the initiation of the accelerated blood clearance of a subsequent dose PEGylated liposome in the ABC phenomenon. PMID- 18160171 TI - Controlling the ABCs of diabetes in clinical practice: a community-based endocrinology practice experience. AB - AIMS: Determine A1C, blood pressure (BP), and total cholesterol (TC) (Diabetes ABCs) control in a community-based endocrinology practice (CBEP) and compare levels to national averages. Additionally, determine patient factors associated with ABC control. METHODS: A retrospective chart audit of 395 consecutive patients seen for diabetes management was conducted for years 2000-2004 to examine levels of control of the ABCs. Multivariate models were used to determine patient factors associated with control. RESULTS: Significantly more patients met the goal of A1C <7% in the CBEP compared to national estimates (CBEP: 47.1% vs. NHANES 1999-2000: 37%, p=0.003). Similar patterns were observed for BP (CBEP: 53.2% vs. NHANES 1999-2000: 35.8%, p<0.0001), TC (CBEP: 82% vs. NHANES 1999-2000: 48.2%, p<0.0001), and all three ABCs (CBEP: 22%, vs. NHANES 1999-2000: 7.3%, p<0.0001). The proportion of patients meeting all three ABC goals in the CBEP increased significantly over time (p<0.0001). Multivariate models demonstrated that patients not needing insulin (p<0.0001), and taking fewer BP (p<0.0001), and cholesterol-lowering medications (p<0.02) were significantly more likely to have ABCs in control. CONCLUSIONS: Attainment of ABC goals is feasible in a CBEP and can be achieved at rates higher than national averages. Attention to factors that affect these goals is warranted. PMID- 18160172 TI - Hepatitis C infection: dying with or from the infection? PMID- 18160173 TI - Calcitonin-derived peptide carriers: mechanisms and application. AB - Among the family of the so-called cell-penetrating peptides (CPP) sequences derived from the native peptide hormone human calcitonin (hCT) have recently proven to translocate different bioactive molecules across cellular membranes. Herein, we give an extensive summary of the development of hCT-derived carrier peptides, beginning with the therapeutic nasal administration of full-length hCT. Hence, N-terminally truncated hCT fragments were investigated and subsequently optimised to extend their field of application. The latest generation of hCT derived carrier peptides are highly effective, branched peptides. The current state of the art is reviewed concerning the structural requirements, mechanistic assumptions and metabolic features of these peptides as well as experiments proving their excellent carrier potential. PMID- 18160174 TI - Lipid-based systems for the enhanced delivery of poorly water soluble drugs. PMID- 18160175 TI - Directions in reintroduction biology. AB - Reintroductions are attempts to return species to parts of their historical ranges where they were extirpated, and might involve release of either captive bred or wild-caught individuals. The poor success rate of reintroductions worldwide has led to frequent calls for greater monitoring, and since 1990 there has been an exponential increase in the number of peer-reviewed publications related to reintroduction. However, these publications have largely been descriptive accounts or have addressed questions retrospectively based on the available data. Here, we advocate a more strategic approach where research and monitoring targets questions that are identified a priori. We propose ten key questions for reintroduction biology, with different questions focusing at the population, metapopulation and ecosystem level. We explain the conceptual framework behind each question, provide suggestions for the best methods to address them, and identify links with the related disciplines of restoration ecology and invasion biology. We conclude by showing how the framework of questions can be used to encourage a more integrated approach to reintroduction biology. PMID- 18160177 TI - Properties of the Kinesin-1 motor DdKif3 from Dictyostelium discoideum. AB - The amoeba Dictyostelium discoideum possesses genes for 13 different kinesins. Here we characterize DdKif3, a member of the Kinesin-1 family. Kinesin-1 motors form homodimers that can move micrometer-long distances on microtubules using the energy derived from ATP hydrolysis. We expressed recombinant motors in Escherichia coli and tested them in different in vitro assays. Full-length and truncated Kif3 motors were active in gliding and ATPase assays. They showed a strong dependence on ionic strength. Like the full-length motor, the truncated DdKif3-592 motor (aa 1-592; comprising motor domain, neck, and partial stalk) reached its maximum speed of around 2.0micrcom s(-1) at a potassium acetate concentration of 200mM. The shortened DdKif3-342 motor (aa 1-342; comprising motor domain, partial neck) showed a high ATP turnover, comparable to that of the fungal Kinesin-1, Nkin. Results from the duty cycle calculations and gliding assays indicate that DdKif3 is a processive motor. A GFP-fusion protein revealed a mainly cytoplasmic localization of DdKif3. Immunofluorescence staining makes an association with the endoplasmic reticulum or mitochondria unlikely. Despite a similar phylogenetic distance to both metazoa and fungi, in terms of its biochemical properties DdKif3 revealed a closer similarity to fungal than animal kinesins. PMID- 18160178 TI - Expression patterns of LmAP2L1 and LmAP2L2 encoding two-APETALA2 domain proteins during somatic embryogenesis and germination of hybrid larch (Larix x marschlinsii). AB - Two APETALA2 domain transcription factors were characterized first in angiosperms, and, recently, in several gymnosperms. These proteins are involved in several processes, from flowering to embryogenesis in Arabidopsis thaliana. We extrapolated this result to hybrid larch (Larixxmarschlinsii Coaz) resulting from a cross between European (Larix decidua) and Japanese (Larix kaempferi) larches. Somatic embryogenesis is well described and controlled for this Pinaceae. We characterized two-AP2 domain genes: LmAP2L1 and LmAP2L2. Phylogenetic analysis confirmed that LmAP2L1 and LmAP2L2 were orthologous to Norway spruce PaAP2L1 and PaAP2L2 and that L1 forms appeared to be specific to Pinaceae. RT-PCR analysis showed that larch APETALA2 was differentially expressed during late somatic embryogenesis and during the first steps of germination. Whereas LmAP2L2 was constitutively expressed during this process, LmAP2L1 expression appeared only during late somatic embryogenesis, when embryos were able to germinate. Further, LmAP2L1 appeared to be the preferentially expressed form during embryo germination. Thus, LmAP2L1 seems to be a valuable molecular marker for hybrid larch late somatic embryogenesis and could play a role during post-embryonic development. PMID- 18160179 TI - Isolation of a novel family of genes related to 2-oxoglutarate-dependent dioxygenases from soybean and analysis of their expression during root nodule senescence. AB - A screen for genes involved in root nodule senescence has led to the isolation of the senescence-associated nodulin 1 (SAN1) multigene family from Glycine max (soybean). The three, tandemly repeated SAN1 genes each have three exons and two introns and are highly conserved. SAN1A and SAN1B code for conceptual proteins of 352 and 353 amino acids, respectively, and share over 83% sequence identity, while SAN1C encodes a truncated protein of 126 amino acids and is likely to be a pseudogene. The SAN1-encoded proteins share sequence similarity and highly conserved motifs with plant 2-oxoglutarate-dependent dioxygenases (2-ODDs), suggesting that they encode 2-ODDs. Analyses of the steady-state mRNA levels of SAN1A and SAN1B during senescence induced by treatment with fixed nitrogen or darkness demonstrate that SAN1A is downregulated during induced senescence. In contrast, SAN1B is upregulated by both treatments. The expression of the SAN1 genes is not restricted to nodules, suggesting that in addition to their function(s) in these organs, they play a more general role in plant metabolism. PMID- 18160180 TI - Identification of ACE-inhibitory peptides in salt-free soy sauce that are transportable across caco-2 cell monolayers. AB - In present study, we aimed to identify angiotensin I-converting enzyme (ACE) inhibitory peptides from a salt-free soy sauce (SFS), a newly developed antihypertensive seasoning obtained by Aspergillus oryzae fermentation of soybean in the absence of salt, which can be transported through caco-2 cell monolayers. Through an Ussing transport investigation of SFS across caco-2 cell monolayers, three di-peptides, Ala-Phe, Phe-Ile and Ile-Phe, were successfully identified from the SFS as transportable inhibitory peptides. Ala-Phe and Ile-Phe, but not Phe-Ile, exhibited ACE-inhibitory activity with IC(50) values of 165.3 microM and 65.8 microM, respectively. Kinetic studies revealed that Ile-Phe (Km: 3.1 mM, P(app): 2.4 x 10(-6) cm/s) exhibited greater affinity toward the transport compared with Ala-Phe (K(m): 48.1 mM, P(app): 1.4 x 10(-6) cm/s) and Phe-Ile (K(m): 12.7 mM, P(app): 1.4 x 10(-6) cm/s). PMID- 18160181 TI - Differential age effects on cerebral blood flow and BOLD response to encoding: associations with cognition and stroke risk. AB - Changes in the cerebrovascular system due to age or disease can significantly alter the blood-oxygenation-level-dependent (BOLD) signal and complicate its interpretation. The simultaneous acquisition of arterial spin labeling (ASL) and BOLD data represents a useful technique to more fully characterize the neurovascular underpinnings of functional brain response to cognition. We conducted a functional magnetic resonance imaging (FMRI) study of episodic memory encoding to investigate whether age is related to cerebral blood flow (CBF) and BOLD response in the medial temporal lobe (MTL). Results demonstrated a significant reduction in resting-state CBF in older compared to young adults. Conversely, older adults showed significantly increased CBF but not BOLD response in the MTL during picture encoding relative to young adults. Correlations between CBF response and cognition were demonstrated whereas associations with BOLD were not observed. Stroke risk was associated with both CBF and BOLD response. Results suggest that aging effects on CBF and BOLD responses to encoding are dissociable and that cerebrovascular alterations contribute to findings of age-related differences. PMID- 18160182 TI - Estrogen modifies arousal but not memory for emotional events in older women. AB - Emotional arousal and the affective content of events influence memory. These effects shift with age such that older people find negative information less arousing and remember proportionately more positive events compared to the young. The emotional enhancement of memory is mediated by medial temporal lobe limbic structures and the prefrontal cortex, which are both affected by sex hormones. We examined whether hormone use (estrogen or estrogen and progesterone) in older women modulated perceptions of valence and arousal, and subsequent memory for emotional images or stories. Their performance was compared to younger women. Hormone use in older women resulted in higher arousal for negative images and stories but memory was not affected. We hypothesize that estrogen modifies the influence of the amygdala and the prefrontal cortex on emotion, but that age related changes in the hippocampus prevent the enhancement of emotional memory in older women. PMID- 18160183 TI - Complete screening for glucocerebrosidase mutations in Parkinson disease patients from Portugal. AB - Mutations in the gene encoding beta-glucocerebrosidase, a lysosomal degrading enzyme, have recently been associated with the development of Parkinson disease. Here we report the results found in a cohort of Portuguese Parkinson disease patients and healthy age-matched controls for mutations in the aforementioned gene. This screening was accomplished by sequencing the complete open-reading frame, as well as intron/exon boundaries, of the glucocerebrosidase gene, in a total of 230 patients and 430 controls. We have found an increased number of Parkinson disease patients presenting mutations in GBA when compared to controls. These results, together with recent literature, clearly suggest a role of glucocerebrosidase in the development of Parkinson disease. PMID- 18160184 TI - [Primary hypertrophic osteoarthropathy in an adolescent]. AB - Hypertrophic osteoarthropathy is a rare syndrome characterized by dysacromelia and periostosis with digital clubbing. Primary form is called pachydermoperiostosis. A case of a primary hypertrophic osteoarthropathy without pachydermia is reported in a 13-year-old boy, which was born of a consanguineous marriage. This case suggested an incomplete form of pachydermoperiostosis with a recessive inheritance. Pachydermoperiostosis is often familial and occurs predominantly in men. This affection is believed to be inherited in an autosomal dominant pattern but some autosomal recessive forms have also been reported. PMID- 18160185 TI - Attachment of the outer membrane lipoprotein (OprI) of Pseudomonas aeruginosa to the mucosal surfaces of the respiratory and digestive tract of chickens. AB - The development of mucosal vaccines requires antigen delivery and adjuvant systems that can efficiently help in presenting vaccine antigens to the mucosal immune system. The outer membrane lipoprotein I (OprI) of Pseudomonas aeruginosa seems to possess both the quality to induce a non-specific immune response (adjuvant effect through its lipid tail) as well as the quality to facilitate uptake of the vaccine antigen by interacting with Toll-like receptor 2/4 (TLR2/4) on antigen-presenting cells (APC) and epithelial cells (adhesion effect). Here, we show for the first time the adhesion of OprI to epithelial cells of the trachea and small intestine of chickens. Adhesion could be seen on cryosections after in vitro as well as after in vivo incubation of the trachea and intestine. This proves the value of OprI as a fusion partner in mucosal protein vaccine development, which is especially important for poultry where mass vaccination is only possible by the respiratory or oral route. PMID- 18160186 TI - Immune responses to polysaccharides: lessons from humans and mice. AB - This review focuses on the immune response to non-conjugated and conjugated polysaccharide vaccines derived from encapsulated pathogens, such as Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis. Special attention is paid to a number of side effects observed following the use of some of these vaccines. For example, we discuss the long-lasting specific refractoriness induced by unconjugated polysaccharides, and the absence of an effective immune response in adults vaccinated with some conjugated vaccines. We argue that studies performed in the mouse model can help to understand those paradoxical effects observed in humans, and the mechanisms underlying such processes. PMID- 18160187 TI - Immunization with the cysteine proteinase Ldccys1 gene from Leishmania (Leishmania) chagasi and the recombinant Ldccys1 protein elicits protective immune responses in a murine model of visceral leishmaniasis. AB - The gene Ldccys1 encoding a cysteine proteinase of 30 kDa from Leishmania (Leishmania) chagasi, as well as the recombinant cysteine proteinase rLdccys1, obtained by cloning and expression of the Ldccys1 gene in the pHIS vector, were used to evaluate their ability to induce immune protective responses in BALB/c mice against L. (L.) chagasi infection. Mice were immunized subcutaneously with rLdccys1 plus Bacille Calmette Guerin (BCG) or Propionibacterium acnes as adjuvants or intramuscularly with a plasmid carrying the Ldccys1 gene (Ldccys1/pcDNA3) and CpG ODN as the adjuvant, followed by a booster with rLdccys1 plus CpG ODN. Two weeks after immunization the animals were challenged with 1 x 10(7) amastigotes of L. (L.) chagasi. Both immunization protocols induced significant protection against L. (L.) chagasi infection as shown by a very low parasite load in the spleen of immunized mice compared to the non-immunized controls. However, DNA immunization was 10-fold more protective than immunization with the recombinant protein. Whereas rLdccys1 induced a significant secretion of IFN-gamma and nitric oxide (NO), animals immunized with the Ldccys1 gene increased the production of IgG2a antibodies, IFN-gamma and NO. These results indicated that protection triggered by the two immunization protocols was correlated to a predominant Th1 response. PMID- 18160188 TI - Evaluation of the Friend Virus model for the development of improved adenovirus vectored anti-retroviral vaccination strategies. AB - We evaluated the suitability of the Friend Virus (FV) model for the development of improved adenovirus vectors for anti-retroviral vaccination using two types of adenovirus vectors, encoding F-MuLV Env and Gag, which differed only in their fiber genes (Ad5 and Ad5F35). Genetically FV-resistant C57BL/6 mice and highly susceptible CB6F1 hybrid mice were vaccinated by either homologous or heterologous prime-boost regimen. After FV challenge, viral loads in the spleens of C57BL/6 mice were reduced approximately 250-fold and were below the detection threshold in >50% of the mice. Vaccination outcome was critically influenced by the route of vector administration. In CB6F1 mice, vaccination resulted in reduced viremia, delayed onset of splenomegaly, and induction of FV-specific T cells as assessed by tetramer staining. Heterologous prime-boost vaccination resulted in significantly higher neutralizing antibody titers, translating into improved immune protection, in contrast to coexpression of cytokines. Our results suggest that the FV model can provide insight into the development of improved adenovirus vectors for HIV-1 vaccination. PMID- 18160189 TI - Intranasal administration of a recombinant adenovirus expressing the norovirus capsid protein stimulates specific humoral, mucosal, and cellular immune responses in mice. AB - Norovirus (NV) is a major cause of acute, epidemic nonbacterial gastroenteritis in individuals of all ages. The immunological mechanism of NV infection and the approaches used to prevent infection remain to be elucidated. In this study, the specific immune responses of BALB/c mice were assessed following intranasal immunization with a recombinant adenovirus vector expressing the genogroup II4 (GGII/4) norovirus capsid protein. Analysis of IgM, IgG, and IgA antibodies specific for the recombinant virus-like particles (VLPs) of NV demonstrated that a high level of humoral immunity developed following immunization. Mucosal immune responses were also detectable in stool, intestinal homogenates, lung homogenates, and lung lavage samples. Specific cellular immune responses were observed in NV VLPs-restimulated splenocytes by ELISPOT and Th1/Th2 cytokine cytometric array (CBA). Serum IgG subclass analysis showed that a balanced Th1- and Th2-like cellular immune response was induced in BALB/c mice following immunization with recombinant adenovirus. These findings demonstrate that the intranasal immunization of a recombinant adenovirus expressing the NV capsid protein is an efficient strategy to stimulate systemic, mucosal, and cellular Th1/Th2 immune responses in mice, and could serve as a novel approach for designing NV vaccines. PMID- 18160190 TI - Determination of bioavailable rhenium fraction in agricultural soils. AB - Rhenium (Re) mobility in agricultural soils was studied in order to obtain information relevant to (99)Tc mobility in soil-to-plant systems. Since water soluble Tc and Re are highly bioavailable, extraction of Re with water was carried out in addition to a total Re determination in the soils. The geometric means of total Re for paddy field, upland field and other soils were 0.34, 0.23, and 0.28 ng g(-1), respectively, while those of water soluble Re (<0.45 microm membrane filterable) were 0.053, 0.015 and 0.008 ng g(-1), respectively. There were no differences for total Re among soil uses; however, the water soluble Re/total Re ratio was significantly higher in paddy field soils (16%) than in other soil uses (6% for upland fields and 3% for other uses). Rhenium mobility and plant availability were higher in paddy fields than in other agricultural fields, and similar phenomena would be expected for (99)Tc. PMID- 18160191 TI - Radionuclides in soils, 1960-2006: a view from the World Congress of Soil Science. PMID- 18160192 TI - Nurse-delivered brief interventions for hazardous drinkers with alcohol-related facial trauma: a prospective randomised controlled trial. AB - AIMS: To assess the impact of two methods of brief nurse-delivered brief interventions in reducing drinking variables in hazardous drinkers with alcohol related facial injuries. METHODS: A randomised controlled trial of two brief interventions involving hazardous drinkers with facial trauma in three Oral and Maxillofacial Surgery outpatient clinics in the West of Scotland; 194 patients were recruited and randomised to have either a nurse-led brief motivational intervention (intervention group) or a leaflet about alcohol misuse (control group). Patients were followed up at 3 and 12 months after the intervention and drinking variables reassessed. RESULTS: A brief motivational intervention for alcohol provided by a nurse was more effective than a leaflet in helping some patients with facial trauma to reduce their alcohol consumption 12 months after the intervention (p<0.05). CONCLUSIONS: Facial trauma in the West of Scotland is strongly associated with alcohol misuse and is a recurrent disease, particularly among those who drink heavily. A nurse-delivered brief motivational intervention is effective in helping patients with high scores in the Alcohol Use Disorders Identification Test to reduce their drinking, and this effect was apparent 12 months after the intervention. PMID- 18160193 TI - An evaluation of the genotoxicity of the antitussive drug Dextromethorphan. AB - Dextromethorphan (DMP) is an effective and widely used antitussive drug. While DMP has over a 50 year safe-marketing history, the only available genotoxicity data was an unpublished, negative Ames assay (Roche). Lack of a complete genotoxicity profile on DMP, specifically covering the chromosomal damage endpoint, prompted a regulatory request for an in vitro chromosome aberration assay. In accordance with EC and CPMP Guidance, we evaluated data for a number of chemicals with a structural relationship to DMP. DMP contains no structural alerts for genotoxicity or carcinogenicity using the Deductive Estimation of Risk from Existing Knowledge (DEREK) software tool, confirming the negative results obtained in the existing Ames assay. This is also consistent with the mostly negative genotoxicity and carcinogenicity data available on structurally related chemicals including morphine, codeine, nalbuphine, buprenorphine, naloxone, hydromorphone, levorphanol, and oxycodone. A state-of-the-science, in vitro chromosome aberration assay was also conducted, which demonstrated a lack of genotoxicity for DMP. The overall weight of evidence for DMP and its structural analogues, supports the conclusion that this class of phenanthrene-based chemicals, and DMP, in particular, are not genotoxic in vitro or in vivo, and do not represent a carcinogenic risk to patients. PMID- 18160194 TI - A multilevel analysis of urban neighborhood socioeconomic disadvantage and health in late life. AB - The associations between neighborhood context and various indicators of health are receiving growing empirical attention, but much of this research is regionally circumscribed or assumes similar effects across the life course. This study utilizes a U.S. national sample to investigate the association between urban neighborhood socioeconomic disadvantage and health specifically among older adults. Data are from 3442 participants aged 70 years and older in the 1993 Asset and Health Dynamics Among the Oldest Old (AHEAD) Study, and the 1990 U.S. Census. Our approach underscores the importance of multiple dimensions of health (self reported physician-diagnosed cardiovascular disease [CVD], functional status, and self-rated health) as well as multiple dimensions of neighborhood disadvantage, which are conceptualized as environmental hazards that may lead to a physiologically consequential stress response. We find that individual-level factors attenuate the association between neighborhood disadvantage and both CVD and functional status, but not self-rated health. Net of covariates, high neighborhood socioeconomic disadvantage is significantly associated with reporting poor health. In late life, neighborhood socioeconomic disadvantage is more consequential to subjective appraisals of health than diagnosed CVD or functional limitations. PMID- 18160195 TI - Emergency department utilization among victims and offenders involved in non lethal violence. AB - The medical literature has focused on violent victimization as a public health concern, examining its correlates and evaluating intervention models. However, the emphasis on victimization in this literature overlooks the strong ties between victimization and offending risks outlined in the criminological literature, which may unnecessarily limit the scope of public health efforts to influence violence in our communities. This study examines whether the similarities observed in the criminological literature are evident in a health care setting. More specifically, do victims and offenders exhibit similar health care utilization patterns? We address this question by comparing the emergency department utilization records, criminal histories, and demographic characteristics of a sample of victims and offenders involved in non-lethal violence in Bernalillo County, New Mexico, USA in 2001. Our results suggest that victims and offenders have similar emergency department utilization patterns, with most visits being for injury. Moreover, most victims seen in the emergency department have criminal records that, in many ways, mirror those of offenders. The results suggest that violence intervention programs in public health settings should target both victims and offenders and capitalize on the overlap across these populations in outlining the long term risks of criminal involvement and motivating individual level change. PMID- 18160196 TI - The hyperglutamatergic hypothesis of autism. PMID- 18160197 TI - Stress at work alters serum brain-derived neurotrophic factor (BDNF) levels and plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) levels in healthy volunteers: BDNF and MHPG as possible biological markers of mental stress? AB - There is growing evidence that blood levels of brain-derived neurotrophic factor (BDNF) and catecholamine, and cytokines are related to not only to depressive, suicidal, and anxious states but also to depression-associated personality traits. Psychological job stress is well known to lead to symptoms of depression and anxiety. In the present study, we examined effects of psychological job stress on serum levels of BDNF and plasma levels of catecholamine metabolites, and cytokines in healthy volunteers (n=106, male/female=42/64, age=36+/-12 yr) working in a hospital setting. The values (mean+/-SD) of scores for stress items in the Stress and Arousal Check List (s-SACL), plasma MHPG levels, and, serum BDNF levels in all participants were 7.2+/-3.3, 5.2+/-3.4 ng/mL, and 23.3+/-14.7 ng/mL, respectively. A negative correlation was found between scores for s-SACL and serum BDNF levels (rho=-0.211, p=0.022). A positive correlation was also found between scores on the s-SACL and plasma levels of 3-methoxy-4 hydroxyphenylglycol (MHPG) (rho=0.416, p=0.01), but not homovanillic acid (HVA). No relationship was found between s-SACL scores and plasma levels of interleukin 6 (IL-6) or tumor necrosis factor alpha (TNFalpha). These results suggest that serum BDNF levels and plasma MHPG levels might be biological markers reflective of psychological job stress in hospital employees. PMID- 18160198 TI - Inhibitory effects of water extract of propolis on doxorubicin-induced somatic mutation and recombination in Drosophila melanogaster. AB - Propolis is a substance produced by honeybees (Apis mellifera L.). Its components are strong antioxidants and free radical scavengers. The aim of this study was to evaluate the protective effects of a water extract of Brazilian green propolis (WEP) combined with the antitumor agent doxorubicin (DXR) on Drosophila melanogaster wing cells through the somatic mutation and recombination test (SMART). Two different crosses were used: The standard (ST) cross and the high bioactivation (HB) cross. The HB cross is characterized by a constitutively enhanced level of cytochrome P450 which leads to an increased sensitivity to a number of promutagens and procarcinogens. Larvae obtained from these two crosses were chronically treated with different concentrations of WEP (12.5,25.0 and 50.0 mg/mL) alone or combined with DXR (0.125 mg/mL). The results obtained with the two different crosses were rather similar. Neither toxicity nor genotoxicity were observed in WEP treated series. Simultaneous treatment with different concentrations of WEP and DXR led to a reduction in the frequency of recombination compared to the treatment with DXR alone. This anti-recombinogenic effect was proportional to the concentrations applied, indicating a dose-response correlation and can be attributed to the powerful scavenger ability of WEP. PMID- 18160200 TI - Non-invasive magnetic resonance imaging-guided focused ultrasound treatment for uterine fibroids - early experience. AB - OBJECTIVE: To describe early results regarding efficacy and safety of magnetic resonance imaging-guided focused ultrasound surgery (MRgFUS) for the treatment of uterine leiomyomas among a population of Japanese women. STUDY DESIGN: In this single site study, 48 Japanese women, over 18 years of age, who were candidates for surgical intervention due to symptomatic fibroids, were recruited. Fifty-five uterine fibroids were treated by MRgFUS and followed up for 12 months. RESULTS: During the 12-month follow-up period, two patients (4%) required surgical interventions, and two patients required additional drug treatment for fibroids. The average reduction in fibroid volume determined by MR imaging at 6 months after treatment was 33%. This volume reduction is correlated with the fibroid's treated volume. No serious complications were recorded during the treatments or follow-up period. CONCLUSION: MRgFUS can safely be used to non-invasively treat symptomatic uterine fibroids and avoid the need for surgical intervention in the short term. With large treatment volumes, MRgFUS also results in significant fibroid shrinkage in a relatively short time. Additional follow-up is needed to determine the long-term durability of this promising non-invasive approach. PMID- 18160201 TI - Pregnancy outcome in patients with patent foramen ovale and cerebral embolism. PMID- 18160202 TI - Absence of gravity-dependent modulation of straight sinus flow velocity in healthy humans. AB - The influence of whole-body positions on the cerebral blood flow in normal subjects is unclear. Blood flow in cerebral veins and sinuses is continuous, pulsatile and proportional to cerebral blood flow. We examined young healthy volunteers to evaluate peak mean flow velocity (vm) in the straight sinus (SS) assessed by transcranial Doppler sonography in predefined variations of the whole body pitch position relative to gravity in the presence of a normal (normocarbia) and an impaired (hypercarbia) cerebral autoregulation. A 2 MHz ultrasound probe was fixed with a headband nearby the protuberantia occipitalis externa. Fifteen subjects were seated in a motorized three-dimensional turntable. Vm-SS, blood pressure and heart rate were monitored in five whole-body pitch positions from upright (0 degrees ) to "20 degrees head-hanging" (110 degrees ): 0, 30, 60, 90 and 110 degrees . The experiment was repeated during the inspiration of 5% CO2. Of 15 subjects, 14 showed reliable ultrasound data; the results of one subject with movement artifacts were excluded. Vm-SS values under normocarbia (hypercarbia) were 23.9 +/- 4.2 cm/s (40.9 +/- 6.7 cm/s) at 0 degrees , 23.1 +/- 5.0 cm/s (38.0 +/- 5.0 cm/s) at 30 degrees , 24.9 +/- 5.1 cm/s (39.9 +/- 3.3 cm/s) at 60 degrees , 29.2 +/- 8.5 cm/s (41.0 +/- 4.7 cm/s) at 90 degrees and 27.0 +/- 11.6 cm/s (43.6 +/- 12.1 cm/s) at 110 degrees . Vm-SS measured under normocarbia (p = 0.09) and hypercarbia (p = 0.25) were not affected while subjects were positioned from upright toward "20 degrees head-hanging", whereas blood pressure and heart rate decreased (p < 0.01). Our results suggest that changes of whole-body position from upright to "20 degrees head-hanging" do not alter cerebral blood flow in healthy subjects. PMID- 18160199 TI - Angiotensin receptor subtype mediated physiologies and behaviors: new discoveries and clinical targets. AB - The renin-angiotensin system (RAS) mediates several classic physiologies including body water and electrolyte homeostasis, blood pressure, cyclicity of reproductive hormones and sexual behaviors, and the regulation of pituitary gland hormones. These functions appear to be mediated by the angiotensin II (AngII)/AT(1) receptor subtype system. More recently, the angiotensin IV (AngIV)/AT(4) receptor subtype system has been implicated in cognitive processing, cerebroprotection, local blood flow, stress, anxiety and depression. There is accumulating evidence to suggest an inhibitory influence by AngII acting at the AT(1) subtype, and a facilitory role by AngIV acting at the AT(4) subtype, on neuronal firing rate, long-term potentiation, associative and spatial learning, and memory. This review initially describes the biochemical pathways that permit synthesis and degradation of active angiotensin peptides and three receptor subtypes (AT(1), AT(2) and AT(4)) thus far characterized. There is vigorous debate concerning the identity of the most recently discovered receptor subtype, AT(4). Descriptions of classic and novel physiologies and behaviors controlled by the RAS are presented. This review concludes with a consideration of the emerging therapeutic applications suggested by these newly discovered functions of the RAS. PMID- 18160203 TI - Variability of middle cerebral artery blood flow with hypercapnia in women. AB - We examined the effect of euoxic hypercapnia on middle cerebral artery (MCA) blood flow velocity waveform parameters in pre- and postmenopausal women by exposing 24 healthy women (12 pre-, 12 postmenopausal) to hypercapnia for 20 min. MCA blood flow velocity was measured continuously by transcranial Doppler ultrasound. The data were run through an algorithm that detected the feature points of the waveforms and then analyzed for statistically significant group differences. The changes in mean blood flow velocity with euoxic hypercapnia were not significant between the two groups. However, certain feature points, particularly the velocity of the reflected shoulder (V(REFLEC)), increased (89.4 +/- 14.6 to 110.0 +/- 20.5 cm/s and 102.3 +/- 14.1 to 125.1 +/- 14.9 cm/s from euoxic eucapnia to euoxic hypercapnia in pre- and postmenopausal women, respectively), as did the augmentation index (79.9 +/- 10.4 to 85.9 +/- 12.6% and 114.7 +/- 12.8 to 119.0 +/- 12.6%) and pulsatility index (0.86 +/- 0.18 to 0.74 +/- 0.15 and 0.71 +/- 0.11 to 0.66 +/- 0.11). Furthermore, while systolic peak velocity (V(SYS)) was the highest point of the waveform in premenopausal women, V(REFLEC) was the highest point for the postmenopausal cohort. The implications of this finding become obvious when calculating pulsatility index (PI), the values of which varied significantly for the postmenopausal women, depending on whether V(SYS) or the absolute maximum was used. These findings suggest that hypercapnia increases blood flow velocity waveform reflections, and that PI calculations, particularly for older age groups, may need to be considered more carefully, since these reflections often exceed the systolic peak velocity. PMID- 18160204 TI - Cholinergic innervation and muscarinic receptors in the human prostate. AB - OBJECTIVE: In light of recent interest in the use of muscarinic receptor antagonists for the treatment of male lower urinary tract symptoms, understanding how such drugs work not only on the bladder but also on the prostate is important. METHODS: A literature review was conducted to identify studies on the cholinergic innervation and presence and function of muscarinic acetylcholine receptors in the human prostate. RESULTS: The available studies demonstrate a dense cholinergic innervation within both stromal and epithelial compartments of the prostate. Concomitantly, the human prostate expresses muscarinic receptors at densities exceeding those of alpha(1)-adrenoceptors. They mainly belong to the M(1) subtype and are found on epithelial cells, but a smaller population of M(2) receptors is found on stromal cells. Both populations have been shown to be functional in signal transduction assays. However, in line with the sparse receptor density on stromal smooth muscle cells, contractile responses of the prostate are only small. Data from prostate cancer cell lines and from botulinum toxin injections into the benign prostate raise the possibility that muscarinic receptors may promote prostatic growth. Animal data suggest that muscarinic receptors may be of primary importance in the genesis of prostatic secretions, but this needs to be confirmed in humans. CONCLUSIONS: Taken together it appears that direct effects on the prostate need to be considered when using muscarinic receptor antagonists in men. They may primarily involve alterations of glandular secretion and prostatic growth. PMID- 18160205 TI - Re: damien pouessel, stephane culine. High frequency of intracerebral hemorrhage in metastatic renal carcinoma patients with brain metastases treated with tyrosine kinase inhibitors targeting the vascular endothelial growth factor receptor. Eur urol 2008;53:376-81. PMID- 18160206 TI - Editorial comment on: impact of gender in renal cell carcinoma: an analysis of the SEER database. PMID- 18160207 TI - Impact of gender in renal cell carcinoma: an analysis of the SEER database. AB - OBJECTIVE: To evaluate gender differences in initial presentation, pathology, and outcomes with renal cell carcinoma (RCC). MATERIALS AND METHODS: The 1973-2004 Surveillance Epidemiology and End Results (SEER) 17-registries database was analyzed for renal tumors from 1988 to 2004 coded as primary site "kidney and renal pelvis." After various exclusions, a final study group of 35,336 cases with complete data was obtained. Demographic variables analyzed included age, sex, and race. Tumor variables included size, stage at diagnosis, grade, and histology. Primary outcome variables included overall and cancer-specific survival. RESULTS: Of the patients, 22,288 were male (63%). Females presented with smaller tumors (5.9 vs. 6.1cm, p<0.0001) of lower grade (p<0.0001). Males had a higher incidence of regional or metastatic spread of renal carcinoma (p<0.0001). Median overall survival from time of diagnosis was 130 mo for females versus 110 mo for males (p<0.0001). In comparisons of males and females, 5-yr cancer-specific survival was 78% versus 81%, and 5-yr overall survival was 65% versus 69% (p<0.0001). On multivariate analysis, cancer-specific survival was similar (HR, 1.00, p=0.960), whereas overall survival was significantly longer for females (HR, 0.92, p<0.0001). Older age at diagnosis, larger tumor size, higher grade, higher SEER historic stage, and sarcomatoid, collecting duct, or "other" histology were related to worse cancer-specific and overall survival. CONCLUSIONS: Men present with larger, higher stage, higher grade RCC than women. Overall survival is better in women, whereas cancer-specific survival is not significantly different. PMID- 18160208 TI - Editorial comment on: impact of gender in renal cell carcinoma: an analysis of the SEER database. PMID- 18160209 TI - Open to debate. The motion: at least 18 cores are necessary to make a prostatic biopsy useful. PMID- 18160210 TI - QSAR using evolved neural networks for the inhibition of mutant PfDHFR by pyrimethamine derivatives. AB - Quantitative structure-activity relationship (QSAR) models were developed for dihydrofolate reductase (DHFR) inhibition by pyrimethamine derivatives using small molecule descriptors derived from MOE and/or QikProp and linear or nonlinear modeling. During this analysis, the best QSAR models were identified when using MOE descriptors and nonlinear models (artificial neural networks) optimized by evolutionary computation. The resulting models can be used to identify key descriptors for DHFR inhibition and are useful for high-throughput screening of novel drug leads. PMID- 18160211 TI - Brainstem tuberculoma in the immunocompetent: case report and literature review. AB - Brainstem tuberculoma is an uncommon entity especially in the immunocompetent adult. The authors describe the case of a 32-year-old immunocompetent woman with no systemic signs or symptoms of tuberculosis, presenting with sudden-onset hemiparesis and diplopia. MRI of brain revealed an enhancing lesion in the midbrain and thalamus. Cerebrospinal fluid examination was normal. Chest imaging was consistent with miliary tuberculosis; however initial expectorated sputum was culture negative. Bronchosopy sputum culture was positive for Mycobacterium tuberculosis. Her PPD was negative. Patient became asymptomatic with treatment for tuberculosis and follow-up MRI showed complete resolution of the lesion a year later. We conclude that a high index of suspicion is essential to make an early diagnosis of intracranial tuberculoma, since often, traditional TB tests like PPD and sputum examinations can be negative. PMID- 18160212 TI - Decreased expression of ING2 gene and its clinicopathological significance in hepatocellular carcinoma. AB - The inhibitor of growth (ING) family member 2 (ING2) is a newly discovered member of ING family that can regulate a wide range of cellular processes including cell growth arrest, apoptosis, and DNA repair. Researches have shown that ING2 can activate p53 and p53-mediated apoptotic pathway involved in the hepatocarcinogenesis. To investigate the role of ING2 in hepatocellular carcinoma (HCC) pathogenesis, we analyzed the correlations between the ING2 expression level and clinicopathologic factors and studied its prognostic role in primary HCC. Using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, ING2 transcription and post-transcription level was found to be downregulated in the majority of tumors compared with matched non-tumors liver tissues (p=0.004 and p=0.014, respectively). The immunohistochemistry data indicated significant reduction of ING2 expression level in 44 of 84 (52.4%) HCC cases. In addition, the expression level of ING2 correlated with tumor size, histopathologic classification, serum AFP (p<0.05). Kaplan-Meier curves demonstrated that patients with reduced ING2 expression were at significantly increased risk for shortened survival time (p=0.009). Using multivariate analysis, ING2 expression was found to be an independent prognostic factor. Our data suggest that ING2 is involved in the progression of HCC, therefore it is considered to be a candidate tumor suppressor gene and its significantly decreased expression in HCC may lead to an unfavorable prognosis. PMID- 18160213 TI - Characterization of the 12q amplicons by high-resolution, oligonucleotide array CGH and expression analyses of a novel liposarcoma cell line. AB - The cytogenetic hallmark of well-differentiated liposarcoma (WDLS) is a giant marker chromosomes containing amplified genes from chromosome 12q13-q15. Here, we have employed SKY and high-resolution 244K oligonucleotide array CGH to characterize rearrangements and amplifications in a new WDLS cell line (GOT3) with a giant marker chromosome derived from chromosomes 12, 1, and X. The most prominent amplifications included 144 genes in 12q11-q21.2, 201 genes in 1q23.3 q44, and six genes in 13q32.1-q32.2. In the 12q amplicons, MDM2 showed the highest level of amplification followed by LYZ, HMGA2 (5'-part), TSPAN8, CNOT2, YEATS4, CDK4, GNS, HELB, and TSFM. Expression analysis of genes from the three major amplicons revealed that several highly amplified potential target genes, including HMGA2, MDM2, YEATS4, CDK4, PKP1, IPO9, and SOX21, were strongly overexpressed. Studies of cell cycle controlling proteins that interact with CDK4 and MDM2 revealed an abnormally strong expression of cyclins D1 and E. The selective high-level amplification of the 5'-part of HMGA2, including the DNA binding domains, suggests that this gene is a major target of amplifications in WDLS. Our results also identify several novel candidate genes of potential pathogenetic and therapeutic importance for WDLS. PMID- 18160214 TI - EGF-IL-18 fusion protein as a potential anti-tumor reagent by induction of immune response and apoptosis in cancer cells. AB - We report here the generation and characterization of EGF-IL-18 fusion protein as an anti-tumor reagent. The epidermal growth factor (EGF) and interleukin-18 (IL 18) fusion protein was shown to induce interferon-gamma (IFNgamma) expression and secretion in KG-1 cells, and to promote PBMNC proliferation. It also stimulated activation of CD4+ T cells, and increased other immune responses. Moreover, EGF IL-18 could induce significant tumor regression in SMMC-7721-xenografted Balb/c nude mice when administered together with peritumoral injection of X-ray irradiated NK-92 cells, and this regression is associated with arresting of the tumor cells in G1 phase and induction of apoptosis. PMID- 18160215 TI - Effects of peptizing conditions on nanometer properties and photocatalytic activity of TiO2 hydrosols prepared by H2TiO3. AB - TiO2 hydrosols were prepared from metatitanic acid (H2TiO3) by chemical precipitation-peptization method under various peptizing conditions. The effects of peptizing conditions on nanosized properties and photocatalytic activity of TiO2 hydrosols were investigated. The crystal structure, crystallinity, particle size distribution, and transparency (T%) of as-obtained hydrosols were characterized by means of X-ray diffraction, transmission electron microscopy, light-scattering size analyzer, and UV-vis transmittance spectra. The results showed that the properties of hydrosols depended on peptizing conditions including a molar ratio of H+/Ti, temperature, and solid content. The photoactivity of TiO2 hydrosols was evaluated in terms of the degradation of rhodamine B (RhB) in aqueous solution, and formaldehyde (HCHO) and methyl mercaptan (CH3SH) in gaseous phase. The results showed that increase in H+/Ti ranging 0.19-0.75 led to the decrease in particle size and the increase in transparency. With increasing of temperature, particle sizes increased while the transparency and photoactivity decreased steadily when the temperature was higher than 65 degrees C. The particle size, transparency and photoactivity of the hydrosols hardly depended on solid content when it was not less than 2%. It should be confirmed that the hydrosols with higher crystallinity, smaller particle size and higher transparency could have the higher photoactivity for the degradation of RhB, CH3SH, and HCHO. In this study, the optimal peptizing conditions were determined to be H+/Ti=0.75, temperature=65 degrees C and solid content=2-6%. PMID- 18160216 TI - Degradation of 4-chlorophenol in TiO2, WO3, SnO2, TiO2/WO3 and TiO2/SnO2 systems. AB - The present study was undertaken to evaluate the degradation performance of 4 chlorophenol (4-CP) using TiO2/WO3 and TiO2/SnO2 systems. A BET surface area analyzer, UV-vis spectroscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD) and electron spectroscopy for chemical analysis (ESCA) were employed to characterize the photocatalyst. The band edge wavelength increased to 475 nm and gap energy decreased to 2.61 eV in the TiO2/WO3 system as compare to the single TiO2. Although the specific surfaces area of TiO2/WO3 decreases due to its larger size as compared to either TiO2 or WO3, the 4-CP degradation efficiency significantly increased as compared to single TiO2 or WO3 system at 435 nm wavelength. The TiO2/WO3 degradation of 4-CP at 369 nm was in fact inhibited. For TiO2/SnO2, the degradation efficiency also suffered at 369 nm, and only slightly increased compared to otherwise hardly 4-CP degraded in single TiO2 or SnO2 system. Since there is a significant accumulation of byproducts, the buildup of these intermediates on the catalyst surface may be responsible for their poor performance. PMID- 18160217 TI - Recovery of surfactant SDS and Cd2+ from permeate in MEUF using a continuous foam fractionator. AB - The purpose of this study was to investigate the use of foam fractionation to recover valuable surfactant (SDS) and metal ion (Cd2+) in the permeate of micellar-enhanced ultrafiltration (MEUF). The effects of operating factors, such as time, air flow rate, feed flow rate, liquid height, foam height, feed surfactant concentration, ethanol concentration and temperature on separation characteristics were studied in the continuous operation. When the concentrations of surfactant (SDS) and metal ion (Cd2+) in the feed solution were 500 mg/L and 10 mg/L, an enrichment ratio of 3.1 was achieved for SDS along with 52% removal fraction, as well as 99.35% Cd2+ was removed, after optimization of different process parameters. As the optimized results, the air and liquid flow rates were 100 L/h and 5 L/h, liquid and foam heights were 45 cm and 66 cm, respectively, sparger pore size was 10 microm. The Cd2+ concentration in the effluent was lower than 0.1mg/L which could meet the integrated wastewater discharge standard (the first grade of national discharge standards in China). PMID- 18160218 TI - Complement activation in the peripheral nervous system following the spinal nerve ligation model of neuropathic pain. AB - Neuroinflammatory and neuroimmune mechanisms, as exemplified by infiltrating immune cells and activation of resident endothelial/glial cells, respectively, are known to be involved in the establishment and maintenance of chronic pain. An immune system pathway that may be involved in the activation of both immune and glial cells is complement. The complement pathway is made up of a large number of distinct plasma proteins which react with one another to opsonize pathogens and induce a series of inflammatory responses to help fight infection. Cleaved products and complexes produced by complement activation are responsible for a range of effects including mediation of immune infiltration, activation of phagocytes, opsonization/lysis of pathogens and injured cells, and production of vasoactive amines such as histamine and serotonin. Gene-expression microarray analysis performed on the rat spinal nerve ligation (SNL) model of neuropathic pain revealed that multiple complement components including the C1 inhibitor, C1q alpha, beta, and gamma, C1r, C1s, C2, C3, C4, C7, and factors B, D, H, and P, were up-regulated while DAF was down-regulated. Regulation of C3 and DAF was confirmed by real-time RT-PCR and in situ hybridization. To test the hypothesis that complement plays a role in neuropathic pain, SNL rats were treated with cobra venom factor (CVF) to deplete plasma of complement component C3. Pain behavior was significantly attenuated in SNL rats treated with CVF as was complement activity at the ipsilateral dorsal root ganglia. Our results suggest the complement pathway might be a novel target for the development of neuropathic pain therapeutics. PMID- 18160219 TI - Hypoalgesia in schizophrenia is independent of antipsychotic drugs: a systematic quantitative review of experimental studies. AB - Diminished sensitivity to pain in schizophrenia has been reported since the early works of Bleuler [Bleuler E. Textbook of psychiatry (trans. Brill HA, 1951). New York: Dover Publications; 1911] and Kraepelin [Kraepelin E. Dementia praecox and paraphrenia. Edinburgh, Scotland: E and S Livingstone; 1919]. Over the last decade, experimental studies have measured pain perception in schizophrenia and produced mixed results. This meta-analysis sought to determine if the scientific literature confirms the hypothesized hypoalgesia in schizophrenia. The search was performed with computerised literature databases. A study was retained in the meta-analysis if: (i) it comprised a group of schizophrenia patients, compared to a control group of healthy volunteers; and (ii) pain was measured via experimental procedures (e.g. thermal, electrical, or mechanical stimuli). Using Comprehensive Meta-Analysis-2, effect size estimates of the differences in pain scores (all pain scores derived from all pain tests) between schizophrenia patients and healthy volunteers were calculated. Eleven studies were included in the meta-analysis. For the composite analysis, a positive, moderate, and significant effect size estimate emerged (N=497; Hedges's g=0.437; p=0.005), suggesting that patients with schizophrenia show a diminished response to experimentally-induced pain. Secondary analyses showed that: (i) drug-free patients also have hypoalgesic responses; and that (ii) sensory thresholds are increased in schizophrenia patients. This meta-analysis substantiates the hypothesis of a diminished pain response in schizophrenia. The study also suggests that hypoalgesia in schizophrenia cannot be solely explained by the effects of antipsychotic drugs, and that it may not be a pain-specific blunted response. Further studies are warranted to determine the clinical and biological correlates, and the social and health consequences, of hypoalgesia in schizophrenia. PMID- 18160220 TI - Effects of tilt on high-resolution ADF-STEM imaging. AB - A study of the effects of small-angle specimen tilt on high-resolution annular dark field images was carried out for scanning transmission electron microscopes with uncorrected and aberration-corrected probes using multislice simulations. The results indicate that even in the cases of specimen tilts of the order of 1 degree a factor of 2 reduction in the contrast of the high-resolution image should be expected. The effect holds for different orientations of the crystal. Calculations also indicate that as the tilted specimen gets thicker the contrast reduction increases. Images simulated with a low-angle annular dark field detector show that tilt effects are more pronounced in this case and suggest that these low-angle detectors can be used to correct specimen tilt during scanning transmission electron microscopes operation. PMID- 18160221 TI - Detection and surveillance for animal trichinellosis in GB. AB - The zoonotic disease trichinellosis is considered one of the re-emerging diseases with surveillance and control methods constantly gaining more importance worldwide. Recent change in European Union (EU) legislation introduces Trichinella-free production, and the possibility of risk-based monitoring for Trichinella in pigs. This has increased the role of wildlife surveillance programmes and their impact on protecting human health as well as highlighted the need for harmonised surveillance protocols and test methods for these infections. A modified digest method, based on the EU reference method for Trichinella testing of pig meat, was used to screen foxes present in Great Britain (England, Scotland and Wales) for trichinellosis. The method was validated using batched pools of 10 g foreleg muscle from up to 20 foxes (maximum amount 200 g). The method gave an average trichinae recovery rate of 71% for spiked samples. Assuming this recovery rate applies to all contaminated samples, then the test sensitivity would be 70% for all tissue samples with 0.1 trichinae per 10 g of foreleg muscle, 99.9% for samples with 1 trichinae per 10 g, and 100% for samples with 2 or more trichinae per 10 g. In two separate studies, conducted between 1999 to 2001 (Smith et al., 2003) and 2003 to 2007, over 3500 wild foxes have been screened for Trichinella with negative results. In the second study reported here, foxes were collected from locations throughout Great Britain using a stratified sampling method based on fox population densities. All work was conducted in compliance with appropriate quality assurance systems, latterly under ISO 9001. Results to date indicate the national prevalence of trichinellosis in foxes is <0.001 based on a 10 g individual sample size, an infection level of 1 larva per gram (l pg) and 95% confidence interval. This, together with no reports of trichinellosis in domesticated pigs, suggests that Britain can be considered a region of negligible risk of trichinellosis. PMID- 18160222 TI - Repeated high dose imidocarb dipropionate treatment did not eliminate Babesia caballi from naturally infected horses as determined by PCR-reverse line blot hybridization. AB - Imidocarb treatment of horses infected with Babesia caballi is supposed to eliminate the infection, but data on the efficacy of this treatment is scarce. The study presented here concerns four Paso Fino horses, which were imported into the island of Curacao on the basis of a piroplasmosis negative complement fixation test (CFT). Upon re-testing with an indirect fluorescent antibody test immediately after arrival in Curacao, two horses appeared to have antibodies to B. caballi and all horses had antibodies to Theileria equi. Subsequent testing with polymerase chain reaction combined with a reverse line blot yielded positive results for both agents in all four horses. Treatment with five consecutive doses of imidocarb dipropionate (4.7 mg/kg BW im q 72 h), temporarily resulted in negative results, but B. caballi and T. equi were detected again in the samples taken at 6 and 18 weeks after completion of the treatment. These results confirm that the CFT is not a suitable test for pre-import testing and that even high dose treatment with imidocarb may not be capable of eliminating B. caballi and T. equi infections from healthy carriers. PMID- 18160223 TI - Prevalence of Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, Bartonella vinsonii berkhoffii, and Rickettsia spp. in dogs from Grenada. AB - To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance. PMID- 18160224 TI - Tumor necrosis factor alpha-mediated nitric oxide production enhances manganese superoxide dismutase nitration and mitochondrial dysfunction in primary neurons: an insight into the role of glial cells. AB - Tumor necrosis factor-alpha (TNF-alpha), a ubiquitous pro-inflammatory cytokine, is an important mediator in the immune-neuroendocrine system that affects the CNS. The present study demonstrates that treatment with TNF-alpha activates microglia to increase TNF-alpha production in primary cultures of glial cells isolated from wild-type (WT) mice and mice deficient in the inducible form of nitric oxide synthase (iNOSKO). However, mitochondrial dysfunction in WT neurons occurs at lower concentrations of TNF-alpha when neurons are directly treated with TNF-alpha or co-cultured with TNF-alpha-treated microglia than iNOSKO neurons similarly treated. Immunofluorescent staining of primary neurons co cultured with TNF-alpha-treated microglia reveals that the antioxidant enzyme in mitochondria, manganese superoxide dismutase (MnSOD), is co-localized with nitrotyrosine in WT but not in iNOSKO primary neuronal cells. Importantly, the percentage of surviving neurons is significantly reduced in WT neurons compared with iNOSKO neurons under identical treatment conditions. Together, the results suggest that TNF-alpha activates microglia to produce high levels of TNF-alpha and that production of nitric oxide (NO) in neurons is an important factor affecting MnSOD nitration and subsequent mitochondrial dysfunction. PMID- 18160225 TI - Time course of blood oxygenation level-dependent signal response after theta burst transcranial magnetic stimulation of the frontal eye field. AB - The aim of the current study was to examine the effect of theta burst repetitive transcranial magnetic stimulation (rTMS) on the blood oxygenation level-dependent (BOLD) activation during repeated functional magnetic resonance imaging (fMRI) measurements. Theta burst rTMS was applied over the right frontal eye field in seven healthy subjects. Subsequently, repeated fMRI measurements were performed during a saccade-fixation task (block design) 5, 20, 35, and 60 min after stimulation. We found that theta burst rTMS induced a strong and long-lasting decrease of the BOLD signal response of the stimulated frontal eye field at 20 and 35 min. Furthermore, less pronounced alterations of the BOLD signal response with different dynamics were found for remote oculomotor areas such as the left frontal eye field, the pre-supplementary eye field, the supplementary eye field, and both parietal eye fields. Recovery of the BOLD signal changes in the anterior remote areas started earlier than in the posterior remote areas. These results show that a) the major inhibitory impact of theta burst rTMS occurs directly in the stimulated area itself, and that b) a lower effect on remote, oculomotor areas can be induced. PMID- 18160226 TI - Opposite effects of shell or core stimulation of the nucleus accumbens on long term potentiation in dentate gyrus of anesthetized rats. AB - Hippocampal long-term potentiation (LTP) is a long-lasting increase in synaptic efficacy which is considered a cellular correlate of learning and memory. It has been shown that both, stimuli with emotional/motivational content and the electrical stimulation the basolateral amygdala, can modulate hippocampal LTP. The nucleus accumbens is part of the ventral striatum and is composed of two main regions: core and shell. Core and shell share a similar cellular composition, but differ in their connectivity with other brain areas. Considering that the nucleus accumbens is related to motivation and that it receives a strong projection from the basolateral amygdala, we have studied the effect of stimulating accumbens shell or core on medial perforant path-granule cells' LTP in anesthetized male Wistar rats. We found that electrical stimulation of the shell enhances the magnitude of LTP while the stimulation of the core completely prevents LTP induction. The stimulation of the accumbens shell or core alone produced no apparent, direct field potential in dentate gyrus. Additionally, the co stimulation of the shell or core with the medial perforant path does not modify the input-output curves obtained using stimulation of the perforant path only. These results demonstrate that electrical stimulation of the accumbens shell or core has a bidirectional effect on LTP induction at the dentate gyrus. PMID- 18160227 TI - Pre-ischemic exercise reduces matrix metalloproteinase-9 expression and ameliorates blood-brain barrier dysfunction in stroke. AB - Exercise reduces ischemia and reperfusion (I/R) injury in the rat stroke model. We investigated whether pre-ischemic exercise ameliorates blood-brain barrier (BBB) dysfunction in stroke by reducing matrix metalloproteinase (MMP)-9 expression and strengthening basal lamina. Adult male Sprague-Dawley rats were subjected to a 30 min exercise program on a treadmill 5 days a week for 3 weeks. Stroke was induced by a 2-h middle cerebral artery (MCA) occlusion using an intraluminal filament in the exercised and non-exercised groups. Brain infarction was measured and neurological deficits were scored. BBB dysfunction was determined by examining brain edema and Evans Blue extravasation. Expression of collagen IV, the major component of basal lamina essential for maintenance of the endothelial permeability barrier, was quantitatively detected by Western blot and immunocytochemistry. Ex vivo techniques were used to compare collagen IV-labeled vessels in response to ischemic insult. Temporal relationship of expression of MMP-9 and its endogenous inhibitor, the tissue inhibitors of metalloproteinase-1 (TIMP-1), was determined by real-time PCR for mRNA and Western blot for protein during reperfusion. Brain edema and Evans Blue leakage were both significantly (P<0.01) reduced after stroke in the exercised group, in association with reduced brain infarct volume and neurological deficits. Western blot analysis indicated that exercise enhanced collagen IV expression and reduced the collagen loss after stroke. Immunocytochemistry demonstrated that collagen IV-labeled vessels were significantly (P<0.01) increased in exercised rats. In the ex vivo study, after exercised brains were incubated with ischemic brain tissue, a significantly (P<0.01) higher level of collagen IV-labeled vessels was observed as compared with non-exercised brains following the same treatment. The ex vivo study also revealed a key role of MMP-9 in exercise-strengthened collagen IV expression against I/R injury. TIMP-1 protein levels were significantly (P<0.01) increased by exercise. Our results indicate that pre-ischemic exercise reduces brain injury by improving BBB function and enhancing basal lamina integrity in stroke. This study suggests that the neuroprotective effect of physical exercise is associated with an imbalance of MMP-9 and TIMP-1 expression. PMID- 18160228 TI - Metastases of well-differentiated thyroid and oral squamous cell carcinoma to the same neck lymph node. PMID- 18160229 TI - [Patient awareness and satisfaction of the Aurore perinatal network]. AB - OBJECTIVES: Assess patient awareness, satisfaction and expectations three years after the implementation of the Aurore perinatal network. METHOD AND MATERIALS: Semi-guided interviews with 60 Aurore network perinatal patients, 20 having undergone in-utero transfers (IUT), 20 whose newborns were transferred to the neonate unit (NNT), and 20 post-delivery without IUT or NNT, between September 2007 and January 2007. RESULTS: Fifteen percent of patients were familiar with the Aurore perinatal network (9/60). The concepts of IUT and NNT were not associated with that of the perinatal network. The information provided helped 85 % of the patients to feel more secure (51/60). For 83 % of the patients with mother-child bond was maintained (50/60). In 90 % of cases, the teams included the father in the healthcare provision process (54/60). Patients had expectations on a greater consideration on the emotional aspect during a transfer. CONCLUSION: Few patients knew the perinatal network. They were globally satisified during a transfer, they insisted on the importance of staff empathy and availability. PMID- 18160230 TI - [Risk of missed diagnosis of 22q11.2 deletion in a fetal cardiac conotruncal malformation when another chromosomal abnormality is detected]. AB - We present a rare case of prenatal diagnosis of two de novo chromosome structural rearrangements including a translocation (1;3) associated with a 22q11.2 deletion. The amniocentesis was performed because the systematic ultrasound examination revealed: right aortic cross with double aortic arch, with normal size of aorta and pulmonary artery. Our report emphasises that 22q11.2 deletion must be looked for when a fetal cardiac conotruncal malformation is diagnosed, even in the presence of another chromosomal abnormality. In prenatal diagnosis, this can have implication for patient management and genetic counselling. PMID- 18160231 TI - [Sexual abuse: epidemiological, clinical aspects and management at gynaecological and obstetrical department of Dakar University Hospital]. AB - OBJECTIVES: To carry out the epidemiological and clinical characteristics of supposed victims of sexual abuse and to evaluate case management. PATIENTS AND METHODS: A prospective study was conducted about cases of presumed sexual abuse received at the gynaecological and obstetrical clinic department of Aristide-le Dantec hospital from January 2003 to May 2005. RESULTS: A total of 55 cases were reported and represented 0.4% of admissions in the clinic during the period of study. Twenty percent of them (20%) were referred on judicial requisition. The mean time between sexual abuse and consultation was 15 days. Victims were 14 years old in average, nulligeste in 96.5% of cases and living in the suburban area of Dakar. The presumed "violenter" was a man of 32 years, belonging to the environment of the victim in 70% of cases (spiritual guide, joint-tenant, friend of the family...). The type of sexual assault was an unprotected genito-genital intercourse in 67.3% of cases. On the clinical plan, 70.9% of patients suffered recent genital traumatism, 54.5% genital examination showed hymeneal lesions. The HIV test was positive in two cases. During the follow-up of the patients, three pregnancies occurred and for only 9.1%, a psychological assistance was proposed. CONCLUSION: Sexual abuses represent a current sociocultural issue. Prevention required large information campaign. Early management is necessary in order to prevent the sexual transmitted diseases and psychological side effects. PMID- 18160232 TI - snoTARGET shows that human orphan snoRNA targets locate close to alternative splice junctions. AB - Among thousands of non-protein-coding RNAs which have been found in humans, a significant group represents snoRNA molecules that guide other types of RNAs to specific chemical modifications, cleavages, or proper folding. Yet, hundreds of mammalian snoRNAs have unknown function and are referred to as "orphan" molecules. In 2006, for the first time, it was shown that a particular orphan snoRNA (HBII-52) plays an important role in the regulation of alternative splicing of the serotonin receptor gene in humans and other mammals. In order to facilitate the investigation of possible involvement of snoRNAs in the regulation of pre-mRNA processing, we developed a new computational web resource, snoTARGET, which searches for possible guiding sites for snoRNAs among the entire set of human and rodent exonic and intronic sequences. Application of snoTARGET for finding possible guiding sites for a number of human and rodent orphan C/D-box snoRNAs showed that another subgroup of these molecules (HBII-85) have statistically elevated guiding preferences toward exons compared to introns. Moreover, these energetically favorable putative targets of HBII-85 snoRNAs are non-randomly associated with genes producing alternatively spliced mRNA isoforms. The snoTARGET resource is freely available at: (http://hsc.utoledo.edu/depts/bioinfo/snotarget.html). PMID- 18160233 TI - Strain differentiating real-time PCR for Mycoplasma gallisepticum live vaccine evaluation studies. AB - Mycoplasma gallisepticum causes respiratory disease and production losses in poultry. Vaccination of poultry with M. gallisepticum live vaccines is an approach to reduce susceptibility to infection and to prevent the economic losses. The development and evaluation of live vaccines usually requires the involvement of several vaccine and challenge strains in the same experimental setup. Our goal was to develop a tool to allow the differentiation between a set of known M. gallisepticum strains in a quantitative manner. We developed 5 real time PCR assays that absolutely differentiated between one of the five commercial and laboratory vaccine strains: F, ts-11, 6/85, K5831, K5054, and the challenge strain R low when tested on in vitro cultures. The assay K5831 vs. R low was also tested on specimens from live birds that were vaccinated with K5831 and challenged with R low, and successfully differentiated between the vaccine and the challenge strains in a quantitative manner. This preliminary in vivo application of the method also shed light on possible protection mechanisms for the M. gallisepticum K5831 vaccine strain. PMID- 18160234 TI - Design and implementation of a database for Brucella melitensis genome annotation. AB - The genome sequences of three Brucella biovars and of some species close to Brucella sp. have become available, leading to new relationship analysis. Moreover, the automatic genome annotation of the pathogenic bacteria Brucella melitensis has been manually corrected by a consortium of experts, leading to 899 modifications of start sites predictions among the 3198 open reading frames (ORFs) examined. This new annotation, coupled with the results of automatic annotation tools of the complete genome sequences of the B. melitensis genome (including BLASTs to 9 genomes close to Brucella), provides numerous data sets related to predicted functions, biochemical properties and phylogenic comparisons. To made these results available, alphaPAGe, a functional auto updatable database of the corrected sequence genome of B. melitensis, has been built, using the entity-relationship (ER) approach and a multi-purpose database structure. A friendly graphical user interface has been designed, and users can carry out different kinds of information by three levels of queries: (1) the basic search use the classical keywords or sequence identifiers; (2) the original advanced search engine allows to combine (by using logical operators) numerous criteria: (a) keywords (textual comparison) related to the pCDS's function, family domains and cellular localization; (b) physico-chemical characteristics (numerical comparison) such as isoelectric point or molecular weight and structural criteria such as the nucleic length or the number of transmembrane helix (TMH); (c) similarity scores with Escherichia coli and 10 species phylogenetically close to B. melitensis; (3) complex queries can be performed by using a SQL field, which allows all queries respecting the database's structure. The database is publicly available through a Web server at the following url: http://www.fundp.ac.be/urbm/bioinfo/aPAGe. PMID- 18160235 TI - Hexagonal boron nitride as a tablet lubricant and a comparison with conventional lubricants. AB - The objective of this study was to investigate the lubrication properties of hexagonal boron nitride (HBN) as a new tablet lubricant and compare it with conventional lubricants such as magnesium stearate (MGST), stearic acid (STAC), and glyceryl behenate (COMP). Tablets were manufactured on an instrumented single station tablet press to monitor lower punch ejection force (LPEF) containing varied lubricants in different ratio (0.5, 1, 2%). Tablet crushing strength, disintegration time and thickness were measured. Tensile strength of compacted tablets were measured by applying a diametrical load across the edge of tablets to determine mechanical strength. The deformation mechanism of tablets was studied during compression from the Heckel plots with or without lubricants. MGST was found to be the most effective lubricant based on LPEF-lubrication concentration profile and LPEF of HBN was found very close to that of MGST. HBN was better than both STAC and COMP. A good lubrication was obtained at 0.5% for MGST and HBN (189 and 195N, respectively). Where COMP and STAC showed 20 and 35% more LPEF compare to that of MGST (239 and 288N, respectively). Even at the concentration of 2% COMP and STAC did not decrease LPEF as much as 0.5% of MGST and HBN. Like all conventional lubricants the higher the concentration of HBN the lower the mechanical properties of tablets because of its hydrophobic character. However, this deterioration was not as pronounced as MGST. HBN had no significant effect on tablet properties. Based on the Heckel plots, it was observed that after the addition of 1% lubricant granules showed less plastic deformation. PMID- 18160236 TI - Ciprofloxacin-encapsulated poly(DL-lactide-co-glycolide) nanoparticles and its antibacterial activity. AB - The aim of this study was to prepare ciprofloxacin HCl (CIP)-encapsulated poly(dl lactide-co-glycolide) (PLGA) copolymer nanoparticles and its antibacterial potential was evaluated with pathogenic bacteria, Escherichia coli (E. coli), in vitro and in vivo. CIP-encapsulated nanoparticles of PLGA were prepared by multiple emulsion solvent evaporation method. PLGA nanoparticles showed spherical shapes with particle sizes around 100-300 nm. Loading efficiency was lower than 50% (w/w) because of water-solubility properties of CIP. At drug release study, CIP showed initial burst effect for 12 h and then continuously released for 2 weeks. At in vitro antibacterial activity test, CIP-encapsulated nanoparticles showed relatively lower antibacterial activity compared to free CIP due to the sustained release characteristics of nanoparticles. However, CIP-encapsulated PLGA nanoparticles (doses: 25 mg CIP/kg of mice) effectively inhibited the growth of bacteria due to the sustained release characteristics of nanoparticles, while free CIP was less effective on the inhibition of bacterial growth. These results indicated that CIP-encapsulated PLGA nanoparticles have superior effectiveness to inhibit the growth of bacteria in vivo. PMID- 18160237 TI - Prediction of suitable amounts of water in fluidized bed granulation of pharmaceutical formulations using corresponding values of components. AB - This study describes application of a newly developed method to the fluidized bed granulation. The method is based on predicting suitable amounts of water to be added to multi-component formulations using the corresponding values of components prior to granulation trials. The range of appropriate amount of water for each component in a model formulation was estimated in our previous studies with a refractive near-infrared (NIR) moisture sensor. Using those values, we calculated the range of suitable amount of water to add for the model pharmaceutical formulation. In this study, we examined the relationship between the amount of water added to the model formulation and the NIR sensor output value. Then, we performed fluidized bed granulation of the model formulation at steady-state moisture content levels under monitoring with NIR sensor, within and beyond the suitable range of added water that was calculated from the corresponding range of each component. For the model formulation, we found that the predicted values for suitable amounts of added water well corresponded to those in the granulation trials, suggesting that this predictive method may be useful in estimating suitable amounts of water to be added to formulations before fluidized bed granulation trials. PMID- 18160238 TI - Validation of an ion mobility spectrometry (IMS) method for the detection of heroin and cocaine on incriminated material. AB - The validation of a qualitative ion mobility spectrometry (IMS) procedure for the detection of trace amounts of heroin and cocaine on incriminated material using a vacuum cleaner for sampling is presented. The limit of detection, the limit of decision, selectivity and robustness were determined. As an approach, robustness was determined using ionizational interferences and matrix effects. By using this simple sampling procedure, a positive result for incriminated clothes needs a contamination of 250ng cocaine and 1000ng heroin, respectively. PMID- 18160239 TI - The determination by gas chromatography with atomic emission detection of total sulfur in fuels used as forensic evidence. AB - In Japan, taxed diesel fuel from non-taxed fuel oil-A is illegally produced by removing coumarin, which is added as a non-taxed marker. The coumarin is removed using concentrated sulfuric acid and this produces a high viscosity and hazardous material, called "sulfuric acid pitch", as a by-product. This compound has a detrimental effect on the environment and is hazardous to humans. The actions have been associated with organized crime with the illegally gained taxes becoming financial bases. To discriminate legal and diesel oil from illegal product, the peak area ratio R(SC), the ratio of total sulfur to carbon (>C(14)), was used. R(SC) is calculated by the total areas of sulfur and carbon (>C(14)) from the gas chromatogram obtained by gas chromatography-atomic emission detection (GC-AED). Sulfur in legal diesel fuels is strictly regulated by a maximum limit, which was 50ppm (and is now 10ppm), but in the preparation of illegal diesel oil, in which coumarin is eliminated, sulfur cannot be removed. Therefore, the R(SC) of fuel oil-A and illegal fuel oil is over 15, whereas those for legal fuel oil and diesel fuel are under 2.0. Furthermore, these ratios do not change in weathering experiments. GC-AED was applied to an actual arson case and was found to be effective for the determination of total sulfur in trace amounts of accelerants detected in fire debris at the arson scene, and hence was effective for the characterization of the ignitable liquids used. PMID- 18160240 TI - [Pediatric salmonellosis at the Tokoin's teaching hospital, Lome (Togo)]. AB - OBJECTIVE: The authors had for aim to describe the epidemiological, clinical, and bacteriological aspects and outcome of pediatric Salmonella enterica, Salmonella septicemia, over the last 10 years. PATIENTS AND METHODS: We analyzed the case history of 132 patients hospitalized for Salmonellasepticemia (positive blood culture) between 1995 and 2004. RESULTS: Salmonellosis accounted for 0.36% of all hospitalizations. The mean age of patients was 5.86 plus or minus 4.06 years, significantly higher in patients with S. ser. Typhi (7.14+/-4.04 years) than in patients with other serotypes (4.95+/-3.8 years). The clinical presentation was severe in many children (with dehydration (34.8%) and emaciation (55.3%)), so HIV was suspected and investigated in 51 patients (38.6%). Eight patients were HIV positive. Three serotypes of S. enterica were predominant: S. ser. Typhi, 55 cases (41.7%), S. ser. Enteritidis, 32 cases (24.2%), and S. ser. Typhimurium, 19 cases (14.4%). The bacterial susceptibility to antibiotics was good for ceftriaxone and ciprofloxacin (100%). But 78.8% of the serotypes were resistant to amoxicillin, 75.4% to chloramphenicol, and 69.4% to cotrimoxazole. The mean duration of hospitalization was 13.7 plus or minus 7.4 days (range 4-34 days). Complications occurred in 15.9% of cases, dominated by digestive bleeding (10.6%), and 6.1% of patients died. PMID- 18160241 TI - Effect of ageing on the regulation of motor unit force in rat medial gastrocnemius muscle. AB - The influence of ageing on the regulation of force through the firing rate (force frequency relationship) and motor unit contractile output were investigated in three types of motor unit (MU): FF, FR and S, in the medial gastrocnemius muscle. A control group of young (5-10 months) Wistar rats was compared to three groups of older (20-21, 24-25 and 28-30 months) animals. The optimal tetanus characterized by the maximum contractile output (force-time area - FTA - per single pulse) was determined. During ageing, the steep part of the force frequency relationship of medial gastrocnemius MUs shifted towards lower stimulation rates. However, in all MU types of the oldest rats, the opposite shift (towards higher rates) was observed. Ageing induced a substantial increase in the maximal FTA per pulse, particularly in S and FF units, but only subtly altered the fusion index of the optimal tetanus of MUs. Moreover, a transient increase in the mean forces of FF MUs was revealed in the groups of 20-21 and 24 25 months rats, and a significant decrease in the fatigue resistance of FR MUs accompanied ageing. These findings increase our understanding of the functional mechanisms responsible for changes in rate coding and alterations in muscle fatigability during ageing. PMID- 18160242 TI - Ultrasonography of symptomatic rotator cuff tears compared with MR imaging and surgery. AB - PURPOSE: To compare the accuracy of ultrasonography and magnetic resonance imaging in the detection of rotator cuff tears. MATERIALS AND METHODS: Ninety-six patients with clinically suspected rotator cuff pathology underwent ultrasonography and magnetic resonance imaging of the shoulder. The findings in 88 patients were compared with arthroscopy or open surgery. RESULTS: Full thickness tear was confirmed in 57 cases, partial-thickness tear in 30 cases and degenerative changes without tear in 1. In all 57 cases of full-thickness tear and in 28 out of 30 cases of partial-thickness tear the supraspinatus tendon was involved. The accuracy in the detection of full-thickness tears was 98 and 100% for ultrasonography and magnetic resonance imaging, respectively. The accuracy in the detection of bursal or articular partial-thickness tears was 87 and 90% for ultrasonography and magnetic resonance imaging, respectively. CONCLUSIONS: In experienced hands ultrasonography should be considered as an accurate modality for the initial investigation of rotator cuff, especially supraspinatus, tears. PMID- 18160243 TI - Detection of cutaneous invasion by malignant head and neck tumors with MDCT. AB - OBJECTIVE: Cutaneous invasion by direct infiltration and metastasis, in malignant head and neck tumors, has a distinct impact on therapeutic options, especially surgical procedures, curative intent, and overall prognosis. Therefore, the purpose of this study was to determine the diagnostic accuracy of MDCT in detecting cutaneous invasion by malignant head and neck tumors. MATERIALS AND METHODS: Nineteen patients with malignant head and neck tumors and clinical suspicion of cutaneous invasion routinely underwent contrast-enhanced 16-row MDCT (Philips MDCT MX 8000) of the region of interest in the axial plane before surgical intervention. The following parameters were used: 16mmx0.75mm detector collimation; 3mm reconstructed slice thickness; 1.5mm increment; 0.75s rotation speed; 120kV, 200mAs; and 100ml non-ionic contrast agent, i.v., with a flow of 2.0ml/s and a scan delay of 50s. The studies were reconstructed with a soft tissue algorithm (W400, L100 HU), and coronal and sagittal planes were also reconstructed. The axial images were retrospectively reviewed in consensus by two radiologists for the evidence of cutaneous invasion blinded to the histological results. The MDCT results were correlated with histology that was obtained by punch biopsy or surgery. RESULTS: MDCT correctly revealed 11 of 11 cases with cutaneous invasion, and correctly excluded 4 of 8 cases without cutaneous invasion. The diagnostic accuracy of MDCT in detecting cutaneous invasion showed a sensitivity of 100%, a specificity of 50%, and an overall accuracy of 79%. CONCLUSION: MDCT reconstructed with a soft tissue algorithm has a good sensitivity and moderate overall accuracy in detecting cutaneous invasion by malignant head and neck tumors. PMID- 18160244 TI - Capillary zone electrophoresis for separation and analysis of hydroxycitric acid and hydroxycitric acid lactone: application to herbal products of Garcinia atroviridis Griff. AB - Capillary zone electrophoresis (CZE) was developed for quantitative determination of hydroxycitric acid and hydroxycitric acid lactone in herbal products of Garcinia atroviridis Griff. Resolution optimization was investigated by varying type, concentration and pH of buffers. Using the pH 9.2 buffer containing 30 mM Na(2)B(4)O(7), 90 mM NaH(2)PO(4) and 0.5 mM tetradecyltrimethyl ammonium bromide, baseline resolution (R(s)>1.5) was found for all analytes. Advantages of the developed CZE method include simple sample preparation, fast analysis time within 5 min and high accuracy and precision. PMID- 18160245 TI - The motherhood choice: a decision aid for women with multiple sclerosis. AB - OBJECTIVE: This randomized controlled trial aimed to evaluate a decision aid for women with multiple sclerosis deciding whether to start or enlarge their families. METHOD: Decision aid materials were mailed to participants, and their response materials returned by mail. One-hundred and ninety-four women with MS agreed to participate and were randomized to receive the DA or not. Participants completed questionnaires measuring decisional-conflict, self-efficacy, knowledge, depression and anxiety as well as their views regarding having children pre- and post-intervention. RESULTS: Women who received the DA had a significant decrease in decisional-conflict, increase in self-efficacy and knowledge of MS and pregnancy and more certainty in their choice than those in the control group. There were no adverse effects on depression or anxiety and women's decisions were not biased in either direction. CONCLUSION: The DA is a useful tool that can be used by women with MS contemplating motherhood. PRACTICE IMPLICATIONS: This DA is used as a resource by the MS Society illustrating how DAs may help with lifestyle choices complicated by illness. It represents a tool adaptable to suit other illnesses where reproductive decision-making may be compromised and should be of interest to health-care professionals who are consulted by women with family planning concerns. PMID- 18160246 TI - Human papillomavirus in esophageal squamous cell carcinoma of the high-risk Kazakh ethnic group in Xinjiang, China. AB - AIMS: To investigate human papillomavirus (HPV) genotype-specific prevalence in the high-risk Kazakh ethnic group with esophageal squamous cell carcinoma (ESCC). METHODS: Sixty-seven Kazakh patients with primary ESCC were studied. From each patient, two tissue samples were collected: one sample of the tumor and one sample of normal esophageal tissue from an area away from the tumor. Tissues were analyzed by INNO-LiPA HPV Genotyping test v2 assay allowing the detection of at least 24 different HPV genotypes. RESULTS: Twenty cancer patients (30%) had HPV DNA detected in collected specimens. Interestingly, 14 patients (21%) had HPV only in the tumor and six (9%) had HPV only in the normal esophageal tissue. Overall, HPV16 was the viral type most frequently detected being present in eight out of 20 positive cases (40%). No correlation between the presence of HPVs and the gender or ESCC grade was observed. CONCLUSION: If the causative factors of esophageal carcinogenesis remain to be firmly established in the Kazakh population, HPV found in 30% of patients might play a role in the etiology of esophageal SCC. PMID- 18160247 TI - Effect of preoperative chemotherapy on liver resection for colorectal liver metastases. AB - AIM: To compare the effects of preoperative chemotherapy on liver parenchyma morphology, as well as morbidity and mortality after liver resection for colorectal liver metastases. METHODS: Prospectively collected data on 173 patients undergoing liver resection for CLM between 1/2003 and 9/2005 was analysed in three groups: A: preoperative oxaliplatin (Ox, n=70); B: other chemotherapeutic agents (OC, n=60); and C: surgery alone without chemotherapy (SA, n=43). Blood transfusion, hospital stay, operative procedure, peak postoperative bilirubin levels, complications and histopathology of the resected liver were compared. RESULTS: Intra-operative blood transfusion requirement (34%) and biliary complications (16%) was higher in patients receiving oxaliplatin based chemotherapy (p=0.01 and p=0.06, respectively). Oxaliplatin-based chemotherapy was also associated with sinusoidal dilatation of mild grade in 52.8% vs. 26.6% and 23.3% patients (p=0.007 and p=0.004) in other groups, respectively. Steatosis was similarly distributed across the study group. Postoperative mortality was 2, 1 and 4 patients, respectively (p=ns). CONCLUSION: Oxaliplatin-based preoperative chemotherapy is associated with vascular alterations in the liver parenchyma without significantly increasing the risk of steatosis, or postoperative morbidity and mortality. PMID- 18160248 TI - [Cranial subdural haematoma after spinal anaesthesia treated by blood patch]. AB - The authors report a case of subdural haematoma after spinal anaesthesia. A 36 year-old woman underwent phlebectomy under spinal anaesthesia. Two days later, she complains of severe headache without neurological signs, not responding to bed rest and analgesics. Magnetic resonance imaging showed a small acute subdural haematoma in the right parieto-occipital region. On the forth day, she was given a blood-patch, which improved rapidly the patient. Recovery was complete. PMID- 18160249 TI - [Sublingual haematoma: an unusual complication of oral anticoagulation therapy: about one case]. AB - Bleeding complications of oral anticoagulant therapy are not rare. The locations for bleeding are most often gastrointestinal, genitourinary and intracranial. Upper airway obstruction remains one of the reported complications. It is produced in most cases by a retropharyngeal haematoma. On the other hand, sublingual haematoma is exceptional and constitutes a problem to secure patient's airway. We report the case of a 16-year-old patient, admitted tenhours after accidental ingestion of 40mg of Sintrom((R)) with major breathing difficulty, haemoptysis and epistaxis. The endotracheal intubation was performed blindly using an Eschmann candle. This rare complication should be prevented by an adapted clinical and biological monitoring. PMID- 18160250 TI - [Use of Eschmann's gum elastic bougie in difficult orotracheal intubation, an audit over eight years of clinical practice]. AB - OBJECTIVES: The objective was to check on the effectiveness of difficult orotracheal intubation by Eschmann's gum elastic bougie in our institution. STUDY DESIGN: Retrospective observation study led for eight years. PATIENTS AND METHODS: All the orotracheal intubations during the inductions in the surgical unit of the hospital of Laon were listed from 1998 to 2005, whether they were achieved in a classical way, with a fibrobronchoscope or with Eschmann's gum elastic bougie. RESULTS: Out of 15,657 intubations, 301 of them were difficult. Eschmann's gum elastic bougie was used 276 times with a rate of success of 99%. CONCLUSION: Our study confirms the figures of publications, particularly the anglo-saxon ones and proves the interest of the Eschmann's gum elastic bougie in orotracheal intubations and this in first intention. PMID- 18160251 TI - [Chemotherapy for prostate cancer]. AB - Chemotherapy treatment for patients with prostate cancer has advanced considerably during the past decade. The first demonstration of the efficacy of palliative chemotherapy in patients with hormone-resistant prostate cancer was followed by FDA approval of mitoxantrone in this setting and by studies showing the usefulness of several different drugs in these patients. Docetaxel became the standard treatment for them. The development of new cytotoxic molecules and targeted therapies as well as the evaluation of the efficacy of docetaxel in earlier stages of prostate cancer, with many ongoing studies, are the current lines of research for improving management of these hormone-refractory patients. PMID- 18160252 TI - [Riedel's thyroiditis: current aspects]. AB - Riedel's thyroiditis is a rare form of chronic thyroiditis characterized by extensive fibrosis of the thyroid gland, also affecting adjacent tissue. Riedel's thyroiditis clinically resembles anaplastic thyroid or lymphoma, with a woody goiter attached to adjacent structures, and compressive. Findings from scintigraphy, ultrasonography, and cytology are rarely conclusive. A surgical biopsy is generally required to rule out a malignant lesion before the diagnosis of Riedel's thyroiditis can be confirmed. The most useful imaging method is computed tomography of the cervical mediastinal region, recommended during the diagnostic process and the follow-up. A thyroidectomy or dissection of the neck is in theory not recommended. Because the pathogenesis of the disease is not yet understood, the therapeutic approach remains empirical and imperfect. Corticosteroids are effective in early stages of the disease, but controversial thereafter. A dose of 10-20mg per day for 4-6 months is recommended, possibly continued thereafter if effective. Tamoxifen is a good alternative at 20 x 2 mg per day, to be reduced if adverse effects occur to 10 x 2 mg per day. There is not yet any consensus about the role of anti-TNFalpha agents and other immunosuppressive treatment for either local or more general forms of multifocal fibrosclerotic diseases. PMID- 18160253 TI - [Stonefish stings: difficult analgesia and notable risk of complications]. AB - INTRODUCTION: Marine activities and water sports in tropical countries entail some dangers. In our emergency department, stonefish (Synancae verrucosa) stings are second only to those by sea urchins among the 400 consultations a year for wounds by marine animals. METHOD: We retrospectively collected data for all emergency department visits for stonefish stings over a 5-year period from 2001 through 2005. RESULTS: The study included 57 patients (42 men; mean age 31.2+/ 15.9 years, range 3-63 years) at 61 consultations. The reason for the emergency department visit was pain, often described as intense. Injuries occurred to the foot in 79% of cases and the hand in 21%. Local signs included a wound (100%), edema (74%), local inflammation (21%), bruising (23%), necrosis (19%), and, in one patient, cellulitis. Pain was reported in 95% of cases. Local care consisted of immersing the stung area in hot water (79%) or in situ lidocaine injection (16%). Analgesics were administered in 75% of the cases, including morphine (54%) and anti-inflammatory drugs in 47%. Other analgesic techniques included ketamine (3.5%), nitrous oxide (3.5%), and local or regional anesthesia (3.5%); 29% of patients received antibiotics. Patients requiring admission (46%) differed from those who did not by a need for more intense analgesia and by greater wound inflammation and necrosis. Three patients required surgery and three others, hyperbaric oxygen therapy. CONCLUSION: Stonefish stings present the risk of local complications. Analgesia is also a major concern for emergency physicians and prophylactic antibiotics must be considered. PMID- 18160254 TI - Toluene-induced hearing loss in phenobarbital treated rats. AB - Exposure to aromatic organic solvents may induce hearing loss in rats, the cochlea being the primary target. The aim of this study which was carried out in rat, was to evaluate the impact of the hepatic metabolism of toluene on its ototoxic potency. To this end, the solvent hepatic metabolism was shifted by treating the rats with 50 mg/kg/d of phenobarbital (PhB), a potent inducer of the microsomal cytochromes P450 system, alcohol and aldehyde dehydrogenases, and glutathione-S-transferases. The two main urinary metabolites of the oxidative and conjugate pathways [hippuric (HA) and benzyl mercapturic acids (BMA) respectively] confirmed the efficacy of the PhB treatment. For the PhB-induced rats, the amount of excreted HA increased by 43% and the amount of BMA by 35%. Auditory function impairments were assessed using auditory-evoked potentials. On completion of the auditory tests, the organs of Corti were dissected to evaluate hair cell losses. The permanent auditory threshold shifts were approximately 15 dB greater in the toluene-exposed rats than in the PhB-induced rats. Both the functional and morphological data confirmed that PhB treatment can decrease the ototoxic potency of toluene. PMID- 18160255 TI - The first five years of the Wnt targetome. AB - The canonical Wnt pathway controls cell differentiation, proliferation and apoptosis by regulating the expression of a high number of target genes. The first target gene of the Wnt pathway was discovered nearly 20 years ago, when analysing gene expression patterns in the Drosophila embryo. Since the year 2002 entire transcriptomes have been screened by microarray analysis in order to identify genes, which are differentially expressed in cells with activated Wnt pathway. Recently, novel genome-based screening methods have been developed, which are less error-prone and independent from RNA. The exemplified methods STAGE (Sequence Tag Analysis Of Genomic Enrichment), ChIP-PET (chromatin immunoprecipitation with paired-end ditag), ChIP-Seq (ChIP followed by direct sequencing) and the bioinformatics approach EEL (Enhancer Element Locator) will be introduced shortly. The high number of potential target genes and regulated functions left questions unanswered, for instance how the Wnt pathway controls such a high number of genes and how it is able to regulate so many different cellular functions. In order to answer these questions we ordered the genes of the published Wnt target screenings according to their functions, and summarized the pathways, which are regulated by the Wnt pathway. This review focuses on the totality of Wnt target genes, the Wnt targetome, which is the clue to understand the manifold roles of the Wnt pathway within the cellular context. PMID- 18160256 TI - A novel Akt/PKB-interacting protein promotes cell adhesion and inhibits familial amyotrophic lateral sclerosis-linked mutant SOD1-induced neuronal death via inhibition of PP2A-mediated dephosphorylation of Akt/PKB. AB - Akt/Protein Kinase B (PKB) family proteins (Akts), consisting of Akt1, 2, and 3, play a crucial role in multiple biological processes. We recently demonstrated that activation of Akt3 by the autosomal-recessive familial amyotrophic lateral sclerosis (ALS)-linked gene 2 (ALS2) product, alsinLF, led to the suppression of motoneuronal death induced by familial ALS-related mutant superoxide dismutase-1 (SOD1). To characterize the mechanism of neuroprotection mediated by Akt3 in detail, we performed a yeast two-hybrid system using Akt3 as a bait and identified BTBD10 as a novel Akt-interacting protein with a BTB/POZ domain. BTBD10 equally binds to any Akt. Overexpression of BTBD10 increased phosphorylation levels of Akts at both Thr(308) and Ser(473) while the reduction of the endogenous BTBD10 level resulted in a decrease in the phosphorylation levels of Akts. In vitro analysis indicated that BTBD10 bound to protein phosphatase 2A (PP2A) and inhibited dephosphorylation of Akts by PP2A. In agreement with BTBD10-mediated upregulation of the Akt phosphorylation levels, enforced expression of BTBD10 led to the suppression of mutant SOD1-induced neuronal death. Furthermore, overexpression of BTBD10 accelerated cell growth by enhancing cell adhesion. Given its ubiquitous expression, BTBD10 appears to behave as a suppressor of cell death including neuronal cell death related to ALS and an enhancer of cell growth via its positive regulation of Akt phosphorylation. PMID- 18160257 TI - Wnt6 induces the specification and epithelialization of F9 embryonal carcinoma cells to primitive endoderm. AB - Epithelial-to-mesenchymal transitions (EMTs) play key roles in the normal development of an organism as well as its demise following the metastasis of a malignant tumour. An EMT during early mouse development results in the differentiation of primitive endoderm into the parietal endoderm that forms part of the parietal yolk sac. In the embryo, primitive endoderm develops from cells in the inner cell mass, but the signals that instruct these cells to become specified and adopt an epithelial fate are poorly understood. The mouse F9 teratocarcinoma cell line, a model that can recapitulate the in vivo primitive to parietal endoderm EMT, has been used extensively to elucidate the signalling cascades involved in extraembryonic endoderm differentiation. Here, we identified Wnt6 as a gene up-regulated in F9 cells in response to RA and show that Wnt6 expressing cells or cells exposed to Wnt6 conditioned media form primitive endoderm. Wnt6 induction of primitive endoderm is accompanied by beta-catenin and Snail1 translocation to the nucleus and the appearance of cytokeratin intermediate filaments. Attenuating glycogen synthase kinase 3 activity using LiCl gave similar results, but the fact that cells de-differentiate when LiCl is removed reveals that other signalling pathways are required to maintain cells as primitive endoderm. Finally, Wnt6-induced primitive endodermal cells were tested to determine their competency to complete the EMT and differentiate into parietal endoderm. Towards that end, results show that up-regulating protein kinase A activity is sufficient to induce markers of parietal endoderm. Together, these findings indicate that undifferentiated F9 cells are responsive to canonical Wnt signalling, which negatively regulates glycogen synthase kinase 3 activity leading to the epithelialization and specification of primitive endoderm competent to receive additional signals required for EMT. Considering the ability of F9 cells to mimic an in vivo EMT, the identification of this Wnt6-beta-catenin Snail signalling cascade has broad implications for understanding EMT mechanisms in embryogenesis and metastasis. PMID- 18160258 TI - Growth and metabolism of infants born to women infected with human immunodeficiency virus and fed acidified whey-adapted starter formulas. AB - OBJECTIVE: To compare the effects of a biologically and chemically acidified formula with or without probiotics with a standard formula on growth of infants negative for human immunodeficiency virus (HIV). METHODS: This was a double masked, randomized, clinical trial. Infants born to consenting HIV-positive women who had decided not to breast-feed before being approached for participating in the study were randomized to receive one of four milk formulas: a chemically acidified formula with or without probiotics (Bifidobacterium lactis), a biologically acidified formula, or a standard whey formula. Infants who subsequently became HIV-positive according to polymerase chain reaction at 6 wk were excluded. Their growth and biochemical status were monitored for 4-6 mo. The z scores at the last visit of infants in each of the four formula groups were compared using analysis of covariance correcting for the z scores at baseline. Blood gases and pH were analyzed using a two-way analysis of variance corrected for center. RESULTS: One hundred thirty-two HIV-negative infants were monitored for growth and biochemical parameters for 4-6 mo. There was an improvement of z scores for all formulas, and there were no differences in weight for age (P = 0.22), length for age (P = 0.56), head circumference for age (P = 0.66), or weight for length (P = 0.13). There were no differences in blood pH and biochemical parameters among the formula groups. CONCLUSION: The growth of infants fed one of the three acidified formulas was not inferior to the standard formula. Growth and metabolism in HIV-negative infants fed the acidified formulas were not affected by the method of milk acidification. PMID- 18160259 TI - Neonatal onset of hot water reflex seizures in monozygotic twins subsequently manifesting episodes of alternating hemiplegia. AB - We report on monozygotic twins with neonatal onset of daily reflex seizures triggered by hot water. Video record during the hot water bathing showed clinical signs consistent with a reflex seizure. The numbers of episodes were markedly reduced when the mother began bathing the children with reduced temperature bath water. At the age of 20 months, the twins developed episodes of paroxysmal disturbances including alternating hemiplegia. These two patients are the youngest reported cases of reflex hot water seizures, and the only reported cases in which reflex hot water seizures subsequently manifested episodes of alternating hemiplegia. PMID- 18160260 TI - The status of spectral EEG abnormality as a diagnostic test for schizophrenia. AB - OBJECTIVE: A literature review was conducted to ascertain whether or not EEG spectral abnormalities are consistent enough to warrant additional effort towards developing them into a clinical diagnostic test for schizophrenia. METHODS: Fifty three papers met criteria for inclusion into the review and 15 were included in a meta-analysis of the degree of significance of EEG deviations as compared to healthy controls. Studies were classified based on a 4-step approach based on guidelines for evaluating the clinical usefulness of a diagnostic test. RESULTS: Our review and meta-analysis revealed that most of the abnormalities are replicated in the expected directions with the most consistent results related to the increased preponderance of slow rhythms in schizophrenia patients. This effect remained consistent in un-medicated patients. Only a small number of studies provided data on the sensitivity and specificity of the findings in differentiating among the psychiatric disorders that frequently appear on the same differential diagnostic list as schizophrenia (Step 3 studies). No multicenter studies using standardized assessment criteria were found (Step 4 studies). CONCLUSIONS: Additional Step 3 and Step 4 studies are needed to draw conclusions on the usefulness of EEG spectral abnormalities as a diagnostic test for schizophrenia. PMID- 18160261 TI - Sensory gating revisited: relation between brain oscillations and auditory evoked potentials in schizophrenia. AB - Disturbances of auditory information processing have repeatedly been shown in schizophrenia. To contribute to a better understanding of the neurophysiological underpinnings of habituation in auditory processing and its disturbance in schizophrenia we used three different approaches to analyze auditory evoked responses, namely phase-locking (PL) analyses, single trial amplitudes, and averaged event-related potentials (P50 and N100). Given that brain oscillations reflect the neuronal correlates of information processing we hypothesized that PL and amplitudes reflect even more essential parts of auditory processing than the averaged ERP responses. In 32 schizophrenia patients and 32 matched controls EEG was continuously recorded using an auditory paired click paradigm. PL of the lower frequency bands (alpha and theta) was significantly reduced in patients whereas no significant differences were present in higher frequencies (gamma and beta). Alpha and theta PL and amplitudes showed a marked increase after the first click and to a minor degree after the second one. This habituation was more prominent in controls whereas in schizophrenia patients the response to both clicks differed only slightly. N100 suppression was significantly reduced in schizophrenia patients whereas no group differences were present with respect to the P50. This corresponded to the finding that gamma mostly contributed to the prediction of the P50 response and theta mostly to the N100 response. Our data showed that analyzing phase and amplitude in single trials provides more information on auditory information processing and reflects differences between schizophrenia patients and controls better than analyzing the averaged ERP responses. PMID- 18160262 TI - Psychometric comparison of computerized and standard administration of the neurocognitive assessment instruments selected by the CATIE and MATRICS consortia among patients with schizophrenia. AB - Neurocognitive assessment is an essential component for clinical trials of candidate "cognitive-enhancing" treatments for schizophrenia. However, manual administration of large, paper-based, neurocognitive batteries is often inefficient, error-prone, and inconsistent across multiple sites. Existing computerized testing systems are also limited both in the assessment instruments available and in the range of impairments that can be accommodated with the subject sitting alone at a single display. Therefore, a dual-display computerized testing system was developed, with funding from the National Institute of Mental Health (NIMH), that integrates (rather than replaces) the examiner for computerized administration of standard neurocognitive assessment batteries. PURPOSE: To compare standard administration versions (SAVs) of tests selected by the NIMH-sponsored CATIE (Clinical Antipsychotic Trials of Intervention Effectiveness) and MATRICS (Measurement and Treatment Research to Improve Cognition in Schizophrenia) consortia, with structurally- and functionally equivalent, computerized administration versions (CAVs). METHOD: 116 outpatients with schizophrenia received both the SAVs and CAVs within one week and again, approximately 30 days later. RESULTS: Intraclass Correlation Coefficient (ICC) comparisons between SAVs and CAVs yielded highly significant measures of absolute agreement for all tests, ranging 0.61-0.95. ICCs for test-retest reliability, ranging 0.56-0.94 for SAVs and 0.59-0.98 for CAVs, were also highly significant for both batteries, though significantly higher for CAVs overall. CONCLUSIONS: The CAVs of the neurocognitive assessment instruments selected by the CATIE and MATRICS consortia are substantially equivalent to antecedent SAVs. Importantly, the increased reliability afforded by computerization highlight the potential for increasing power, thereby decreasing sample size requirements, for clinical evaluations of putative "cognitive-enhancing" treatments. PMID- 18160263 TI - The incidence of infectious diseases after renal transplantation: a single-centre experience. AB - This single-centre study was designed to investigate the incidence of infections and their causative pathogens during the first three months after renal transplantation (RTx) in patients who had undergone the procedure in 2005 (n=174). We compared this group of patients with a previous one (1998-2000, n=437). In 2005, infection was diagnosed in 82 patients (47%). Symptomatic lower urinary tract infection (UTI) was present in 43 patients (25%), pyelonephritis in 15 (8.6%), and urosepsis in 7 (4%). Wound infection developed in 21 patients (12%), cytomegalovirus (CMV) disease in 15 (8.6%), and pneumonia in 5 (3%). The most frequent pathogens in UTI were Klebsiella pneumoniae and Enterococcus faecalis. Pathogens of wound infection included Staphylococcus coagulase negative and K. pneumoniae. Pneumonia was frequently caused by Mycoplasma pneumophila. Compared with the previous group, we noted decreases in the total number of infections (77.7 vs. 47%, P<0.001), pneumonia (8.5 vs. 3%, P<0.02) and UTI (33.3 vs. 24.7%, P<0.05). We observed an increased incidence of multiresistant Klebsiella. Based on these results, we have changed our scheme of antibiotic prophylaxis and the algorithms of antibiotic treatment. We reduced the use of antibiotics with an adverse epidemiological effect (quinolones, third-generation cephalosporins) and increased the use of relatively safe antibiotics (penicillins, aminopenicillins, with and without beta-lactam inhibitors). PMID- 18160264 TI - Fungal biofilm inhibition by a component naturally produced by Candida albicans yeasts growing as a biofilm. AB - The purpose of this work was to demonstrate production by the Candida yeasts growing as a biofilm of a compound that reduces candidal biofilm growth. An in vitro model of Candida albicans biofilm associated with silicone catheters was used. Supernatant medium recovered from 24-h-old C. albicans (ATCC 3153) biofilm (S24h) was added during the course of new biofilm formation by four C. albicans strains (182, 444, 2091 or ATCC 3153). Addition of S24h to adherent C. albicans cells induced an inhibition of biofilm growth (P<0.001) but displayed no activity on established biofilms. Ultrafiltration and purification assays demonstrated that the antagonistic molecule is hydrophilic and <3000Da in size. PMID- 18160265 TI - Is experimental endotoxaemia in humans suitable for simulating antibiotic pharmacokinetics in sepsis and septic shock? A pilot study using the model compound fosfomycin. PMID- 18160266 TI - Antibiotic resistance of motile aeromonads in indoor catfish and eel farms in the southern part of The Netherlands. AB - The prevalence and degree of antibiotic resistance in catfish and eel farms in the southern part of The Netherlands was examined using motile aeromonads as indicator bacteria. A total of 29 water samples were collected, originating from six catfish farms, one catfish hatchery and three eel farms, and were plated on an Aeromonas-selective agar with and without antibiotics. From each plate, one colony was screened for presumptive motile aeromonads and tested for antibiotic susceptibility. The prevalence of resistance was as follows: ampicillin and oxytetracycline 100%; sulfamethoxazole 24%; trimethoprim 3%; and ciprofloxacin and chloramphenicol 0%. The majority of samples showed a high degree of oxytetracycline resistance, implicating fish farms as a major reservoir of oxytetracycline resistance genes. This reservoir might form a risk for human health and has major consequences for the effectiveness of this antibiotic in the treatment of infectious diseases in fish. PMID- 18160267 TI - Awareness of metabolic concerns in patients with bipolar disorder: a survey of European psychiatrists. AB - PURPOSE: An online survey of European psychiatrists assessed awareness of the metabolic syndrome and its influence on the management of bipolar disorder. MATERIALS AND METHODS: Psychiatrists in the United Kingdom, France, Germany, Spain, and Italy were surveyed from April to June 2006. Eligibility criteria w ere 4-30 years in practice, >or=50% of time in direct patient care, had seen >or=10 bipolar patients in the preceding month. Aggregate data were weighted to represent the practicing physician population per country. RESULTS: Of 718 respondents, 56% had diagnosed metabolic syndrome. Respondents reported that metabolic syndrome prevalence was higher in bipolar patients (25%) than in the general population (20%). Seventy-two percent felt that metabolic syndrome poses significant health risks, warranting monitoring/treatment, and were most concerned with the bipolar medication adverse effects of weight gain, cognitive impairment, and glucose intolerance. Survey respondents recognized clear differences among psychotropic agents in the propensity to induce metabolic adverse effects. Sixty-five percent of respondents indicated that they had made interviewing and monitoring changes in the past three years as a result of metabolic concerns. CONCLUSIONS: European psychiatrists view metabolic syndrome as highly prevalent in the general population and in bipolar patients; two-thirds have changed their management of bipolar patients because of metabolic health concerns. PMID- 18160268 TI - The receptor protein tyrosine phosphatase PTP69D antagonizes Abl tyrosine kinase to guide axons in Drosophila. AB - During Drosophila embryogenesis, both the cytoplasmic Abelson tyrosine kinase (Abl) and the membrane bound tyrosine phosphatase PTP69D are required for proper guidance of CNS and motor axons. We provide evidence that PTP69D modulates signaling by Abl and its antagonist, Ena. An Abl loss-of function mutation dominantly suppresses most Ptp69D mutant phenotypes including larval/pupal lethality and CNS and motor axon defects, while increased Abl and decreased Ena expression dramatically increase the expressivity of Ptp69D axonal defects. In contrast, Ptp69D mutations do not affect Abl mutant phenotypes. These results support the hypothesis that PTP69D antagonizes the Abl/Ena genetic pathway, perhaps as an upstream regulator. We also find that mutation of the gene encoding the cytoplasmic Src64B tyrosine kinase exacerbates Ptp69D phenotypes, suggesting that two different cytoplasmic tyrosine kinases, Abl and Src64B, modify PTP69D mediated axon patterning in quite different ways. PMID- 18160270 TI - Effect of pH on cadmium and lead binding by extracellular polymeric substances (EPS) extracted from environmental bacterial strains. AB - Cd and Pb binding by extracellular polymeric substances (EPS) extracted from seven environmental bacterial strains were investigated as a function of pH. The study was carried out by using a polarographic method in the stripping mercury dropping electrode (SMDE) mode which is able to determine labile metal in solution containing soluble ligands such as EPS. The results obtained provide evidence for the presence of a pH-sorption/desorption edge for Cd and Pb by EPS. Kurbatov's model gives information on the mechanisms involved through the determination of "relative complexation constants" (operationally defined) and the number of exchanged protons. This model shows that proton exchange with metals is not the only mechanism involved in metal biosorption by bacterial EPS. The position of the pH-sorption edge curves and the "relative complexation constants" show that Pb displays a greater affinity for EPS than Cd. For a given metal, the seven EPS investigated did not display differences regarding the binding strength. This work suggests that a wide range of products of bacterial species isolated from activated sludge exhibit similar binding behaviours for Cd or Pb, which may potentially simplify the modelling of the distribution and the speciation of metals in bacteria-bearing natural systems. PMID- 18160269 TI - H,K-ATPase protein localization and Kir4.1 function reveal concordance of three axes during early determination of left-right asymmetry. AB - Consistent laterality is a fascinating problem, and study of the Xenopus embryo has led to molecular characterization of extremely early steps in left-right patterning: bioelectrical signals produced by ion pumps functioning upstream of asymmetric gene expression. Here, we reveal a number of novel aspects of the H+/K+-ATPase module in chick and frog embryos. Maternal H+/K+-ATPase subunits are asymmetrically localized along the left-right, dorso-ventral, and animal-vegetal axes during the first cleavage stages, in a process dependent on cytoskeletal organization. Using a reporter domain fused to molecular motors, we show that the cytoskeleton of the early frog embryo can provide asymmetric, directional information for subcellular transport along all three axes. Moreover, we show that the Kir4.1 potassium channel, while symmetrically expressed in a dynamic fashion during early cleavages, is required for normal LR asymmetry of frog embryos. Thus, Kir4.1 is an ideal candidate for the K+ ion exit path needed to allow the electroneutral H+/K+-ATPase to generate voltage gradients. In the chick embryo, we show that H+/K+-ATPase and Kir4.1 are expressed in the primitive streak, and that the known requirement for H+/K+-ATPase function in chick asymmetry does not function through effects on the circumferential expression pattern of Connexin43. These data provide details crucial for the mechanistic modeling of the physiological events linking subcellular processes to large-scale patterning and suggest a model where the early cytoskeleton sets up asymmetric ion flux along the left-right axis as a system of planar polarity functioning orthogonal to the apical-basal polarity of the early blastomeres. PMID- 18160271 TI - Stevens-Johnson syndrome after treatment with capecitabine. PMID- 18160272 TI - Antigen presentation by monocytes and monocyte-derived cells. AB - Monocytes are circulating mononuclear phagocytes with a fundamental capacity to differentiate into macrophages. This differentiation can, in the presence of the right environmental cues, be re-directed instead to dendritic cells (DCs). Recent advances have been made in understanding the role of monocytes and their derivatives in presenting antigen to drive immune responses, and we review this topic herein. We briefly discuss the heterogeneity of monocytes in the blood and subsequently raise the possibility that one of the major monocyte phenotypes in the blood corresponds with a population of 'blood DCs' previously proposed to drive T-independent antibody reactions in the spleen. Then we evaluate the role of monocytes in T-dependent immunity, considering their role in acquiring antigens for presentation before exiting the bloodstream and their ability to differentiate into macrophages versus antigen-presenting DCs. Finally, we review recent literature on the role of monocyte-derived cells in cross-presentation and discuss the possibility that monocyte-derived cells participate critically in processing antigen for cross-priming, even if they do not present that antigen to T cells themselves. PMID- 18160273 TI - Leaching behaviour of hazardous demolition waste. AB - Demolition wastes are generally disposed of in unlined landfills for inert waste. However, demolition wastes are not just inert wastes. Indeed, a small fraction of demolition waste contains components that are hazardous to human health and the environment, e.g., lead-based paint, mercury-contained in fluorescent lamps, treated wood, and asbestos. The objective of this study is to evaluate the release potential of pollutants contained in these hazardous components when they are mixed with inert wastes in unlined landfills. After identification of the different building products which can contain hazardous elements and which can be potentially pollutant in landfill scenario, we performed leaching tests using three different lysimeters: one lysimeter containing only inert wastes and two lysimeters containing inert wastes mixed with hazardous demolition wastes. The leachates from these lysimeters were analysed (heavy metals, chlorides, sulphates fluoride, DOC (Dissolved Organic Carbon), phenol index, and PAH). Finally, we compared concentrations and cumulative releases of elements in leachates with the limits values of European regulation for the acceptance of inert wastes at landfill. Results indicate that limit values are exceeded for some elements. We also performed a percolation column test with only demolition hazardous wastes to evaluate the specific contribution of these wastes in the observed releases. PMID- 18160274 TI - Nanotoxicity: the growing need for in vivo study. AB - Nanotoxicology is emerging as an important subdiscipline of nanotechnology. Nanotoxicology refers to the study of the interactions of nanostructures with biological systems with an emphasis on elucidating the relationship between the physical and chemical properties (e.g. size, shape, surface chemistry, composition, and aggregation) of nanostructures with induction of toxic biological responses. In the past five years, a majority of nanotoxicity research has focused on cell culture systems; however, the data from these studies could be misleading and will require verification from animal experiments. In vivo systems are extremely complicated and the interactions of the nanostructures with biological components, such as proteins and cells, could lead to unique biodistribution, clearance, immune response, and metabolism. An understanding of the relationship between the physical and chemical properties of the nanostructure and their in vivo behavior would provide a basis for assessing toxic response and more importantly could lead to predictive models for assessing toxicity. In this review article, we describe the assumptions and challenges in the nanotoxicity field and provide a rationale for in vivo animal studies to assess nanotoxicity. PMID- 18160275 TI - New insight into the recognition of branched DNA structure by junction-resolving enzymes. AB - Junction-resolving enzymes are nucleases that exhibit structural selectivity for the four-way (Holliday) junction in DNA. In general, these enzymes both recognize and distort the structure of the junction. New insight into the molecular recognition processes has been provided by two recent co-crystal structures of resolving enzymes bound to four-way DNA junctions in highly contrasting ways. T4 endonuclease VII binds the junction in an open conformation to an approximately flat binding surface whereas T7 endonuclease I envelops the junction, which retains a much more three-dimensional structure. Both proteins make contacts with the DNA backbone over an extensive area in order to generate structural specificity. The comparison highlights the versatility of Holliday junction resolution, and extracts some general principles of recognition. PMID- 18160276 TI - HIV-1 protease folding and the design of drugs which do not create resistance. AB - Human immunodeficiency virus type 1 (HIV-1) protease (PR) plays an essential role in the life cycle of the virus. Consequently, its inhibition can control acquired immunodeficiency syndrome (AIDS). Any pharmacological treatment targeting the active site of the protease is known to generate escape mutants. On the other hand, if a drug targets a site crucial for the correct folding of the protease, mutations affecting this region would denaturate the protein and thus will not be expressed. We review the progress in our understanding of the folding of the protease, which has been instrumental in the design of a (non-conventional) folding inhibitor. The transferability of these results to other proteins testify to the universality of the folding-inhibition scenario for the design of leads of drugs which are unlikely to generate resistance. PMID- 18160277 TI - Coarse-grained models of protein folding: toy models or predictive tools? AB - Coarse-grained models are emerging as a practical alternative to all-atom simulations for the characterization of protein folding mechanisms over long time scales. While a decade ago minimalist toy models were mainly designed to test general hypotheses on the principles regulating protein folding, the latest coarse-grained models are increasingly realistic and can be used to characterize quantitatively the detailed folding mechanism of specific proteins. The ability of such models to reproduce the essential features of folding dynamics suggests that each single atomic degree of freedom is not by itself particularly relevant to folding and supports a statistical mechanical approach to characterize folding transitions. When combined with more refined models and with experimental studies, the systematic investigation of protein systems and complexes using coarse-grained models can advance our theoretical understanding of the actual organizing principles that emerge from the complex network of interactions among protein atomic constituents. PMID- 18160278 TI - Custom prosthetic reconstruction for proximal tibial osteosarcoma with proximal tibiofibular joint involved. AB - BACKGROUND: The therapeutic results of managing proximal tibial osteosarcoma have been dramatically improved because of the recent advancements in imaging, chemotherapy, and surgical techniques. Further, the prognosis of patients with proximal tibial osteosarcoma is improved and the chance of survival with limb salvage surgery is increased. Among the limb salvage procedures, endoprosthetic reconstruction is now preferred to other methods such as allograft, composite allograft prosthesis, or arthrodesis. However, the treatment for proximal tibial osteosarcoma with proximal tibiofibular joint involved has not been reported. OBJECTIVE: The report of the preliminary results of custom prosthetic reconstruction for proximal tibial osteosarcoma with proximal tibiofibular joint involved following the primary tumor resection. The oncological results, functional outcomes, and complications were assessed in this present study. MATERIALS AND METHOD: Eleven patients with osteosarcoma of the proximal tibia and proximal tibiofibular joint were studied in 1995-2005, at Qilu Hospital of the Shandong University. Seven cases were males and four cases were females, with a mean age of 17 years (range 14-23). The surgical stage of all of the patients was Stage IIB, according to the Enneking Surgical Staging System. After neo-adjuvant chemotherapy, all of the patients underwent en bloc resection. This procedure included the proximal tibiofibular joint and the upper end of the fibula, followed by custom prosthetic replacement. The extensor mechanism was repaired by reattachment of the patellar tendon to the slot in the tibial component, with reinforcement by autologous bone-graft and suturing of the patellar tendon to a medial gastrocnemius rotation flap. The medial gastrocnemius rotation flap was then used to cover the implants. RESULTS: During the mean follow-up time of 47 months (range 12-96), six patients lived free of the disease, and five patients experienced various complications. One patient was still alive yet affected by the disease; one patient had an amputation due to local recurrence; and three died of pulmonary metastases. Several early complications had occurred amongst those five patients: one developed skin necrosis; two experienced transient palsy of the common peroneal nerve; and two developed deep vein thrombus of the lower limb. The mean Musculoskeletal Tumor Society score was 70% (range 55-86%), the mean postoperative range of motion was 85 degrees (range 0-120 degrees ), and the mean extension lag was 11 degrees (range 0-20 degrees ). CONCLUSION: Custom prosthetic reconstruction could yield as much satisfactory results as neoadjuvant chemotherapy and limb salvage surgery, for patients with osteosarcoma in proximal tibia and proximal tibiofibular joint. During the present study, most patients had good or excellent motor functioning with few incidences of complications. However, more attention should have been given to both the quality of function and long-term survival of patients. PMID- 18160279 TI - Critical analysis of application of generalized distance function for optimization of important variables for esterase synthesis by Saccharomyces cerevisiae. AB - The generalized distance function approach is employed to obtain a suitable near optimal conditions of variables. The optimal values of variables (medium constituents, microbiological parameters, and process parameters) have been evaluated separately using single responses (either specific esterase activity or cell mass) as per central-composite-design and multi-responses following generalized distance function approach. The optimal conditions (medium composition (gl(-1)): dextrose, 13.43; peptone, 7.285; yeast extract, 2.55; and malt extract, 1.695; microbiological parameters: slant age, 39.9h; inoculum age, 9.6 h; and number of cells, 1.49 x 10(8) numbers ml(-1); process conditions: temperature, 29.9 degrees C; and pH, 6.2) obtained by generalized distance approach can be considered as a 'near optimal' solution of interactive multi response systems of intracellular esterase synthesis by Saccharomyces cerevisiae. PMID- 18160280 TI - Olive stone an attractive source of bioactive and valuable compounds. AB - The olive stone and seed are an important byproduct generated in the olive oil extraction and pitted table olive industries. As a lignocellulosic material, the hemicellulose, cellulose and lignin are the main components of olive stone as wells as protein, fat, phenols, free sugars and poliols composition. The main use of this biomass is as combustion to produce electric energy or heat. Other uses such as activated carbon, furfural production, plastic filled, abrasive and cosmetic or other potential uses such as biosorbent, animal feed or resin formation have been cited. In this article, an overview of the characterization and main uses of olive stone and seed are described for the first time. Also, this review discusses the potential use of this material based on each component. In this way, a new approach to the olive stone and seed by pretreating with a steam explosion followed by chemical fractionation is described. PMID- 18160281 TI - Design and SAR of selective T-type calcium channel antagonists containing a biaryl sulfonamide core. AB - T-type calcium channel antagonists were designed using a protocol involving the program SPROUT and constrained by a ComFA-based pharmacophore model. Scaffolds generated by SPROUT were evaluated based on their ability to be translated into structures that were synthetically tractable. From this exercise, a novel series of potent and selective T-type channel antagonists containing a biaryl sulfonamide core were discovered. PMID- 18160282 TI - Discovery of novel orally active ureido NPY Y5 receptor antagonists. AB - We have derived a novel series of neuropeptide Y (NPY) Y5 receptor antagonists from the biphenylurea 3. Cyclohexylurea 21c, a member of the series, is a potent NPY Y5 receptor antagonist that exhibits excellent pharmacokinetic parameters in rats and dogs. On chronic oral administration to diet-induced obese rats, 21c displayed an anti-obesity profile, causing a modest reduction in food intake, a significant decrease in body weight gain, a decrease in adipose mass, and an increase in lean tissue mass. PMID- 18160283 TI - Targeting ACE and ECE with dual acting inhibitors. AB - A series of urea analogues related to SA6817 and a GSK phosphonic acid with reported ACE inhibitory activity were prepared and tested for dual ACE and ECE activities. Although excellent ACE and NEP inhibition was achieved, only modest ECE inhibition was observed with one analogue. PMID- 18160284 TI - From rigid cyclic templates to conformationally stabilized acyclic scaffolds. Part II: Acyclic replacements for the (3S)-3-benzylpiperidine in a series of potent CCR3 antagonists. AB - Conformational analysis of the 3-benzylpiperidine in CCR3 antagonist clinical candidate 1 (BMS-639623) predicts that the benzylpiperidine may be replaced by acyclic, conformationally stabilized, anti-1,2-disubstituted phenethyl- and phenpropylamines. Ab initio calculations, enantioselective syntheses, and evaluation in CCR3 binding and chemotaxis assays of anti-1-methyl-2 hydroxyphenethyl- and phenpropylamine-containing CCR3 antagonists support this conformational correlation. PMID- 18160285 TI - Synthesis and antimicrobial activity of novel C-linked imidazole glycoconjugates. AB - Novel C-linked imidazole derivatives have been synthesized in good to excellent yields and characterized by analytical and spectral analysis. Four of the newly synthesized compounds exhibited moderate antibacterial activity. PMID- 18160286 TI - Three-component, one-pot synthesis of novel 2,4-substituted 5 azolylthiopyrimidine library for screening against anti-influenza virus A. AB - A novel one-pot synthesis of 2,4-substituted 5-azolylthiopyrimidines is achieved by sequential Michael-addition of 3-iodochromones with mercaptoazole (or mercaptotriazoles) and then condensation with a variety of amidines. Compound A(1)B(6)C(1) exhibits a potent anti-influenza virus A activity with an IC(50) value of 21.56 mg/mL and SI value of 9. PMID- 18160287 TI - Design and synthesis of 4-[(s-triazin-2-ylamino)methyl]-N-(2-aminophenyl) benzamides and their analogues as a novel class of histone deacetylase inhibitors. AB - Inhibition of histone deacetylases (HDAC) is emerging as a new strategy in human cancer therapy. The synthesis and biological evaluation of a variety of 4 (heteroarylaminomethyl)-N-(2-aminophenyl)-benzamides is presented herein. From the different series bearing a six-membered heteroaromatic ring studied, the s triazine series showed the best HDAC1 enzyme and in vitro anti-proliferative activities with IC(50) values below micromolar range. Some of these compounds can also significantly reduce tumor growth in human tumor xenograft models in mice. PMID- 18160288 TI - Synthetic study of VLA-4/VCAM-1 inhibitors: synthesis and structure-activity relationship of piperazinylphenylalanine derivatives. AB - To improve the poor pharmacokinetic characteristics of VLA-4 inhibitors, novel piperazinylphenylalanine derivatives were designed. This structure is expected to improve physicochemical properties by increasing overall basicity. By changing components at the 4-position of piperazine and the terminal group of the amido bond, 12t was found to be the most potent of this series of compounds. In addition, dichlorobenzoyl derivative 12aa exhibited better oral availability and showed efficacy in an in vivo model after oral administration. PMID- 18160289 TI - 1-Aminoindanes as novel motif with potential atypical antipsychotic properties. AB - As part of an on-going effort to investigate the chemical space requirements for D(2)/5-HT(2A) receptor antagonists as atypical antipsychotics, new 1-aminoindanes were synthesized. The replacement of the heterocycle (oxindole) in ziprasidone with a carbocycle (indane) was well tolerated and was found to retain binding affinities for dopamine D(2), serotonin 5-HT(2A), and serotonin 5-HT(1A). Such compounds hold promise as a new chemical motif with atypical antipsychotic properties for the treatment of schizophrenia and related disorders. PMID- 18160290 TI - 155th ENMC workshop: polymerase gamma and disorders of mitochondrial DNA synthesis, 21-23 September 2007, Naarden, The Netherlands. PMID- 18160291 TI - Sporadic inclusion body myositis: a continuing puzzle. AB - There is now compelling evidence that sporadic inclusion body myositis (sIBM) is a muscle-specific autoimmune disease in which both T and B-cells play a part and in which both cytotoxic muscle fibre necrosis and degeneration occur. However the factors responsible for breakdown of immune tolerance and the nature of the target antigens expressed by muscle fibres remain unknown. Genetic factors are known to contribute to susceptibility, in particular MHC haplotyes which may influence antigenic presentation, and could also operate through genetic variations in muscle fibre constituents or immune effector mechanisms. Viral infection may act as a trigger mechanism, as in cases of HIV-associated sIBM. Our understanding of the mechanisms leading to the degenerative changes in muscle fibres is still incomplete. Protein misfolding and proteasomal dysfunction rather than defective transcriptional control is likely to underlie the abnormal accumulation of multiple proteins in the muscle fibre inclusions. However, aberrant transcription is thought to be the basis for the accumulation of potentially toxic mutant protein forms (e.g. UBB(+1)). The origin of the multiple clonally expanded somatic mtDNA mutations in COX-negative segments of muscle fibres remains uncertain but may be linked to the effects of oxidative stress. It is proposed that the disproportionate involvement of certain muscles in sIBM may be due to the existence of muscle group-specific transcriptomes which are differentially affected by the disease process and that the male predominance of the disease may indicate the influence of genes preferentially expressed in males. There is a need to develop better animal models of sIBM in which the relationship between the inflammatory and degenerative components of the disease as well as the gender difference in susceptibility and differential vulnerability of different muscle groups can be more critically investigated. PMID- 18160292 TI - The peroxin loss-of-function mutation abstinence by mutual consent disrupts male female gametophyte recognition. AB - In eukaryotes, fertilization relies on complex and specialized mechanisms that achieve the precise delivery of the male gamete to the female gamete and their subsequent union [1-4]. In flowering plants, the haploid male gametophyte or pollen tube (PT) [5] carries two nonmotile sperm cells to the female gametophyte (FG) or embryo sac [6] during a long assisted journey through the maternal tissues [7-10]. In Arabidopsis, typically one PT reaches one of the two synergids of the FG (Figure 1A), where it terminates its growth and delivers the sperm cells, a poorly understood process called pollen-tube reception. Here, we report the isolation and characterization of the Arabidopsis mutant abstinence by mutual consent (amc). Interestingly, pollen-tube reception is impaired only when an amc pollen tube reaches an amc female gametophyte, resulting in pollen-tube overgrowth and completely preventing sperm discharge and the development of homozygous mutants. Moreover, we show that AMC is strongly and transiently expressed in both male and female gametophytes during fertilization and that AMC functions in gametophytes as a peroxin essential for protein import into peroxisomes. These findings show that peroxisomes play an unexpected key role in gametophyte recognition and implicate a diffusible signal emanating from either gametophyte that is required for pollen-tube discharge. PMID- 18160293 TI - Oligocene CO2 decline promoted C4 photosynthesis in grasses. AB - C4 photosynthesis is an adaptation derived from the more common C3 photosynthetic pathway that confers a higher productivity under warm temperature and low atmospheric CO2 concentration [1, 2]. C4 evolution has been seen as a consequence of past atmospheric CO2 decline, such as the abrupt CO2 fall 32-25 million years ago (Mya) [3-6]. This relationship has never been tested rigorously, mainly because of a lack of accurate estimates of divergence times for the different C4 lineages [3]. In this study, we inferred a large phylogenetic tree for the grass family and estimated, through Bayesian molecular dating, the ages of the 17 to 18 independent grass C4 lineages. The first transition from C3 to C4 photosynthesis occurred in the Chloridoideae subfamily, 32.0-25.0 Mya. The link between CO2 decrease and transition to C4 photosynthesis was tested by a novel maximum likelihood approach. We showed that the model incorporating the atmospheric CO2 levels was significantly better than the null model, supporting the importance of CO2 decline on C4 photosynthesis evolvability. This finding is relevant for understanding the origin of C4 photosynthesis in grasses, which is one of the most successful ecological and evolutionary innovations in plant history. PMID- 18160294 TI - The role of the FH1 domain and profilin in formin-mediated actin-filament elongation and nucleation. AB - BACKGROUND: Formin proteins nucleate actin filaments de novo and stay associated with the growing barbed end. Whereas the formin-homology (FH) 2 domains mediate processive association, the FH1 domains-in concert with the actin-monomer-binding protein profilin-increase the rate of barbed-end elongation. The mechanism by which this effect is achieved is not well understood. RESULTS: We used total internal reflection fluorescence microscopy to measure the effect of profilin on the elongation of single actin filaments associated with FH1FH2 constructs (derived from the formin Bni1p from S. cerevisiae) with FH1 domains containing one to eight profilin-binding polyproline tracks. Over a large range of profilin concentrations (0.5-25 microM), the rate of barbed-end elongation increases with the number of polyproline tracks in the FH1 domain. The binding of profilin-actin to the FH1 domain is the rate-limiting step (up to rates of at least 88 s(-1)) in FH1-mediated transfer of actin subunits to the barbed end. Dissociation of formins from barbed ends growing in the presence of profilin is proportional to the elongation rate. Profilin profoundly inhibits nucleation by FH2 and FH1FH2 constructs, but profilin-actin bound to FH1 might contribute weakly to nucleation. CONCLUSIONS: To achieve fast elongation, formin FH1 domains bind profilin-actin complexes and deliver them rapidly to the barbed end associated with the FH2 domain. Because subunit addition promotes dissociation of FH2 domains from growing barbed ends, FH2 domains must pass through a state that is prone to dissociation during each cycle of actin subunit addition coupled to formin translocation. PMID- 18160295 TI - Evolution and functional diversity of jellyfish opsins. AB - Cnidaria are the most basal animal phylum possessing complex eyes [1]. Their eyes predominantly use ciliary photoreceptor cells (c-PRCs) like vertebrates, whereas insect eyes use rhabdomeric photoreceptor cells (r-PRCs) [1-4]. These two cell types show not only different cytoarchitectures but distinct phototransduction cascades, which are triggered by the respective types of opsins (e.g., [5]), ciliary opsins (c-opsins) and rhabdomeric opsins (r-opsins) [6]. Recent reports suggested that the c- and r-PRCs and their respective opsins diverged at least before the deuterostome-protostome split [7-9]. To study the earlier evolution of animal PRCs and opsins, we investigated two hydrozoan jellyfishes. We report here the first-characterized cnidarian opsins. Molecular phylogeny revealed that the cloned 20 jellyfish opsins, together with all the opsins from a hydra and some from a sea anemone, are more closely related to the c-opsins than to any other major opsin subfamily, indicating that the divergence of c- and r-opsins antedates the Cnidaria-Bilateria split. Possible scenarios of animal PRC evolution are discussed. Furthermore, Cladonema opsins show several distinct tissue- and stage-specific expression patterns. The expression of specific opsins in the eyes suggests a role in vision, whereas that in the gonads suggests a role in light-controlled release of gametes. PMID- 18160296 TI - C-type lectin receptors in antifungal immunity. AB - Fungal infections represent a significant health burden, especially in immunocompromised individuals, yet many of the underlying immunological mechanisms involved in the recognition and control of these pathogens are unclear. The identification of the Toll-like receptors (TLRs) has shed new insights on innate microbial recognition and the initiation of immune responses; however, recent evidence indicates that the 'non-TLR' receptors also have a significant role in these processes, particularly in antifungal immunity. Of interest are members of the C-type lectin-receptor family, including the mannose receptor, dendritic cell-specific intercellular adhesion molecule-3 (ICAM-3) grabbing non-integrin (DC-SIGN), Dectin-1, Dectin-2 and the collectins. Here, we review the roles of each of these receptors, describing how they contribute to fungal recognition, uptake and killing and also participate in the induction and/or modulation of the host immune response. PMID- 18160297 TI - Oligo(tyrosine sulfate)s as heparin pentasaccharide mimic: evaluation by surface noncovalent affinity mass spectrometry. AB - Since the discovery of anti-HIV activity in oligo(tyrosine sulfate)s in our laboratory, we have been interested in their potential as heparin pentasaccharide mimics. In this study, we investigated their interactions with synthetic heparin binding peptides, derived from human antithrombin III (hAT III) and heparin interacting protein (HIP), using surface noncovalent affinity mass spectrometry. We compared binding affinities to those heparin-binding peptides between oligo(tyrosine sulfate)s and several known sulfated compounds and found that oligo(tyrosine sulfate)s bind to hAT III (123-139) more strongly than a heparin derived hexasaccharide dp6. Moreover, we found longer oligo(tyrosine sulfate) has higher binding affinity to hAT III (123-139). PMID- 18160298 TI - Novel nitroheterocyclic hypoxic markers for solid tumor: synthesis and biological evaluation. AB - Based on the principle that the nitro-group can quench the fluorescence and can be reduced under hypoxic conditions, several novel nitroheterocyclic compounds without 2-nitroimidazole as potential hypoxic markers were prepared. Although they were synthesized from the same matrix, nitrosubstituted acenaphtho[1,2 b]quinoxaline, these compounds exhibited quite different fluorescence changes when they were differently nitrosubstituted. Their evaluation for imaging tumor hypoxia was carried out in V79 cells in vitro by Fluorescence Microplate Reader. After 3.5 h, the hypoxic-oxic fluorescence differential incubated with A1, A4, and A5 in V79 cells could reach 6, 9, and 11 times differential fluorescence between oxic and hypoxic cells separately, which are suitable for further evaluation as probes for hypoxic cells in tumors in vivo. PMID- 18160299 TI - Synthetic derivatives of the alpha- and beta-amyrin triterpenes and their antinociceptive properties. AB - Fifteen different derivatives of an alpha- and beta-amyrin mixture were synthesized by acylation with appropriate anhydride or acid chlorides and oxidation in the presence of tert-butyl chromate or PCC. The molecular structures of the obtained compounds were confirmed by means of IR and (1)H NMR spectra. The compounds were screened for antinociceptive activity using the acetic acid pain model. The 3-O-acyl derivatives alpha- and beta-amyrin propionate 4, alpha- and beta-amyrin hexanoate 6, and alpha- and beta-amyrin octanoate 7 were found to be the most active compounds of the series. In addition, we also have found that alpha- and beta-amyrin octanoate 7 was able to reduce acetic acid-induced abdominal constriction when administered by oral route. Furthermore, this compound reduced the nociceptive response induced by intraplantar injection of formalin. PMID- 18160300 TI - Gene expression profiling of the different stages of Arabidopsis thaliana trichome development on the single cell level. AB - Leaf hairs (trichomes) of Arabidopsis thaliana are a model system for studying cell development, differentiation and cell cycle regulation. To exploit this model system with ultimate spatial resolution we applied single cell sampling, thus avoiding the averaging effect induced by complex tissue mixtures. In particular, we analysed gene expression profiles of two selected stages of the developing trichome: trichome initial cells and mature trichomes, as well as pavement cells. Ten single cells per sample were collected by glass microcapillaries and used for the generation of radioactive probes for subsequent hybridization to nylon filters representing approximately 8000 genes of A. thaliana. Functional categorization of genes transcribed in trichome initials, mature trichomes and pavement cells demonstrated involvement of these surface cells in the stress response. In silico promoter analysis of genes preferentially expressed in trichome initials revealed enrichment in MYB-binding sites and presence of elements involved in hormonal, metal, sulphur response and cell cycle regulation. Three candidate genes preferentially expressed in trichome initials were selected for further analysis: At3g16980 (putative RNA polymerase II), At5g15230 (GASA4) and At4g27260 (GH3.5, WES1). Promoter:GUS studies confirmed expression of the putative RNA polymerase II and the gibberellin responsive GASA4 in trichome initials and partially in mature trichomes. Functional implication of the three selected candidates in trichome development and hence in cell cycle regulation in A. thaliana is discussed. We suggest that these genes are involved in differentiation and initiation of endocycling during trichome development. PMID- 18160301 TI - Multicentric analysis of performance after major lung resections by using the European Society Objective Score (ESOS). AB - OBJECTIVE: Outcome endpoints are still the most widely used indicators of performance. However, they need to be risk-adjusted in order to be reliable instruments of audit. Recently, the European Society Objective Score (ESOS) was developed from the online European Thoracic Surgery Database as an audit tool. In this study, we applied for the first time the ESOS.01 to assess the performance of three European thoracic surgery units during three successive years of activity. METHODS: This study is a retrospective analysis performed on prospective databases. We analysed 695 patients submitted to pneumonectomy (117) or lobectomy (578) for lung neoplasm at three European dedicated thoracic surgery units (unit A 264 patients, unit B 262, unit C 169) from January 2004 through December 2006. Qualified thoracic surgeons performed all the operations. No patients in this series were in the original ESOS development set. ESOS.01 was used to estimate the risk of in-hospital mortality in all patients. Observed and predicted mortality rates were then compared within each unit by the z-test. RESULTS: Cumulative observed mortality rates in units A, B and C were 2.3% (six cases), 2.7% (seven cases) and 4.1% (seven cases), respectively. We were not able to find statistically significant differences between observed and ESOS-predicted mortality rates. The comparison of risk-adjusted mortality rates between units did not show significant differences (unit A 3.9%, unit B 3.3%, unit C 5.6%). CONCLUSIONS: The use of ESOS.01 revealed that the performances of all units were in line with the predicted ones during each period under analysis and did not differ between each other. The results of our study warrant future efforts to refine the ESOS model and to develop other risk-adjusted outcome indicators with the aim to establish European benchmarks of performance. PMID- 18160302 TI - Three-dimensional numerical simulation of blood flow in the aortic arch during cardiopulmonary bypass. AB - OBJECTIVE: To better understand the mechanism of stroke during cardiopulmonary bypass, it is necessary to obtain information on the location of turbulence, wall pressure, and flow distribution within the aortic arch. METHODS: Blood flow was numerically simulated using the finite element method in the following representative case: a curved arterial cannula was inserted into the anterior wall of the distal ascending aorta 2 cm below the orifice of brachiocephalic artery. Perfusion was performed, with a bypass flow index of 2.5l min(-1) m(-2). Computational grids, consisting of 1,493,297 tetrahedral elements, were generated. RESULTS: The highest wall pressure (3104.8 Pa) was observed at the superior-posterior wall of the aorta below the orifice of the brachiocephalic artery where jet flow impingement occurred. The maximum wall shear stress was 25.1 Pa. High velocity vortex started below the orifice of the brachiocephalic artery. The turbulent flows continued along the posterior wall and then mainly flowed off into the left subclavian artery. Therefore, in the present case, an embolic event in the territory of the left subclavian artery could occur if a plaque was present at the superior-posterior wall of the aorta below the orifice of the brachiocephalic artery. The flow rates in each of the branches were 132, 613, 175, and 821 ml/min for the right subclavian, right common carotid, left common carotid, and left subclavian artery, respectively. CONCLUSION: This study confirmed that blood flow during cardiopulmonary bypass can be simulated and visualized. Computational fluid dynamics could be applied in the future to assess an individual's risk of stroke. Further multiple representative cases need to be simulated. PMID- 18160303 TI - Weekly docetaxel versus CMF as adjuvant chemotherapy for elderly breast cancer patients: safety data from the multicentre phase 3 randomised ELDA trial. AB - Within an ongoing multicentre phase 3 randomised trial (ELDA, cancertrials.gov ID: NCT00331097), early breast cancer patients, 65-79 years old, with average to high risk of recurrence, are randomly assigned to receive CMF (cyclophosphamide 600 mg/m2, methotrexate 40 mg/m2, fluorouracil 600 mg/m2, days 1-8) or docetaxel (35 mg/m2 days 1-8-15), every 4 weeks. Here we report an unplanned safety analysis prompted by an amendment introducing creatinine clearance as a tool to adjust methotrexate dose. Before such change, 101 patients with a median age of 70 were randomly assigned CMF (53 patients) or docetaxel (48 patients). At least one grades 3-4 toxic event of any type was reported in 40 (75.5%) and 19 (39.6%) patients with CMF and docetaxel, respectively (p=0.0002). Grades 3-4 hematological events were observed in 37 (69.8%) vs. 4 (8.3%) cases (p<0.0001) and grades 3-4 non-hematological toxicity in 12 (22.6%) vs. 15 (31.2%) patients (p=0.11), with CMF and docetaxel, respectively. A higher incidence of anemia, neutropenia, thrombocytopenia and febrile neutropenia was reported with CMF. Constipation, mucositis, nausea and vomiting were more common with CMF; diarrhoea, abdominal pain, dysgeusia, neuropathy and liver toxicity were more frequent with docetaxel. No significant interaction was found between the occurrence of severe toxicity and baseline variables, including creatinine clearance and geriatric activity scales. In conclusion, weekly docetaxel appears to be less toxic than CMF in terms of hematological toxicity. PMID- 18160305 TI - Focus honoring R. Graham Cooks, recipient of the 2006 ASMS award for distinguished contribution in mass spectrometry. PMID- 18160304 TI - Absence of endogenous interleukin-10 enhanced organ dysfunction and mortality associated to zymosan-induced multiple organ dysfunction syndrome. AB - Experimental evidence suggests that interleukin (IL)-10 plays a pivotal role in generalized inflammation. Here we investigate the effects of IL-10 gene deletion on the acute phase of the multiple organ dysfunction syndrome (MODS) caused by zymosan in the mouse. MODS was induced by zymosan administration (500 mg/kg, suspended in saline solution, i.p.) in IL-10 wild-type and knockout mice; sham groups were treated with vehicle. Mice were sacrificed 18 h after zymosan or saline administration. In another set of experiments, animals were monitored for 12 days to assess systemic toxicity and survival rate. Mice lacking IL-10 displayed increased peritoneal exudate volume and leukocytes. Also, we observed a significant increase in myeloperoxidase activity and lipid peroxidation in ileum and lung tissues, as well as augmented levels of TNF-alpha, IL-1beta and nitrogen derived species in the plasma. With regard to organ injury, absence of IL-10 enhanced the renal, hepatocellular and pancreatic dysfunction caused by zymosan administration. All of these parameters significantly influenced the systemic toxicity and the overall survival at 12 days, which was significantly lower in IL 10 knockout mice. Therefore, this study demonstrates that the absence of endogenous IL-10 enhances the MODS induced by zymosan in mice. PMID- 18160306 TI - Recent advancements towards the derivation of immune-compatible patient-specific human embryonic stem cell lines. AB - The derivation of human embryonic stem cell lines from blastocyst stage embryos, first achieved almost a decade ago, demonstrated the potential to prepare virtually unlimited numbers of therapeutically beneficial cells in vitro. Assuming that large-scale production of differentiated cells is attainable, it is imperative to develop strategies to prevent immune responses towards the grafted cells following transplantation. In this review, I will discuss recent advances in the production of pluripotent cell lines using three emerging techniques: somatic cell nuclear transfer into enucleated oocytes and zygotes, parthenogenetic activation of unfertilized oocytes and induction of pluripotency in somatic cells. Importantly, if these techniques can be harnessed for generating patient-specific pluripotent cell lines, then immunological processes are expected to be low or completely absent. PMID- 18160307 TI - Yersinia pestis YopD 150-287 fragment is partially unfolded in the native state. AB - Yersinia pestis, a human and animal pathogen, uses the type III secretion system (T3SS) for delivering virulence factors and effectors into the host cells. The system is conserved in animal pathogens and is hypothesized to deliver the virulence factors directly from bacterial to mammalian cells through a pore composed of YopB and YopD translocation proteins. The YopB and YopD translocator proteins must be delivered first to form a functional pore in the mammalian cell. The criteria by which Yersinia selects the two proteins for initial delivery are not known and we hypothesized that the extensive binding by the chaperone and partial unfolding of the unbound region may be the criteria for selection. The YopB and YopD translocator proteins, unlike other effectors, have a common chaperone SycD, which binds through multiple regions. Due to the small size of the pore, we hypothesized that many of the transported virulence factors, translocators YopB and YopD included, are delivered in a partially unfolded state stabilized by binding to specific chaperones. The YopD protein binds the chaperone through amino acid (a.a.) 53-149 and a.a. 278-292 regions but biophysical characterization of YopD has not been possible due to the lack of an expression system for soluble, large fragments of the protein. In our present work, we demonstrated that the YopD 150-287 peptide fragment, almost the full soluble C-terminal part, including the non-interacting peptide fragment YopD 150 277, was partially unfolded in its native state by a combination of biophysical methods: circular dichroism, quasi-elastic light scattering, chemical unfolding and 8-anilino-1-naphthalene sulfonate (ANS) binding. The secondary structure of the peptide converted easily between alpha-helical and random coil states at neutral pH, and the alpha-helical state was almost fully recovered by lowering the temperature to 263 K. The current results suggest that YopD 150-287 peptide may have the postulated transport-competent state in its native form. PMID- 18160308 TI - Baculovirus-mediated expression, purification, and characterization of a fully activated catalytic kinase domain construct of the 70 kDa 40S ribosomal protein S6 kinase-1 alphaII isoform (S6K1alphaII). AB - S6K1alphaII is a member of the AGC subfamily of serine-threonine protein kinases, whereby catalytic activation requires dual phosphorylation of critical residues in the conserved T-loop (T229) and hydrophobic motif (HM; T389) regions of its catalytic kinase domain [S6K1alphaII(DeltaAID); deletion of C-terminal autoinhibitory domain residues 399-502]. With regard to mimicking the synergistic effect of full dual site phosphorylation, baculovirus-mediated expression and affinity purification of the His(6)-S6K1alphaII(DeltaAID)-T229E,T389E double mutant from Sf9 insect cells yielded enzyme with compromised activity. Higher activity preparations were generated using the Sf9 purified His(6) S6K1alphaII(DeltaAID)-T389E single mutant isoform, which was in vitro phosphorylated by the upstream T229 kinase, PDK1 ( approximately 75 nmol/min/mg). Most significantly, we report that the His(6)-S6K1alphaII(DeltaAID)-T389E construct was generated in its most highly active form (250 nmol/min/mg) by baculovirus-mediated expression and purification from Sf9 insect cells that were coinfected with recombinant baculovirus expressing the catalytic kinase domain of PDK1 [His(6)-PDK1(DeltaPH)]. Approximately equal amounts of fully activated His(6)-S6K1alphaII(DeltaAID)-T389E (5+/-1 mg) and His(6)-PDK1(DeltaPH) (8+/-2 mg) were His(6) affinity co-purified 60 h after initial coinfection of 200 mL of Sf9 insect cells (2x10(6) cells/mL), which were resolved by MonoQ anion exchange chromatography. ESI-TOF mass spectrometry, MonoQ anion exchange chromatography, and kinetic assays showed His(6)-PDK1(DeltaPH) to phosphorylate T229 to approximately 100% after co-expression in Sf9 insect cells as compared to approximately 50% under in vitro conditions, raising interest to mechanistic components not fully achieved in the in vitro reaction. Generation of fully activated S6K1 will facilitate more rigorous analysis of its structure and mechanism. PMID- 18160309 TI - GMP production and characterization of the bivalent anti-human T cell immunotoxin, A-dmDT390-bisFv(UCHT1) for phase I/II clinical trials. AB - The bivalent anti-T cell immunotoxin, A-dmDT390-bisFv(UCHT1), was developed for treatment of T-cell leukemia, autoimmune diseases and tolerance induction for transplantation. To obtain clinical grade bivalent anti-T cell immunotoxin for phase I/II clinical trials, a single batch of 120 L bioreactor culture was performed using the Pichia pastoris mutEF2JC307-8(2) strain expressing the bivalent anti-T cell immunotoxin. After 162 h induction of the culture by methanol, the culture medium was harvested by a 0.1 microm hollow-fiber microfiltration step. The recombinant protein was purified by a 3-step purification procedure (Butyl 650 M capturing step, borate anion exchange step and final Poros anion exchange step). The final material was filter sterilized, aseptically vialed, and stored at -80 degrees C. Expression level was 207 mg/L of culture supernatant and the final production yield was 69.6% or 144.2mg/L of culture supernatant. The final product was characterized by multiple assays. Vialed product was sterile. The drug concentration was 0.8 mg/mL in 150 mM NaCl, 5% glycerol, 1mM EDTA, and 5mM Tris (pH 8.0). Purity by SDS-PAGE was 98%. Aggregates by Superdex 200 HPLC were <1%. Potency revealed a 20 h IC(50) of 17f M on Jurkat cells. Endotoxin level was 0.02 U/mg. Chemical and biologic assays confirmed the purity, composition, and functional activities of the molecule. The drug did not react with tested frozen human tissue sections except for T cells. LD(10) in mice was between 500 and 75 0microg/kg. There was no evidence of loss of solubility, proteolysis, aggregation, or loss of potency over 1.5 year at -80 degrees C. The scalable synthesis of this protein drug should be useful for production for phase I/II clinical trials and can be applicable for other diphtheria toxin fusion drugs for clinical development. PMID- 18160310 TI - A prophenoloxidase from the Chinese mitten crab Eriocheir sinensis: gene cloning, expression and activity analysis. AB - Prophenoloxidase (proPO) is a conserved copper-containing enzyme that plays important roles in immune response of crustaceans and insects. In the present study, the full-length cDNA of a prophenoloxidase (designated EsproPO) was cloned from haemocytes of Chinese mitten crab Eriocheir sinensis by expressed sequence tag (EST) and PCR techniques. The isolated 3549bp full-length cDNA of EsproPO contained a 2040bp open reading frame (ORF) encoding a putative proPO protein of 679 amino acids, a 5'-untranslated region (UTR) of 68bp, and a long 3'-UTR of 1441bp. Two putative copper-binding sites, a proteolytic activation site, and a complement-like motif (GCGWPQHM) were identified in the deduced amino acid sequence of EsproPO. Homology analysis revealed that EsproPO was highly similar to other proPOs from crustaceans with identities from 52% to 68%. The conserved domains and motifs, and higher similarity with other proPOs suggested that EsproPO was a member of the proPO family. The mRNA expression of EsproPO and PO specific activities in the tissues of hepatopancreas, gill, gonad, muscle, heart, eye and haemocytes were measured by quantitative real-time PCR and colorimetric assay, respectively. The mRNA transcripts of EsproPO and PO specific activities could be detected in all the examined tissues with the highest level both in hepatopancreas. Three peaks of EsproPO mRNA expression were recorded at 2h, 12h and 48h in haemocytes of Chinese mitten crab post Vibrio anguillarum challenge, which was consistent with the temporal profile of PO specific activity. The mRNA expression pattern and the activity fluctuation of EsproPO post V. anguillarum stimulation indicated that it was potentially involved in the acute response against invading bacteria in Chinese mitten crab. PMID- 18160311 TI - Activity of deep abdominal muscles increases during submaximal flexion and extension efforts but antagonist co-contraction remains unchanged. AB - Lumbo-pelvic stability relies, amongst other factors, on co-contraction of the lumbo-pelvic muscles. However, during submaximal trunk flexion and extension efforts, co-contraction of antagonist muscles is limited. It was predicted that activity of the deeper lumbo-pelvic muscles that are often excluded from analysis (transversus abdominis (TrA) and the deep fascicles of multifidus (DM)), would increase with load in each direction. In eleven healthy subjects, electromyographic activity (EMG) was recorded from eight trunk muscles using surface and fine-wire electrodes. Subjects performed isometric flexion and extension efforts to submaximal loads of 50, 100, 150 and 200 N and a maximal voluntary contraction (MVC). Loading tasks were then repeated in trials in which subjects knew that the load would release at an unpredictable time. Compared to the starting position, EMG of all muscles, except DM, increased during MVC efforts in both directions. During the flexion and extension submaximal tasks, there was no increased co-contraction of antagonist muscles. However, TrA EMG increased in both directions. In the unpredictable trials, EMG of all lumbo pelvic muscles except TrA was decreased. These findings provide further support for a contribution of TrA to lumbo-pelvic stability. In submaximal tasks, TrA activation may enhance stability as a strategy to improve trunk stiffness without requiring a concurrent increase in activity of the larger torque producing trunk muscles. PMID- 18160312 TI - MMSE scores correlate with local ventricular enlargement in the spectrum from cognitively normal to Alzheimer disease. AB - In this work, we aimed at correlating focal atrophy in periventricular structures with cognitive function, in the spectrum from healthy subjects to severe Alzheimer disease: 28 subjects with normal cognition and 84 patients presenting various degrees of cognitive impairment were included in the study. The cognitive level of each subject was assessed with the Mini-Mental State Examination (MMSE). Atrophy in periventricular structures was inferred by modeling and analyzing local shape variations of brain ventricles: for a given subject, we distinguished between the severity of atrophy, estimated as local enlargement (in mm) of the ventricular surface relative to an average normal subject, and the extent of atrophy, defined as the percentage of the ventricular surface (global or per anatomical region) significantly different from an average control. Linear regression across subjects was performed to evaluate the correlation between atrophy and MMSE score. The severity of atrophy showed good correlation with MMSE score in the left thalamus, the left temporal horn, the left corona radiata, and the right caudate nuclei. The extent of atrophy showed no significant correlations. In conclusion, the MMSE scores correlate with localized depth of atrophy in well-defined periventricular structures. PMID- 18160313 TI - Teratogenic effects of antiepileptic drugs. AB - The use of older generation antiepileptic drugs (AEDs) during pregnancy is known to be associated with a two- to threefold increased risk of birth defects in the offspring and possible also other adverse outcomes in the exposed infant. Much less has been known about newer generation AEDs in this respect. Recent studies based on national registries as well as specific epilepsy and pregnancy registries are beginning to provide information on comparative teratogenic effects of different AEDs. Hence, the prevalence of birth defects appears to be higher with exposure to valproate compared with carbamazepine and possibly also in comparison with lamotrigine. Further studies based on larger cohorts are needed to compare AEDs at different dosages and to analyse the possible impact of confounding factors. Furthermore, data is insufficient to assess the human teratogenic potential of other newer generation AEDs than lamotrigine. Retrospective and a few small prospective studies suggest that exposure to valproate also might be associated with a lower verbal IQ at school age, but further prospective studies are needed to draw firm conclusions. PMID- 18160314 TI - Auxin and cytoskeletal organization in algae. AB - Hormones affect growth and alter the cytoskeleton suggesting that hormones and the cytoskeleton interact with each other. The cytoskeleton of ancestral algae such as Chara showed similar sensitivity to auxin as higher plants, even in generative structures but the sensitivity differed between IAA and alpha-NAA and presumably other auxins. The ability of cells to elongate depends on microtubule organization during the transition from disorganized to perpendicular to longitudinal organization of the cytoskeleton. Because of the many functions of the cytoskeleton it is possible that its composition is influenced by selective gene expression and adaptation to growth regulators. Co-localization of microtubules and F-actin change at a high temporal and spatial scale. High resolution measurements of mRNA expression indicate rapid turnover that may affect the composition of the cytoskeleton. PMID- 18160315 TI - Reactivated memory of an inhibitory avoidance response in mice is sensitive to a nitric oxide synthase inhibitor. AB - It is accepted that once consolidation is completed memory becomes permanent. However, it has also been suggested that reactivation (retrieval) of the original memory, again, makes it sensitive to the same treatments that affect memory consolidation when given after training. Previous results demonstrated that the immediate post-training intraperitoneal administration of N(omega)-nitro-l arginine methyl ester (L-NAME), a non-specific inhibitor of nitric oxide synthase (NOS), impairs retention test performance of a one-trial step-through inhibitory avoidance response in adult mice. The effect of L-NAME on retention was attributed to an action on memory consolidation of the original learning. For the first time, we report that the administration of L-NAME after the first retention test (memory reactivation) of the inhibitory avoidance response impairs retention performance over six consecutive days. This impairment effect is dose-and-time dependent and could not be attributed to a retrieval deficit since a mild footshock did not reinstate the original avoidance response and no spontaneous recovery was observed at least 21 days after training. Further support for a storage deficit interpretation as opposed to a retrieval deficit was obtained from the fact that L-NAME's effects after retrieval were not due to state dependency. The impairment effect of L-NAME was dependent on the age of the original memory. That is, there was an inverse correlation between the susceptibility of the memory trace when reactivated and the time elapsed between training and the first retrieval session. We suggest an action of L-NAME on memory reactivation-induced processes that are different from memory extinction of the original learning extending the biological significance of nitric oxide on memory. PMID- 18160316 TI - What does 'best medical therapy' really mean? AB - Carotid disease is like any other atherosclerotic manifestation, a condition, which may induce thrombosis, in this case with subsequent cerebral ischemia. Carotid endarterectomy has proven effective in preventing ipsilateral stroke, however, the studies providing the evidence were conducted before the use of statins, newer antiplatelet and antihypertensive drugs, and at a time when less emphasis was on lifestyle modification. Therefore, it is likely that, not only would all patients with carotid stenosis benefit from modern medical treatment, in addition, some patients could have similar risk reduction to that of endarterectomy, were these effective preventive drugs used systematically, as recommended, in this patient group. This article reviews the evidence that is available concerning medical therapy for patients with carotid stenosis, with special emphasis on antiplatelet and statin therapy. An example on how this treatment may be organised is given. PMID- 18160317 TI - Hepatic iron concentration, fibrosis and response to venesection associated with the A77D and V162del "loss of function" mutations in ferroportin disease. AB - Ferroportin disease is an autosomal dominant form of hemochromatosis associated with siderosis in cells of the mononuclear phagocyte system and, to varying degrees, in hepatocytes. Ferroportin was investigated as a candidate gene in two pedigrees with hyperferritinaemia and siderosis in mononuclear phagocytes. The entire ferroportin coding region was sequenced and hepatic iron concentration, histology and response to treatment were determined. The results were compared with previously reported cases. The A77D mutation was detected in patient 1, his father (patient 2) and his brother (patient 3), who had portal fibrosis. The V162del mutation was detected in patient 4, who developed anemia after the third weekly venesection. While the disease is rare, A77D and V162del are the most common ferroportin mutations in Caucasians. The spectrum of clinical expression of these two mutations was reviewed in all cases described to date. These mutations were associated with fibrosis in about a third of cases. For A77D and V162del, this analysis confirms that the threshold hepatic iron concentration for development of fibrosis may be higher than for classical hemochromatosis. These two mutations, which both decreased iron export in cell culture studies, give rise to similar patterns of clinical expression and morbidity, although the highest hepatic iron concentrations have been observed with A77D. It is important for clinicians to consider ferroportin disease in cases where there are features of iron overload unrelated to HFE, autosomal dominant inheritance and/or iron deposition in mononuclear phagocytes. PMID- 18160318 TI - Number of pain sites is associated with demographic, lifestyle, and health related factors in the general population. AB - Research on widespread pain often relies upon case definitions bounded by duration limits or "cut-offs." In clinical reality, however, there are no natural cut-off points between localized and widespread pain. Rather, pain is best represented by a continuum of "widespreadness" from localized pain to pain spread across the body. The objective of this paper was to describe the number of pain sites (NPS) reported in a population study and its association with demographic, lifestyle, and health-related factors. Using a cross-sectional design, the Standardized Nordic Questionnaire was used to measure musculoskeletal pain among seven age groups in Ullensaker, Norway (n=2926). Results showed that women reported a higher mean NPS than men. A higher NPS was also found for individuals who were separated or divorced, undergoing rehabilitation, or who had a disability pension. Additionally, greater NPS was reported by smokers, individuals with less physical activity, and a higher BMI. A strong linear relationship was found between NPS and reduction in overall health, sleep quality, and psychological health. Results from a multivariate linear regression analysis showed that overall health, sleep quality, and gender demonstrated the strongest associations with increasing NPS, accounting for 31.4% of the variance. Our study indicates that the straightforward and simple method of counting the NPS could be important in managing the complex problem of musculoskeletal pain. PMID- 18160319 TI - A comparison of the relationship between depression, perceived disability, and physical performance in persons with chronic pain. AB - This study examined the relationships between self-report of depressive symptoms, perceived disability, and physical performance among 267 persons with chronic pain. Prior research has reported a relationship between depression and disability using self-report measures. However, self-report instruments may be prone to biases associated with depression as depressed persons with pain may have an exaggerated negative view of their level of function. In addition, we examined whether the relationship between depression and functional activity was mediated by physiologic effort (as measured by heart rate). The results indicated that self-report of depressive symptoms (using the Center for Epidemiological Studies-Depression Scale (CES-D)) was significantly correlated with self-report of disability on the Quebec Back Pain Disability Scale (QBPDS) and physical performance on the Progressive Isoinertial Lifting Evaluation (PILE). Regression analyses revealed that depression assessed by the CES-D significantly contributed to the prediction of QBPDS scores and PILE performance even when controlling for age, gender, site of pain, and pain intensity. The magnitude of the relationships between depression and self-report and functional activity were similar, suggesting that a self-report bias associated with depression is not responsible for an observed relationship between depression and disability. Physiologic effort partially mediated the relationship between depression and physical performance. The findings further highlight the importance of depression in the experience of chronic pain. PMID- 18160320 TI - Effect of preoperative stability of alignment on outcome of strabismus surgery for infantile esotropia. AB - INTRODUCTION: Some strabismus surgeons wait until the angle of deviation stabilizes prior to operating on patients with infantile esotropia. This study evaluated whether a preoperative increase in the angle of deviation affected surgical outcomes. METHODS: This was a retrospective comparative case series in which records of patients with infantile esotropia, who had surgery performed before age 2 years and who were followed for at least 2 years postoperatively, were reviewed to identify two groups for comparison: the first had /=10(Delta) increase in the angle of deviation prior to surgery. In the latter group, surgery was performed for the angle present at the time of surgery. Outcomes were considered successful if the patients had microtropias. RESULTS: Fifteen patients were identified in each group. The interval between the first office visit and time of surgery was the same in both groups (mean, 2 months). Outcomes were successful in 11 of 15 (73%) in each group (no significant difference, p = 0.659). CONCLUSIONS: There was no difference in outcome between children whose angles of deviation were stable and those whose angles increased prior to surgery. This indicates that strabismus surgery does not need to be delayed while waiting for the angle of deviation to stabilize. Surgical correction may therefore be achieved at an earlier age, which may have a beneficial effect on outcome. PMID- 18160321 TI - 6-MBOA affects testis size, but not delayed-type hypersensitivity, in white footed mice (Peromyscus leucopus). AB - Many rodents use day length to time reproduction to occur when resources are abundant, but some species also use supplementary environmental cues. One supplementary cue is the plant-derived compound, 6-methoxy-2-benzoxazolinone (6 MBOA). Most rodents grow their gonads in response to 6-MBOA in their diets, but it is presently unknown whether they also use 6-MBOA to adjust other aspects of physiology, specifically their immune systems. 6-MBOA is structurally similar to melatonin, and seasonal changes in rodent immune activities are often mediated by melatonin. We therefore predicted that white-footed mice (Peromyscus leucopus), which breed seasonally and are reproductively sensitive to melatonin, would adjust their immune systems when fed 6-MBOA. 6-MBOA treated mice in long day lengths regressed their testes to a greater extent than mice fed a standard diet, or mice kept in short day lengths and fed 6-MBOA or a standard diet. One type of immune activity (delayed-type hypersensitivity) was not affected by 6-MBOA, however, although responses were greater in short versus long day mice. In sum, P. leucopus responded reproductively to 6-MBOA, although differently than other species; immune activity was unaffected. Other aspects of the immune system, especially in herbivorous rodents, may be affected by 6-MBOA and thus warrant further study. PMID- 18160322 TI - Intracellular cholesterol modifies the ERAD of glucocerebrosidase in Gaucher disease patients. AB - Gaucher disease (GD) is a lysosomal storage disorder resulting from an inborn reduced activity or deficiency of glucocerebrosidase due mainly to mutations in the glucocerebrosidase gene. We have recently shown that mutant glucocerebrosidase variants present variable degrees of endoplasmic reticulum (ER) retention and undergo ER associated degradation (ERAD) in the proteasomes. The degree of ERAD is one of the factors that determine GD severity. In order to define what factors affect the ERAD process of glucocerebrosidase in GD, we focused on two brothers with GD, carrying the same mutations but presenting extremely different clinical manifestations. One is mildly affected while the other developed severe GD with nervous system complications. Our results strongly indicated that both brothers presented variable degrees of ERAD, which was more extensive in the severely affected brother. Measurement of cholesterol demonstrated high intracellular levels in cells that derived from the severely affected brother. Growing the cells in cholesterol depleted medium led to lessening in the degree of ERAD in cells that derived from the severely affected brother and thus to improvement in stabilization, maturation, lysosomal localization and activity of the mutant glucocerebrosidase variants. The same effect was achieved by treating the cells with the HMG CoA reductase inhibitor mevastatin. None of the treatments had a significant effect on glucocerebrosidase properties in normal cells or in cells that derived from the mildly affected brother, indicating that intracellular cholesterol is one of the factors that affect the ERAD process of glucocerebrosidase and may influence the severity of GD. PMID- 18160323 TI - Oncocytoma in the mandibular salivary gland of a cat. AB - A cat was referred for investigation of a soft tissue mass caudal to the left mandible. Initial investigations suggested a malignant salivary gland tumour, and the mass was removed by extracapsular resection of the mandibular gland. Histopathology showed an oncocytoma within the salivary gland. An oncocytoma is a neoplastic transformation of oncocytes. Oncocytes are cells with a small nucleus and intense eosinophilic granular cytoplasm due to numerous mitochondria, which proliferate during ageing in exocrine and endocrine glandular tissues. Physiological proliferation occurs next to oncocytosis, oncocytoma, and oncocytic carcinoma. This is the first report of an oncocytoma in a feline mandibular salivary gland, and the first report of long-term survival after surgical removal. PMID- 18160324 TI - PGF2alpha-associated vascular smooth muscle hypertrophy is ROS dependent and involves the activation of mTOR, p70S6k, and PTEN. AB - Prostaglandin F2alpha (PGF2alpha) increases reactive oxygen species (ROS) and induces vascular smooth muscle cell (VSMC) hypertrophy by largely unknown mechanism(s). To investigate the signaling events governing PGF2alpha-induced VSMC hypertrophy we examined the ability of the PGF2alpha analog, fluprostenol to elicit phosphorylation of Akt, the mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase (p70S6k), glycogen synthase kinase-3beta (GSK-3beta), phosphatase and tensin homolog (PTEN), extracellular signal-regulated kinase 1/2 (ERK1/2) and Jun N-terminal kinase (JNK) in growth arrested A7r5 VSMC. Fluprostenol-induced hypertrophy was associated with increased ROS, mTOR translocation from the nucleus to the cytoplasm, along with Akt, mTOR, GSK-3beta, PTEN and ERK1/2 but not JNK phosphorylation. Whereas inhibition of phosphatidylinositol 3-kinase (PI3K) by LY-294002 blocked fluprostenol-induced changes in total protein content, pre-treatment with rapamycin or with the MEK1/2 inhibitor U0126 did not. Taken together, these findings suggest that fluprostenol induced changes in A7r5 hypertrophy involve mTOR translocation and occur through PI3K-dependent mechanisms. PMID- 18160325 TI - Editorial by the editor-in-chief of microbes and infection. PMID- 18160326 TI - The paradox of infection and inflammation: friends or foes? The dilemma of biological usage in high endemic tuberculous areas. PMID- 18160327 TI - Insight into the selectivity of arsenic trioxide for acute promyelocytic leukemia cells by characterizing Saccharomyces cerevisiae deletion strains that are sensitive or resistant to the metalloid. AB - The genome-wide set of Saccharomyces cerevisiae deletion strains provides the opportunity to analyze how other organisms may respond to toxic agents. Since arsenic trioxide selectively kills human acute promyelocytic leukemia (APL) cells by a poorly understood mechanism we screened the yeast deletion strains for sensitivity or resistance. In addition to confirming mutants previously identified as sensitive to sodium arsenite, a large number of additional genes, and cellular processes, were required for arsenic trioxide tolerance. Of the 4546 mutants, 7.6% were more sensitive to arsenic trioxide than the wild type, while 1.5% was more resistant. IC50 values for all sensitive and resistant mutants were determined. Prominent as sensitive was that missing the MAP kinase, Hog1. The most resistant lacked the plasma-membrane glycerol and arsenite transporter, Fps1. Hog1 and Fps1 control the response to osmotic stress in yeast by regulating glycerol production and plasma membrane flux, respectively. We therefore tested whether APL cells have impaired osmoregulation. The APL cell line NB4 did not produce glycerol in response to osmotic stress and underwent apoptotic cell death. Moreover, the glycerol content of NB4 and differentiated NB4 cells correlated with the level of arsenic trioxide uptake and the sensitivity of the cells. Additionally, NB4 cells accumulated more arsenic trioxide than non-APL cells and were more sensitive. These findings demonstrate the usefulness of the S. cerevisiae deletion set and show that the selectivity of arsenic trioxide for APL cells relates, at least in part, to impaired osmoregulation and control of uptake of the drug. PMID- 18160328 TI - Clioquinol inhibits peroxide-mediated toxicity through up-regulation of phosphoinositol-3-kinase and inhibition of p53 activity. AB - A growing body of evidence supports a central role for biometals in neurodegenerative disorders. Biometals induce oxidative stress through the generation of reactive oxygen species and contribute to neuronal cell dysfunction in Alzheimer's disease (AD), prion disorders and Parkinson's disease (PD). Therapies based on modulation of biometal metabolism are currently being developed and the metal ligand, 5-chloro-7-iodo-8-hydroxyquinoline (clioquinol or CQ) has been investigated for the treatment of AD. CQ has also shown therapeutic benefits in an animal model of PD. However, little is known about the neuroprotective processes of CQ in vivo. In this study, we examined the effect of CQ in BE(2)-M17 human neuroblastoma cells exposed to increased oxidative stress (hydrogen peroxide (H2O2) treatment). Although CQ alone induced a moderate toxic effect on cells, when added to H2O2-treated M17 cells, CQ induced a significant inhibition of H2O2 toxicity. This correlated with up-regulation of phosphoinositol-3-kinase (PI3K) activity in CQ-treated cells. The protective action of CQ was not observed in murine N2a neuroblastoma cells treated with H2O2 and this cell-line did not reveal CQ-mediated increases in PI3K activation. The protective effect was specific for CQ and was not induced by a number of different metal ligands. Inhibition of PI3K activity with LY294002 prevented CQ protection against H2O2 toxicity, demonstrating a crucial role for CQ activation of PI3K in protection against oxidative stress. Furthermore, CQ inhibited H2O2 mediated up-regulation of p53 activity in the M17 cells and this was dependent on PI3K activation. Our studies demonstrate that in human M17 cells, CQ can protect against oxidative stress by activating the PI3K-dependent survival pathway and blocking p53-mediated cell death. These findings have important implications for the development of protective metal ligand-based therapies for treatment of disorders involving oxidative stress. PMID- 18160329 TI - Post-genomics dissection of seed dormancy and germination. AB - Studies of genome expression in Arabidopsis have provided important new information about mechanisms controlling germination and suggest new avenues to explore. Unexpectedly, changes in transcription and protein metabolism were observed in the 'dry'(1) quiescent seed state, suggesting that careful controls are required for seed expression-profiling experiments. Changes in the transcriptome following seed imbibition suggest a dynamic relationship between RNAs 'stored' from seed maturation, and synthesis of new RNAs related to post imbibition germinating- or dormant-seed states. Recent post-genomics approaches suggest that RNA translation or post-translation are the major levels of control for germination completion and that transcriptome changes might reflect alteration in dormancy status or enhancement of germination vigour and effects on post-germination functions that relate to seedling growth. PMID- 18160330 TI - Decoding genes with coexpression networks and metabolomics - 'majority report by precogs'. AB - Following the sequencing of whole genomes of model plants, high-throughput decoding of gene function is a major challenge in modern plant biology. In view of remarkable technical advances in transcriptomics and metabolomics, integrated analysis of these 'omics' by data-mining informatics is an excellent tool for prediction and identification of gene function, particularly for genes involved in complicated metabolic pathways. The availability of Arabidopsis public transcriptome datasets containing data of >1000 microarrays reinforces the potential for prediction of gene function by transcriptome coexpression analysis. Here, we review the strategy of combining transcriptome and metabolome as a powerful technology for studying the functional genomics of model plants and also crop and medicinal plants. PMID- 18160331 TI - Enhanced polarizing microscopy as a new tool in aneuploidy research in oocytes. AB - Chromosomal non-disjunction in female meiosis gives rise to reduced fertility and trisomy in humans. Human oocytes, especially from aged women, appear especially susceptible to non-disjunction. The oocyte spindle is crucial for high fidelity of chromosome segregation at meiotic divisions, and alterations in spindle morphology are therefore indicators of adverse conditions during oocyte development that may result in meiotic aneuploidy. In the past, oocytes had to be fixed for spindle analysis, precluding direct non-invasive identification of aneugens and adverse maturation conditions that affect spindle integrity and chromosome behaviour. Aneuploidy research for detection of spindle aberrations was therefore mainly focused on in vivo or in vitro exposed, fixed animal oocytes or cytogenetic analysis of spread oocytes. Orientation independent enhanced polarizing microscopy with nearly circularly polarized light and electronically controlled liquid crystal compensator optics is a new tool to study spindle morphology non-invasively in vivo for qualitative as well as quantitative analysis. Image generation by polarization microscopy depends on the intrinsic optical properties of the spindle with its paracrystalline microtubule lattice. When polarized light passes through such a lattice it induces a splitting of the beam and shift in the plane of vibration and retardation of light (termed birefringence and retardance). Studies of animal oocytes and follicle-cell denuded human oocytes fertilized by intracytoplasmic sperm injection for assisted conception have demonstrated the safety and efficacy of enhanced polarization microscopy. The method can be employed in aneuploidy research for non-invasive dose-response studies to detect spindle aberrations, for instance, in combination with cytogenetic analysis. Due to the non-invasive nature of the technique it may be employed in routine analysis of human oocytes to assess risks by lifestyle factors, and occupational and adverse environmental exposures. PMID- 18160332 TI - Solar-simulated light-exposed benzo[a]pyrene induces phosphorylation of histone H2AX. AB - Polycyclic aromatic hydrocarbons (PAHs), wide-spread mutagenic and carcinogenic environmental pollutants, are consistently exposed to sunlight in the environment. The exposure causes structural change, resulting in the generation of a variety of photomodified products having different bioactivities compared with the parent compounds. In this study, we found that benzo[a]pyrene (BaP) exposed to solar-simulated light (SSL)-induced phosphorylation of histone H2AX (gamma-H2AX), which was recently identified as an early event after the induction of DNA double strand breaks (DSBs). Although BaP itself did not produce gamma H2AX, SSL-exposed BaP significantly generated gamma-H2AX depending on the period of exposure. Furthermore, we revealed that reactive oxygen species produced by the SSL-exposed BaP mainly contributed to the generation of gamma-H2AX. The appearance of gamma-H2AX means the induction of the most serious form of DNA damage, DSBs, suggesting the potential risk of carcinogenesis. PMID- 18160333 TI - DNA-damaging activity and mutagenicity of 16 newly synthesized thiazolo[5,4 a]acridine derivatives with high photo-inducible cytotoxicity. AB - The discovery of the potent anticancer properties of natural alkaloids in the pyrido-thiazolo-acridine series has suggested that thiazolo-acridine derivatives could be of great interest. In a continuous attempt to develop DNA-binding molecules and DNA photo-cleavers, 16 new thiazolo[5,4-a]acridines were synthesized and studied for their photo-inducible DNA-intercalative, cytotoxic and mutagenic activities, by use of the DNA methyl-green bioassay, the Alamar Blue viability assay and the Salmonella mutagenicity test using strains TA97a and TA98 with and without metabolic activation and photo-activation. Without photo activation, one compound showed a DNA-intercalative activity in the DNA major groove while three compounds displayed intercalating properties after photo activation. In the dark, four molecules possessed cytotoxic activities against a THP1 acute monocytic leukemia cell line while 15 derivatives displayed photo inducible cytotoxic activity against this cell line. All compounds were mutagenic in strain TA97a with metabolic activation (+S9mix) and 15 molecules were mutagenic in strain TA98 without activation (-S9mix). Study of the quantitative structure-activity relationships (QSAR) from the Salmonella mutagenicity data revealed that several descriptors could describe cytotoxic and mutagenic activities after photo-activation. From the results of the mutagenicity test, four compounds with elevated mutagenic activities were selected for additional experiments. Their capacities to induce single-strand breaks (SSB) and chromosome damaging effects were monitored by the comet and the micronucleus assays in normal human keratinocytes. Comparison of the minimal genotoxic concentrations showed that two compounds possessed higher capacities to induce SSB after photo activation. In the micronucleus assay, three molecules were able to induce high numbers of micronuclei following photo-activation. Overall, the results of this study confirm that acridines are predominantly genotoxic via a DNA-intercalating mechanism in the dark, while DNA-adducts were probably induced following photo activation. PMID- 18160334 TI - Unexpected DNA damage caused by polycyclic aromatic hydrocarbons under standard laboratory conditions. AB - The genotoxicity of 15 polycyclic aromatic hydrocarbons was determined with the alkaline version of the comet assay employing V79 lung fibroblasts of the Chinese hamster as target cells. These cells lack the enzymes necessary to convert PAHs to DNA-binding metabolites. Surprisingly, 11 PAHs, i.e., benzo[a]pyrene (BaP), benz[a]anthracene, 7,12-dimethylbenz[a]anthracene, 3-methylcholanthrene, fluoranthene, anthanthrene, 11H-benzo[b]fluorene, dibenz[a,h]anthracene, pyrene, benzo[ghi]perylene and benzo[e]pyrene caused DNA strand breaks even without external metabolic activation, while naphthalene, anthracene, phenanthrene and naphthacene were inactive. When the comet assay was performed in the dark or when yellow fluorescent lamps were used for illumination the DNA-damaging effect of the 11 PAHs disappeared. White fluorescent lamps exhibit emission maxima at 334.1, 365.0, 404.7, and 435.8 nm representing spectral lines of mercury. In the case of yellow fluorescent lamps these emissions were absent. Obviously, under standard laboratory illumination many PAHs are photo-activated, resulting in DNA damaging species. This feature of PAHs should be taken into account when these compounds are employed for the initiation of skin cancer. The genotoxicity of BaP that is metabolically activated in V79 cells stably expressing human cytochrome P450-dependent monooxygenase (CYP1A1) as well as human epoxide hydrolase (V79 hCYP1A1-mEH) could not be detected with the comet assay performed under yellow light. Likewise the DNA-damaging effect of r-7,t-8-dihydroxy-t-9,10-epoxy 7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BaPDE) observed with the comet assay was only weak. However, upon inhibition of nucleotide excision repair (NER), which is responsible for the removal of stable DNA adducts caused by anti-BaPDE, the tail moment rose 3.4-fold in the case of BaP and 12.9-fold in the case of anti-BaPDE. These results indicate that the genotoxicity of BaP and probably of other compounds producing stable DNA adducts are reliably detected with the comet assay only when NER is inhibited. PMID- 18160335 TI - Validation of an automatic comet assay analysis system integrating the curve fitting of combined comet intensity profiles. AB - In recent years, the single-cell gel electrophoresis (comet) assay has become a reference technique for the assessment of DNA fragmentation both in vitro and in vivo at the cellular level. In order to improve the throughput of genotoxicity screening, development of fully automated systems is clearly a must. This would allow us to increase processing time and to avoid subjectivity brought about by frequent manual settings required for the 'classical' analysis systems. To validate a fully automatic system developed in our laboratory, different experiments were conducted in vitro on murine P388D1 cells with increasing doses of ethyl methanesulfonate (up to 5 mM), thus covering a large range of DNA damage (up to 80% of DNA in the tail). The present study (1) validates our 'in house' fully automatic system versus a widely used semi-automatic commercial system for the image-analysis step, and versus the human eye for the image acquisition step, (2) shows that computing tail DNA a posteriori on the basis of a curve fitting concept that combines intensity profiles [G. Dehon, P. Bogaerts, P. Duez, L. Catoire, J. Dubois, Curve fitting of combined comet intensity profiles: a new global concept to quantify DNA damage by the comet assay, Chemom. Intell. Lab. Syst. 73 (2004) 235-243] gives results not significantly different from the 'classical' approach but is much more accurate and easy to undertake and (3) demonstrates that, with these increased performances, the number of comets to be scored can be reduced to a minimum of 20 comets per slide without sacrificing statistical reliability. PMID- 18160336 TI - A survey of malarial infection in endemic areas of Savannakhet province, Lao PDR and comparative diagnostic efficiencies of Giemsa staining, acridine orange staining, and semi-nested multiplex PCR. AB - Malaria remains one of the most important parasitic diseases in Lao PDR, especially in forested rural areas. Knowing the rate of infection using highly sensitive and specific methods, and the factors related to malarial infection, may be helpful in reducing the infection and mortality rates. We aimed to study the malarial infection rate by comparing three detection methods, i.e., Giemsa staining, acridine orange (AO) staining and semi-nested multiplex PCR. The study also included some factors related to malarial infection in the endemic areas of Savannakhet province, Lao PDR. The respective malarial infection rates by Giemsa staining, AO staining and semi-nested multiplex PCR in Houy Jang vs. Keng Thong villages were 13.1 vs. 20.8, 16.2 vs. 25.4 and 20.8 vs. 30.8%. The infection rate among children not over 10 years of age was higher than infection rate among the older ages (p=0.002, Z-test for two proportions). The higher infection rates by semi-nested multiplex PCR over Giemsa and AO staining suggest the existence of many subclinical cases with low level parasitemia, undetected by microscopic techniques. We found no mixed infections using Giemsa or AO staining, but using semi-nested multiplex PCR we found 1.2% (3/260) mixed P. falciparum and P. vivax infections, suggesting that semi-nested multiplex PCR is suitable for detecting malarial infection from endemic areas whose cases may have low parasitemia and/or mixed infection. The factors significantly related to malarial infection from 260 questionnaires were: (1) children and young adults, (2) not having lived in the area more than 5 years, and (3) not using a mosquito net over the bed, indicating an increased risk of new residents of contracting malaria and a need to promote bed nets. PMID- 18160337 TI - Substituted tertiary phosphine Ru(II) organometallics: catalytic utility on the hydrolysis of etofibrate in pharmaceuticals. AB - Some new organometallics of ruthenium(II) of the type [RuCl2(COD)(CO)L] (1a-f) and [RuCl2(COD)L2] (2a-f) (where L is substituted tertiary phosphines), have been synthesized by using precursors [RuCl2(COD)(CO)(CH3CN)] (1) and [RuCl2(COD)(CH3CN)2] (2) with the substituted tertiary phosphine ligands in 1:1 and 1:2 molar ratio. The organometallics (2a-f) have been further reacted with carbonmonoxide to produce compounds of the type [RuCl2(CO)L2] (3a-f). These compounds were characterized by elemental analysis, IR, NMR (1H, 13C and 31P), mass and electronic spectral data. The catalytic activity of all these organometallics were studied and found that they are efficient catalysts for hydrolysis of etofibrate. The hydrolyzed product was separated by column chromatography and the percent yields are found in the range of 98.6-99.1%. PMID- 18160338 TI - Role of coordinated metal ions on the orientation of phthalocyanine based coatings. AB - In the present work, we have investigated the molecular orientation of phthalocyanine films deposited on polycrystalline gold. Three films built from the following molecules are investigated: phthalocyanine (H2Pc), cobalt phthalocyanine (CoPc) and copper phthalocyanine (CuPc). The films are prepared by spin coating and drop casting methods. Orientation analysis has been performed using polarization dependent Fourier transform infrared (FTIR) spectroscopy using transmission and grazing angle reflectance mode. The FTIR study suggests that each phthalocyanine film contains both alpha- and beta-phases. H2Pc based films demonstrate deposition method dependence on the molecular orientation, while the CuPc and CoPc films preserve their molecular orientation independent of deposition method. Grazing angle analysis also suggests that CoPc films show negligible preferred orientation irrespective of film deposition methods. In literature, the band at 878 cm(-1) in CuPc has been assigned to out-of-plane bending of C-H. Our grazing angle experiments suggest that this band cannot be assigned to out-of-plane bending vibrations of C-H. Accurate band assignments are also described here for the phthalocyanine system. PMID- 18160339 TI - Vibrational spectra and quantum chemical calculations of 3,4-diaminobenzoic acid. AB - The Fourier Transform Raman and Fourier Transform infrared spectra of 3,4 diaminobenzoic acid (3,4-DABA) were recorded in the solid phase. Geometry optimizations were done without any constraint and harmonic-vibrational wave numbers and several thermodynamic parameters were calculated for the minimum energy conformer at ab initio and DFT levels invoking 6-311++G(d,p) basis set. The results were compared with the experimental values with the help of specific scaling procedures, the observed vibrational wavenumbers in FT-IR and FT-Raman spectra were analyzed and assigned to different normal modes of the molecule. Most of the modes have wavenumbers in the expected range, the error obtained was in general very low. The appropriate theoretical spectrograms for the IR and Raman spectra of the title molecule were also constructed. PMID- 18160340 TI - C-reactive protein, diastolic dysfunction, and risk of heart failure in patients with coronary disease: Heart and Soul Study. AB - BACKGROUND: High-sensitivity C-reactive protein (CRP) is an inflammatory marker that predicts coronary heart disease (CHD) and, in recent studies, incident heart failure (HF). Whether the association of inflammation with incident HF is explained by worse baseline left ventricular dysfunction or by underlying CHD is unknown. METHODS AND RESULTS: Serum CRP was measured in a cohort of 985 outpatients with established CHD from the Heart and Soul Study. During 3 years of follow-up, 15% of the participants with elevated CRP levels (>3 mg/L) were hospitalised for HF, compared with 7% of those with CRP 100 mg/d increases bleeding complications without any additional reduction in cardiovascular events. In summary, although aspirin is recommended for every patient with coronary artery disease, its optimal dose for patients who underwent stent implantation is still not clear. PMID- 18160362 TI - [Use of aspirin in patients taking angiotensin converting enzyme inhibitors]. AB - Aspirin and angiotensin converting enzyme (ACE) inhibitors both have beneficial effects on prognosis in patients with cardiovascular disease. For this reason, they are usually prescribed together. Some of the beneficial effects of ACE inhibitors are thought to be due to reduced degradation of bradykinin. Bradykinin enhances nitric oxide and vasodilatory prostaglandins. Theoretically, aspirin, which inhibits cyclooxygenase enzyme, may reduce bradykinin mediated prostaglandin synthesis and blunt the beneficial effects of ACE inhibitors, when used together. Although some hemodynamic studies support this hypothesis, clinical studies have conflicting results. In this article, we reviewed the possible interaction between aspirin and ACE inhibitors in light of literature findings. PMID- 18160363 TI - [A current problem in atherothrombotic diseases--aspirin resistance: definition, mechanisms, determination with laboratory tests and clinical implications]. AB - Aspirin (acetylsalicylic acid) is a powerful antiplatelet agent used in prevention of atherothrombotic vascular events. However, antiplatelet effect of aspirin is not uniform and some patients could not benefit from aspirin. These patients are clinically called as aspirin resistant or aspirin non-responders. Aspirin resistance could be determined by: bleeding time, optical aggregometry, PFA-100 (Platelet Function Analyzer), Ultegra-RPFA (Rapid Platelet Function Assay), activated aggregation time, whole blood aggregometry, platelet aggregate ratio, flow cytometry, measurements of platelet surface proteins and blood or urine thromboxane B2 levels. Mechanisms of aspirin resistance have not been elucidated yet. There is evidence that aspirin resistance increases clinical cardiovascular events. Adequate additional therapies may reduce atherothrombotic risks and major cardiovascular events rate in aspirin resistant subjects. However, we need further studies to decrease major cardiovascular events risk in aspirin resistant subjects and to optimize antiplatelet therapy. PMID- 18160364 TI - Aspirin and gastrointestinal toxicity. AB - Aspirin is the main actor in primary and secondary preventive treatments in cardiovascular diseases. However, it has several side effects including gastrointestinal toxicity (peptic ulcer formation, bleeding). Although lower doses are relatively safe, we should keep in mind that even the lowest dose may cause gastrointestinal bleeding. Gastrointestinal toxicity profile does not differ between conventional and enteric-coated aspirin use. In patients who have cardiovascular disease but are at high risk for gastrointestinal bleeding, eradication of Helicobacter pylori infection and concurrent proton pump inhibitor therapy may help to reduce the risk of gastrointestinal toxicity. PMID- 18160367 TI - The sexual double standard in African American adolescent women's sexual risk reduction socialization. AB - This study explored the sexual double standard (SDS) (in which males are afforded more freedom and power than females in heterosexual interactions) in African American mothers' sexual messages to sons and daughters. We used a convenience sample of 129 African American adolescents, aged 14 to 17 years, and their mothers who reported SDS attitudes. Qualitative analyses revealed gender differences based on an SDS in mothers' sexual risk reduction socialization. Mothers typically took a proactive approach with sons and a neutral or prohibitive approach with daughters. Findings provide directions for socially relevant programs for African American parents, schools, and communities. PMID- 18160368 TI - AIDS cases among women who reported sex with a bisexual man, 2000-2004--United States. AB - INTRODUCTION: Some HIV-infected men who have sex with men also have sex with women. Additionally, some women do not know that they are or have been in a sexual relationship with a bisexual man. Knowledge of their male partner's risks for HIV infection is crucial if reductions in HIV/AIDS are to occur among women. METHODS: We examined AIDS diagnosed cases reported to CDC from the 50 states and the District of Columbia, 2000 through 2004, in women aged 13 years and older. Cases were analyzed by transmission category, race/ethnicity, age at diagnosis, and geographic region, and data were adjusted for missing risk factor information and reporting delays. RESULTS: From 2000 through 2004, an estimated 1,576 women (from a total of 35,376 women reported with HIV from heterosexual contact and diagnosed with AIDS) reported sexual contact with a bisexual man (BSXM) as their primary risk factor for HIV infection. Non-Hispanic blacks accounted for the majority (62.8%) of cases, followed by non- Hispanic whites (20.5%) and Hispanics (14.8%). The average AIDS rate attributed to sex with a BSXM differed significantly by race/ethnicity (p < 0.01), with the rate for non-Hispanic black women 13 times the rate for non-Hispanic whites and 4 times the rate for Hispanics. Sexual contact with a BSXM accounted for 6.3% of AIDS cases among non- Hispanic white women with heterosexually acquired HIV compared to 4.4% among Hispanics and 4.0% among non-Hispanic blacks. CONCLUSIONS: The proportion of AIDS cases among women attributed to sex with a BSXM was similar across races/ethnicities; however, rates were highest among non-Hispanic black women. Because some women were unaware of their male partner's risk for HIV infection, the number of women with AIDS who had a bisexual partner was probably underreported. HIV prevention programs should provide information on risks of sex with BSXM, as many women may not be fully aware of their risks for acquiring HIV infection. PMID- 18160369 TI - Correlates of consistent condom use among HIV-positive African American women. AB - INTRODUCTION: African American women are the fastest growing group at risk of contracting HIV, as over 68% of women diagnosed with HIV are African American. The present study used social-cognitive theory and the theory of gender and power to identify correlates of consistent condom use among African American women living with HIV. METHODS: We recruited 366 women, 18-50 years of age and living with HIV from several HIV/AIDS clinics in the southeastern United States. The majority of women, 84.2% (N = 308), were African American. Women completed a baseline interview assessing sociodemographic, psychosocial, and partner-related factors potentially associated with consistent condom use, defined as reported use of condoms during every vaginal sex episode with a male sexual partner in the past 30 days. RESULTS: Among those HIV-positive African American women reporting use of a condom in the past 30 days (n = 234, 70%), consistent condom use was reported by 67.1% of women. This rate was lower (51.6%) among women having an HIV positive male sexual partner; the rate was higher (74.1%) among women having an HIV-negative male sexual partner. Compared to women who were inconsistent condom users, women with HIV were more likely to use condoms if they: had high partner communication self-efficacy (OR = 7.77, 95% CI = 3.3-18.6, p = .001), had a HIV negative male sex partner (OR = 4.62, 95% CI = 1.9-11.5, p = 0.001), had low partner-related barriers to condom use (OR = 4.68, 95% CI = 1.8-12.2, p = 0.001), and had perceived peer norms supportive of condom use (OR = 2.38, 95% CI = 1.0 5.7, p = 0.03). CONCLUSION: These findings suggest that HIV interventions may enhance consistent condom use among African American women living with HIV by targeting women's self-efficacy to communicate with their partners and women's perception of personal and partner-related barriers to condom use. PMID- 18160370 TI - Demographic correlates of HIV testing, high-risk behaviors, and condom/STD consultation among a multi-ethnic sample of women. AB - BACKGROUND: In the United States, HIV rates are disproportionately high among black women, with 78% of cases attributed to heterosexual transmission. This analysis examined HIV testing, high risk behaviors and condom use consultation for the prevention of sexually transmitted diseases (STD) among a diverse sample of women. METHODS: Secondary analyses were conducted using data from the 2004 Behavioral Risk Factor Surveillance System (BRFSS). This dataset included a sample of 43,550 (12% black, 79% white, and 9% Hispanic) women aged 18 to 49 years, living in the United States. Both bivariate and multivariate statistical analyses were conducted. RESULTS: Multivariate logistic regression analyses revealed that ethnic group differences existed for past-year HIV test, high-risk behaviors, and condom use consultation even after we controlled for age, education,income, employment status, and region of residence. Black women were three times more likely than white women to report a past-year HIV test and Hispanic women were 50% more likely than white women and report a past-year HIV test. Women who reported receiving condom use consultations were most likely to be Hispanic, 30 to 34 years old, and residents of the northeast. CONCLUSIONS: Initiatives to promote HIV testing appeared to have affected HIV testing behaviors among black women. However, increased HIV testing among black women was not associated with an increased likelihood of condom use counseling by a healthcare professional. More emphasis on HIV counseling is warranted to assist with HIV risk reduction and to complement HIV testing initiatives. PMID- 18160371 TI - HIV risk reduction in rural African American women who use cocaine. AB - Research has demonstrated associations between cocaine use and sexually transmitted infections (STIs), including Human Immunodeficiency Virus (HIV). More research has been conducted among the sub-population of women, with less focus on rural African American women. Investigation of the social context that influences rural African American women's behavior that places them at risk for STI/HIV has received limited attention. The results of one component of the comprehensive study presented here focused on perceived STI/HIV risk reduction strategies of U.S. southern rural African American women who used cocaine. This ethnographic study of 30 rural African American women using cocaine was conducted over four years. Respondents described HIV risk reduction strategies and their motives for using them. Condoms were used more with casual partners (80%) than primary partners (16%) and with partners who were less known, less trusted, young, or physically or sexually "dirty." Condoms were also used to prevent pregnancy (60%) more than to prevent STI/HIV (40%). HIV testing patterns varied, as did the motives for seeking testing. The sexual behavior of the women in this study exposed them to increased risk of STI/HIV. Although many perceived that they were using sexual risk reduction strategies, these strategies were not scientifically sound risk reduction strategies. Consequently, their exposure to STI/HIV continued as they used ineffective strategies both in assessing partner risk and in implementing risk reduction behaviors. PMID- 18160372 TI - HIV discrimination and the health of women living with HIV. AB - Women living with HIV are especially vulnerable to discrimination because of the stigma associated with the disease, as well as their race, gender and class status. To investigate the association between self-reported HIV discrimination and health outcomes among African- American and white women living with HIV, 366 women living with HIV were recruited from HIV/AIDS clinics in Georgia and Alabama. In this cross-sectional study, participants completed an interview that assessed self-reported HIV discrimination and depressive symptomatology, suicidal ideation, self-esteem, stress, quality of life, sexual health and HIV/AIDS related health care seeking. Nearly a sixth of the sample reported experiencing HIV discrimination. Women reporting HIV discrimination had higher mean scores for stress, suicidal ideation, depressive symptoms, number of unprotected sexual episodes; they had lower mean scores for self-esteem, and quality of life, and were more likely to have not sought medical care for HIV/AIDS. In race-specific analyses, none of the relationships between HIV discrimination and health outcomes were significant for white women. African-American women who reported HIV discrimination had higher mean scores for stress, suicidal ideation, depressive symptoms, number of unprotected sexual episodes; they had lower mean scores for self-esteem, and quality of life, and were more likely not to have sought medical care for HIV/AIDS. The findings indicated that HIV discrimination adversely affects women's mental, sexual and physical health. However, separate race-specific analyses indicated that compared to white women, African-American women were markedly more likely to experience the adverse affects of HIV discrimination. Eradication of HIV discrimination remains an important public health priority. PMID- 18160373 TI - The role of spirituality in sustaining the psychological well-being of HIV positive black women. AB - Historically, spirituality has been an instrumental component to the survival of Black women. In an era when the HIV epidemic disproportionately compromises their health, it is imperative to explore spirituality's role in sustaining the psychological health of Black women living with HIV. This study examined the relationship between spirituality and self-reported depression among Black women living with HIV. A sample of 308 HIV-positive Black women were recruited from HIV/ AIDS clinics in the Southeastern United States. Participants completed an interview assessing demographics, quality of life, depression, coping, and spirituality. A hierarchical multiple regression was used to determine the association between spirituality and depression. The results suggest that in our sample, spirituality accounted for a small, yet significant proportion of variance in reducing depressive symptoms, above and beyond variance accounted for by demographic variables and other theoretically important psychosocial factors. In light of these findings, future studies with HIV-positive Black women should assess spirituality as a salient factor affecting psychological health. Developing interventions that address spirituality may serve to enhance women's psychological adjustment to living with HIV. PMID- 18160374 TI - The Exchange coalition--identifying and addressing structural factors affecting HIV prevention in African American women on a national basis. AB - INTRODUCTION: From 1992 to 2002, African American women accounted for almost 70% of all newly diagnosed cases of HIV among women (CDC, 2002). Systemic issues such as poverty, domestic violence, mental illness, and limited access to culturally competent HIV/ health care services facilitate high infection rates among these women (OMH, 2005) and render many prevention/treatment messages ineffective. In response, Community Education Group (CEG) formed "The Exchange"-a national advocacy group comprised representatives from various organizations and agencies. To inform the groups formation CEG conducted a formative survey at three conferences to assess interest and need of such a group. METHODS: Using a convenience sample, CEG surveyed care providers, health specialist, and social/behavioral scientists from various organizations at three conferences about: (1) primary issues facing African American women at risk of and living with HIV/AIDS, (2) their interest in building a coalition with organizations with differing foci, and (3) what they believe to be the top health issues facing these women. RESULTS: Of the 1,186 surveys, 96.7% believed a coalition would provide effective advocacy, 77.3% were willing to build a coalition, and the top three issues affecting rates of infection were health care (62.8%), access to affordable treatment (50%) and unknown partner risk (47%). CONCLUSION: Survey respondents felt that a coalition of organizations with differing foci to address the systemic conditions of African American women is needed if we are to have an effect on their rates of HIV/AIDS. PMID- 18160375 TI - The use of epidemiologic and other data in selecting behavioral HIV prevention interventions for African American women. AB - We describe a "research to practice" method by which public health policymakers and HIV prevention service providers can integrate the findings of national surveillance with other sources of public health data. We suggest developing a comprehensive risk profile, based on multiple sources of data, to inform the selection and implementation of evidence- based behavioral interventions (EBIs) for African-American women. PMID- 18160376 TI - The SISTA pilot project: understanding the training and technical assistance needs of community-based organizations implementing HIV prevention interventions for African American women--implications for a capacity building strategy. AB - The disproportionate rates of HIV/AIDS among African American women in the U.S. signify the ongoing need for targeted HIV prevention interventions. Additionally, building the capacity of service providers to sustain prevention efforts is a major concern. The Centers for Disease Control and Prevention (CDC) conducted a pilot project to disseminate the Sisters Informing Sisters about Topics on AIDS (SISTA), an HIV prevention intervention designed for African American women. The project was to inform the diffusion process and examine the training and technical assistance needs of participating community-based organizations. Results demonstrated a need for extensive pre-planning and skills-building prior to implementation. PMID- 18160378 TI - Experiences and perceptions of intimate partner violence among older Chinese immigrants. AB - This study examines the experiences and perceptions of domestic violence among older Chinese immigrants, using data from a larger sample of Chinese Americans who reside in a major metropolitan area in the West Coast (Yick, 2000). A subset of respondents 50 years and older who are foreign-born (n = 77) were selected to (1) examine older Chinese immigrants' experiences with physical abuse by a spouse, (2) explore the definitions and perceptions of intimate partner violence among older Chinese, and (3) examine the relationship between acculturation, victimization experiences, and perceptions of domestic violence among this population. Findings indicate that close to 7% of the women and 6% of the men have experienced minor physical violence by their spouses during the past 12 months. Gender and acculturation were associated with perceptions and attitudes towards domestic violence. PMID- 18160379 TI - Perceptions of elder physical abuse in the courtroom: the influence of hearsay witness testimony. AB - A community sample of 226 participants (41% men, 59% women) who answered questions about a trial summary that manipulated what type of witness (a 45-year old hearsay [second-hand information based on the report of a crime victim] witness, a 75-year-old hearsay witness, or the 75-year-old victim) presented an allegation of elder physical abuse. Overall, participants who read the testimony of a 45-year-old hearsay witness had higher conviction rates than participants who read the testimony of the 75-year-old hearsay witness or the elder victim. Additionally, participants who had previously been victims of abuse and/or who had positive attitudes toward elderly people rendered more pro-prosecution verdicts. Finally, as the age of the participant increased so did conviction rates. PMID- 18160380 TI - Evaluation of adult protective services: perspectives of community professionals. AB - Due to challenges associated with client characteristics, funding, and the expectation of poor outcomes, few studies have evaluated the services provided by Adult Protective Services (APS). The article reports community-based professionals' satisfaction with APS when investigating cases of suspected elder abuse. Results from a 12-item quantitative survey indicate respondents were most satisfied with the intake process, followed by assessment, and case management. Satisfaction was higher than expected, though areas for improvement are identified. This survey and its results should serve as a starting point for more extensive evaluation methodologies needed to enhance the delivery of services provided by APS. PMID- 18160381 TI - Searching for elder abuse: a systematic review of database citations. AB - The aim of this review was to describe trends and patterns in elder abuse literature and research. All citations in PUB MED, CINAHL, and PsycINFO databases located using the search term "elder abuse" were retrieved (3,059 citations) and 2,418 unique references were identified. Using manifest content analysis, non research material (1,986 references) was sorted by type and research references (34 dissertations and 398 research articles) were categorized. Research article references most commonly: (1) investigated prevalence, typology, and definitions of elder abuse; (2) utilized quantitative methodology; (3) had first author affiliations to medicine, nursing, and social sciences; and (4) sampled populations of professionals, personnel, cases, or charts. The USA was most frequently listed of 24 identified countries of origin. Elder abuse research shows lack of diversity, slow increase of qualitative research, limited involvement of older persons and family members as participants, and minimal research from developing countries. This review contributes to knowledge valuable for researchers and experts planning future studies or elder abuse projects. PMID- 18160382 TI - Elder abuse and neglect in an urban chinese population. AB - This study aims to investigate the prevalence of elder abuse and neglect in an urban Chinese population. A cross-sectional study was performed in a major urban medical center in NanJing, China. A total of 412 participants completed the survey and 145 (35%) participants screened positive for elder abuse and neglect. The mean age of the victims was 69 years and 59% were male. Caregiver neglect was the most common form of abuse, followed by financial exploitation, psychological abuse, physical abuse, sexual abuse, and abandonment. Thirty-six percent of the victims suffered multiple forms of abuse and neglect. In the logistical regression analyses of the data, female gender, lower education and lower income were demographic risk factors associated with elder abuse and neglect. A better understanding of these and additional risk factors associated with elder abuse and neglect in older Chinese people is needed. PMID- 18160383 TI - Elderly prisoners: a growing and forgotten group within correctional systems vulnerable to elder abuse. AB - There are over 2.1 million people incarcerated in the nation's jails and prisons. Additionally, close to 600,000 prisoners are released annually into communities across the country. Many prisoners and those released from prisons are elderly. The purpose of this article is to examine the systemic abuse and neglect experienced by elderly prisoners while they are incarcerated and when they are released from prison. Most correctional systems have inadequate resources, processes, and personnel to manage the elderly population inside and outside of prisons. In addition to providing a definition of "elderly prisoner," two specific problems-prison health care and prisoner re-entry-are examined in the article. The article concludes with recommendations for both policy and research on how best we can further understand and address the multiple needs and concerns faced by elderly prisoners. PMID- 18160384 TI - Nursing home abuse report and investigation legislation. AB - Nursing home complaint investigation procedures are established by each state's respective legislation. The purpose of this study was to determine the state statutes that had text addressing nursing home report and investigation of abuse and to describe the content of that text. All states and the District of Columbia have statutes addressing the licensure and re-certification of nursing homes. Fourteen of these licensure and re-certification of nursing home state statutes have text specifically addressing the topic of abuse reports and investigation in the nursing homes. Main concepts discussed in those sections were reporting requirements, mandatory reporting, and definitions. PMID- 18160385 TI - The vulnerablility for elder abuse among a sample of custodial grandfathers: an exploratory study. AB - Older adults have been known to make sacrifices in their caregiving roles. Gerontology literature on custodial grandparents has primarily focused on grandmothers and the challenges they face when they assume primary care for grandchildren. Little is known about the risks that older men face when they become custodial grandparents. This article highlights types and warning signs of abuse, exploitation and neglect. Exploratory study was undertaken with a racially diverse group of custodial grandfathers to fill a gap in the literature about the vulnerability for elder abuse, exploitation and neglect as expressed by older Black, Latino and White custodial grandfathers. To provide a more inclusive understanding of elder abuse, areas of vulnerability were identified for consideration by practitioners, educators and researchers. The implications of this research point to the need to rethink elder abuse assessment, prevention and intervention strategies with older men. PMID- 18160386 TI - Analysis and treatment of self-neglectful behaviors in three elderly female patients. AB - Three cases of self-neglect in elderly female patients are presented. Ranging in age from 84 to 92, subjects suffered from chronic, non-terminal illnesses, and engaged in such behaviors as refusal to eat, non-adherence to a prescribed therapeutic regimen, and extreme social withdrawal.One lived in the community and two resided in assisted living facilities in Louisville, Kentucky. Initial interviews strongly suggested that the patients' engagement in self-neglect was an inappropriate yet purposeful response to aversive events or other negative stimuli. Goals of treatment were to (1) identify specific factors which precipitated patient use of potentially life-threatening behavior, and (2) employ behavioral methods of intervention in response to the identified precipitators. Outcomes were moderately positive. Behavioral interventions reduced engagement in self-neglectful behaviors for each of the participants to a clinically significant degree. Positive results were maintained during the three-month period following treatment termination. PMID- 18160387 TI - Neonatal and postneonatal mortality by maternal education--a population-based study of trends in the Nordic countries, 1981-2000. AB - BACKGROUND: This study examined changes in the educational gradients in neonatal and postneonatal mortality over a 20-year period in the four largest Nordic countries. METHODS: The study populations were all live-born singleton infants with gestational age of at least 22 weeks from 1981 to 2000 (Finland 1987-2000). Information on births and infant deaths from the Medical Birth Registries was linked to information from census statistics. Numbers of eligible live-births were: Denmark 1 179 831, Finland 834 299 (1987-2000), Norway 1 017 168 and Sweden 1 971 645. Differences in mortality between education groups were estimated as risk differences (RD), relative risks (RR) and index of inequality ratio (RII). RESULTS: Overall, rates of infant mortality were in Denmark 5.9 per 1000 live births, in Finland 4.2 (1987-2000), in Norway 5.3 and in Sweden 4.7. Overall the mortality decreased in all educational groups, and the educational level increased in the study period. The time-trends differed between neonatal and postneonatal death. For neonatal death, both the absolute and relative educational differences decreased in Finland and Sweden, increased in Denmark, whereas in Norway a decrease in absolute differences and a slight increase in relative differences occurred. For postneonatal death, the relative educational differences increased in all countries, whereas the absolute differences decreased. CONCLUSIONS: All educational groups experienced a decline in infant mortality during the period under study. Still, the inverse association between maternal education and RR of postneonatal death has become more pronounced in all Nordic countries. PMID- 18160388 TI - An investigation of the growing number of deaths of unidentified people in Russia. AB - BACKGROUND: We examined mortality among working-age Russian men whose identity could not be determined, focusing on where and how they died. METHODS: Employing micro-data from deaths that occurred in Izhevsk (Ural region) between June 2004 and September 2005, we analysed the characteristics of decedent men aged 25-54 (n = 2158). Differences between completely identified (n = 1699) and unidentified deaths (n = 282) were compared via logistic regression. Data on all deaths in Russia in 2002 were used for supplemental comparisons. RESULTS: We found that relative to identified men, unidentified men were at a higher risk of death from exposure to natural cold, violence, alcoholic cardiomyopathy, acute respiratory infections and poisonings. Our results also revealed that alcohol played an important role in the mortality of unidentified men. The places and causes of death among these unidentified men provide substantial evidence of their homelessness and social isolation. CONCLUSION: The increase in deaths among unidentified men of working-age indicates the emergence of a health threat associated with homelessness and social marginalization. This vulnerable group is exposed to different levels and causes of mortality compared with the larger population and represent a new challenge that requires serious and immediate scholarly attention and policy responses. PMID- 18160389 TI - Cost effectiveness of hepatitis B vaccination strategies in Ireland: an economic evaluation. AB - BACKGROUND: In accordance with World Health Organization recommendations, many European countries have introduced universal hepatitis B vaccination policies. The UK and Ireland are exceptions. In this study, we conducted an economic evaluation of a universal infant hepatitis B vaccination programme, using a six component vaccine, compared with the current selective strategy of vaccinating high-risk infants with a monovalent hepatitis B vaccine. METHODS: A cost effectiveness analysis was conducted using a Markov model. The perspective of the analysis was the Irish Health Service Executive. Unit cost and resource utilization data were derived from expert clinical opinion, published sources, diagnosis-related group costs for hospital admissions and local cost estimates for medical fees and laboratory investigations. A full probabilistic sensitivity analysis was undertaken. Both costs and outcomes were modelled over a period of 80 years and discounted at 3.5%. RESULTS: Assuming an incidence of acute hepatitis B virus (HBV) infection in Ireland of 8.4 per 100,000 population, the incremental cost effectiveness ratio ranged from euro10,992/life years gained (LYG) to euro67 200/LYG, at the lowest and highest price estimates for the six component vaccine, respectively. The cost effectiveness of universal versus selective hepatitis B vaccination was sensitive to the risk of acute HBV infection, the cost of the universal infant vaccination programme and the discount rate. CONCLUSION: At a cost of euro29.00 per dose of the six-component vaccine, universal infant hepatitis B vaccination is cost effective at euro37 018/LYG. This compares favourably with other preventive programmes in Ireland. PMID- 18160390 TI - Health visitors and breastfeeding support: influence of knowledge and self efficacy. AB - BACKGROUND: Little is known about what influences health visitors' breastfeeding support. The objective was to describe health visitors' breastfeeding experiences, beliefs, knowledge and self-efficacy in breastfeeding guidance and determine the impact of a training course on these factors, and how they were reflected in practice. METHODS: A randomized intervention study enrolled 52 health visitors in the intervention group and 57 in the comparison group. The intervention group participated in an 18-hour pre-study training course that focused on knowledge about lactation and how to guide the mother to learn the mechanisms of breastfeeding. Data were collected through self-administered questionnaires before the intervention and after the follow-up period. One hundred and six (97%) health visitors and 1302 (82%) mothers responded. RESULTS: At baseline no substantial differences were seen between the two groups on years since education, own breastfeeding experiences, beliefs or self-efficacy in breastfeeding guidance except that health visitors in the intervention group, who had completed the course, demonstrated significantly higher scores on knowledge questions (P < 0.01). After the intervention health visitors in the intervention group reported significantly higher self-efficacy in guidance on three of five breastfeeding problems (P < 0.01). Mothers in the intervention group reported having received more support than mothers in the comparison group. CONCLUSION: An interactive course increased the health visitors' knowledge of breastfeeding practice. After the intervention period the health visitors in the intervention group had increased their self-efficacy in helping mothers with common breastfeeding problems. The mothers in the intervention group reported more informational and instrumental breastfeeding support. PMID- 18160391 TI - Material deprivation and health-related dysfunction in older Dutch people: findings from the SMILE study. AB - BACKGROUND: Even in generally wealthy Western countries material deprivation and poverty are not uncommon. There is, however, little data on the prevalence of material deprivation and its associations with health-related dysfunction in older people. METHODS: Cross-sectional data from the SMILE study were used to examine the prevalence of material deprivation and the associations between material deprivation and health-related dysfunction in persons aged 55 years and older (n > 4000). Material deprivation was measured with a comprehensive questionnaire assessing seven subdomains referring to current and anticipated financial problems and poverty in childhood. Health-related dysfunction was measured using the SF36-based physical and mental components. In addition, self reported heart disease was examined as an indicator of health-related dysfunction as well. RESULTS: Almost 29% of subjects experienced at least one financial problem. Those reporting material deprivation had more than twice the risk of physical (OR = 2.22; 95% CI: 1.72-2.86) and mental (OR = 2.34; 95% CI: 1.84-2.97) dysfunction compared with non-deprived persons. A slightly weaker association was found when self-reported heart disease was used as an outcome variable (OR = 1.74; 95% CI: 1.40-2.15). Although odds ratios were generally higher for diseased older persons, no significant interaction effect between chronic disease and material deprivation subscales was found. CONCLUSIONS: Material deprivation in the Netherlands is not uncommon and is strongly related to both mental and physical dysfunction, and therefore needs further attention in public health policy. Longitudinal research is necessary to clarify the causal nature of our results and to develop appropriate interventions. PMID- 18160392 TI - Prescribing patterns for upper respiratory tract infections in general practice in France and in the Netherlands. AB - BACKGROUND: France and the Netherlands are often presented as two contrasting countries with regard to drug prescriptions and consumption. This study aimed to analyse general practitioners' (GP's) prescription patterns for upper respiratory tract infections (URTI). METHODS: Data on diagnoses and prescriptions were derived from two databases recording daily electronic medical patient files: the 'Societe Francaise de Medecine Generale' database (SFMG-DB) and the Dutch Landelijk Informatie Netwerk Huisatsenzorg database (LINH-DB). Logit regression models were developed to estimate and compare prescription patterns in both countries. We carried out a study including all the patients consulting for URTI in 2003. RESULTS: French GPs had more URTI patients than their Dutch counterparts (372.1 URTI patients/GP versus 181.3). They prescribed higher volumes of URTI medications (3.55 per patient/year versus 0.82). Striking differences were observed in analgesic and symptomatic prescriptions (0.84 per patient/year versus 0.12 and 1.01 per patient/year versus 0.21, respectively). We did not observe important discrepancies in volume of antibiotic prescriptions (0.29 per patient/year in France versus 0.32). After adjustment for patient characteristics, the logit model showed that prescription patterns for antibiotic were quite similar and associated with a diagnosis of acute tonsillitis. CONCLUSION: The analysis per consultation in this study did not highlight important differences in antibiotic prescribing volumes and patterns. But symptomatic and analgesic prescriptions were significantly higher in the French database. This can be explained by differences in help-seeking behaviour, medication perception, status of OTC medications and remuneration system. PMID- 18160393 TI - Reporting of diabetes on death certificates of 1872 people with type 2 diabetes in Tayside, Scotland. AB - BACKGROUND: It has been suggested that diabetes is under-recorded on death certificates. METHODS: We examined the death certificates of 1,872 people with type 2 diabetes in Tayside, Scotland, to determine how frequently diabetes was recorded. RESULTS: Diabetes was mentioned on the certificates of 42.8% and was the underlying cause of death for 6.4%. There was mention of diabetes for 51.3% of the 811 people for whom cardiovascular disease was the underlying cause of death. Being male was associated with less frequent mention of diabetes, with more frequent mention associated with increasing duration of diabetes, increasing age and underlying cardiovascular cause of death. CONCLUSIONS: This study highlights the limitations of using routine mortality data for monitoring the burden of diabetes in populations. PMID- 18160394 TI - Structure of the bacteriophage phi KZ lytic transglycosylase gp144. AB - Lytic transglycosylases are enzymes that act on the peptidoglycan of bacterial cell walls. They cleave the glycosidic linkage between N-acetylmuramoyl and N acetylglucosaminyl residues with the concomitant formation of a 1,6 anhydromuramoyl product. The x-ray structure of the lytic transglycosylase gp144 from the Pseudomonas bacteriophage phi KZ has been determined to 2.5-A resolution. This protein is probably employed by the bacteriophage in the late stage of the virus reproduction cycle to destroy the bacterial cell wall to release the phage progeny. phi KZ gp144 is a 260-residue alpha-helical protein composed of a 70-residue N-terminal cell wall-binding domain and a C-terminal catalytic domain. The fold of the N-terminal domain is similar to the peptidoglycan-binding domain from Streptomyces albus G D-Ala-D-Ala carboxypeptidase and to the N-terminal prodomain of human metalloproteinases that act on extracellular matrices. The C-terminal catalytic domain of gp144 has a structural similarity to the catalytic domain of the transglycosylase Slt70 from Escherichia coli and to lysozymes. The gp144 catalytic domain has an elongated groove that can bind at least five sugar residues at sites A-E. As in other lysozymes, the peptidoglycan cleavage (catalyzed by Glu 115 in gp144) occurs between sugar-binding subsites D and E. The x-ray structure of the phi KZ transglycosylase complexed with the chitotetraose (N-acetylglucosamine)(4) has been determined to 2.6-A resolution. The N-acetylglucosamine residues of the chitotetraose bind in sites A-D. PMID- 18160396 TI - Mitochondrial targeting of adenomatous polyposis coli protein is stimulated by truncating cancer mutations: regulation of Bcl-2 and implications for cell survival. AB - The adenomatous polyposis coli (APC) protein tumor suppressor is mutated in the majority of colon cancers. Most APC gene mutations cause deletion of the C terminus and disrupt APC regulation of beta-catenin turnover, microtubule dynamics, and chromosome segregation. Truncated APC mutant peptides may also gain unique properties, not exhibited by wild-type APC, which contribute to tumor cell survival and proliferation. Here we report a differential subcellular localization pattern for wild-type and mutant APC. A pool of APC truncation mutants was detected at mitochondria by cellular fractionation and confocal microscopy. In contrast, wild-type APC located poorly at mitochondria. Similar results were observed for endogenous and stably induced forms of APC, with the shortest N-terminal mutant peptides (N750, N853, N1309, N1337) displaying the strongest mitochondrial staining. The knock down of mutant APC(N1337) in SW480 tumor cells caused an increase in apoptosis and mitochondrial membrane permeability, and this correlated with reduced Bcl-2 protein levels in mitochondrial fractions. Interestingly, the silencing of APC did not alter expression of beta-catenin or the apoptotic regulatory factors Bax, Bcl-xL, or survivin. APC formed a complex with Bcl-2 in mitochondrial fractions, and this may contribute to the APC-dependent regulation of Bcl-2. We propose that a subset of cancer mutations induce APC mitochondrial localization and that APC regulation of Bcl-2 at mitochondria may contribute to tumor cell survival. PMID- 18160395 TI - Apolipoprotein AII is a regulator of very low density lipoprotein metabolism and insulin resistance. AB - Apolipoprotein AII (apoAII) transgenic (apoAIItg) mice exhibit several traits associated with the insulin resistance (IR) syndrome, including IR, obesity, and a marked hypertriglyceridemia. Because treatment of the apoAIItg mice with rosiglitazone ameliorated the IR and hypertriglyceridemia, we hypothesized that the hypertriglyceridemia was due largely to overproduction of very low density lipoprotein (VLDL) by the liver, a normal response to chronically elevated insulin and glucose. We now report in vivo and in vitro studies that indicate that hepatic fatty acid oxidation was reduced and lipogenesis increased, resulting in a 25% increase in triglyceride secretion in the apoAIItg mice. In addition, we observed that hydrolysis of triglycerides from both chylomicrons and VLDL was significantly reduced in the apoAIItg mice, further contributing to the hypertriglyceridemia. This is a direct, acute effect, because when mouse apoAII was injected into mice, plasma triglyceride concentrations were significantly increased within 4 h. VLDL from both control and apoAIItg mice contained significant amounts of apoAII, with approximately 4 times more apoAII on apoAIItg VLDL. ApoAII was shown to transfer spontaneously from high density lipoprotein (HDL) to VLDL in vitro, resulting in VLDL that was a poorer substrate for hydrolysis by lipoprotein lipase. These results indicate that one function of apoAII is to regulate the metabolism of triglyceride-rich lipoproteins, with HDL serving as a plasma reservoir of apoAII that is transferred to the triglyceride rich lipoproteins in much the same way as VLDL and chylomicrons acquire most of their apoCs from HDL. PMID- 18160397 TI - CD147 inhibits the nuclear factor of activated T-cells by impairing Vav1 and Rac1 downstream signaling. AB - CD147 is a transmembrane protein that plays crucial roles in the development and function of the reproductive, visual, and nervous systems. CD147 also exerts positive and negative actions in T-cells by still obscure mechanisms. In this study, we have analyzed the expression, localization, and function of CD147 during T-cell receptor signaling responses. We show here that CD147 is an integral component of the T-cell immune synapse and that its overexpression leads to the inhibition of NF-AT (nuclear factor of activated T-cells) activity induced by Vav1, a Rac1 exchange factor. This inhibitory activity is mediated by the CD147 intracellular tail and is totally independent of its extracellular or transmembrane regions. The molecular dissection of the influence of CD147 on the Vav1 pathway indicates that its inhibitory action takes place downstream of Vav1 and Rac1 but upstream of the serine/threonine kinases JNK and Pak1. The interference of CD147 with these pathways is highly specific because the overexpression of CD147 does not affect the activity of other GDP/GTP exchange factors or the stimulation of the ERK cascade. Finally, we show that the CD147 knockdown in Jurkat cells promotes higher levels of NF-AT stimulation and Pak1 phosphorylation upon T-cell receptor cross-linking. Instead, the lack of CD147 does not affect other signaling cascades that participate in the same cellular response. Taken together, these results indicate that CD147, via the selective inhibition of specific downstream elements of the Vav1/Rac1 route, contributes to the negative regulation of T-cell responses. PMID- 18160398 TI - Stimulus-dependent regulation of the phagocyte NADPH oxidase by a VAV1, Rac1, and PAK1 signaling axis. AB - The p21-activated kinase-1 (PAK1) is best known for its role in the regulation of cytoskeletal and transcriptional signaling pathways. We show here in the microglia cell line Ra2 that PAK1 regulates NADPH oxidase (NOX-2) activity in a stimulus-specific manner. Thus, conditional expression of PAK1 dominant-positive mutants enhanced, whereas dominant-negative mutants inhibited, NADPH oxidase mediated superoxide generation following formyl-methionyl-leucylphenylalanine or phorbol 12-myristate 13-acetate stimulation. Both Rac1 and the GTP exchange factor VAV1 were required as upstream signaling proteins in the formyl-methionyl leucyl-phenylalanine-induced activation of endogenous PAK1. In contrast, PAK1 mutants had no effect on superoxide generation downstream of FcgammaR signaling during phagocytosis of IgG-immune complexes. We further present evidence that the effect of PAK1 on the respiratory burst is mediated through phosphorylation of p47(Phox), and we show that expression of a p47(Phox) (S303D/S304D/S320D) mutant, which mimics phosphorylation by PAK1, induced basal superoxide generation in vivo. In contrast PAK1 substrates LIMK-1 or RhoGDI are not likely to contribute to the PAK1 effect on NADPH oxidase activation. Collectively, our findings define a VAV1-Rac1-PAK1 signaling axis in mononuclear phagocytes regulating superoxide production in a stimulus-dependent manner. PMID- 18160399 TI - The SBF- and MBF-associated protein Msa1 is required for proper timing of G1 specific transcription in Saccharomyces cerevisiae. AB - In the budding yeast Saccharomyces cerevisiae, cell cycle initiation is prompted during G(1) phase by Cln3/cyclin-dependent protein kinase-mediated transcriptional activation of G(1)-specific genes. A recent screening performed to reveal novel interactors of SCB-binding factor (SBF) and MCB-binding factor (MBF) identified, in addition to the SBF-specific repressor Whi5 and the MBF specific corepressor Nrm1, a pair of homologous proteins, Msa1 and Msa2 (encoded by YOR066w and YKR077w), as interactors of SBF and MBF, respectively. MSA1 is expressed periodically during the cell cycle with peak mRNA levels occurring at the late M/early G(1) phase and peak protein levels occurring in early G(1). Msa1 associates with SBF- and MBF-regulated target promoters consistent with a role in G(1)-specific transcriptional regulation. Msa1 affects cell cycle initiation by advancing the timing of transcription of G(1)-specific genes. Msa1 binds to SBF- and MBF-regulated promoters and binding is maximal during the G(1) phase. Binding depends upon the cognate transcription factor. Msa1 overexpression advances the timing of SBF-dependent transcription and budding, whereas depletion delays both indicators of cell cycle initiation. Similar effects on MBF-regulated transcription are observed. Based upon these results, we conclude that Msa1 acts to advance the timing of G(1)-specific transcription and cell cycle initiation. PMID- 18160400 TI - Hes6 controls cell proliferation via interaction with cAMP-response element binding protein-binding protein in the promyelocytic leukemia nuclear body. AB - Hes6 is a basic helix-loop-helix transcription factor that functions in the differentiation of pluripotent progenitor cells and during tumorigenesis. However, the molecular mechanism for its function is largely unknown. Here we show that Hes6 is a component of the promyelocytic leukemia nuclear body (PML-NB) complex in the nuclei and that Hes6 inhibits cell proliferation through induction of p21 cyclin-dependent kinase inhibitor. We further show that Hes6 directly interacts with CREB-binding protein (CBP), one of the key components of PML-NB, via its basic domain. This association is critical for p21 induction through multiple mechanisms, including chromatin remodeling and p53 acetylation. Taken together, these results suggest that the Hes6-CBP complex in PML-NB may influence the proliferation of cells via p53-dependent and -independent pathways. PMID- 18160401 TI - Structure analysis of bone morphogenetic protein-2 type I receptor complexes reveals a mechanism of receptor inactivation in juvenile polyposis syndrome. AB - Bone morphogenetic proteins regulate many developmental processes during embryogenesis as well as tissue homeostasis in the adult. Signaling of bone morphogenetic proteins (BMPs) is accomplished by binding to two types of serine/threonine kinase transmembrane receptors termed type I and type II. Because a large number of ligands signal through a limited number of receptors, ligand-receptor interaction in the BMP superfamily is highly promiscuous, with a ligand binding to various receptors and a receptor binding many different BMP ligands. In this study we investigate the interaction of BMP-2 with its two high affinity type I receptors, BMP receptors IA (BMPR-IA) and BMPR-IB. Interestingly, 50% of the residues in the BMP-2 binding epitope of the BMPR-IA receptor are exchanged in BMPR-IB without a decrease in binding affinity or specificity for BMP-2. Our structural and functional analyses show that promiscuous binding of BMP-2 to both type I receptors is achieved by inherent backbone and side-chain flexibility as well as by variable hydration of the ligand-receptor interface enabling the BMP-2 surface to adapt to different receptor geometries. Despite the high degree of amino acid variability found in BMPR-IA and BMPR-IB binding equally to BMP-2, three single point missense mutations in the ectodomain of BMPR IA cannot be tolerated. In juvenile polyposis syndrome these mutations have been shown to inactivate BMPR-IA. On the basis of our biochemical and biophysical analyses, we can show that the mutations, which are located outside the ligand binding epitope, alter the local or global fold of the receptor, thereby inactivating BMPR-IA and causing a loss of the BMP-2 tumor suppressor function in colon epithelial cells. PMID- 18160402 TI - Group A streptococcal cysteine protease degrades C3 (C3b) and contributes to evasion of innate immunity. AB - A relative lack of neutrophils around Streptococcus pyogenes is observed in streptococcal toxic shock syndrome (STSS). Because the bacteria spread rapidly into various organs in STSS, we speculated that S. pyogenes is equipped with molecules to evade the host innate immune system. Complement C3b opsonizes the pathogen to facilitate phagocytosis, and a complex of C3b converts C5 into anaphylatoxin. Because we found that C3 (C3b) is degraded in sera from patients with STSS, we investigated the mechanism of C3 (C3b) degradation by S. pyogenes. We incubated human C3b or serum with recombinant SpeB (rSpeB), a wild-type S. pyogenes strain isolated from an STSS patient or its isogenic DeltaspeB mutant and examined the supernatant by Western blotting with anti-human C3b. Western blot and Biacore analyses revealed that rSpeB and wild-type S. pyogenes rapidly degrade C3b. Additionally, C3 (C3b) was not detected in sera collected from infected areas of STSS patients. Furthermore, the survival rate in human blood and in mice was lower for the DeltaspeB mutant than the wild-type strain. Histopathological observations demonstrated that neutrophils were recruited to and phagocytosed the DeltaspeB mutant, whereas with the wild-type strain, few neutrophils migrated to the site of infection, and the bacteria spread along the fascia. We observed the degradation of C3 (C3b) in sera from STSS patients and the degradation of C3 (C3b) by rSpeB. This suggests that SpeB contributes to the escape of S. pyogenes from phagocytosis at the site of initial infection, allowing it to invade host tissues during severe infections. PMID- 18160403 TI - Ferritin L and H subunits are differentially regulated on a post-transcriptional level. AB - Ferritin plays an important role in the storage and release of iron, an element utilized in cellular processes such as respiration, gene regulation, and DNA replication and repair. Ferritin in animals is composed of 24 ferritin L (FTL) and ferritin H (FTH) subunits in ratios that vary in different cell types. Because the subunits are not functionally interchangeable, both L and H units are critical for maintaining iron homeostasis and protecting against iron overload. FTL and FTH are regulated primarily at a post-transcriptional level in response to cellular iron concentrations. Individual regulation of FTL and FTH is of much interest, and although transcriptional differences between FTL and FTH have been shown, differences in their post-transcriptional regulation have not been evaluated. We report here that FTL and FTH are differentially regulated in 1% oxygen on a post-transcriptional level. We have designed a quantitative assay system sensitive enough to detect differences between FTL and FTH iron regulatory elements (IREs) that a standard electrophoretic mobility shift assay does not. The FTL IRE is the primary responder in the presence of an iron donor in hypoxic conditions, and this response is reflected in endogenous FTL protein levels. These results provide evidence that FTL and FTH subunits respond independently to cellular iron concentrations and underscore the importance of evaluating FTL and FTH IREs separately. PMID- 18160404 TI - Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica. AB - The helminth parasite Fasciola hepatica secretes cysteine proteases to facilitate tissue invasion, migration, and development within the mammalian host. The major proteases cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) were recombinantly produced and biochemically characterized. By using site-directed mutagenesis, we show that residues at position 67 and 205, which lie within the S2 pocket of the active site, are critical in determining the substrate and inhibitor specificity. FheCL1 exhibits a broader specificity and a higher substrate turnover rate compared with FheCL2. However, FheCL2 can efficiently cleave substrates with a Pro in the P2 position and degrade collagen within the triple helices at physiological pH, an activity that among cysteine proteases has only been reported for human cathepsin K. The 1.4-A three-dimensional structure of the FheCL1 was determined by x-ray crystallography, and the three-dimensional structure of FheCL2 was constructed via homology-based modeling. Analysis and comparison of these structures and our biochemical data with those of human cathepsins L and K provided an interpretation of the substrate-recognition mechanisms of these major parasite proteases. Furthermore, our studies suggest that a configuration involving residue 67 and the "gatekeeper" residues 157 and 158 situated at the entrance of the active site pocket create a topology that endows FheCL2 with its unusual collagenolytic activity. The emergence of a specialized collagenolytic function in Fasciola likely contributes to the success of this tissue-invasive parasite. PMID- 18160405 TI - X-ray structure of the complete ABC enzyme ABCE1 from Pyrococcus abyssi. AB - The ATP binding cassette enzyme ABCE1 (also known as RNase-L (ribonuclease L) inhibitor, Pixie, and HP68), one of the evolutionary most sequence-conserved enzymes, functions in translation initiation, ribosome biogenesis, and human immunodeficiency virus capsid assembly. However, its structural mechanism and biochemical role in these processes have not been revealed. We determined the crystal structure of Pyrococcus abyssi ABCE1 in complex with Mg(2+) and ADP to 2.8A resolution. ABCE1 consists of four structural domains. Two nucleotide binding domains are arranged in a head-to-tail orientation by a hinge domain, suggesting that these domains undergo the characteristic tweezers-like powerstroke of ABC enzymes. In contrast to all other known ABC enzymes, ABCE1 has a N-terminal iron-sulfur-cluster (FeS) domain. The FeS domain contains two [4Fe 4S] clusters and is structurally highly related to bacterial-type ferredoxins. However, one cluster is coordinated by an unusual CX(4)CX(3/4)C triad. Surprisingly, intimate interactions of the FeS domain with the adenine and ribose binding Y-loop on nucleotide binding domain 1 suggest a linkage between FeS domain function and ATP-induced conformational control of the ABC tandem cassette. The structure substantially expands the functional architecture of ABC enzymes and raises the possibility that ABCE1 is a chemomechanical engine linked to a redox process. PMID- 18160406 TI - Crystal structure of lactadherin C2 domain at 1.7A resolution with mutational and computational analyses of its membrane-binding motif. AB - Lactadherin is a phosphatidyl-L-serine (Ptd-L-Ser)-binding protein that decorates membranes of milk fat globules. The major Ptd-l-Ser binding function of lactadherin has been localized to its C2 domain, which shares homology with the C2 domains of blood coagulation factor VIII and factor V. Correlating with this homology, purified lactadherin competes efficiently with factors VIII and V for Ptd-L-Ser binding sites, functioning as a potent anticoagulant. We have determined the crystal structure of the lactadherin C2 domain (Lact-C2) at 1.7A resolution. The bovine Lact-C2 structure has a beta-barrel core that is homologous with the factor VIII C2 (fVIII-C2) and factor V C2 (fV-C2) domains. Two loops at the end of the beta-barrel, designated spikes 1 and 3, display four water-exposed hydrophobic amino acids, reminiscent of the membrane-interactive residues of fVIII-C2 and fV-C2. In contrast to the corresponding loops in fVIII C2 and fV-C2, spike 1 of Lact-C2 adopts a hairpin turn in which the 7-residue loop is stabilized by internal hydrogen bonds. Further, central glycine residues in two membrane-interactive loops may enhance conformability of Lact-C2 to membrane binding sites. Mutagenesis studies confirmed a membrane-interactive role for the hydrophobic and/or Gly residues of both spike 1 and spike 3. Substitution of spike 1 of fVIII-C2 into Lact-C2 also diminished binding. Computational ligand docking studies identified two prospective Ptd-l-Ser interaction sites. These results identify two membrane-interactive loops of Lact-C2 and provide a structural basis for the more efficient phospholipid binding of lactadherin as compared with factor VIII and factor V. PMID- 18160408 TI - FlyBase: integration and improvements to query tools. AB - FlyBase (http://flybase.org) is the primary resource for molecular and genetic information on the Drosophilidae. The database serves researchers of diverse backgrounds and interests, and offers several different query tools to provide efficient access to the data available and facilitate the discovery of significant relationships within the database. Recently, FlyBase has developed Interactions Browser and enhanced GBrowse, which are graphical query tools, and made improvements to the search tools QuickSearch and QueryBuilder. Furthermore, these search tools have been integrated with Batch Download and new analysis tools through a more flexible search results list, providing powerful ways of exploring the data in FlyBase. PMID- 18160407 TI - The role of DNA damage repair in aging of adult stem cells. AB - DNA repair maintains genomic stability and the loss of DNA repair capacity results in genetic instability that may lead to a decline of cellular function. Adult stem cells are extremely important in the long-term maintenance of tissues throughout life. They regenerate and renew tissues in response to damage and replace senescent terminally differentiated cells that no longer function. Oxidative stress, toxic byproducts, reduced mitochondrial function and external exposures all damage DNA through base modification or mis-incorporation and result in DNA damage. As in most cells, this damage may limit the survival of the stem cell population affecting tissue regeneration and even longevity. This review examines the hypothesis that an age-related loss of DNA damage repair pathways poses a significant threat to stem cell survival and longevity. Normal stem cells appear to have strict control of gene expression and DNA replication whereas stem cells with loss of DNA repair may have altered patterns of proliferation, quiescence and differentiation. Furthermore, stem cells with loss of DNA repair may be susceptible to malignant transformation either directly or through the emergence of cancer-prone stem cells. Human diseases and animal models of loss of DNA repair provide longitudinal analysis of DNA repair processes in stem cell populations and may provide links to the physiology of aging. PMID- 18160409 TI - The cell cycle DB: a systems biology approach to cell cycle analysis. AB - The cell cycle database is a biological resource that collects the most relevant information related to genes and proteins involved in human and yeast cell cycle processes. The database, which is accessible at the web site http://www.itb.cnr.it/cellcycle, has been developed in a systems biology context, since it also stores the cell cycle mathematical models published in the recent years, with the possibility to simulate them directly. The aim of our resource is to give an exhaustive view of the cell cycle process starting from its building blocks, genes and proteins, toward the pathway they create, represented by the models. PMID- 18160410 TI - PepCyber:P~PEP: a database of human protein protein interactions mediated by phosphoprotein-binding domains. AB - Phosphoprotein-binding domains (PPBDs) mediate many important cellular and molecular processes. Ten PPBDs have been known to exist in the human proteome, namely, 14-3-3, BRCT, C2, FHA, MH2, PBD, PTB, SH2, WD-40 and WW. PepCyber:P approximately PEP is a newly constructed database specialized in documenting human PPBD-containing proteins and PPBD-mediated interactions. Our motivation is to provide the research community with a rich information source emphasizing the reported, experimentally validated data for specific PPBD-PPEP interactions. This information is not only useful for designing, comparing and validating the relevant experiments, but it also serves as a knowledge-base for computationally constructing systems signaling pathways and networks. PepCyber:P approximately PEP is accessible through the URL, http://www.pepcyber.org/PPEP/. The current release of the database contains 7044 PPBD-mediated interactions involving 337 PPBD-containing proteins and 1123 substrate proteins. PMID- 18160411 TI - DNA mimicry by a high-affinity anti-NF-kappaB RNA aptamer. AB - The binding of RNA molecules to proteins or other ligands can require extensive RNA folding to create an induced fit. Understanding the generality of this principle involves comparing structures of RNA before and after complex formation. Here we report the NMR solution structure of a 29-nt RNA aptamer whose crystal structure had previously been determined in complex with its transcription factor target, the p50(2) form of NF-kappaB. The RNA aptamer internal loop structure has pre-organized features that are also found in the complex, including non-canonical base pairing and cross-strand base stacking. Remarkably, the free RNA aptamer structure possesses a major groove that more closely resembles B-form DNA than RNA. Upon protein binding, changes in RNA structure include the kinking of the internal loop and distortion of the terminal tetraloop. Thus, complex formation involves both pre-formed and induced fit binding interactions. The high affinity of the NF-kappaB transcription factor for this RNA aptamer may largely be due to the structural pre-organization of the RNA that results in its ability to mimic DNA. PMID- 18160412 TI - ATR protects the genome against CGG.CCG-repeat expansion in Fragile X premutation mice. AB - Fragile X mental retardation syndrome is a repeat expansion disease caused by expansion of a CGG.CCG-repeat tract in the 5' UTR of the FMR1 gene. In humans, small expansions occur more frequently on paternal transmission while large expansions are exclusively maternal in origin. It has been suggested that expansion is the result of aberrant DNA replication, repair or recombination. To distinguish amongst these possibilities we crossed mice containing 120 CGG.CCG repeats in the 5' UTR of the mouse Fmr1 gene to mice with mutations in ATR, a protein important in the cellular response to stalled replication forks and bulky DNA lesions. We show here that ATR heterozygosity results in increased expansion rates of maternally, but not paternally, transmitted alleles. In addition, age related somatic expansions occurred in mice of both genders that were not seen in ATR wild-type animals. Some ATR-sensitive expansion occurs in postmitotic cells including haploid gametes suggesting that aberrant DNA repair is responsible. Our data suggest that two mechanisms of repeat expansion exist that may explain the small and large expansions seen in humans. In addition, our data provide an explanation for the maternal bias of large expansions in humans and the lower incidence of these expansions in mice. PMID- 18160413 TI - Phytohormone abscisic acid control RNA-dependent RNA polymerase 6 gene expression and post-transcriptional gene silencing in rice cells. AB - RNA-dependent RNA polymerase 6 (RDR6) catalyses dsRNA synthesis for post transcriptional gene silencing (PTGS)-associated amplification and the generation of endogeneous siRNAs involved in developmental determinations or stress responses. The functional importance of RDR6 in PTGS led us to examine its connection to the cellular regulatory network by analyzing the hormonal responses of RDR6 gene expression in a cultured cell system. Delivery of dsRNA, prepared in vitro, into cultured rice (Oryza sativa cv. Japonica Dongjin) cells successfully silenced the target isocitrate lyase (ICL) transcripts. Silencing was transient in the absence of abscisic acid (ABA), while it became persistent in the presence of ABA in growth medium. A transcription assay of the OsRDR6 promoter showed that it was positively regulated by ABA. OsRDR6-dependent siRNA(ICL) generation was also significantly up-regulated by ABA. The results showed that, among the five rice OsRDR isogenes, only OsRDR6 was responsible for the observed ABA-mediated amplification and silencing of ICL transcripts. We propose that ABA modulates PTGS through the transcriptional control of the OsRDR6 gene. PMID- 18160414 TI - Influence of the incorporation of a cyclohexenyl nucleic acid (CeNA) residue onto the sequence d(CGCGAATTCGCG). AB - Cyclohexene nucleic acids (CeNA), which are characterized by the presence of a cyclohexene moiety instead of a natural (deoxy)ribose sugar, are known to increase the thermal and enzymatic stability when incorporated in RNA oligonucleotides. As it has been demonstrated that even a single cyclohexenyl nucleoside, when incorporated in an oligonucleotide, can have a profound effect on the biological activity of the oligonucleotide, further research is warranted to study the complex of such oligonucleotides with target proteins. In order to analyse the influence of CeNA residues onto the helix conformation and hydration of natural nucleic acid structures, a cyclohexenyl-adenine building block (xAr) was incorporated into the Dickerson sequence CGCGA(xAr)TTCGCG. The crystal structure of this sequence determined to a resolution of 1.90 A. The global helix belongs to the B-type family and shows a water spine, which is partially broken up by the apolar cyclohexene residue. The cyclohexene ring adopts the (2)E conformation allowing a better incorporation of the residue in the dodecamer sequence. The crystal packing is stabilized by cobalt hexamine residues and belongs to space group P222(1), never before reported for nucleic acids. PMID- 18160415 TI - Proteomic screen defines the hepatocyte nuclear factor 1alpha-binding partners and identifies HMGB1 as a new cofactor of HNF1alpha. AB - Hepatocyte nuclear factor (HNF)-1alpha is one of the liver-enriched transcription factors involved in many tissue-specific expressions of hepatic genes. The molecular mechanisms for determining HNF1alpha-mediated transactivation have not been explained fully. To identify unknown proteins that interact with HNF1alpha, we developed a co-IP-MS strategy to search HNF1alpha interactions, and high mobility group protein-B1 (HMGB1), a chromosomal protein, was identified as a novel HNF1alpha-interacting protein. In vitro glutathione S-transferase pull-down and in vivo co-immunoprecipitation studies confirmed an interaction between HMGB1 and HNF1alpha. The protein-protein interaction was mediated through the HMG box domains of HMGB1 and the homeodomain of HNF1alpha. Furthermore, electrophoretic mobility shift assay and chromatin-immunoprecipitation assay demonstrated that HMGB1 was recruited to endogenous HNF1alpha-responsive promoters and enhanced HNF1alpha binding to its cognate DNA sequences. Moreover, luciferase reporter analyses showed that HMGB1 potentiated the transcriptional activities of HNF1alpha in cultured cells, and downregulation of HMGB1 by RNA interference specifically affected the HNF1alpha-dependent gene expression in HepG2 cell. Taken together, these findings raise the intriguing possibility that HMGB1 is a new cofactor of HNF1alpha and participates in HNF1alpha-mediated transcription regulation through protein-protein interaction. PMID- 18160416 TI - Subjects expressing the glucose-6-phosphate dehydrogenase deficient phenotype experience a lower cardiovascular mortality. PMID- 18160417 TI - Cardiovascular risk factors in adult patients with multisystem Langerhans-cell histiocytosis: evidence of glucose metabolism abnormalities. AB - BACKGROUND: Langerhans-cell histiocytosis (LCH) is a rare disease with features of chronic inflammation and it may also induce hypopituitarism, conditions associated with an increased risk of cardiovascular diseases. AIM: Cardiovascular and metabolic risk profile investigation in multisystem LCH patients with and without anterior pituitary deficiency. DESIGN: Prospective, observational study. METHODS: Fourteen adult patients with LCH, 7 with and 7 without anterior pituitary deficiency, and 42 controls matched for age, body mass index (BMI) and smoking. Cardiovascular risk factors were estimated in all subjects: glucose and lipid profile, mathematical indices of insulin resistance (IR), blood pressure, structural arterial and functional endothelial properties (intima-media thickness, brachial artery flow-mediated dilatation). Cardiovascular risk factors were estimated in the three groups studied; the effect of disease activity and/or treatment was also determined in patients with LCH. RESULTS: Ten patients had diabetes insipidus, and 7 anterior pituitary hormone deficiencies: 8 patients had active disease and 11 had received systemic treatment. No difference was observed between the study groups in vascular parameters, in lipid profile or in blood pressure. However, the insulin resistance index GIR was decreased in patients with LCH without anterior pituitary deficiency compared to controls (P = 0.033). Three patients had impaired glucose tolerance and one diabetes mellitus type 2. These patients were older and had active disease; there was no association with hypopituitarism and/or previous treatment. CONCLUSION: Adults patients with LCH have abnormalities of glucose metabolism that tend to occur in patients with active disease, and may be a consequence of the pro-inflammatory state. PMID- 18160418 TI - Paediatric systemic lupus erythematosus: prognostic impact of antiphospholipid antibodies. AB - OBJECTIVES: The aim of our study was to investigate the prognostic impact of aPL in paediatric onset systemic lupus erythematosus (p-SLE). METHODS: This retrospective study included 56 patients with p-SLE. Chi2-test, Fisher's exact test, incidence rate ratio and Kaplan-Meier survival curves were used to compare aPL-positive and aPL-negative patients considering the value of SDI (Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index for SLE) at the end of follow-up, the occurrence of thromboses, organ system involvements and need for immunosuppressive treatment in addition to corticosteroids. RESULTS: Anti-cardiolipin antibodies and lupus anticoagulants were detected in 27 (49%) and 19 (35%) patients, respectively. These aPL were frequently transient or intermittent (10 and 15 cases, respectively), and only rarely persistent over time (five cases). The risk of thrombosis was significantly higher (odds ratio = 6.42) and occurred earlier in the presence of aPL, especially if aPL were persistent (P < 0.05). The association between aPL and neurological, renal, haematological manifestations or need for immunosuppressive treatment was not statistically significant. After a mean follow-up of 7.2 yrs, 30 patients (54.5%) had an SDI score > or = 1. The risk of damage (SDI > or = 1) in aPL-positive patients was three times higher than in aPL negative patients (P < 0.05). Four of the six fatal cases occurred in the aPL positive group. CONCLUSIONS: The presence of aPL in p-SLE could represent not only a risk factor for thrombosis but also a poor prognostic factor overall. PMID- 18160419 TI - Discrepancies between the EULAR response criteria and the NICE guidelines for continuation of anti-TNF therapy in RA: a cause for concern? AB - OBJECTIVES: A discrepancy exists between the National Institute for Health and Clinical Excellence (NICE) guidelines for continuation of TNF therapy in RA and EULAR response criteria. We performed a retrospective study of patients starting anti-TNF therapy to establish how many NICE non-responders would have met EULAR response criteria, and whether this may increase. METHOD: We calculated the percentage of NICE non-responders who would have met EULAR moderate response criteria. We then compared the mean decrease in disease activity score (DAS28) for patients with low and high baseline scores. We analysed trends for treating RA in Derby with anti-TNF to address whether we were treating less active disease over time. RESULTS: At 3 months (n = 271 patients), 7.7% of NICE non-responders would have met EULAR moderate response criteria. At 6 months (n = 240 patients) this was 23.7%. Patients starting with a higher DAS28 had a significantly greater absolute drop in score. The mean decrease between the 1st and 3rd tertiles of patients divided by baseline DAS28 was significant at 3 and 6 months (P < 0.001). Derby rheumatologists were treating less active RA over time. Comparing the mean DAS28 baseline between the 1st and 3rd tertiles of patients divided by anti-TNF commencement date was significant (P < 0.001). CONCLUSIONS: A significant minority of NICE non-responders would fall within the moderate EULAR response criteria. This is likely to increase in future due to the increasing tendency to initiate anti-TNF in patients with less active disease. Consequently, NICE guidelines should be brought in line with EULAR response criteria. PMID- 18160420 TI - Role of pathogenic auto-antibody production by Toll-like receptor 9 of B cells in active systemic lupus erythematosus. AB - OBJECTIVES: Toll-like receptor 9 (TLR9) is a pattern-associated receptor functioning in innate immunity that may be involved in the recognition of self antigens and the production of pathogenic auto-antibodies. Therefore, we examined the expression of TLR9 in systemic lupus erythematosus (SLE) to determine whether TLR9 is involved in the production of pathogenic auto-antibodies. METHODS: B cells were collected from patients with active SLE, and subjected to analysis of the TLR9 molecule using flow cytometry fluorescence activated cell sorting (FACS) and TLR9 mRNA by reverse-transcriptase polymerase chain reaction. SLE B cells were stimulated with CpG-ODN, and subsequent cytokine and anti-dsDNA antibody production was measured by enzyme-linked immunosorbent assay. RESULTS: The expression and mRNA level of TLR9 on B cells was up-regulated in SLE patients, and SLE disease activity index (SLEDAI) and CH50 were correlated with TLR9 expression on CD20+ B cells. Moreover, TLR9-CpG interaction enhanced the production of anti-dsDNA antibody and IL-10. CONCLUSIONS: The present study demonstrated that higher expression of TLR9 on peripheral blood B cells from patients with active SLE was significantly correlated with CH50 and SLEDAI to TLR9, and induced the production of anti-dsDNA antibody and IL-10 by TLR9-CpG ligation. These results suggest that an abnormality of innate immunity plays a crucial role in the pathology of SLE, and that blockade of CpG-TLR9 interaction may be a new therapeutic approach for SLE. PMID- 18160421 TI - Anti-Saccharomyces cerevisiae antibodies (ASCA) in spondyloarthropathies: a reassessment. AB - OBJECTIVES: Seronegative spondyloarthropathies, especially ankylosing spondylitis (AS), is shown to be associated with inflammatory bowel disease. Anti Saccharomyces cerevisiae antibodies (ASCA) is a valid serological marker for Crohn's disease. Presence of ASCA is controversial in AS. In this study, we aimed to investigate the prevalence of ASCA in spondyloarthropathies and its relationship with disease activity and severity. METHODS: One hundred and seventy five patients with AS, 47 patients with undifferentiated spondyloarthropathy (uSpA) and 103 healthy controls (HCs) were studied. All patients were questioned for demographic features and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores. Radiological damage is assessed by Bath Ankylosing Spondylitis Radiology Index (BASRI) and modified Stroke Ankylosing Spondylitis Spinal Score (mSASSS). ASCA levels were measured with standard ELISA kits. RESULTS: There was an overall increased prevalence of ASCA IgA in AS and uSpA compared with HCs (20.6 and 19.1% vs 5.8%, P = 0.0008 and P = 0.02, respectively). No association was observed between ASCA positivity and erythrocyte sedimentation rate, C reactive protein levels and BASDAI scores. However, ASCA-positive patients had higher BASRI scores [median BASRI: 7 (2-12) vs 6 (2-12); P = 0.037]. Although not reaching significance, they also had reduced chest expansion and higher Bath Ankylosing Spondylitis Functional Index (BASFI) scores. ASCA-positive AS patients also required anti-tumour necrosis factor therapy more frequently (P = 0.006). CONCLUSIONS: ASCA IgA seems to be more prevalent in AS and uSpA. ASCA can also be a marker of radiological damage and a more severe course in AS. PMID- 18160422 TI - Tonotopic control of auditory thalamus frequency tuning by reticular thalamic neurons. AB - GABAergic cells of the thalamic reticular nucleus (TRN) can potentially exert strong control over transmission of information through thalamus to the cerebral cortex. Anatomical studies have shown that the reticulo-thalamic connections are spatially organized in the visual, somatosensory, and auditory systems. However, the issue of how inhibitory input from TRN controls the functional properties of thalamic relay cells and whether this control follows topographic rules remains largely unknown. Here we assessed the consequences of increasing or decreasing the activity of small ensembles of TRN neurons on the receptive field properties of medial geniculate (MG) neurons. For each MG cell, the frequency tuning curve and the rate-level function were tested before, during, and after microiontophoretic applications of GABA, or of glutamate, in the auditory sector of the TRN. For 66 MG cells tested during potent pharmacological control of TRN activity, group data did not reveal any significant effects. However, for a population of 20/66 cells (all but 1 recorded in the ventral, tonotopic, division), the breadth of tuning, the frequency selectivity and the acoustic threshold were significantly modified in the directions expected from removing, or reinforcing, a dominant inhibitory input onto MG cells. Such effects occurred only when the distance between the characteristic frequency of the recorded ventral MG cell and that of the TRN cells at the ejection site was <0.25 octaves; they never occurred for larger distances. This relationship indicates that the functional interactions between TRN cells and ventral MG cells rely on precise topographic connections. PMID- 18160423 TI - Morphology, intrinsic membrane properties, and rotation-evoked responses of trochlear motoneurons in the turtle. AB - Intrinsic properties and rotation-evoked responses of trochlear motoneurons were investigated in the turtle using an in vitro preparation consisting of the brain stem with attached temporal bones that retain functional semicircular canals. Motoneurons were divided into two classes based on intrinsic properties. The first class exhibited higher impedance (123.0 +/- 11.0 MOmega), wider spikes (0.99 +/- 0.05 ms), a single spike afterhyperpolarization (AHP), little or no spike frequency adaptation (SFA), and anomalous rectification, characterized by an initial "sag" in membrane potential in response to hyperpolarizing current injection. The second class exhibited lower impedance (21.8 +/- 2.5 MOmega), narrower spikes (0.74 +/- 0.03 ms), a double AHP, substantial SFA, and little or no rectification. Vestibular responses were evoked by horizontal sinusoidal rotation (1/12-1/3 Hz; peak velocity: 30-100 degrees /s). Spiking in higher impedance cells was recruited earlier in the response and exhibited a more limited dynamic range relative to that of lower impedance cells. Spiking evoked by injecting depolarizing current during rotation was blocked during contraversive motion and was consistent with a shunting inhibition. No morphological features were identified in neurobiotin-filled cells that correlated with the two physiological classes. Recovered motoneurons were multipolar but exhibited a less-complex dendritic morphology than ocular motoneurons of similarly sized mammals. The two physiologically defined cell classes have homologues in other vertebrates, suggesting that intrinsic membrane properties play an important role in oculomotor processing. PMID- 18160424 TI - TRPM5-expressing solitary chemosensory cells respond to odorous irritants. AB - Inhaled airborne irritants elicit sensory responses in trigeminal nerves innervating the nasal epithelium, leading to protective reflexes. The sensory mechanisms involved in the detection of odorous irritants are poorly understood. We identified a large population of solitary chemosensory cells expressing the transient receptor potential channel M5 (TRPM5) using transgenic mice where the promoter of TRPM5 drives the expression of green fluorescent protein (GFP). Most of these solitary chemosensory cells lie in the anterior nasal cavity. These GFP labeled solitary chemosensory cells exhibited immunoreactivity for synaptobrevin 2, a vesicle-associated membrane protein important for synaptic transmission. Concomitantly, we found trigeminal nerve fibers apposed closely to the solitary chemosensory cells, indicating potential transmission of sensory information to trigeminal fibers. In addition, stimulation of the nasal cavity with high concentrations (0.5-5 mM) of a variety of odorants elicited event-related potentials (ERPs) in areas rich in TRPM5-expressing solitary chemosensory cells. Furthermore, odorous chemicals and trigeminal stimuli induced changes in intracellular Ca(2+) levels in isolated TRPM5-expressing solitary chemosensory cells in a concentration-dependent manner. Together, our data show that the TRPM5 expressing cells respond to a variety of chemicals at high exposure levels typical of irritants and are positioned in the nasal cavity appropriately to monitor inhaled air quality. PMID- 18160425 TI - Inferring spike trains from local field potentials. AB - We investigated whether it is possible to infer spike trains solely on the basis of the underlying local field potentials (LFPs). Using support vector machines and linear regression models, we found that in the primary visual cortex (V1) of monkeys, spikes can indeed be inferred from LFPs, at least with moderate success. Although there is a considerable degree of variation across electrodes, the low frequency structure in spike trains (in the 100-ms range) can be inferred with reasonable accuracy, whereas exact spike positions are not reliably predicted. Two kinds of features of the LFP are exploited for prediction: the frequency power of bands in the high gamma-range (40-90 Hz) and information contained in low-frequency oscillations (<10 Hz), where both phase and power modulations are informative. Information analysis revealed that both features code (mainly) independent aspects of the spike-to-LFP relationship, with the low-frequency LFP phase coding for temporally clustered spiking activity. Although both features and prediction quality are similar during seminatural movie stimuli and spontaneous activity, prediction performance during spontaneous activity degrades much more slowly with increasing electrode distance. The general trend of data obtained with anesthetized animals is qualitatively mirrored in that of a more limited data set recorded in V1 of non-anesthetized monkeys. In contrast to the cortical field potentials, thalamic LFPs (e.g., LFPs derived from recordings in the dorsal lateral geniculate nucleus) hold no useful information for predicting spiking activity. PMID- 18160426 TI - Do corticomotoneuronal cells predict target muscle EMG activity? AB - Data from two rhesus macaques were used to investigate the pattern of cortical cell activation during reach-to-grasp movements in relation to the corresponding activation pattern of the cell's facilitated target muscles. The presence of postspike facilitation (PSpF) in spike-triggered averages (SpTAs) of electromyographic (EMG) activity was used to identify cortical neurons with excitatory synaptic linkages with motoneurons. EMG activity from 22 to 24 muscles of the forelimb was recorded together with the activity of M1 cortical neurons. The extent of covariation was characterized by 1) identifying the task segment containing the cell and target muscle activity peaks, 2) quantifying the timing and overlap between corticomotoneuronal (CM) cell and EMG peaks, and 3) applying Pearson correlation analysis to plots of CM cell firing rate versus EMG activity of the cell's facilitated muscles. At least one firing rate peak, for nearly all (95%) CM cells tested, matched a corresponding peak in the EMG activity of the cell's target muscles. Although some individual CM cells had very strong correlations with target muscles, overall, substantial disparities were common. We also investigated correlations for ensembles of CM cells sharing the same target muscle. The ensemble population activity of even a small number of CM cells influencing the same target muscle produced a relatively good match (r >/= 0.8) to target muscle EMG activity. Our results provide evidence in support of the notion that corticomotoneuronal output from primary motor cortex encodes movement in a framework of muscle-based parameters, specifically muscle activation patterns as reflected in EMG activity. PMID- 18160427 TI - The oscillation score: an efficient method for estimating oscillation strength in neuronal activity. AB - We present a method that estimates the strength of neuronal oscillations at the cellular level, relying on autocorrelation histograms computed on spike trains. The method delivers a number, termed oscillation score, that estimates the degree to which a neuron is oscillating in a given frequency band. Moreover, it can also reliably identify the oscillation frequency and strength in the given band, independently of the oscillation in other frequency bands, and thus it can handle superimposed oscillations on multiple scales (theta, alpha, beta, gamma, etc.). The method is relatively simple and fast. It can cope with a low number of spikes, converging exponentially fast with the number of spikes, to a stable estimation of the oscillation strength. It thus lends itself to the analysis of spike-sorted single-unit activity from electrophysiological recordings. We show that the method performs well on experimental data recorded from cat visual cortex and also compares favorably to other methods. In addition, we provide a measure, termed confidence score, that determines the stability of the oscillation score estimate over trials. PMID- 18160428 TI - Bcl-xL inhibitor ABT-737 reveals a dual role for Bcl-xL in synaptic transmission. AB - A role for BCL-xL in regulating neuronal activity is suggested by its dramatic effects on synaptic function and mitochondrial channel activity. When recombinant BCL-xL is injected into the giant presynaptic terminal of squid stellate ganglion or applied directly to mitochondrial outer membranes within the living terminal, it potentiates synaptic transmission acutely, and it produces mitochondrial channel activity. The squid, however, is a genetically intractable model, making it difficult to apply genetic tools in squid to explore the role of endogenous BCL-xL in synaptic function. Therefore the small molecule inhibitor ABT-737, a mimetic of the BH3-only protein BAD, binding to the BH3-binding domain pocket, was tested in squid, revealing a dual role for BCL-xL. ABT-737 slowed recovery of synaptic responses after repetitive synaptic activity, indicating that endogenous BCL-xL is necessary for timely recovery of rapidly firing synapses. Unexpectedly, however, ABT-737 also protected neurons from hypoxia-induced synaptic rundown and from increased permeability of the mitochondrial outer membrane during hypoxia. This implies that endogenous BCL-xL or a modified form of BCL-xL, such as the N truncated, proteolytic, pro-apoptotic cleavage product, DeltaN BCL-xL, contributes to injurious responses of the hypoxic synapse. To determine if ABT 737 is also an inhibitor of DeltaN BCL-xL, recombinant DeltaN BCL-xL protein was injected into the synapse. ABT-737 potently inhibited synaptic rundown induced by recombinant DeltaN BCL-xL. These observations support the possibility that endogenous proteolysis or a functionally equivalent modification of BCL-xL is responsible for the deleterious effects of hypoxia on synaptic activity. PMID- 18160429 TI - Human immunodeficiency virus type 1 Vpr induces cell cycle G2 arrest through Srk1/MK2-mediated phosphorylation of Cdc25. AB - Human immunodeficiency virus type 1 (HIV-1) Vpr induces cell cycle G(2) arrest in fission yeast (Schizosaccharomyces pombe) and mammalian cells, suggesting the cellular pathway(s) targeted by Vpr is conserved among eukaryotes. Our previous studies in fission yeast demonstrated that Vpr induces G(2) arrest in part through inhibition of Cdc25, a Cdc2-specific phosphatase that promotes G(2)/M transition. The goal of this study was to further elucidate molecular mechanism underlying the inhibitory effect of Vpr on Cdc25. We show here that, similar to the DNA checkpoint controls, expression of vpr promotes subcellular relocalization of Cdc25 from nuclear to cytoplasm and thereby prevents activation of Cdc2 by Cdc25. Vpr-induced nuclear exclusion of Cdc25 appears to depend on the serine/threonine phosphorylation of Cdc25 and the presence of Rad24/14-3-3 protein, since amino acid substitutions of the nine possible phosphorylation sites of Cdc25 with Ala (9A) or deletion of the rad24 gene abolished nuclear exclusion induced by Vpr. Interestingly, Vpr is still able to promote Cdc25 nuclear export in mutants defective in the checkpoints (rad3 and chk1/cds1), the kinases that are normally required for Cdc25 phosphorylation and nuclear exclusion of Cdc25, suggesting that others kinase(s) might modulate phosphorylation of Cdc25 for the Vpr-induced G(2) arrest. We report here that this kinase is Srk1. Deletion of the srk1 gene blocks the nuclear exclusion of Cdc25 caused by Vpr. Overexpression of srk1 induces cell elongation, an indication of cell cycle G(2) delay, in a similar fashion to Vpr; however, no additive effect of cell elongation was observed when srk1 and vpr were coexpressed, indicating Srk1 and Vpr are likely affecting the cell cycle G(2)/M transition through the same cellular pathway. Immunoprecipitation further shows that Vpr and Srk1 are part of the same protein complex. Consistent with our findings in fission yeast, depletion of the MK2 gene, a human homologue of Srk1, either by small interfering RNA or an MK2 inhibitor suppresses Vpr-induced cell cycle G(2) arrest in mammalian cells. Collectively, our data suggest that Vpr induces cell cycle G(2) arrest at least in part through a Srk1/MK2-mediated mechanism. PMID- 18160430 TI - Genetic determinants of Sindbis virus mosquito infection are associated with a highly conserved alphavirus and flavivirus envelope sequence. AB - Wild-type Sindbis virus (SINV) strain MRE16 efficiently infects Aedes aegypti midgut epithelial cells (MEC), but laboratory-derived neurovirulent SINV strain TE/5'2J infects MEC poorly. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of MRE16 and TE/5'2J differ at 60 residue sites. To identify the genetic determinants of MEC infection of MRE16, the TE/5'2J virus genome was altered to contain either domain chimeras or more focused nucleotide substitutions of MRE16. The growth patterns of derived viruses in cell culture were determined, as were the midgut infection rates (MIR) in A. aegypti mosquitoes. The results showed that substitutions of MRE16 E2 aa 95 to 96 and 116 to 119 into the TE/5'2J virus increased MIR both independently and in combination with each other. In addition, a unique PPF/.GDS amino acid motif was located between these two sites that was found to be a highly conserved sequence among alphaviruses and flaviviruses but not other arboviruses. PMID- 18160431 TI - Hepatitis C virus core protein upregulates serine phosphorylation of insulin receptor substrate-1 and impairs the downstream akt/protein kinase B signaling pathway for insulin resistance. AB - Chronic hepatitis C virus (HCV) infection has a significantly increased prevalence of type 2 diabetes mellitus (T2DM). Insulin resistance is a critical component of T2DM pathogenesis. Several mechanisms are likely to be involved in the pathogenesis of HCV-related insulin resistance. Since we and others have previously observed that HCV core protein activates c-Jun N-terminal kinase (JNK) and mitogen-activated protein kinase, we examined the contribution of these pathways to insulin resistance in hepatocytes. Our experimental findings suggest that HCV core protein alone or in the presence of other viral proteins increases Ser(312) phosphorylation of the insulin receptor substrate-1 (IRS-1). Hepatocytes infected with cell culture-grown HCV genotype 1a or 2a displayed a significant increase in the Ser(473) phosphorylation status of the Ser/Thr kinase protein kinase B (Akt/PKB), while Thr(308) phosphorylation was not significantly altered. HCV core protein-mediated Ser(312) phosphorylation of IRS-1 was inhibited by JNK (SP600125) and phosphatidylinositol-3 kinase (LY294002) inhibitors. A functional assay also suggested that hepatocytes expressing HCV core protein alone or infected with cell culture-grown HCV exhibited a suppression of 2-deoxy-d [(3)H]glucose uptake. Inhibition of the JNK signaling pathway significantly restored glucose uptake despite HCV core expression in hepatocytes. Taken together, our results demonstrated that HCV core protein increases IRS-1 phosphorylation at Ser(312) which may contribute in part to the mechanism of insulin resistance. PMID- 18160432 TI - BK virus as a cofactor in the etiology of prostate cancer in its early stages. AB - Prostate cancer has been projected to cause almost 10% of all male cancer deaths in the United States in 2007. The incidence of mutations in the tumor suppressor genes Rb1 and p53, especially in the early stages of the disease, is low compared to those for other cancers. This has led to the hypothesis that a human virus such as BK virus (BKV), which establishes a persistent subclinical infection in the urinary tract and encodes oncoproteins that interfere with these tumor suppressor pathways, is involved. Previously, we detected BKV DNA in the epithelial cells of benign and proliferative inflammatory atrophy ducts of cancerous prostate specimens. In the present report, we demonstrate that BKV is present at a much lower frequency in noncancerous prostates. Additionally, in normal prostates, T-antigen (TAg) expression is observed only in specimens harboring proliferative inflammatory atrophy and prostatic intraepithelial neoplasia. We further demonstrate that the p53 gene from atrophic cells expressing TAg is wild type, whereas tumor cells expressing detectable nuclear p53 contain a mix of wild-type and mutant p53 genes, suggesting that TAg may inactivate p53 in the atrophic cells. Our results point toward a role for BKV in early prostate cancer progression. PMID- 18160433 TI - NKG2D receptor signaling enhances cytolytic activity by virus-specific CD8+ T cells: evidence for a protective role in virus-induced encephalitis. AB - Inoculation with the neurotropic JHM strain of mouse hepatitis virus (JHMV) into the central nervous system (CNS) of mice results in an acute encephalitis associated with an immune-mediated demyelinating disease. During acute disease, infiltrating CD8(+) T cells secrete gamma interferon (IFN-gamma) that controls replication in oligodendrocytes, while infected astrocytes and microglia are susceptible to perforin-mediated lysis. The present study was undertaken to reveal the functional contributions of the activating NKG2D receptor in host defense and disease following JHMV infection. NKG2D ligands RAE-1, MULT1, and H60 were expressed within the CNS following JHMV infection. The immunophenotyping of infiltrating cells revealed that NKG2D was expressed on approximately 90% of infiltrating CD8(+) T cells during acute and chronic disease. Blocking NKG2D following JHMV infection resulted in increased mortality that correlated with increased viral titers within the CNS. Anti-NKG2D treatment did not alter T-cell infiltration into the CNS or the generation of virus-specific CD8(+) T cells, and the expression of IFN-gamma was not affected. However, cytotoxic T-lymphocyte (CTL) activity was dependent on NKG2D expression, because anti-NKG2D treatment resulted in a dramatic reduction in lytic activity by virus-specific CD8(+) T cells. Blocking NKG2D during chronic disease did not affect either T-cell or macrophage infiltration or the severity of demyelination, indicating that NKG2D does not contribute to virus-induced demyelination. These findings demonstrate a functional role for NKG2D in host defense during acute viral encephalitis by selectively enhancing CTL activity by infiltrating virus-specific CD8(+) T cells. PMID- 18160434 TI - cis- and trans-acting functions of brome mosaic virus protein 1a in genomic RNA1 replication. AB - RNA viruses employ a combination of mechanisms to regulate their gene expression and replication. Brome mosaic virus (BMV) is a tripartite positive-strand RNA virus used to study the requirements for virus infection. BMV genomic RNA1 encodes protein 1a, which contains a methyltransferase (MT) domain and a helicase domain that are required for replication. 1a forms a complex with the 2a RNA dependent RNA polymerase for the replication and transcription of all BMV RNAs. RNA1 expressed with 2a from Agrobacterium-based vectors can result in RNA1 replication in Nicotiana benthamiana. A mutation in the 1a translation initiation codon significantly decreased RNA1 accumulation even when wild-type (WT) 1a and 2a were provided in trans. Therefore, efficient RNA1 replication requires 1a translation from RNA1 in cis, indicating a linkage between replication and translation. Mutation analyses showed that the full-length 1a protein was required for efficient RNA1 replication, not just the process of translation. Three RNA1s with mutations in the 1a MT domain could be partially rescued by WT 1a expressed in trans, indicating that the cis-acting function of 1a was retained. Furthermore, an RNA motif in the 5'-untranslated region of RNA1, named the B box, was required for 1a to function in cis and in trans for BMV RNA accumulation. The B box is required for the formation of the replication factory (M. Schwartz, J. Chen, M. Janda, M. Sullivan, J. den Boon, and P. Ahlquist, Mol. Cell 9:505-514, 2002). Results in this work demonstrate a linkage between BMV RNA1 translation and replication. PMID- 18160435 TI - Alpha/beta interferon inhibits cap-dependent translation of viral but not cellular mRNA by a PKR-independent mechanism. AB - The alpha/beta interferon (IFN-alpha/beta) response is critical for host protection against disseminated replication of many viruses, primarily due to the transcriptional upregulation of genes encoding antiviral proteins. Previously, we determined that infection of mice with Sindbis virus (SB) could be converted from asymptomatic to rapidly fatal by elimination of this response (K. D. Ryman et al., J. Virol. 74:3366-3378, 2000). Probing of the specific antiviral proteins important for IFN-mediated control of virus replication indicated that the double stranded RNA-dependent protein kinase, PKR, exerted some early antiviral effects prior to IFN-alpha/beta signaling; however, the ability of IFN-alpha/beta to inhibit SB and protect mice from clinical disease was essentially undiminished in the absence of PKR, RNase L, and Mx proteins (K. D. Ryman et al., Viral Immunol. 15:53-76, 2002). One characteristic of the PKR/RNase L/Mx-independent antiviral effect was a blockage of viral protein accumulation early after infection (K. D. Ryman et al., J. Virol. 79:1487-1499, 2005). We show here that IFN-alpha/beta priming induces a PKR-independent activity that inhibits m(7)G cap-dependent translation at a step after association of cap-binding factors and the small ribosome subunit but before formation of the 80S ribosome. Furthermore, the activity targets mRNAs that enter across the cytoplasmic membrane, but nucleus transcribed RNAs are relatively unaffected. Therefore, this IFN-alpha/beta induced antiviral activity represents a mechanism through which IFN-alpha/beta exposed cells are defended against viruses that enter the cytoplasm, while preserving essential host activities, including the expression of antiviral and stress-responsive genes. PMID- 18160436 TI - Temporal association of the herpes simplex virus genome with histone proteins during a lytic infection. AB - Previous work has determined that there are nucleosomes on the herpes simplex virus (HSV) genome during a lytic infection but that they are not arranged in an equally spaced array like in cellular DNA. However, like in cellular DNA, the promoter regions of several viral genes have been shown to be associated with nucleosomes containing modified histone proteins that are generally found associated with actively transcribed genes. Furthermore, it has been found that the association of modified histones with the HSV genome can be detected at the earliest times postinfection (1 h postinfection) and increases up to 3 h postinfection. However from 3 h to 6 h postinfection (the late phase of the replication cycle), the association decreases. In this study we have examined histone association with promoter regions of all kinetic classes of genes. This was done over the time course of an infection in Sy5y cells using sucrose gradient sedimentation, bromodeoxyuridine labeling, chromatin immunoprecipitation assays, Western blot analysis, trypsin and DNase digestion, and quantitative real time PCR. Because no histones were detected inside HSV type 1 capsids, the viral genome probably starts to associate with histones after being transported from infecting virions into the host nucleus. Promoter regions of all gene classes (immediate early, early, and late) bind with histone proteins at the start of viral gene expression. However, after viral DNA replication initiates, histones appear not to associate with newly synthesized viral genomes. PMID- 18160437 TI - RNA structure modulates splicing efficiency at the human immunodeficiency virus type 1 major splice donor. AB - The untranslated leader of the human immunodeficiency virus type 1 (HIV-1) RNA genome encodes essential sequence and structural motifs that control various replication steps. The 5' splice site or splice donor (SD) is embedded in a semistable hairpin, but the function of this structure is unknown. We stabilized this SD hairpin by creating an additional base pair and demonstrated a severe HIV 1 replication defect. A splicing defect was apparent in RNA analyses of virus infected cells and cells transfected with appropriate reporter constructs. We selected multiple virus revertants in search for interesting second-site escape pathways. Most revertants acquired an additional mutation that modulated the stability of the mutant SD hairpin. One revertant acquired a single nucleotide change in the upstream DIS hairpin. We demonstrate that a novel SD site is created by this upstream mutation, which obviously reduces the number of leader nucleotides that are included in spliced HIV-1 transcripts. These results suggest a novel role of RNA structure in the regulation of HIV-1 splicing. PMID- 18160438 TI - Human butyrate-induced transcript 1 interacts with hepatitis C virus NS5A and regulates viral replication. AB - Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is required for the replication of the viral genome and is involved in several host signaling pathways. To gain further insight into the functional role of NS5A in HCV replication, we screened human cDNA libraries by a yeast two-hybrid system using NS5A as the bait and identified human butyrate-induced transcript 1 (hB-ind1) as a novel NS5A-binding protein. Endogenously and exogenously expressed hB-ind1 was coimmunoprecipitated with NS5A of various genotypes through the coiled-coil domain of hB-ind1. The small interfering RNA (siRNA)-mediated knockdown of hB ind1 in human hepatoma cell lines suppressed the replication of HCV RNA replicons and the production of infectious particles of HCV genotype 2a strain JFH1. Furthermore, these reductions were canceled by the expression of an siRNA resistant hB-ind1 mutant. Among the NS5A-binding host proteins involved in HCV replication, hB-ind1 exhibited binding with FKBP8, and hB-ind1 interacted with Hsp90 through the FxxW motif in its N-terminal p23 homology domain. The impairment of the replication of HCV RNA replicons and of the production of infectious particles of JFH1 virus in the hB-ind1 knockdown cell lines was not reversed by the expression of an siRNA-resistant hB-ind1 mutant in which the FxxW motif was replaced by AxxA. These results suggest that hB-ind1 plays a crucial role in HCV RNA replication and the propagation of JFH1 virus through interaction with viral and host proteins. PMID- 18160439 TI - High level of PD-1 expression on hepatitis C virus (HCV)-specific CD8+ and CD4+ T cells during acute HCV infection, irrespective of clinical outcome. AB - We monitored expression of PD-1 (a mediator of T-cell exhaustion and viral persistence) on hepatitis C virus (HCV)-specific CD8(+) and CD4(+) T cells from blood and liver during acute and chronic infections and after the resolved infection stage. PD-1 expression on HCV-specific T cells was high early in acute infection irrespective of clinical outcome, and most cells continued to express PD-1 in resolved and chronic stages of infection; intrahepatic expression levels were especially high. Our results suggest that an analysis of PD-1 expression alone is not sufficient to predict infection outcome or to determine T-cell functionality in HCV infection. PMID- 18160440 TI - Ex vivo-generated CD36+ erythroid progenitors are highly permissive to human parvovirus B19 replication. AB - The pathogenic parvovirus B19 (B19V) has an extreme tropism for human erythroid progenitor cells. In vitro, only a few erythroid leukemic cell lines (JK-1 and KU812Ep6) or megakaryoblastoid cell lines (UT7/Epo and UT7/Epo-S1) with erythroid characteristics support B19V replication, but these cells are only semipermissive. By using recent advances in generating large numbers of human erythroid progenitor cells (EPCs) ex vivo from hematopoietic stem cells (HSCs), we produced a pure population of CD36(+) EPCs expanded and differentiated from CD34(+) HSCs and assessed the CD36(+) EPCs for their permissiveness to B19V infection. Over more than 3 weeks, cells grown in serum-free medium expanded more than 800,000-fold, and 87 to 96% of the CD36(+) EPCs were positive for globoside, the cellular receptor for B19V. Immunofluorescence (IF) staining showed that about 77% of the CD36(+) EPCs were positive for B19V infection, while about 9% of UT7/Epo-S1 cells were B19V positive. Viral DNA detected by real-time PCR increased by more than 3 logs in CD36(+) EPCs; the increase was 1 log in UT7/Epo S1 cells. Due to the extensive permissivity of CD36(+) EPCs, we significantly improved the sensitivity of detection of infectious B19V by real-time reverse transcription-PCR and IF staining 100- and 1,000-fold, respectively, which is greater than the sensitivity of UT7/Epo-S1 cell-based methods. This is the first description of an ex vivo method to produce large numbers of EPCs that are highly permissive to B19V infection and replication, offering a cellular system that mimics in vivo infection with this pathogenic human virus. PMID- 18160441 TI - Replication of ICP0-null mutant herpes simplex virus type 1 is restricted by both PML and Sp100. AB - Herpes simplex virus type 1 (HSV-1) mutants that fail to express the viral immediate-early protein ICP0 have a pronounced defect in viral gene expression and plaque formation in limited-passage human fibroblasts. ICP0 is a RING finger E3 ubiquitin ligase that induces the degradation of several cellular proteins. PML, the organizer of cellular nuclear substructures known as PML nuclear bodies or ND10, is one of the most notable proteins that is targeted by ICP0. Depletion of PML from human fibroblasts increases ICP0-null mutant HSV-1 gene expression, but not to wild-type levels. In this study, we report that depletion of Sp100, another major ND10 protein, results in a similar increase in ICP0-null mutant gene expression and that simultaneous depletion of both proteins complements the mutant virus to a greater degree. Although chromatin assembly and modification undoubtedly play major roles in the regulation of HSV-1 infection, we found that inhibition of histone deacetylase activity with trichostatin A was unable to complement the defect of ICP0-null mutant HSV-1 in either normal or PML-depleted human fibroblasts. These data lend further weight to the hypothesis that ND10 play an important role in the regulation of HSV-1 gene expression. PMID- 18160442 TI - Combination of an antiviral drug and immunomodulation against hepadnaviral infection in the woodchuck model. AB - The essential role of multispecific immune responses for the control of hepatitis B virus (HBV) infection implies the need of multimodal therapeutic strategies for chronic HBV infection, including antiviral chemotherapy and immunomodulation. This hypothesis was tested in the woodchuck model by a combination of lamivudine pretreatment and subsequent immunizations of woodchucks chronically infected with woodchuck hepatitis virus. The immunizations were performed with DNA vaccines or antigen-antibody immune complexes (IC)/DNA vaccines. Immunizations with IC/DNA vaccines led to an anti-woodchuck hepatitis virus surface antibody response and significant reductions of viral load and antigenemia, suggesting that such a strategy may be effective against chronic HBV infection. PMID- 18160443 TI - Functional analysis of potential carboxy-terminal cleavage sites of tick-borne encephalitis virus capsid protein. AB - The mature capsid protein C of flaviviruses is generated through the proteolytic cleavage of the precursor polyprotein by the viral NS2B/3 protease. This cleavage is a prerequisite for the subsequent processing of the viral surface protein prM, and the concerted progression of these events plays a key role in the process of the assembly of infectious virions. Protein C of tick-borne encephalitis virus (TBEV) contains two amino acid sequence motifs within the carboxy-terminal region that match the canonical NS2B/3 recognition site. Site-specific mutagenesis in the context of the full-length TBEV genome was used to investigate the in vivo cleavage specificity of the viral protease in this functionally important domain. The results indicate that the downstream site is necessary and sufficient for efficient cleavage and virion assembly; in contrast, the upstream site is dispensable and placed in a structural context that renders it largely inaccessible to the viral protease. Mutants with impaired C-prM cleavage generally exhibited a significantly increased cytotoxicity. In spite of the clear preference of the protease for only one of the two naturally occurring motifs, the enzyme was unexpectedly tolerant to both the presence of a noncanonical threonine residue at position P2 and the position of cleavage relative to the adjacent internal prM signal sequence. The insertion of three amino acid residues downstream of the cleavage site did not change the viral phenotype. Thus, this study further illuminates the specificity of the TBEV protease and reveals that the carboxy-terminal region of protein C has a remarkable functional flexibility in its role in the assembly of infectious virions. PMID- 18160444 TI - Nuclear egress and envelopment of herpes simplex virus capsids analyzed with dual color fluorescence HSV1(17+). AB - To analyze the assembly of herpes simplex virus type 1 (HSV1) by triple-label fluorescence microscopy, we generated a bacterial artificial chromosome (BAC) and inserted eukaryotic Cre recombinase, as well as beta-galactosidase expression cassettes. When the BAC pHSV1(17(+))blueLox was transfected back into eukaryotic cells, the Cre recombinase excised the BAC sequences, which had been flanked with loxP sites, from the viral genome, leading to HSV1(17(+))blueLox. We then tagged the capsid protein VP26 and the envelope protein glycoprotein D (gD) with fluorescent protein domains to obtain HSV1(17(+))blueLox-GFPVP26-gDRFP and RFPVP26-gDGFP. All HSV1 BACs had variations in the a-sequences and lost the oriL but were fully infectious. The tagged proteins behaved as their corresponding wild type, and were incorporated into virions. Fluorescent gD first accumulated in cytoplasmic membranes but was later also detected in the endoplasmic reticulum and the plasma membrane. Initially, cytoplasmic capsids did not colocalize with viral glycoproteins, indicating that they were naked, cytosolic capsids. As the infection progressed, they were enveloped and colocalized with the viral membrane proteins. We then analyzed the subcellular distribution of capsids, envelope proteins, and nuclear pores during a synchronous infection. Although the nuclear pore network had changed in ca. 20% of the cells, an HSV1-induced reorganization of the nuclear pore architecture was not required for efficient nuclear egress of capsids. Our data are consistent with an HSV1 assembly model involving primary envelopment of nuclear capsids at the inner nuclear membrane and primary fusion to transfer capsids into the cytosol, followed by their secondary envelopment on cytoplasmic membranes. PMID- 18160445 TI - The PDZ binding motif of human papillomavirus type 16 E6 induces PTPN13 loss, which allows anchorage-independent growth and synergizes with ras for invasive growth. AB - The human papillomavirus (HPV) oncogene E6 has been shown to perform multiple functions (p53 degradation, telomerase activation, etc.) that play a role in oncogenic transformation. Beyond known E6 functions, an undefined mechanism that allows cellular invasion requires the E6 PDZ binding motif (PDZBM). Here, we show that HPV type 16 (HPV16) E6 interacts with and induces loss of a protein tyrosine phosphatase (PTPN13) in a PDZBM-dependent manner. PTPN13 loss induced either by the presence of E6 or by a short hairpin RNA strategy allows for anchorage independent growth (AIG) and synergy with a known oncogene, Ras(v12), resulting in invasive growth in vivo. Restoring PTPN13 expression reverses AIG in cells lacking PTPN13. A genomic analysis of colorectal carcinoma has identified an association between PTPN13 loss-of-function mutations and aberrant Ras signaling. Our findings support this correlation and provide methods for further evaluation of the mechanisms by which PTPN13 loss/Ras expression leads to invasive growth, the results of which will be important for treatment of HPV-related and non-HPV cancer. PMID- 18160446 TI - Novel approach to the development of effective H5N1 influenza A virus vaccines: use of M2 cytoplasmic tail mutants. AB - Outbreaks of highly pathogenic H5N1 influenza viruses in avian species began in Asia and have since spread to other continents. Concern regarding the pandemic potential of these viruses in humans is clearly warranted, and there is an urgent need to develop effective vaccines against them. Previously, we and others demonstrated that deletions of the M2 cytoplasmic tail caused a growth defect in A/WSN/33 (H1N1) influenza A virus in vitro (K. Iwatsuki-Horimoto, T. Horimoto, T. Noda, M. Kiso, J. Maeda, S. Watanabe, Y. Muramoto, K. Fujii, and Y. Kawaoka, J. Virol. 80:5233-5240, 2006; M. F. McCown and A. Pekosz, J. Virol. 79:3595-3605, 2005; M. F. McCown and A. Pekosz, J. Virol. 80:8178-8189, 2006). We therefore tested the feasibility of using M2 tail mutants as live attenuated vaccines against H5N1 virus. First we generated a series of highly pathogenic H5N1 (A/Vietnam/1203/04 [VN1203]) M2 cytoplasmic tail deletion mutants and examined their growth properties in vitro and in vivo. We found that one mutant, which contains an 11-amino-acid deletion from the C terminus (M2del11 virus), grew as well as the wild-type virus but replicated in mice less efficiently. We then generated a recombinant VN1203M2del11 virus whose hemagglutinin (HA) gene was modified by replacing sequences at the cleavage site with those of an avirulent type of HA (M2del11-HAavir virus). This M2del11-HAavir virus protected mice against challenge with lethal doses of homologous (VN1203; clade 1) and antigenically distinct heterologous (A/Indonesia/7/2005; clade 2) H5N1 viruses. Our results suggest that M2 cytoplasmic tail mutants have potential as live attenuated vaccines against H5N1 influenza viruses. PMID- 18160447 TI - Novel multiplex PCR assay for simultaneous identification of community-associated methicillin-resistant Staphylococcus aureus strains USA300 and USA400 and detection of mecA and Panton-Valentine leukocidin genes, with discrimination of Staphylococcus aureus from coagulase-negative staphylococci. AB - We developed a novel multiplex PCR assay for rapid identification and discrimination of the USA300 and USA400 strains and concomitant detection of Panton-Valentine leukocidin genes, with simultaneous discrimination of methicillin-resistant Staphylococcus aureus strains from methicillin-susceptible S. aureus strains, S. aureus strains from coagulase-negative staphylococci, and staphylococci from other bacteria. PMID- 18160448 TI - Novel loop-mediated isothermal amplification method for detection of the JP2 clone of Aggregatibacter actinomycetemcomitans in subgingival plaque. AB - We developed a loop-mediated isothermal amplification method that detects the JP2 clone of Aggregatibacter actinomycetemcomitans, which induces aggressive periodontitis in adolescents of North and West African descents. Being independent of special equipment, this specific and sensitive method offers significant advantages for screening of patients on a population basis and in clinical settings. PMID- 18160449 TI - Rapid diagnosis of vivax malaria by the SD Bioline Malaria Antigen test when thrombocytopenia is present. AB - An easy and reliable diagnostic method for malaria is highly desirable. We examined the recently introduced SD Bioline Malaria Antigen test, which detects Plasmodium lactate dehydrogenase, with the additional aid of the presence or absence of thrombocytopenia to diagnose vivax malaria. We enrolled 732 patients with clinically suspected malaria in an area where vivax malaria is endemic. We performed microscopic examination of thin film, applied the SD Bioline Malaria Antigen test, and checked platelet counts. One hundred ninety-five patients were smear positive for vivax malaria. The sensitivity of the SD Bioline Malaria Antigen test was 96.4%, and its specificity was 98.9%. We found that 95.4% of malaria patients had thrombocytopenia, and the proportion with malaria increased as platelet counts decreased. A positive SD Bioline Malaria Antigen test when thrombocytopenia was present showed a 100% positive predictive value for vivax malaria. In conclusion, the SD Bioline Malaria Antigen test is a rapid and accurate diagnostic method for vivax malaria, and a platelet count can facilitate a rapid diagnosis of malaria. PMID- 18160450 TI - Identification of clinical coryneform bacterial isolates: comparison of biochemical methods and sequence analysis of 16S rRNA and rpoB genes. AB - We compared the relative levels of effectiveness of three commercial identification kits and three nucleic acid amplification tests for the identification of coryneform bacteria by testing 50 diverse isolates, including 12 well-characterized control strains and 38 organisms obtained from pediatric oncology patients at our institution. Between 33.3 and 75.0% of control strains were correctly identified to the species level by phenotypic systems or nucleic acid amplification assays. The most sensitive tests were the API Coryne system and amplification and sequencing of the 16S rRNA gene using primers optimized for coryneform bacteria, which correctly identified 9 of 12 control isolates to the species level, and all strains with a high-confidence call were correctly identified. Organisms not correctly identified were species not included in the test kit databases or not producing a pattern of reactions included in kit databases or which could not be differentiated among several genospecies based on reaction patterns. Nucleic acid amplification assays had limited abilities to identify some bacteria to the species level, and comparison of sequence homologies was complicated by the inclusion of allele sequences obtained from uncultivated and uncharacterized strains in databases. The utility of rpoB genotyping was limited by the small number of representative gene sequences that are currently available for comparison. The correlation between identifications produced by different classification systems was poor, particularly for clinical isolates. PMID- 18160451 TI - Preparation of armored RNA as a control for multiplex real-time reverse transcription-PCR detection of influenza virus and severe acute respiratory syndrome coronavirus. AB - The common respiratory viruses, including influenza A, influenza B, and newly emerging severe acute respiratory syndrome (SARS) viruses, may cause similar clinical symptoms. Therefore, differential diagnosis of these virus pathogens is frequently required for single clinical samples. In addition, there is an urgent need for noninfectious and stable RNA standards and controls for multivirus detection. In this study, reverse transcription-PCR (RT-PCR) targeting of the RNAs of influenza A and influenza B viruses and SARS coronavirus was performed, and the resulting products were spliced into a fragment which was packaged into armored RNA for use as a noninfectious, quantifiable synthetic substitute. Furthermore, in the present study we developed a multiplex real-time RT-PCR assay in which the armored RNA was used as an external positive control and the three RNA viruses could be detected simultaneously in a single reaction mix. The detection limit of the multiplex real-time PCR was 10 copies/microl of armored RNA. PMID- 18160452 TI - DNA microarray based on arrayed-primer extension technique for identification of pathogenic fungi responsible for invasive and superficial mycoses. AB - An oligonucleotide microarray based on the arrayed-primer extension (APEX) technique has been developed to simultaneously identify pathogenic fungi frequently isolated from invasive and superficial infections. Species-specific oligonucleotide probes complementary to the internal transcribed spacer 1 and 2 (ITS1 and ITS2) region were designed for 24 species belonging to 10 genera, including Candida species (Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida tropicalis, Candida kefyr, Candida krusei, Candida guilliermondii, Candida lusitaniae, Candida metapsilosis, Candida orthopsilosis, Candida parapsilosis, and Candida pulcherrima), Cryptococcus neoformans, Aspergillus species (Aspergillus fumigatus and Aspergillus terreus), Trichophyton species (Trichophyton rubrum and Trichophyton tonsurans), Trichosporon cutaneum, Epidermophyton floccosum, Fusarium solani, Microsporum canis, Penicillium marneffei, and Saccharomyces cerevisiae. The microarray was tested for its specificity with a panel of reference and blinded clinical isolates. The APEX technique was proven to be highly discriminative, leading to unequivocal identification of each species, including the highly related ones C. parapsilosis, C. orthopsilosis, and C. metapsilosis. Because of the satisfactory basic performance traits obtained, such as reproducibility, specificity, and unambiguous interpretation of the results, this new system represents a reliable method of potential use in clinical laboratories for parallel one-shot detection and identification of the most common pathogenic fungi. PMID- 18160453 TI - Determination of accessory gene patterns predicts the same relatedness among strains of Streptococcus pneumoniae as sequencing of housekeeping genes does and represents a novel approach in molecular epidemiology. AB - Relatedness between isolates of Streptococcus pneumoniae can be determined from sequences of multiple genes belonging to the core genome (multilocus sequence typing [MLST]), but these do not provide information on gene content that may affect the potential of isolates to cause invasive pneumococcal disease. Gene content data, obtained using microarrays, were gathered for 40 clinical isolates of 12 serotypes belonging to 30 multilocus sequence types. We found that sequence variations in housekeeping genes assessed by MLST correlated well with whole genome microarray analyses identifying the presence/absence of accessory genes/regions. However, isolates belonging to the same clonal complex, as determined by MLST, may not have identical gene contents, potentially affecting virulence. We found fewer intraclonal (same MLST sequence type) differences associated with pneumococcal serotypes of high invasive disease potential, i.e., serotypes rarely found among carriers compared to serotypes frequently found in carriage. Molecular typing of pneumococci based on the presence/absence of 25 genes localized to accessory regions shows the same relatedness among pneumococcal strains as MLST does. We conclude that molecular typing of pneumococci based on variation in the nucleotide sequences of parts of housekeeping genes (MLST) correlates with the presence/absence of genes in the accessory part of the genome. This covariation is likely due to the fact that both sequence variations and gene content variations are created primarily by recombination events in pneumococci. PMID- 18160454 TI - Comparative study of virulence traits of Escherichia coli clinical isolates causing early and late neonatal sepsis. AB - Neonatal meningitis and septicemia caused by Escherichia coli are still major health problems in industrialized countries. Forty-seven E. coli strains causing neonatal sepsis were analyzed. Twenty-two and 25 strains caused early (detected from 0 to 3 days after birth) and late (detected from 4 to 28 days after birth) infections, respectively. Only the ibeA gene was significantly more prevalent in the strains causing early infections. PMID- 18160455 TI - Acinetobacter septicus sp. nov. association with a nosocomial outbreak of bacteremia in a neonatal intensive care unit. AB - Acinetobacter species other than Acinetobacter baumannii have rarely been reported to be associated with nosocomial outbreaks of bloodstream infections. Within a period of 1 week, seven Acinetobacter-like isolates were recovered from peripheral blood and catheter specimens of five patients at a neonatal intensive care unit (NICU) in a tertiary hospital in Turkey. All five patients had placement of central venous catheters and had received total parenteral nutrition before the onset of bacteremia. Two of the five patients died. Medical devices, tap water, aerators, water samples, various surfaces, intravenous fluids, and the hands of health care workers in the NICU were sampled and were culture negative for the bacterium. All seven of the isolates had identical biochemical reactions, antimicrobial susceptibility results, and pulsed-field gel electrophoresis patterns, indicating a clonal nosocomial outbreak. A panel of standard biochemical reaction profiles and three phenotypic commercial identification systems failed to identify these isolates. Phenotypically, the isolate differed from Acinetobacter ursingii by its hemolysis on sheep blood agar and its negative citrate utilization. Sequences of the full 16S rRNA gene, which contained at least three different gene copies with polymorphic sequences between nucleotide positions 70 and 206, were determined from the first recovered isolate. The complete 1,529- to 1,531-bp 16S rRNA gene sequences and partial 801-bp rpoB gene sequences had similarities of 99.5% and 97.2%, respectively, to an A. ursingii isolate. The DNA-DNA similarities of the strain against the type strain of A. ursingii were 64.7 and 68.7%, which were lower than the recommended threshold value of 70% for the definition of bacterial species. These data indicate that a novel Acinetobacter organism caused the nosocomial outbreak of bacteremia in the NICU unit. We propose the designation of Acinetobacter septicus sp. nov. for these isolates, with isolate AK001 as the type strain. PMID- 18160456 TI - Contribution of the (1-->3)-beta-D-glucan assay for diagnosis of invasive fungal infections. AB - Diagnosis of invasive fungal infection (IFI) remains a challenge. A retrospective study was performed on 279 patients at three French university hospitals to evaluate the performance of the (1-->3)-beta-D-glucan assay (BG assay; Fungitell; Associates of Cape Cod, Inc.) for the diagnosis of IFI. The results of one serum per subject were analyzed for 117 patients who had probable or proven IFI according to the European Organization for Research and Treatment of Cancer criteria (70 invasive pulmonary aspergilloses [IPA], 27 fungal bloodstream infections, and 20 Pneumocystis jiroveci pneumonias), 40 blood donors, and 122 patients who were hospitalized in hematology wards or intensive care units and were at risk for IFI but in whom IFI had not been diagnosed. For the overall IFI diagnosis, the BG assay had 77.8% sensitivity and specificities of 92.5 and 70.5% for blood donors and patients at risk, respectively. The assay was positive in 48 patients with IPA (68%), in 23 with bloodstream infections (85.2%), and in all who had P. jiroveci pneumonias (100%), and the false-positive rate varied depending on the controls used. It allowed a higher rate of detection among IPA patients compared to the galactomannan enzyme-linked immunosorbent assay (ELISA) (48 versus 39 patients, respectively) and among candidemia patients compared to the mannan ELISA (20 versus 11 patients, respectively). This assay therefore appears to be useful in the diagnosis of IFI, particularly for serum analysis of pneumocystosis pneumonia patients, but further studies are needed to evaluate false-positive rates and its future role in IFI diagnosis. PMID- 18160457 TI - The exercise dose response: key lessons from the past. PMID- 18160458 TI - Acylation-stimulating protein deficiency and altered adipose tissue in alternative complement pathway knockout mice. AB - Acylation-stimulating protein (C3adesArg/ASP) is an adipokine that acts on its receptor C5L2 to stimulate triglyceride (TG) synthesis in adipose tissue. The present study investigated ASP levels in mouse models of obesity and leanness and the effect of ASP deficiency in C3 knockout (C3KO) mice on adipose tissue morphology. Plasma ASP levels in wild-type (WT) mice correlated positively with plasma nonesterified fatty acids (NEFA) (R = 0.664, P < 0.001) and total cholesterol (R = 0.515, P < 0.001). Plasma ASP was increased by 85% in obese ob/ob leptin-deficient mice and decreased in lean diacylglycerol acyltransferase 1 (DGAT1) KO mice (-54%) and C/EBPalpha(beta/beta) transgenic mice (-70%) compared with WT. Mice lacking alternative complement factor B or adipsin (FBKO or ADKO), required for ASP production, were also ASP deficient. Both FBKO and C3KO mice had delayed postprandial TG and NEFA clearance on low-fat (LF) and high fat (HF) diets, suggesting that lack of ASP, not C3, drives the metabolic phenotype. Adipocyte size distribution in C3KO mice was polarized (increased number of both small and large cells), with decreased adipsin expression (-33% gonadal HF), DGAT1 expression (-31% to -50%) and DGAT activity (-41%). Overall, a reduction/deficiency in ASP is associated with an antiadipogenic state and ASP may provide a target for controlling fat storage. PMID- 18160459 TI - Generation and characterization of two novel mouse models exhibiting the phenotypes of the metabolic syndrome: Apob48-/-Lepob/ob mice devoid of ApoE or Ldlr. AB - The metabolic syndrome is a group of disorders including obesity, insulin resistance, atherogenic dyslipidemia, hyperglycemia, and hypertension. To date, few animal models have been described to recapitulate the phenotypes of the syndrome. In this study, we generated and characterized two lines of triple knockout mice that are deficient in either apolipoprotein E (Apoe(-/-)) or low density lipoprotein receptor (Ldlr(-/-)) and express no leptin (Lep(ob/ob)) or apolipoprotein B-48 but exclusively apolipoprotein B-100 (Apob(100/100)). These two lines are referred to as Apoe triple-knockout-Apoe 3KO (Apoe(-/ )Apob(100/100)Lep(ob/ob)) and Ldlr triple-knockout-Ldlr 3KO (Ldlr(-/ )Apob(100/100)Lep(ob/ob)) mice. Both lines develop obesity, hyperinsulinemia, hyperlipidemia, hypertension, and atherosclerosis. However, only Apoe 3KO mice are hyperglycemic and glucose intolerant and are more obese than Ldlr 3KO mice. To evaluate the utility of these lines as pharmacological models, we treated both with leptin and found that leptin therapy ameliorated most metabolic derangements. Leptin was more effective in improving glucose tolerance in Ldlr 3KO than Apoe 3KO animals. The reduction of plasma cholesterol by leptin in Ldlr 3KO mice can be accounted for by its suppressive effect on food intake. However, in Apoe 3KO mice, leptin further reduced plasma cholesterol independently of its effect on food intake, and this improvement correlated with a smaller plaque lesion area. These effects suggest a direct role of leptin in modulating VLDL levels and, likewise, the lesion areas in VLDL-enriched animals. These two lines of mice represent new models with features of the metabolic syndrome and will be useful in testing therapies targeted for combating the human condition. PMID- 18160460 TI - Patterns of inheritance of constitutional delay of growth and puberty in families of adolescent girls and boys referred to specialist pediatric care. AB - CONTEXT AND OBJECTIVE: Constitutional delay of growth and puberty (CDGP), more commonly observed in boys than girls, often has a familial background. We characterized the occurrence of CDGP in relatives of CDGP patients to elucidate the mechanisms influencing timing of puberty. PARTICIPANTS AND DESIGN: We identified 492 subjects with CDGP from hospital records of two pediatric clinics in Finland; 95 male and 29 female subjects and their first-degree relatives participated. In family members, CDGP was defined by use of growth charts (growth spurt taking place 2 sd beyond the mean). One third of the families was expanded to include also second-degree relatives with an interview-based assessment of pubertal timing. RESULTS: Of males, 80%, and of female probands, 75% had first degree relatives with CDGP. Of all probands, 45% had one parent (unilineal families) and 32% had two parents affected. In 2% of the families, only siblings were affected. The prevalence of CDGP in male first-degree relatives was only slightly higher than in female relatives: 79 of 148 (53%) vs. 64 of 164 (39%), respectively (P=0.01); male to female ratio was 1.2:1. In 74% of extended unilineal pedigrees (17 of 23), the inheritance pattern of CDGP was consistent with autosomal dominant inheritance. CONCLUSIONS: CDGP clusters in families. Although its inheritance likely is complex, some predisposing genetic factors may have a dominant effect. CDGP was almost as common in male and female relatives of the CDGP subjects seen at specialist care, challenging the view of a marked overall male preponderance of CDGP. PMID- 18160461 TI - Effects of graded doses of testosterone on erythropoiesis in healthy young and older men. AB - CONTEXT: Erythrocytosis is a dose-limiting adverse effect of testosterone therapy, especially in older men. OBJECTIVE: Our objective was to compare the dose-related changes in hemoglobin and hematocrit in young and older men and determine whether age-related differences in erythropoietic response to testosterone can be explained by changes in erythropoietin and soluble transferrin receptor (sTfR) levels. DESIGN: We conducted a secondary analysis of data from a testosterone dose-response study in young and older men who received long-acting GnRH agonist monthly plus one of five weekly doses of testosterone enanthate (25, 50, 125, 300, or 600 mg im) for 20 wk. SETTING: The study took place at a General Clinical Research Center. PARTICIPANTS: Participants included 60 older men aged 60-75 yr and 61 young men aged 19-35 yr. OUTCOME MEASURES: Outcome measures included hematocrit and hemoglobin and serum erythropoietin and sTfR levels. RESULTS: Hemoglobin and hematocrit increased significantly in a linear, dose-dependent fashion in both young and older men in response to graded doses of testosterone (P<0.0001). The increases in hemoglobin and hematocrit were significantly greater in older than young men. There was no significant difference in percent change from baseline in erythropoietin or sTfR levels across groups in either young or older men. Changes in erythropoietin or sTfR levels were not significantly correlated with changes in total or free testosterone levels. CONCLUSIONS: Testosterone has a dose-dependent stimulatory effect on erythropoiesis in men that is more pronounced in older men. The testosterone-induced rise in hemoglobin and hematocrit and age-related differences in response to testosterone therapy may be mediated by factors other than erythropoietin and sTfR. PMID- 18160462 TI - Effects of previous antiresorptive therapy on the bone mineral density response to two years of teriparatide treatment in postmenopausal women with osteoporosis. AB - INTRODUCTION: EUROFORS was a 2-yr prospective, randomized trial of postmenopausal women with established osteoporosis, designed to investigate various sequential treatments after teriparatide 20 microg/d for 1 yr. The present secondary analysis examined the effects of 2 yr of open-label teriparatide in women previously treated with antiresorptive drugs for at least 1 yr. METHODS: A subgroup of 245 women with osteoporosis who had 2 yr of teriparatide treatment were stratified by previous predominant antiresorptive treatment into four groups: alendronate (n=107), risedronate (n=59), etidronate (n=30), and non bisphosphonate (n=49). Bone mineral density (BMD) at the lumbar spine and hip was determined after 6, 12, 18, and 24 months, and bone formation markers were measured after 1 and 6 months. RESULTS: Significant increases in bone formation markers occurred in all groups after 1 month of teriparatide treatment. Lumbar spine BMD increased at all visits, whereas a transient decrease in hip BMD, which was subsequently reversed, was observed in all groups. BMD responses were similar in all previous antiresorptive groups. Previous etidronate users showed a higher increase at the spine but not at the hip BMD. Duration of previous antiresorptive therapy and lag time between stopping previous therapy and starting teriparatide did not affect the BMD response at any skeletal site. Treatment-emergent adverse events were similar to those reported in treatment-naive postmenopausal women with osteoporosis treated with teriparatide. CONCLUSIONS: Teriparatide induces positive effects on BMD and markers of bone formation in postmenopausal women with established osteoporosis, regardless of previous long-term exposure to antiresorptive therapies. PMID- 18160463 TI - Reduced high-molecular-weight adiponectin and elevated high-sensitivity C reactive protein are synergistic risk factors for metabolic syndrome in a large scale middle-aged to elderly population: the Shimanami Health Promoting Program Study. AB - OBJECTIVE: In Western countries, one of the most important modifiable targets for the prevention of cardiovascular diseases is metabolic syndrome. Adiponectin is an adipose tissue-specific plasma protein that inversely associates with metabolic syndrome. Among several molecular isoforms, high-molecular-weight (HMW) complex is considered the active form. Increased serum high-sensitivity C reactive protein (hsCRP) concentration also associates with metabolic syndrome, and adiponectin could modulate plasma C-reactive protein levels. Here, through cross-sectional investigation, we investigated whether reduced HMW adiponectin and increased hsCRP levels in plasma are synergistically associated with metabolic syndrome. Measurement of HMW complex of adiponectin is one of the novelties of this study. DESIGN: We analyzed 1845 community-dwelling middle-aged to elderly subjects (62+/-13 yr). Plasma HMW adiponectin levels were measured by ELISA. Clinical parameters were obtained from the subjects' personal health records, evaluated at their annual medical check-up. RESULTS: Each component of metabolic syndrome, except for raised blood pressure, showed significantly lower plasma HMW adiponectin concentrations for both men and women (P<0.001). In contrast, plasma hsCRP levels were significantly higher in subjects with metabolic disorders (P<0.001). After adjusting for other confounding factors, HMW adiponectin [log normalized, odds ratio 0.084 (95% confidence interval 0.050 0.142), P<0.001] and hsCRP [3.009 (2.175-4.163), P<0.001] were identified as independent determinants of metabolic syndrome. In addition to the direct associations, we also observed a synergistic effect between these two molecules (F=11.8, P<0.001). CONCLUSIONS: Reduced HMW adiponectin and elevated hsCRP are synergistically associated with the accumulation of metabolic disorders. The combination of these markers would be useful for identifying at-risk populations. PMID- 18160464 TI - Higher serum thyroid stimulating hormone level in thyroid nodule patients is associated with greater risks of differentiated thyroid cancer and advanced tumor stage. AB - CONTEXT: TSH is a known thyroid growth factor, but the pathogenic role of TSH in thyroid oncogenesis is unclear. OBJECTIVE: The aim was to examine the relationship between preoperative TSH and differentiated thyroid cancer (DTC). DESIGN: The design was a retrospective cohort. SETTING, PARTICIPANTS: Between May 1994 and January 2007, 1198 patients underwent thyroid surgery at a single hospital. Data from the 843 patients with preoperative serum TSH concentration were recorded. MAIN OUTCOME MEASURES: Serum TSH concentration was measured with a sensitive assay. Diagnoses of DTC vs. benign thyroid disease were based on surgical pathology reports. RESULTS: Twenty-nine percent of patients (241 of 843) had DTC on final pathology. On both univariate and multivariable analyses, risk of malignancy correlated with higher TSH level (P=0.007). The likelihood of malignancy was 16% (nine of 55) when TSH was less than 0.06 mIU/liter vs. 52% (15 of 29) when 5.00 mIU/liter or greater (P=0.001). When TSH was between 0.40 and 1.39 mIU/liter, the likelihood of malignancy was 25% (85 of 347) vs. 35% (109 of 308) when TSH was between 1.40 and 4.99 mIU/liter (P=0.002). The mean TSH was 4.9+/-1.5 mIU/liter in patients with stage III/IV disease vs. 2.1+/-0.2 mIU/liter in patients with stage I/II disease (P=0.002). CONCLUSIONS: The likelihood of thyroid cancer increases with higher serum TSH concentration. Even within normal TSH ranges, a TSH level above the population mean is associated with significantly greater likelihood of thyroid cancer than a TSH below the mean. Shown for the first time, higher TSH level is associated with advanced stage DTC. PMID- 18160465 TI - The exon 3-deleted/full-length growth hormone receptor polymorphism did not influence growth response to growth hormone therapy over two years in prepubertal short children born at term with adequate weight and length for gestational age. AB - CONTEXT: Consensus is lacking as to whether the exon 3-deleted (d3)/full-length (fl) GH receptor (GHR) polymorphism is associated with responsiveness to GH therapy. OBJECTIVE: Our objective was to evaluate, in short, prepubertal, appropriate-for-gestational age (AGA) patients, 2-yr growth response to GH therapy (31.7+/-3.5 microg/kg.d) according to exon 3-deleted/full-length GHR genotypes. DESIGN: We conducted a retrospective study. PATIENTS: We studied 106 short AGA children, 58 boys and 48 girls, 7.8+/-2.3 yr, (d3/d3 n=18, d3/fl n=42, and fl/fl n=46). The GH response to two provocative stimuli were under 10 ng/ml in 65 and one or both over 10 ng/ml in 41 patients. MAIN OUTCOME MEASURES: Patients were followed by a single clinical team and remained prepubertal during the study. The exon 3-deleted/full-length GHR genotypes were determined and analyzed in the same hospital. RESULTS: Growth velocity significantly (P<0.0001) increased during the first and second years of therapy, as did height sd score (SDS). These increases were similar in each exon 3-deleted/full-length GHR genotype. Total 2-yr height gain (SDS) did not differ statistically among genotypes: 15.5+/-2.2 cm and 1.2+/-0.5 SDS in d3/d3, 15.9+/-2.0 cm and 1.3+/-0.4 SDS in d3/fl, and 15.4+/-2.1 cm and 1.1+/-0.3 SDS in fl/fl. No significant differences among the three genotypes were found in both sexes or in patients with different GH peak response to provocative stimuli for these parameters. An analysis of previously published studies was also performed. CONCLUSIONS: These results confirm in AGA patients those previously found by us and others in small for-gestational-age patients and suggest that neither sex nor GH peaks after provocative stimuli might influence significantly the responsiveness to GH therapy according to the exon 3-deleted/full-length GHR genotypes. PMID- 18160466 TI - A functional common polymorphism in the vitamin D-responsive element of the GH1 promoter contributes to isolated growth hormone deficiency. AB - CONTEXT: Causal mutations have been detected only in a minority of isolated GH deficiency (IGHD) patients. Idiopathic IGHD might be the result of the interaction between several low-penetrance genetic factors and the environment. OBJECTIVE: The aim of this study was to test the contribution to IGHD of genetic variations in the GH1 gene regulatory regions. DESIGN AND PATIENTS: A case control association study was performed including 118 sporadic IGHD patients with a nonsevere phenotype (height -4/-1 sd score and partial GH deficiency) and two control groups, normal stature (n=200) and short-stature individuals with normal GH secretion (n=113). Seven single-nucleotide polymorphisms in the GH1 promoter, one in the IVS4 region, and two in the locus control region were analyzed. RESULTS: The -57T allele within the vitamin D-responsive element showed a positive significant association when comparing patients with normal (P=0.006) or short stature (P=0.0011) controls. The genotype -57TT showed an odds ratio of 2.93 (1.44-5.99) and 2.99 (1.42-6.31), respectively. The functional relevance of the -57 variation was demonstrated by the luciferase assay in the presence of vitamin D. The vitamin D-induced inhibition of luciferase activity was significantly (P=0.012) stronger for the promoter haplotype carrying the associated variation -57T [haplotype #1 (hp#1)] with respect to hp#2, bearing 57G. Replacement of the T with a G at -57 on hp#1 abolished the repression, demonstrating that the T at position -57 is necessary to determine the greater vitamin D-induced inhibitory effect of hp#1. EMSA experiments showed a different band-shift pattern of the T and G sequences. CONCLUSION: The common -57G-->T polymorphism contributes to IGHD susceptibility, indicating that it may have a multifactorial etiology. PMID- 18160467 TI - Bone mineral density changes during the menopause transition in a multiethnic cohort of women. AB - CONTEXT: Rates of bone loss across the menopause transition and factors associated with variation in menopausal bone loss are poorly understood. OBJECTIVE: Our objective was to assess rates of bone loss at each stage of the transition and examine major factors that modify those rates. DESIGN, SETTING, AND PARTICIPANTS: We conducted a longitudinal cohort study of 1902 African American, Caucasian, Chinese, or Japanese women participating in The Study of Women's Health Across the Nation. Women were pre- or early perimenopausal at baseline. OUTCOME MEASURE: We assessed bone mineral density (BMD) of the lumbar spine and total hip across a maximum of six annual visits. RESULTS: There was little change in BMD during the pre- or early perimenopause. BMD declined substantially in the late perimenopause, with an average loss of 0.018 and 0.010 g/cm2.yr from the spine and hip, respectively (P<0.001 for both). In the postmenopause, rates of loss from the spine and hip were 0.022 and 0.013 g/cm2.yr, respectively (P<0.001 for both). During the late peri- and postmenopause, bone loss was approximately 35-55% slower in women in the top vs. the bottom tertile of body weight. Apparent ethnic differences in rates of spine bone loss were largely explained by differences in body weight. CONCLUSIONS: Bone loss accelerates substantially in the late perimenopause and continues at a similar pace in the first postmenopausal years. Body weight is a major determinant of the rate of menopausal BMD loss, whereas ethnicity, per se, is not. Healthcare providers should consider this information when deciding when to screen women for osteoporosis. PMID- 18160468 TI - Attenuated morning salivary cortisol concentrations in a population-based study of persons with chronic fatigue syndrome and well controls. AB - CONTEXT: A substantial body of research on the pathophysiology of chronic fatigue syndrome (CFS) has focused on hypothalamic-pituitary-adrenal axis dysregulation. The cortisol awakening response has received particular attention as a marker of hypothalamic-pituitary-adrenal axis dysregulation. OBJECTIVE: The objective of the current study was to evaluate morning salivary cortisol profiles in persons with CFS and well controls identified from the general population. DESIGN AND SETTING: We conducted a case-control study at an outpatient research clinic. CASES AND OTHER PARTICIPANTS: We screened a sample of 19,381 residents of Georgia and identified those with CFS and a matched sample of well controls. Seventy-five medication-free CFS cases and 110 medication-free well controls provided complete sets of saliva samples. MAIN OUTCOME MEASURES: We assessed free cortisol concentrations in saliva collected on a regular workday immediately upon awakening and 30 and 60 min after awakening. RESULTS: There was a significant interaction effect, indicating different profiles of cortisol concentrations over time between groups, with the CFS group showing an attenuated morning cortisol profile. Notably, we observed a sex difference in this effect. Women with CFS exhibited significantly attenuated morning cortisol profiles compared with well women. In contrast, cortisol profiles were similar in men with CFS and male controls. CONCLUSIONS: CFS was associated with an attenuated morning cortisol response, but the effect was limited to women. Our results suggest that a sex difference in hypocortisolism may contribute to increased risk of CFS in women. PMID- 18160469 TI - Common and rare alleles in apolipoprotein B contribute to plasma levels of low density lipoprotein cholesterol in the general population. AB - CONTEXT: We have previously shown that rare mutations in the apolipoprotein B gene (APOB) may result in not only severe hypercholesterolemia and ischemic heart disease but also hypocholesterolemia. Despite this, common single-nucleotide polymorphisms (SNPs) in APOB have not convincingly been demonstrated to affect low-density lipoprotein (LDL) cholesterol levels. OBJECTIVE: We tested the hypothesis that nonsynonymous SNPs in three important functional domains of APOB and APOB tag SNPs predict levels of LDL cholesterol and apolipoprotein B and risk of ischemic heart disease. DESIGN: This was a prospective study with 25 yr 100% follow up, The Copenhagen City Heart Study. SETTING: The study was conducted in the Danish general population. PARTICIPANTS: Participants included 9185 women and men aged 20-80+ yr. MAIN OUTCOME MEASURES: Levels of LDL cholesterol and apolipoprotein B and risk of ischemic heart disease and myocardial infarction were measured. The hypothesis was formulated before genotyping. RESULTS: We genotyped 9185 individuals for APOB T71I (minor allele frequency: 0.33), Ivs4+171c>a (0.14), A591V (0.47), Ivs18+379a>c (0.30), Ivs18+1708g>t (0.45), T2488Tc>t (0.48), P2712L (0.21), R3611Q (0.09), E4154K (0.17), and N4311S (0.21). SNPs were associated with increases (T71I, Ivs181708g>t, T2488Tc>t, R3611) or decreases (Ivs4+171c>a, A591V, Ivs18+379a>c, P2712L, E4154, N4311S) in LDL cholesterol from -4.7 to +8.2% (-0.28 to 0.30 mmol/liter; P/=72 and loss in height were significantly more common among men with fractures of the hip than in those with fractures elsewhere. PMID- 18160504 TI - Wide en bloc extra-articular excision of the elbow for sarcoma with complex reconstruction. AB - We describe a consecutive series of five patients with bone or soft-tissue sarcomas of the elbow and intra-articular extension treated by complex soft tissue, allograft bone and prosthetic joint replacement after wide extra articular en bloc excision. All had a pedicled myocutaneous latissimus dorsi rotation flap for soft-tissue cover and reconstruction of the triceps. Wide negative surgical margins were obtained in all five patients. No local wound complications or infections were seen. There were no local recurrences at a mean follow-up of 60 months (20 to 105). The functional results were excellent in four patients and good in one. Longer term follow-up is necessary to confirm the durability of the elbow reconstruction. PMID- 18160505 TI - The three-pin modified 'Harrington' procedure for advanced metastatic destruction of the acetabulum. AB - Pathological fractures due to metastasis with destruction of the acetabulum and central dislocation of the hip present a difficult surgical challenge. We describe a series using a single technique in which a stable and long-lasting reconstruction was obtained using standard primary hip replacement implants augmented by strong, fully-threaded steel rods with cement and steel mesh, where required. Between 1997 and 2006, 19 patients with a mean age of 66 years (48 to 83) were treated using a modified Harrington technique. Acetabular destruction was graded as Harrington class II in six cases and class III in 13. Reconstruction was achieved using three 6.5 mm rods inserted through a separate incision in the iliac crest followed by augmentation with cement and a conventional cemented Charnley or Exeter primary hip replacement. There were no peri-operative deaths. At the final follow-up (mean 25 months (5 to 110)) one rod had fractured and one construct required revision. Of the 18 patients who did not require revision, 13 had died. The mean time to death was 16 months (5 to 55). The mean follow-up of the five survivors was 31 months (18 to 47). There were no cases of dislocation, deep infection or injury to a nerve, the blood vessels or the bladder. PMID- 18160506 TI - Morphometric changes in the acetabulum after Dega osteotomy in patients with cerebral palsy. AB - We carried out a morphometric analysis of the acetabulum following Dega osteotomy in patients with cerebral palsy using three-dimensional CT. We assessed 17 acetabula in 12 patients with instability of the hip. A Dega osteotomy and varus derotation femoral osteotomy were performed in all 17 hips. Three-dimensional CT scans were taken before and approximately one year after operation. Acetabular cover was evaluated using anterosuperior, superolateral and posterosuperior acetabular indices, and the change in the acetabular volume was calculated. Inter and intra-observer reliability was assessed using the intraclass correlation coefficient. After the osteotomy, the anterosuperior, superolateral and posterosuperior cover had improved significantly towards the value seen in a control group. The mean acetabular volume increased by 68%. PMID- 18160507 TI - Changing incidence of slipped capital femoral epiphysis: a relationship with obesity? AB - Obesity is thought to be an aetiological factor for slipped capital femoral epiphysis (SCFE). We analysed changes in the incidence of SCFE in Scotland over the last two decades. During this period rates of childhood obesity have risen substantially and evidence for a relationship between these changes and the incidence of SCFE was sought. We found that the incidence of SCFE increased from 3.78 per 100,000 children in 1981 to 9.66 per 100,000 in 2000 (R(2) = 0.715): a two and a half times increase over two decades. It was seen at a younger age, with a fall in the mean age at diagnosis from 13.4 to 12.6 years for boys (p = 0.007) and 12.2 to 11.6 for girls (p = 0.047). More children under eight years old were seen with SCFE in Scotland in the decade to 2000 than in the previous decade (p = 0.002, R(2) = 0.346). A close correlation was observed between rising childhood obesity over the last 20 years in Scotland and an increasing incidence of SCFE. PMID- 18160508 TI - Recurrent intravascular papillary endothelial hyperplasia of the right middle finger treated with radiation therapy. AB - We describe a case of recurrent intravascular papillary endothelial hyperplasia involving the middle finger which was successfully-treated with photon and proton radiotherapy following two previous surgical excisions. PMID- 18160509 TI - Cyclical sciatica: endometriosis of the sciatic nerve. AB - We describe a case of sciatic endometriosis in a 25-year-old woman diagnosed by MRI and histology with no evidence of intrapelvic disease. The presentation, diagnosis and management of this rare condition are described. Early diagnosis and treatment are important to prevent irreversible damage to the sciatic nerve. PMID- 18160510 TI - Reattachment of the distal tendon of biceps: factors affecting the failure strength of the repair. AB - Operative fixation is the treatment of choice for a rupture of the distal tendon of biceps. A variety of techniques have been described including transosseous sutures and suture anchors. The poor quality of the bone of the radial tuberosity might affect the load to failure of the tendon repair in early rehabilitation. The aim of this study was to determine the loads to failure of different techniques of fixation and to investigate their association with the bone mineral density of the radial tuberosity. Peripheral quantitative computed tomography was carried out to measure the trabecular and cortical bone mineral density of the radial tuberosity in 40 cadaver specimens. The loads to failure in four different techniques of fixation were determined. The Endobutton-based method showed the highest failure load at 270 N (sd 22) (p < 0.05). The mean failure load of the transosseous suture technique was 210 N (sd 66) and that of the TwinFix-QuickT 5.0 mm was 57 N (sd 22), significantly lower than those of all other repairs (p < 0.05). No significant correlation was seen between bone mineral density and loads to failure. The transosseous technique is an easy and cost-saving procedure for fixation of the distal biceps tendon. TwinFix-QuickT 5.0 mm had significantly lower failure loads, which might affect early rehabilitation, particularly in older patients. PMID- 18160511 TI - Quantification of stem-cement interfacial gaps: in vitro CT analysis of Charnley Kerboul and Lubinus SPII femoral hip implants. AB - Interfacial defects between the cement mantle and a hip implant may arise from constrained shrinkage of the cement or from air introduced during insertion of the stem. Shrinkage-induced interfacial porosity consists of small pores randomly located around the stem, whereas introduced interfacial gaps are large, individual and less uniformly distributed areas of stem-cement separation. Using a validated CT-based technique, we investigated the extent, morphology and distribution of interfacial gaps for two types of stem, the Charnley-Kerboul and the Lubinus SPII, and for two techniques of implantation, line-to-line and undersized. The interfacial gaps were variable and involved a mean of 6.43% (sd 8.99) of the surface of the stem. Neither the type of implant nor the technique of implantation had a significant effect on the regions of the gaps, which occurred more often over the flat areas of the implant than along the corners of the stems, and were more common proximally than distally for Charnley-Kerboul stems cemented line-to-line. Interfacial defects could have a major effect on the stability and survival of the implant. PMID- 18160512 TI - Sealing the skin barrier around transcutaneous implants: in vitro study of keratinocyte proliferation and adhesion in response to surface modifications of titanium alloy. AB - Conventional amputation prostheses rely on the attachment of the socket to the stump, which may lead to soft-tissue complications. Intraosseous transcutaneous amputation prostheses (ITAPs) allow direct loading of the skeleton, but their success is limited by infection resulting from breaching of the skin at the interface with the implant. Keratinocytes provide the skin's primary barrier function, while hemidesmosomes mediate their attachment to natural ITAP analogues. Keratinocytes must attach directly to the surface of the implant. We have assessed the proliferation, morphology and attachment of keratinocytes to four titaniumalloy surfaces in order to determine the optimal topography in vitro. We used immunolocalisation of adhesion complex components, scanning electron microscopy and transmission electron microscopy to assess cell parameters. We have shown that the proliferation, morphology and attachment of keratinocytes are affected by the surface topography of the biomaterials used to support their growth. Smoother surfaces improved adhesion. We postulate that a smooth topography at the point of epithelium-ITAP contact could increase attachment in vivo, producing an effective barrier of infection. PMID- 18160513 TI - In vivo measurement of tissue metabolism in tendons of the rotator cuff. PMID- 18160514 TI - Enhanced production of phospholipase C and perfringolysin O (alpha and theta toxins) in a gatifloxacin-resistant strain of Clostridium perfringens. AB - Clostridium perfringens-induced gas gangrene is mediated by potent extracellular toxins, especially alpha toxin (a phospholipase C [PLC]) and theta toxin (perfringolysin O [PFO], a thiol-activated cytolysin); and antibiotic-induced suppression of toxin synthesis is an important clinical goal. The production of PLC and PFO by a gatifloxacin-induced, fluoroquinolone-resistant mutant strain of C. perfringens, strain 10G, carrying a stable mutation in DNA gyrase was compared with that of the wild-type (WT) parent strain. Zymography (with sheep red blood cell and egg yolk overlays) and time course analysis [with hydrolysis of egg yolk lecithin and O-(4 nitrophenyl-phosphoryl)choline] demonstrated that strain 10G produced more PLC and PFO than the WT strain. Increased toxin production in strain 10G was not related either to differences in growth characteristics between the wild-type and the mutant strain or to nonsynonymous polymorphisms in PLC, PFO, or their known regulatory proteins. Increased PLC and PFO production by strain 10G was associated with increased cytotoxic activity for HT-29 human adenocarcinoma cells and with increased platelet-neutrophil aggregate formation. Four other gatifloxacin-induced gyrase mutants did not show increased toxin production, suggesting that gatifloxacin resistance was not always associated with increased toxin production in all strains of C. perfringens. This is the first report of increased toxin production in a fluoroquinolone-resistant strain of C. perfringens. PMID- 18160515 TI - Fitness of Streptococcus pneumoniae fluoroquinolone-resistant strains with topoisomerase IV recombinant genes. AB - The low prevalence of ciprofloxacin-resistant (Cp r) Streptococcus pneumoniae isolates carrying recombinant topoisomerase IV genes could be attributed to a fitness cost imposed by the horizontal transfer, which often implies the acquisition of larger-than-normal parE-parC intergenic regions. A study of the transcription of these genes and of the fitness cost for 24 isogenic Cp r strains was performed. Six first-level transformants were obtained either with PCR products containing the parC quinolone resistance-determining regions (QRDRs) of S. pneumoniae Cp r mutants with point mutations or with a PCR product that includes parE-QRDR-ant-parC-QRDR from a Cp r Streptococcus mitis isolate. The latter yielded two strains, T6 and T11, carrying parC-QRDR and parE-QRDR-ant-parC QRDR, respectively. These first-level transformants were used as recipients in further transformations with the gyrA-QRDR PCR products to obtain 18 second-level transformants. In addition, strain Tr7 (which contains the GyrA E85K change) was used. Reverse transcription-PCR experiments showed that parE and parC were cotranscribed in R6, T6, and T11; and a single promoter located upstream of parE was identified in R6 by primer extension. The fitness of the transformants was estimated by pairwise competition with R6 in both one-cycle and two-cycle experiments. In the one-cycle experiments, most strains carrying the GyrA E85K change showed a fitness cost; the exception was recombinant T14. In the two-cycle experiments, a fitness cost was observed in most first-level transformants carrying the ParC changes S79F, S79Y, and D83Y and the GyrA E85K change; the exceptions were recombinants T6 and T11. The results suggest that there is no impediment due to a fitness cost for the spread of recombinant Cp r S. pneumoniae isolates, since some recombinants (T6, T11, and T14) exhibited an ability to compensate for the cost. PMID- 18160516 TI - Treatment of experimental anthrax with recombinant capsule depolymerase. AB - Bacillus anthracis produces an antiphagocytic gamma-linked poly-D-glutamic acid capsule that is required for virulence. Capsule depolymerase (CapD) is a membrane associated poly-gamma-glutamate-specific depolymerase encoded on the B. anthracis capsule plasmid, pX02, that is reported to contribute to virulence by anchoring the capsule to the peptidoglycan and partially degrading high-molecular-weight capsule from the bacterial surface. We previously demonstrated that treatment with CapD effectively removes the capsule from anthrax bacilli, rendering them susceptible to phagocytic killing in vitro. Here we report that CapD promoted in vivo phagocytic killing of B. anthracis bacilli by mouse peritoneal neutrophils and that parenteral administration of CapD protected mice in two models of anthrax infection. CapD conferred significant protection compared with controls when coinjected with encapsulated bacilli from fully virulent B. anthracis Ames or the nontoxigenic encapsulated strain Delta Ames and when injected 10 min after infection with encapsulated bacilli from B. anthracis Ames. Protection was also observed when CapD was administered 30 h after infection with B. anthracis Delta Ames spores, while significant protection could not be demonstrated following challenge with B. anthracis Ames spores. These data support the proposed role of capsule in B. anthracis virulence and suggest that strategies to target anthrax bacilli for neutrophil killing may lead to novel postexposure therapies. PMID- 18160517 TI - Urine bactericidal activity against Escherichia coli isolates exhibiting different resistance phenotypes/genotypes in an in vitro pharmacodynamic model simulating urine concentrations obtained after oral administration of a 400 milligram single dose of cefditoren-pivoxil. AB - Activity of simulated cefditoren urinary concentrations was determined against seven Escherichia coli isolates. Bactericidal activity was obtained from 4 to 24 h against TEM-1 (penicillinase production/hyperproduction), TEM-34 (IRT-6), and TEM-116 (extended-spectrum beta-lactamase [ESBL]) and from 6 to 8 h against SHV/TEM-116 (ESBL) but never against SHV/TEM-1 (ESBL). Extension of bactericidal activity depended on the resistance genotype/phenotype tested. PMID- 18160518 TI - Microbicidal properties and cytocidal selectivity of rhesus macaque theta defensins. AB - Rhesus macaque theta-defensins (RTDs) are unique macrocyclic antimicrobial peptides. The three RTDs (RTD 1-3), isolated from macaque leukocytes, have broad spectrum antimicrobial activities in vitro and share certain structural features with acyclic porcine protegrins, which are microbicidal peptides of the cathelicidin family. To understand the structural features that confer the respective cytocidal properties to theta-defensins and protegrins, we determined and compared the biological properties of RTD 1-3 and protegrin 1 (PG-1) in assays for antimicrobial activity, bacterial membrane permeabilization, and toxicity to human cells. RTD 1-3 and PG-1 had similar microbicidal potencies against Escherichia coli, Staphylococcus aureus, and Candida albicans in low ionic-strength (10 mM) buffers at pH 7.4. The inclusion of physiologic sodium chloride partially inhibited the microbicidal activities of the RTDs, and the degree of inhibition depended on the buffer used in the assay. Similarly, the inclusion of 10% normal human serum partially antagonized the bactericidal activities of all four peptides. In contrast, the microbicidal activities of PG-1 and RTD 1-3 against E. coli were unaffected by physiologic concentrations of calcium chloride and magnesium chloride. Treatment of E. coli ML35 cells with RTD 1-3 or PG-1 rapidly rendered the bacteria permeable to omicron-nitrophenyl-beta-D galactopyranoside, and this was accompanied by the rapid entry of the RTDs. Finally, although PG-1 was toxic to human fibroblasts and caused a marked lysis of erythrocytes, the RTDs were not cytotoxic or hemolytic. Thus, compared to PG 1, RTD 1-3 possess substantially greater cytocidal selectivity against microbes. Surprisingly, the low cytotoxicity of the RTDs did not depend on the peptides' cyclic conformation. PMID- 18160519 TI - Effects of Escherichia coli lipopolysaccharide on telithromycin pharmacokinetics in rats: inhibition of metabolism via CYP3A. AB - It has been reported that telithromycin is metabolized primarily via hepatic microsomal cytochrome P450 (CYP) 3A1/2 in rats and that the expression of hepatic and intestinal CYP3A decreases in rats pretreated with Escherichia coli lipopolysaccharide (ECLPS rats; an animal model of inflammation). Thus, it is possible that the area under the plasma concentration-time curve from 0 h to infinity (AUC 0-infinity) of intravenous and oral telithromycin is greater for ECLPS rats than for the controls. To assess this, the pharmacokinetic parameters of telithromycin were compared after intravenous and oral administration (50 mg/kg). After intravenous administration of telithromycin, the AUC 0-infinity was significantly greater (by 83.4%) in ECLPS rats due to a significantly lower nonrenal clearance (by 44.5%) than in the controls. This may have been due to a significantly decreased hepatic metabolism of telithromycin in ECLPS rats. After oral administration of telithromycin, the AUC 0-infinity in ECLPS rats was also significantly greater (by 140%) than in the controls and the increase was considerably greater than the 83.4% increase after intravenous administration. This could have been due to a decrease in intestinal metabolism in addition to a decreased hepatic metabolism of telithromycin in ECLPS rats. PMID- 18160520 TI - E240V substitution increases catalytic efficiency toward ceftazidime in a new natural TEM-type extended-spectrum beta-lactamase, TEM-149, from Enterobacter aerogenes and Serratia marcescens clinical isolates. AB - The aim of this study was to characterize a novel extended-spectrum beta lactamase that belongs to the TEM family, the TEM-149 enzyme, and that was isolated from the urine of two hospitalized patients from different hospitals in southern Italy. The peculiarity of this enzyme was the finding of a valine residue at position 240. The array of amino acid substitutions found in TEM-149 was as follows: E104K, R164S, M182T, and E240V. A reversion of a threonine residue at position 182 was also performed to create a new mutant, TEM-149 T182M, in order to assess the contribution of this substitution on the kinetic profile and the stability of TEM-149. The bla TEM-149 and bla TEM-149/T182M genes were cloned into pBC-SK, and the corresponding enzymes were purified from recombinant Escherichia coli HB101 by the same procedure. Both enzymes hydrolyzed all beta lactams tested, with a preference for ceftazidime, which was found to be the best substrate. By comparison of the kinetic parameters of the TEM-149 and the TEM-149 T182M enzymes, a reduction of the catalytic efficiency for the TEM-149 T182M mutant was observed against all substrates tested except benzylpenicillin, cefotaxime, and aztreonam. Tazobactam, clavulanic acid, and sulbactam were good inhibitors of the TEM-149 beta-lactamase. PMID- 18160521 TI - The naphthyridinone GSK364735 is a novel, potent human immunodeficiency virus type 1 integrase inhibitor and antiretroviral. AB - The naphthyridinone GSK364735 potently inhibited recombinant human immunodeficiency virus type 1 (HIV-1) integrase in a strand transfer assay (mean 50% inhibitory concentration +/- standard deviation, 8 +/- 2 nM). As expected based on the structure of the drug, it bound competitively with another two-metal binding inhibitor (Kd [binding constant], 6 +/- 4 nM). In a number of different cellular assays, GSK364735 inhibited HIV replication with potency at nanomolar concentrations (e.g., in peripheral blood mononuclear cells and MT-4 cells, 50% effective concentrations were 1.2 +/- 0.4 and 5 +/- 1 nM, respectively), with selectivity indexes of antiviral activity versus in-assay cytotoxicity of at least 2,200. When human serum was added, the antiviral potency decreased (e.g., a 35-fold decrease in the presence of 100% human serum was calculated by extrapolation from the results of the MT-4 cell assay). In cellular assays, GSK364735 blocked viral DNA integration, with a concomitant increase in two-long terminal-repeat circles. As expected, this integrase inhibitor was equally active against wild-type viruses and mutant viruses resistant to approved drugs targeting either reverse transcriptase or protease. In contrast, some but not all viruses resistant to other integrase inhibitors were resistant to GSK364735. When virus was passaged in the presence of the inhibitor, we identified resistance mutations within the integrase active site that were the same as or similar to mutations arising in response to other two-metal binding inhibitors. Finally, either additive or synergistic effects were observed when GSK364735 was tested in combination with approved antiretrovirals (i.e., no antagonistic effects were seen). Thus, based on all the data, GSK364735 exerted potent antiviral activity through the inhibition of viral DNA integration by interacting at the two-metal binding site within the catalytic center of HIV integrase. PMID- 18160522 TI - Fluoroquinolone-resistant mutants of Burkholderia cepacia. AB - Fluoroquinolone-resistant Burkholderia cepacia mutants were selected on ciprofloxacin. The rate of mutation in gyrA was estimated to be 9.6 x 10(-11) mutations per division. Mutations in gyrA conferred 12- to 64-fold increases in MIC, and an additional parC mutation conferred a large increase in MIC (>256 fold). Growth rate, biofilm formation, and survival in water and during drying were not impaired in strains containing single gyrA mutations. Double mutants were impaired only in growth rate (0.85, relative to the susceptible parent). PMID- 18160523 TI - Highly variable penicillin resistance determinants PBP 2x, PBP 2b, and PBP 1a in isolates of two Streptococcus pneumoniae clonal groups, Poland 23F-16 and Poland 6B-20. AB - Penicillin-binding proteins (PBPs) in representatives of two Streptococcus pneumoniae clonal groups that are prevalent in Poland, Poland 23F-16 and Poland 6B-20, were investigated by PBP profile analysis, antibody reactivity pattern analysis, and DNA sequence analysis of the transpeptidase (TP) domain-encoding regions of the pbp2x, pbp2b, and pbp1a genes. The isolates differed in their MICs of beta-lactam antibiotics. The majority of the 6B isolates were intermediately susceptible to penicillin (penicillin MICs, 0.12 to 0.5 microg/ml), whereas all 23F isolates were penicillin resistant (MICs, >or=2 microg/ml). The 6B isolates investigated had the same sequence type (ST), determined by multilocus sequence typing, as the Poland 6B-20 reference strain (ST315), but in the 23F group, isolates with three distinct single-locus variants (SLVs) in the ddl gene (ST173, ST272, and ST1506) were included. None of the isolates showed an identical PBP profile after labeling with Bocillin FL and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and only one pair of 6B isolates and one pair of 23F isolates (ST173 and ST272) each contained an identical combination of PBP 2x, PBP 2b, and PBP 1a TP domains. Some 23F isolates contained PBP 3 with an apparently higher electrophoretic mobility, and this feature also did not correlate with their STs. The data document a highly variable pool of PBP genes as a result of multiple gene transfer and recombination events within and between different clonal groups. PMID- 18160524 TI - Tipranavir (TPV) genotypic inhibitory quotient predicts virological response at 48 weeks to TPV-based salvage regimens. AB - The virological response (VR) to a tipranavir-ritonavir (TPV-RTV)-based regimen had been shown to be associated with a number of mutations in the protease gene, the use of enfuvirtide (T20), and the TPV phenotypic inhibitory quotient (IQ). The role of the TPV genotypic IQ (gIQ) has not yet been fully investigated. The aim of our study was to evaluate the relationship between the TPV gIQ and the VR at 48 weeks to TPV-based salvage regimens. Patients placed on regimens containing two nucleoside reverse transcriptase inhibitors plus TPV-RTV 500/200 mg twice a day with or without T20 were prospectively studied. Regular follow-up was performed over the study period. VR, considered a viral load (VL) decrease of >or=1 log unit and/or the achievement of <50 copies/ml with no VL rebound of >0.5 log unit compared to the maximal VL decrease at week 48, was assessed. Thirty eight patients who had received multiple drugs were included. At week 48 the VL decrease was -1.48 (interquartile range [IQR], -2.88 to -0.48), 15 patients (39.5%) had VLs of <50 copies/ml, and the CD4+ cell count increase was 37 cells/mm3 (IQR, -30 to +175). Twenty subjects (52.6%) achieved VRs. The TPV gIQ and optimized background score (OBS) were independently associated with higher VL decreases. The TPV gIQ and OBS were also independent predictors of a VR at week 48. TPV gIQ and OBS cutoff values of 14,500 and 2, respectively, were associated with a higher rate of VR. The TPV gIQ was shown to be able to predict the VR at 48 weeks to TPV-containing salvage regimens better than the TPV trough concentration or TPV-associated mutations alone. A possible TPV gIQ cutoff value of 14,500 for reaching a VR at week 48 was suggested. Further studies are needed in order to evaluate the calculation of TPV gIQ as a new tool for the optimization of TPV-based salvage therapy. PMID- 18160525 TI - Bobel-24 activity against Cryptosporidium parvum in cell culture and in a SCID mouse model. AB - The anticryptosporidial activity of Bobel-24 (2,4,6-triiodophenol) was studied for the first time, resulting in a reduction of the in vitro growth of Cryptosporidium of up to 99.6%. In a SCID mouse model of chronic cryptosporidiosis, significant differences (P < 0.05) in oocyst shedding were observed in animals treated with 125 mg/kg/day. These results merit further investigation of Bobel-24 as a chemotherapeutic option for cryptosporidiosis. PMID- 18160526 TI - Assessment of serum (1->3)-beta-D-glucan concentration as a measure of disease burden in a murine model of invasive pulmonary aspergillosis. AB - Serum (1-->3)-beta-D-glucan concentrations were serially measured in the presence and absence of antifungal therapy in a murine model of invasive pulmonary aspergillosis. Serum (1-->3)-beta-D-glucan was detected early during the course of infection, and reductions in this biomarker were associated with improved survival in animals treated with antifungal agents. PMID- 18160527 TI - Elective cesarean delivery affects gut maturation and delays microbial colonization but does not increase necrotizing enterocolitis in preterm pigs. AB - Although preterm birth and formula feeding increase the risk of necrotizing enterocolitis (NEC), the influences of cesarean section (CS) and vaginal delivery (VD) are unknown. Therefore, gut characteristics and NEC incidence and severity were evaluated in preterm pigs (92% gestation) delivered by CS or VD. An initial study showed that newborn CS pigs (n = 6) had decreased gastric acid secretion, absorption of intact proteins, activity of brush-border enzymes and pancreatic hydrolases, plasma cortisol, rectal temperature, and changes in blood chemistry, indicating impaired respiratory function, compared with VD littermates (n = 6). In a second experiment, preterm CS (n = 16) and VD (n = 16) pigs were given total parenteral nutrition (36 h) then fed porcine colostrum (VD-COL, n = 6; CS-COL, n = 6) or infant milk formula (VD-FORM, n = 10; CS-FORM, n = 10) for 2 days. Across delivery, FORM pigs showed significantly higher NEC incidence, tissue proinflammatory cytokines (IFN-gamma and IL-6), Clostridium colonization, and impaired intestinal function, compared with COL pigs. NEC incidence was equal for CS (6/16) and VD (6/16) pigs, CS pigs had decreased bacterial diversity and density, higher villus heights, and increased brush-border enzyme activities (lactase, aminopeptidases) compared with VD pigs. In particular, VD-FORM pigs showed reduced mucosal proportions, reduced lactase and aminopeptidases, and increased proinflammatory cytokine IL-6 compared with CS-FORM (P < 0.06). Despite the initial improvement of intestinal and metabolic functions following VD, gut function, and inflammation were similar, or more negatively affected in VD neonates than CS neonates. Both delivery modes exhibited positive and negative influences on the preterm gut, which may explain the similar NEC incidence. PMID- 18160528 TI - Adverse effects of pneumoperitoneum on renal function: involvement of the endothelin and nitric oxide systems. AB - Increased intra-abdominal pressure (IAP) during laparoscopy adversely affects kidney function. The mechanism underlying this phenomenon is largely unknown. This study was designed to investigate the involvement of endothelin (ET)-1 and nitric oxide (NO) systems in IAP-induced renal dysfunction. Rats were subjected to IAP of 14 mmHg for 1 h, followed by a deflation for 60 min (recovery). Four additional groups were pretreated with 1) ABT-627, an ET(A) antagonist; 2) A 192621, an ET(B) antagonist; 3) nitroglycerine; and 4) N(G)-nitro-L-arginine methyl ester, a NO synthase inhibitor, before IAP. Urine flow rate (V), absolute Na+ excretion (U(Na)V), glomerular filtration rate (GFR), and renal plasma flow (RPF) were determined. Significant reductions in kidney function and hemodynamics were observed when IAP was applied. V decreased from 8.1 +/- 1.0 to 5.8 +/- 0.5 microl/min, U(Na)V from 1.08 +/- 0.31 to 0.43 +/- 0.10 microeq/min, GFR from 1.84 +/- 0.12 to 1.05 +/- 0.06 ml/min (-46.9 +/- 2.7% from baseline), and RPF from 8.62 +/- 0.87 to 3.82 +/- 0.16 ml/min (-54 +/- 3.5% from baseline). When the animals were pretreated with either ABT-627 or A-192621, given alone or combined, the adverse effects of IAP on GFR, RPF, V, and U(Na)V were significantly augmented. When the animals were pretreated with nitroglycerine, the adverse effects of pneumoperitoneum on GFR and RPF were substantially improved. In contrast, pretreatment with N(G)-nitro-L-arginine methyl ester remarkably aggravated pneumoperitoneum-induced renal dysfunction. In conclusion, decreased renal excretory function and hypofiltration are induced by increased IAP. These effects are related to impairment of renal hemodynamics and could be partially ameliorated by pretreatment with nitroglycerine and aggravated by NO and ET blockade. PMID- 18160529 TI - Characterization of mice lacking the gene for cholecystokinin. AB - CCK acts peripherally as a satiating peptide released during meals in response to lipid feeding and centrally functions in the modulation of feeding, exploratory, and memory activities. The present study determined metabolic parameters, food intake, anxiety-like behaviors, and cognitive function in mice lacking the CCK gene. We studied intestinal fat absorption, body composition, and food intake of CCK knockout (CCK-KO) mice by using the noninvasive measurement of intestinal fat absorption along with quantitative magnetic resonance (QMR) imaging and the DietMax system, respectively. Additionally, exploratory and memory capacities were assessed by monitoring running wheel activity and conducting elevated plus maze and Morris water-maze tests with these mice. Compared with wild-type (WT) littermate controls, CCK-KO mice had normal food intake, fat absorption, body weight, and body mass. CCK-KO mice ate more food than control animals during the light period and less food during the dark period. Energy expenditure was unchanged between the genotypes; however, CCK-KO mice displayed greater fatty acid oxidation. CCK-KO mice were as active as WT animals in the running wheel test. CCK-KO mice spent more time in the closed arms of an elevated plus-maze, indicative of increased anxiety. Additionally, CCK-KO mice exhibited attenuated performance in a passive avoidance task and impaired spatial memory in the Morris water maze test. We conclude that CCK is involved in metabolic rate and is important for memory and exploration. CCK is intimately involved in multiple processes related to cognitive function and food intake regulation. PMID- 18160530 TI - Neuroregulation of nonexercise activity thermogenesis and obesity resistance. AB - High levels of spontaneous physical activity in lean people and the nonexercise activity thermogenesis (NEAT) derived from that activity appear to protect lean people from obesity during caloric challenge, while obesity in humans is characterized by dramatically reduced spontaneous physical activity. We have similarly demonstrated that obesity-resistant rats have significantly greater spontaneous physical activity than obesity-prone rats, and that spontaneous physical activity predicts body weight gain. Although the energetic cost of activity varies between types of activity and may be regulated, individual level of spontaneous physical activity is important in determining propensity for obesity. We review the current status of knowledge about the brain mechanisms involved in controlling the level of spontaneous physical activity and the NEAT so generated. Focus is on potential neural mediators of spontaneous physical activity and NEAT, including orexin A (also known as hypocretin 1), agouti related protein, ghrelin, and neuromedin U, in addition to brief mention of neuropeptide Y, corticotrophin releasing hormone, cholecystokinin, estrogen, leptin, and dopamine effects on spontaneous physical activity. We further review evidence that strain differences in orexin stimulation pathways for spontaneous physical activity and NEAT appear to track with the body weight phenotype, thus providing a potential mechanistic explanation for reduced activity and weight gain. PMID- 18160531 TI - A further observation of the structural changes of microvessels in the extensor digitorum longus muscle of the aged rat. AB - We further examined the structural changes of microvessels in the extensor digitorum longus muscle of the aged (18 months) rat. Muscle bundles in this aged muscle constantly consisted of numerous large muscle fibers 50-60 mum in diameter and a few small muscle fibers <30 mum in diameter. Neuromuscular junctions (NMJs) in large muscle fibers often showed degenerative figures, thus degenerating muscle fibers. On the other hand, NMJs in small muscle fibers were mainly characterized by sparse and short junctional folds, being possibly in the course of regeneration. In some muscle bundles, the extracellular matrix was a little widened. Microvascular networks from arterioles to venules via capillaries seemed to vary in structural features between muscle bundles. In addition to the normal microvascular network consisting of microvessels with a round or oval vascular lumen during their course, two different types of microvascular networks were found. One type was characterized by the constriction of arterioles, capillaries and venules, probably representing a degenerative process of the microvascular network. In fact, uneven and compressed scaffolds of basal laminae of capillaries were often observed around these constricted microvessels. The other type consisted of arterioles and capillaries with an irregular slit-like vascular lumen and venules with a round or oval vascular lumen, and these capillaries had thick or two-layered basal laminae, being probably in the course of remodeling of the microvascular network. From these findings, it is suggested that the constriction and/or contraction of microvessels by smooth muscle cells and pericytes may be involved in the degeneration and remodeling of the microvascular network in the muscle bundles following degeneration and regeneration of the muscle fibers. PMID- 18160532 TI - The thyroid hormone mimetic compound KB2115 lowers plasma LDL cholesterol and stimulates bile acid synthesis without cardiac effects in humans. AB - Atherosclerotic cardiovascular disease is a major problem despite the availability of drugs that influence major risk factors. New treatments are needed, and there is growing interest in therapies that may have multiple actions. Thyroid hormone modulates several cardiovascular risk factors and delays atherosclerosis progression in humans. However, use of thyroid hormone is limited by side effects, especially in the heart. To overcome this limitation, pharmacologically selective thyromimetics that mimic metabolic effects of thyroid hormone and bypass side effects are under development. In animal models, such thyromimetics have been shown to stimulate cholesterol elimination through LDL and HDL pathways and decrease body weight without eliciting side effects. We report here studies on a selective thyromimetic [KB2115; (3-[[3,5-dibromo-4-[4 hydroxy-3-(1-methylethyl)-phenoxy]-phenyl]-amino]-3-oxopropanoic acid)] in humans. In moderately overweight and hypercholesterolemic subjects KB2115 was found to be safe and well tolerated and elicited up to a 40% lowering of total and LDL cholesterol after 14 days of treatment. Bile acid synthesis was stimulated without evidence of increased cholesterol production, indicating that KB2115 induced net cholesterol excretion. KB2115 did not provoke detectable effects on the heart, suggesting that the pharmacological selectivity observed in animal models translates to humans. Thus, selective thyromimetics deserve further study as agents to treat dyslipidemia and other risk factors for atherosclerosis. PMID- 18160533 TI - Effect of aging on regional cerebral blood flow responses associated with osmotic thirst and its satiation by water drinking: a PET study. AB - Levels of thirst and ad libitum drinking decrease with advancing age, making older people vulnerable to dehydration. This study investigated age-related changes in brain responses to thirst and drinking in healthy men. Thirst was induced with hypertonic infusions (3.1 ml/kg 0.51M NaCl) in young (Y) and older (O) subjects. Regional cerebral blood flow (rCBF) was measured with positron emission tomography (PET). Thirst activations were identified by correlating rCBF with thirst ratings. Average rCBF was measured from regions of interest (ROI) corresponding to activation clusters in each group. The effects of drinking were examined by correlating volume of water drunk with changes in ROI rCBF from maximum thirst to postdrinking. There were increases in blood osmolality (Y, 2.8 +/- 1.8%; O, 2.2 +/- 1.4%) and thirst ratings (Y, 3.1 +/- 2.1; O, 3.7 +/- 2.8) from baseline to the end of the hypertonic infusion. Older subjects drank less water (1.9 +/- 1.6 ml/kg) than younger subjects (3.9 +/- 1.9 ml/kg). Thirst related activation was evident in S1/M1, prefrontal cortex, anterior midcingulate cortex (aMCC), premotor cortex, and superior temporal gyrus in both groups. Postdrinking changes of rCBF in the aMCC correlated with drinking volumes in both groups. There was a greater reduction in aMCC rCBF relative to water drunk in the older group. Aging is associated with changes in satiation that militate against adequate hydration in response to hyperosmolarity, although it is unclear whether these alterations are due to changes in primary afferent inflow or higher cortical functioning. PMID- 18160534 TI - Multiroute memories in desert ants. AB - When offered a permanent food source, central Australian desert ants, Melophorus bagoti, develop individually distinct, view-based foraging routes, which they retrace with amazing accuracy during each foraging trip. Using a particular channel setup connected to an artificial feeder, we trained M. bagoti ants to either two or three inward routes that led through different parts of their maze like foraging grounds. Here, we show that ants are able to adopt multiple habitual paths in succession and that they preserve initially acquired route memories even after they have been trained to new routes. Individual ants differ in the consistency with which they run along habitual pathways. However, those ants that follow constant paths retain their route-specific memories for at least 5 days of suspended foraging, which suggests that even multiple route memories, once acquired, are preserved over the entire lifetime of a forager. PMID- 18160535 TI - Three-dimensional cellular microarray for high-throughput toxicology assays. AB - We have developed a miniaturized 3D cell-culture array (the Data Analysis Toxicology Assay Chip or DataChip) for high-throughput toxicity screening of drug candidates and their cytochrome P450-generated metabolites. The DataChip consists of human cells encapsulated in collagen or alginate gels (as small as 20 nl) arrayed on a functionalized glass slide for spatially addressable screening against multiple compounds. A single DataChip containing 1,080 individual cell cultures, used in conjunction with the complementary human P450-containing microarray (the Metabolizing Enzyme Toxicology Assay Chip or MetaChip), simultaneously provided IC(50) values for nine compounds and their metabolites from CYP1A2, CYP2D6, and CYP3A4 and a mixture of the three P450s designed to emulate the human liver. Similar responses were obtained with the DataChip and conventional 96-well plate assays, demonstrating that the near 2,000-fold miniaturization does not influence the cytotoxicity response. The DataChip may therefore enable toxicity analyses of drug candidates and their metabolites at throughputs compatible with the availability of compounds at early-stage drug discovery. PMID- 18160536 TI - Evolutionarily conserved gene family important for fat storage. AB - The ability to store fat in the form of cytoplasmic triglyceride droplets is conserved from Saccharomyces cerevisiae to humans. Although much is known regarding the composition and catabolism of lipid droplets, the molecular components necessary for the biogenesis of lipid droplets have remained obscure. Here we report the characterization of a conserved gene family important for lipid droplet formation named fat-inducing transcript (FIT). FIT1 and FIT2 are endoplasmic reticulum resident membrane proteins that induce lipid droplet accumulation in cell culture and when expressed in mouse liver. shRNA silencing of FIT2 in 3T3-LI adipocytes prevents accumulation of lipid droplets, and depletion of FIT2 in zebrafish blocks diet-induced accumulation of lipid droplets in the intestine and liver, highlighting an important role for FIT2 in lipid droplet formation in vivo. Together these studies identify and characterize a conserved gene family that is important in the fundamental process of storing fat. PMID- 18160537 TI - p53 and TFIIEalpha share a common binding site on the Tfb1/p62 subunit of TFIIH. AB - The general transcription factor IIH is recruited to the transcription preinitiation complex through an interaction between its p62/Tfb1 subunit and the alpha-subunit of the general transcription factor IIE (TFIIEalpha). We have determined that the acidic carboxyl terminus of TFIIEalpha (TFIIEalpha(336-439)) directly binds the amino-terminal PH domain of p62/Tfb1 with nanomolar affinity. NMR mapping and mutagenesis studies demonstrate that the TFIIEalpha binding site on p62/Tfb1 is identical to the binding site for the second transactivation domain of p53 (p53 TAD2). In addition, we demonstrate that TFIIEalpha(336-439) is capable of competing with p53 for a common binding site on p62/Tfb1 and that TFIIEalpha(336-439) and the diphosphorylated form (pS46/pT55) of p53 TAD2 have similar binding constants. NMR structural studies reveal that TFIIEalpha(336-439) contains a small domain (residues 395-433) folded in a novel betabetaalphaalphaalpha topology. NMR mapping studies demonstrate that two unstructured regions (residues 377-393 and residues 433-439) located on either side of the folded domain appear to be required for TFIIEalpha(336-439) binding to p62/Tfb1 and that these two unstructured regions are held close to each other in three-dimensional space by the novel structured domain. We also demonstrate that, like p53, TFIIEalpha(336-439) can activate transcription in vivo. These results point to an important interplay between the general transcription factor TFIIEalpha and the tumor suppressor protein p53 in regulating transcriptional activation that may be modulated by the phosphorylation status of p53. PMID- 18160538 TI - The re-emerging role of the intestinal microflora in critical illness and inflammation: why the gut hypothesis of sepsis syndrome will not go away. AB - Recent advances in the ability to genetically interrogate microbial communities within the intestinal tract of humans have revealed many striking findings. That there may be as many as 300 unculturable and unclassified microbes within the human intestinal tract opens the possibility that yet-unidentified microbes may play a role in various human diseases [( 1) ]. Technologically, the regional and spatial aspects of intestinal microbial communities can now be better appreciated by emerging genetic and in vivo imaging systems using a bioinformatics approach [( 2) ]. Finally, in situ PCR of tissues and blood now allows the detection of microbes at concentrations that would otherwise remain undetected by culture alone [( 3) ]. In the aggregate, these studies have empowered clinicians to readdress the issue of how our microbial partners are affected by extreme states of physiologic stress and antibiotic use through the course of critical illness. The role of microbes in systemic inflammatory states, such as systemic inflammatory response syndrome, as well as in primary intestinal mucosal diseases, such as necrotizing enterocolitis, inflammatory bowel disease, and ischemia-reperfusion injury, can now be more completely defined, and the microbial genes that mediate the immune activation during these disorders can be identified. The 2008 roadmap initiative at the National Institutes of Health to fully define the human microbiome is further testament to the power of this technology and the importance of understanding how intestinal microbes, their genes, and their gene products affect the course of human disease and inflammation. PMID- 18160540 TI - The role of epithelial Toll-like receptor signaling in the pathogenesis of intestinal inflammation. AB - Emerging evidence suggests that the innate immune system, comprised of Toll-like receptors (TLRs) and their associated molecules, plays a pivotal role in the regulation of intestinal inflammation and in the response to invading pathogens. Although TLRs are thought to have predominantly beneficial effects in pathogen recognition and bacterial clearance by leukocytes, their dysregulation and unique signaling effects within intestinal epithelia in the setting of inflammation may have devastating consequences. For instance, activation of TLR4 in enterocytes leads to an inhibition of enterocyte migration and proliferation as well as the induction of enterocyte apoptosis-factors that would be expected to promote intestinal injury while inhibiting intestinal repair. TLR signaling has been shown to be abnormal in several intestinal inflammatory diseases, including Crohn's disease, ulcerative colitis, and necrotizing enterocolitis. This review serves to examine the evidence regarding the patterns of expression and signaling of TLRs in the intestinal mucosa at basal levels and during physiologic stressors to gain insights into the pathogenesis of intestinal inflammation. We conclude that the data reviewed suggest that epithelial TLR signaling-acting in concert with TLR signaling by leukocytes-participates in the development of intestinal inflammation. We further conclude that the evidence reviewed provides a rationale for the development of novel, epithelial-specific, TLR-based agents in the management of diseases of intestinal inflammation. PMID- 18160539 TI - A2A receptors in inflammation and injury: lessons learned from transgenic animals. AB - Adenosine regulates the function of the innate and adaptive immune systems through targeting virtually every cell type that is involved in orchestrating an immune/inflammatory response. Of the four adenosine receptors (A(1), A(2A), A(2B), A(3)), A(2A) receptors have taken center stage as the primary anti inflammatory effectors of extracellular adenosine. This broad, anti-inflammatory effect of A(2A) receptor activation is a result of the predominant expression of A(2A) receptors on monocytes/macrophages, dendritic cells, mast cells, neutrophils, endothelial cells, eosinophils, epithelial cells, as well as lymphocytes, NK cells, and NKT cells. A(2A) receptor activation inhibits early and late events occurring during an immune response, which include antigen presentation, costimulation, immune cell trafficking, immune cell proliferation, proinflammatory cytokine production, and cytotoxicity. In addition to limiting inflammation, A(2A) receptors participate in tissue remodeling and reparation. Consistent with their multifaceted, immunoregulatory action on immune cells, A(2A) receptors have been shown to impact the course of a wide spectrum of ischemic, autoimmune, infectious, and allergic diseases. Here, we review the regulatory roles of A(2A) receptors in immune/inflammatory diseases of various organs, including heart, lung, gut, liver, kidney, joints, and brain, as well as the role of A(2A) receptors in regulating multiple organ failure and sepsis. PMID- 18160541 TI - Walking habits in elderly widows. AB - Walking habits were studied in 3 groups of elderly widows. The average walking time per week was calculated from interviews or questionnaires. There was in a small studied group a tendency for walking time to be lower at 3 and 12 months after loss than at 4 or 5 years. An increased odds ratio was demonstrated in larger groups of widows for walking less than 120 minutes per week in those who "did not feel healthy," or who had "musculoskeletal health problems," or "cardiovascular health problems." Widows from a population-based study also showed increased odds ratio for not walking as long with "lack of friends" and "not being active in associations." This was not found in married women from the population study. Our results indicate that newly bereaved women may reduce their physical activity, and that the change in exercise habits may be associated with reduced perception of being healthy and a decreased social network. PMID- 18160542 TI - A poor prognosis: guide or misleading? AB - Patients admitted to a palliative care unit have a limited life expectancy. Sometimes, however, the accuracy of the prognosis is compromised by atypical progress of a disease or incorrect assumptions. If a predicted short life expectancy proves to be longer, the increased need of care may result in chronic admission and in a significantly reduced quality of life. A given poor prognosis may be mistakenly adhered to and might lead to insufficient diagnostic procedures or treatment. However, palliative care requires a patient-oriented and active approach. To be aware of possible incomplete diagnostics is an important step in preventing adverse consequences of a wrong prognosis. PMID- 18160543 TI - Mini-Suffering State Examination: suffering and survival of end-of-life cancer patients in a hospice setting. AB - The objective of this study was to evaluate suffering of end-of-life cancer patients during their final hospice stay and the possible interrelations with survival. A cohort of consecutive patients hospitalized in an inpatient cancer hospice of a tertiary hospital was used for this study. Patients were evaluated upon admission and once weekly by the Mini-Suffering State Examination (MSSE). The distribution of patients for each MSSE score group, changes in MSSE scores (total and for each item of scale), and MSSE interrelation with clinical parameters associated with patients' condition during hospitalization were studied. In addition, the interrelation of MSSE score and survival was examined. Documentation of a high suffering level by the MSSE scale helps in identifying end-of-life cancer patients expected to benefit from enrollment in a hospice setting, provides quantitative information about various symptom dimensions, and may improve medical management and facilitate communication between these patients and caregivers. PMID- 18160544 TI - Confronting death: perceptions of a good death in adults with lung cancer. AB - Investigations regarding patients' concerns about death have focused on the importance of autonomy, resolution of concerns, family relationships, and religiosity, and relied on data from physicians, nurses, family members, and healthy older adults. Few studies have focused on patients with diseases that have short-term survival rates. This study examined lung cancer patients' perceptions of a good death. One hundred lung cancer patients answered open-ended questions about what a good death was and completed measures assessing coping, spirituality, religious coping, and life satisfaction. Content analysis revealed 4 themes describing a good death: (a) during sleep, (b) pain-free, (c) peaceful, and (d) quick. These findings have implications for those caring for terminally ill patients as the 4 themes differ from those derived from studies of more heterogeneous patients, their families, and health care providers. PMID- 18160545 TI - A UK perspective on worldwide inadequacies in palliative care training: a short postgraduate course is proposed. AB - A chronological literature review illustrates how undergraduate and postgraduate education and training in the care of the dying and bereaved is inadequate worldwide. This is despite the foundation of the modern hospice movement in the United Kingdom in 1967 and its wider dissemination as a specialty in 1985. This situation has implications for those doctors working in both primary and secondary care, and this paper describes a 3-day course which has been successfully run in the West Midlands, UK, since 1997 for family physicians in training. A pre-course survey of 250, with a response rate of 54%, in 2003 revealed that 100% of respondents felt that they needed further training, and 51.5% said that they had had no previous training in palliative care. PMID- 18160546 TI - Documentation in palliative care: nursing documentation in a palliative care unit -a pilot study. AB - Palliative care seeks to enhance quality of life in the face of death by addressing the physical, psychological, social, and spiritual needs of patients with advanced disease. The purpose of this paper is to explore whether palliative patients' needs, nursing actions, and evaluation can be identified in the nursing documentation. Data consisted of reviews of patients' case records in a palliative care unit. Data were analyzed using content analysis and counting frequency of keywords used from the Well-being Integrity Prevention and Safety (VIPS) model, followed by an inductive analysis of the case record documentation aiming to identify palliative care components. The result shows that the documentation revealed physical care, especially pain, more frequently than other needs. Nursing documentation focuses on identification more than on nursing actions and evaluation. PMID- 18160547 TI - Identification of quality indicators of end-of-life cancer care from medical chart review using a modified Delphi method in Japan. AB - End-of life care is one of the principle components of cancer care. Measurement of the quality of care provided for end-of-life cancer patients is an important issue. The aim of this study was to identify the quality indicators (QIs) for end of-life cancer care for Japanese patients using a medical chart review. A modified Delphi method for the development of QIs was adopted. Seventeen multi professional specialists participated by rating the appropriateness and feasibility of potential QIs. Thirty QIs for end-of-life cancer care were ultimately identified within 4 domains: (1) symptom control, (2) decision-making and preference of care, (3) family care, and (4) psychosocial and spiritual concerns. These QIs will be useful for monitoring and evaluating end-of-life care for Japanese cancer patients. The QIs are feasible for use in any clinical setting and cover a comprehensive area in accordance with the World Health Organization's (WHO) definition of palliative care including physical, psychosocial, and spiritual concerns. PMID- 18160548 TI - Periodic lateralized epileptiform discharges in La Crosse encephalitis, a worrisome subgroup: clinical presentation, electroencephalogram (EEG) patterns, and long-term neurologic outcome. AB - La Crosse virus encephalitis is the most common mosquito-borne virus in children in the United States. La Crosse virus encephalitis has emerged as a significant health concern due to its potential for acute morbidity, including seizures, alterations in mental status, and, in rare cases, death, as well as the potential for chronic morbidity, including, epilepsy and cognitive and behavioral disorders. The aim of this study is to provide a clinical description of the largest series of children reported with periodic lateralizing epileptiform discharges (PLEDS) associated with La Cross virus encephalitis with reference to their clinical course, seizure type, electroencephalogram (EEG) patterns, and 2- and 10-year long-term neurologic outcome. In addition, to evaluate whether this subset of children may indeed have more severe disease than children with La Crosse virus encephalitis without PLEDS, comparisons are made between the 2 groups on specific variables. All patients presented with fever and disorientation; 6 of the 9 (66%) presented with seizures. PLEDS localized to the temporal lobe in 7 patients (77%). The children with PLEDS had longer intensive care unit stays (6.5 +/- 2.4 vs 3.2 +/- 1.9; P < .0001), a higher rate of intubation (88% vs 20%; P < .001), and a higher rate of cerebral herniation (1%; P < .05) than children with La Crosse virus encephalitis without PLEDS. Follow-up data on the subset with PLEDS also suggest a relatively high rate of epilepsy and behavioral difficulties with hyperactivity symptoms, memory deficits, and school difficulties. The implications for recognition, management, and follow-up of this worrisome subset of patients with La Crosse virus encephalitis are discussed. PMID- 18160549 TI - Characterization of autism in young children with tuberous sclerosis complex. AB - Both cognitive impairment and autism are common in the tuberous sclerosis complex, but the relationship between the 2 diagnoses has not been formally explored. The authors evaluated 20 clinic-referred children with tuberous sclerosis complex at ages 18, 24, 36, and 60 months and classified them as autism, autism spectrum disorder, or normal on the basis of the Autism Diagnostic Observation Schedule. Using the Mullen Scale of Early Learning, cognitive function in each subgroup was assessed. The authors then analyzed the subscores of the Autism Diagnostic Observation Schedule in children with autism. Children with autism showed significantly more global cognitive impairment than those without autism. In addition, all children had some baseline cognitive impairment and the majority had deficits in play scores. The authors conclude that clinic referred children with tuberous sclerosis complex and autism are at considerable risk for cognitive impairment. These characteristics may help to guide more tailored services for these high-risk children. PMID- 18160550 TI - A pilot study on cord blood levels of erythropoietin and its relationship to periventricular leukomalacia in preterm infants. AB - This article reports the results of a study on the relationship between cord blood levels of erythropoietin and periventricular leukomalacia. Cord blood was obtained from 19 infants with gestational age between 27 and 32 weeks. Cystic periventricular leukomalacia was seen in 4 of them. Erythropoietin levels were not different between those with and those without periventricular leukomalacia. PMID- 18160551 TI - Enhancement of muscle activity by electrical stimulation in cerebral palsy: a case-control study. AB - The objectives of this study were to compare the effects of low-intensity electrical stimulation of the quadriceps muscle in children with cerebral palsy in the following 2 modes: reconditioning by long-term training of the muscle versus real-time assist to the muscle during motion. To evaluate the force enhancement in the assist mode, we developed a method to dissociate the volitional and the induced components from the total electromyographic signal. The study group, including 5 children with cerebral palsy (mean age, 3.3 years; 0.4 SD), underwent 2 testing sessions: 1 before and 1 after 3-month training by electrical stimulation. Each session included 2 series of trials: 1 with electrical stimulation, as an orthotic assist, and 1 without electrical stimulation. The tests included flexion-extension movements of the knee at a self selected pace. The results showed that, compared to before training, there was a significant increase in the average motion velocity and a decrease in motion jerk and in knee torque after training in both the electrical stimulation- assisted and -unassisted modes. Of special interest was the significant decrease in quadriceps-hamstrings co-contraction following training by electrical stimulation but not during electrical stimulation-assisted motion. The results obtained for the group with cerebral palsy were statistically different from those of the control group, but this difference decreased after long-term training by electrical stimulation. It was concluded that, in children with cerebral palsy, electrical stimulation is more beneficial in long-term training than when used as a real-time motion assist. Although muscle strength is not affected, more centrally controlled attributes such as co-contraction are improved. PMID- 18160552 TI - Brain tumors in children: initial symptoms and their influence on the time span between symptom onset and diagnosis. AB - Brain tumors are the most common solid tumor entity in childhood. Symptoms are often unspecific, depending not only on the localization of the tumor, but also on the age of the child. The aim of this study was to detect factors influencing the time span between the occurrence of symptoms and the diagnosis to alert health professionals to the early symptoms of pediatric brain tumors. The records of 245 consecutive patients treated for brain tumors between 1980 and 2004 at the neuropediatric department of the University of Muenster were analyzed regarding their primary symptoms, tumor location, entity, and, in 151 cases, the primary electroencephalogram findings. The median time span between symptom onset and diagnosis in our study was 24 days. Multivariate analysis showed a significant influence of 6 parameters on the interval between symptom onset and diagnosis. An additional symptom had a significant influence on the time span between symptom onset and diagnosis in the univariate analysis. The findings that several symptoms influence the interval between symptom onset and diagnosis emphasize the necessity to systematically inquire about the key symptoms of brain tumors. The challenge for every consultant is to decide in which cases cerebral imaging is appropriate. As the most frequent symptoms are unspecific and often underestimated, a detailed anamnesis is crucial to detect possible brain tumor patients. In doubtful cases, a systematic interrogation regarding the catalogue of symptoms can be helpful. PMID- 18160553 TI - Parasagittal lesions and ulegyria in hypoxic-ischemic encephalopathy: neuroimaging findings and review of the pathogenesis. AB - Hypoxic-ischemic brain injury is a very important neurological problem of the perinatal period and a major cause of chronic disability later in childhood. The subsequent neurological deficits are a variety of motor defects-especially spasticity but also choreoathetosis, dystonia and ataxia, often grouped together as "cerebral palsy," mental retardation, and seizures. The gestational age determines the neuropathology of the brain injury. One of the patterns of hypoxic ischemic encephalopathy, typically affecting full-term infants, consists of parasagittal lesions and ulegyria. The aim of this study is to describe the magnetic resonance imaging (MRI) features and discuss the "suggested" pathogenetic mechanisms of this pattern, which affects the cortex and the white matter in a mainly parasagittal distribution; in this type of brain injury, the damage usually involves the deeper sulcal portion while sparing the apex, thus resulting in the so-called mushroom gyri characteristic ulegyric pattern. We discuss the MRI findings of parasagittal lesions and ulegyria in the brain examinations of 14 patients with a clinical history of perinatal hypoxia/anoxia presenting with mental retardation, seizures, and cerebral palsy. Differential diagnosis from polymicrogyria is discussed. PMID- 18160554 TI - Predictive value of sequential electroencephalogram (EEG) in neonates with seizures and its relation to neurological outcome. AB - The aim of this study was to evaluate the relationship of sequential neonatal electroencephalography (EEG) and neurological outcome in neonates with seizures to identify polysomnographic features predictive of outcome. Sequential EEGs recordings of 58 neonates that belonged to 2 historical cohorts of newborns with seizures from the same neonatal intensive care unit and who had follow-up at the Neurodevelopment Clinic of the Hospital Sao Lucas, Pontificia Universidade Catolica do Rio Grande do Sul (PUCRS) in Porto Alegre, Brazil, were analyzed and classified into 4 groups: normal-normal, abnormal-normal, abnormal-abnormal, normal-abnormal. In patients with more than 2 recordings, during the neonatal period, the first EEG was compared with the following more abnormal. A total of 58 pairs of 2 sequential EEGs were analyzed. Considering the first EEG, a statistically significant difference was observed between the relationship of the result of this exam, if it was abnormal, with developmental delay (P = .030) and postnatal death (P = .030). Abnormal background activity was also related to neurodevelopment delay (P = .041). EEG sequences abnormal-abnormal and normal abnormal significantly correlated to the outcome epilepsy ( P = .015). Abnormal sequential background activity was associated with neurodevelopment delay (P = .006) and epilepsy (P = .041). The burst suppression pattern when present in any EEG correlated with epilepsy (P = .013) and postnatal death (P = .034). Sequential abnormal background patterns in the first and second EEG increased the risk for epilepsy (relative risk [RR] = 1.8; 95% confidence interval [CI] = 1.03 3.0) and neurodevelopment delay (RR = 2.20; 95% CI = 1.3-3.0). Abnormal background activity only in the second electroencephalogram increased the risk for neurodevelopment delay (RR = 2.20; 95% CI = 1.3-3.0). All the neonates (n = 33) with seizures related to probable hypoxic ischemic encephalopathy had abnormalities in the first EEG (P = .030). Postnatal epilepsy was diagnosed in 24 infants (41.4%). Five (20.8%) presented West syndrome, 7 (29.2%) focal symptomatic epilepsy, 6 (25%) generalized symptomatic epilepsy, 2 (8.3%) early myoclonic encephalopathy, 1 (4.2%) early infantile epileptic encephalopathy, and in 3, the epileptic syndrome was undefined (12.5%). All infants (n = 5) with West syndrome had some degree of neurodevelopment delay. In conclusion, our findings suggest that sequential EEG in neonates with seizures has more predictive value to estimate the outcomes of neurodevelopment delay, epilepsy, and postnatal death than a single EEG recording. The abnormal background activity in even 1 EEG of the sequential recordings was more significant to determine neurological outcome than abnormal ictal activity or abnormalities in the organization of sleep state. PMID- 18160555 TI - Hemorrhagic shock and encephalopathy syndrome: clinical course and neurological outcome. AB - The syndrome of hemorrhagic shock and encephalopathy is associated with an acute onset of diarrhea, followed by shock, disseminated intravascular coagulopathy, multiorgan failure, and encephalopathy. The etiology of this syndrome is unknown, and despite intensive treatment, the outcome is often fatal or associated with severe neurological sequelae. Two infants aged 6 and 9 months were admitted with this syndrome within a 24-hour time interval. The authors hereby present the clinical course and a prospective evaluation of the neurological outcome. A review of the literature regarding this infrequent syndrome is presented. PMID- 18160556 TI - Neurodevelopmental screening in toddlers and early preschoolers with sickle cell disease. AB - Sickle cell disease is associated with an elevated risk for neurologic complications beginning in early childhood. Detecting higher-risk cases with developmental screening instruments may be a cost-effective method for identifying young children in need of more frequent or intensive assessment. We evaluated the validity of the Denver II test as a tool to detect lower levels of developmental attainment and their association with neurologic risk in 50 young children with sickle cell disease. Children with suspect Denver II outcomes showed lower scores for functional communication skills, had lower hematocrit percentage, higher mean velocities on transcranial Doppler ultrasound imaging, and were more likely to have had preterm birth. Validity of age equivalencies from specific Denver II areas was demonstrated for Language and Fine Motor scores, suggesting the instrument could be used to index children's developmental levels in these domains. The Denver II may be a useful behavioral screening tool for neurodevelopmental risk in sickle cell disease. PMID- 18160557 TI - A quantitative magnetic resonance imaging analysis of the cerebellar deficit hypothesis of dyslexia. AB - Recent evidence suggests that the primary source of dysfunction in dyslexia is the cerebellum. To examine the cerebellar deficit hypothesis of dyslexia, 20 children with dyslexia and 20 children without dyslexia were assessed using neuropsychological testing and quantitative magnetic resonance imaging. Results demonstrated that the volumes of both hemispheres and the vermis were not statistically significantly different between groups. However, children without dyslexia demonstrated greater rightward cerebellar hemisphere asymmetry. The relationship between cerebellar morphologic structure and phonological processing was assessed. For children without dyslexia, bilateral hemisphere volume moderately correlated with phonological awareness and phonological short-term memory. Hemisphere asymmetry moderately correlated with rapid naming errors, and the anterior vermis volume moderately correlated with phonological awareness. For children with dyslexia, the only statistically significant correlation was between rapid naming errors and the left hemisphere volume. Evidence suggests that atypical cerebellar morphologic structure may have a role in dyslexia for a subgroup of individuals. Although children with and without attention deficit/hyperactivity disorder did not differ in cerebellar morphologic structure, the anterior vermis volume moderately correlated with inattention, hyperactivity, and impulsivity, while the right hemisphere volume moderately correlated with inattention and hyperactivity. Our findings provide mixed support for the cerebellar deficit hypothesis of dyslexia. Although cerebellar morphologic structure is atypical in some individuals with dyslexia, it is inconsistently related to cognitive or motor dysfunction. In our sample, cerebellar morphologic structure may be related to about one-third of cases of dyslexia. Hence, dyslexia may be best accounted for by a combination of cortical and cerebellar contributions. PMID- 18160558 TI - Intractable gelastic seizures during infancy: ictal positron emission tomography (PET) demonstrating epileptiform activity within the hypothalamic hamartoma. AB - Gelastic seizures comprise a very rare form of epilepsy. They present with recurrent bursts of laughter voices without mirth and are most commonly associated with the evolution of a hypothalamic hamartoma. The purpose of this article is to describe the second reported ictal fluorodeoxyglucose-positron emission tomography study in a unique case of an infant with intractable gelastic seizures since the neonatal period associated with a hypothalamic hamartoma. The patient presented at 4 months old with recurrent, almost persistent, gelastic seizures consisting of laughter bouts without mirth. The seizures were noticeable at the first week of life and increased in frequency to last up to 12 hours, namely status gelasticus. These gelastic fits were accompanied with focal motor seizures, including unilateral right-eye blinking and mouth twitching. Developmental mile-stones were intact for age. Magnetic resonance imaging of the cortex demonstrated a large hypothalamic hamartoma within the third ventricle, hampering cerebrovascular fluid drainage of the lateral ventricles. An electroencephalography was nondiagnostic. Ictal fluorodeoxyglucose-positron emission tomography demonstrated a large circumscribed hypermetabolic region within the location of the hypothalamic hamartoma, representing localized intense epileptiform activity. The infant became instantly free of all seizure types given minute doses of oral benzodiazepine (clonazepam) and remains completely controlled after 12 months. Her overall development remains intact. This ictal fluorodeoxyglucose-positron emission tomography is the second reported study verifying that the main source of the epileptic activity inducing gelastic seizures originates from the hypothalamic hamartoma itself; therefore, a complementary fluorodeoxyglucose-positron emission tomography study should be considered in any patient presenting with intractable gelastic seizures, especially in those associated with hypothalamic hamartoma, in order to localize the region of epileptiform activity amenable to surgical resection if intensive drug therapy fails. PMID- 18160559 TI - Epidemiological study of autism spectrum disorder in China. AB - The object of this study was to investigate the epidemiologic pattern of autism spectrum disorder in Chinese children. An autism spectrum disorder registry has been established in Hong Kong since 1986 by collecting data in a single center (the only university-affiliated child assessment center in Hong Kong). Since 1997, inpatient data from all public hospitals under the Hospital Authority have been stored in a central computerized program and retrieved from the Clinical Data Analysis and Reporting System. Clinical data have also been retrieved through the Clinical Data Analysis and Reporting System to ensure the completion of the registry, and these suspected cases have undergone the same diagnostic evaluation for autism spectrum disorder, as some of the new autism cases might be hospitalized in the public hospital. The incidence and prevalence of autism spectrum disorder have been calculated for the period of 1986 to 2005 using the population statistics available in the government for children less than 15 years old in Hong Kong. This study has included 4 247 206 person-years from 1986 to 2005 for children less than 15 years old and 1 174 322 person-years for those less than 5 years old in Hong Kong. Altogether, 645 children 0 to 4 years old with diagnoses of autism spectrum disorder were identified from 1986 to 2005. The estimated incidence of autism spectrum disorder was 5.49 per 10 000. The prevalence was 16.1 per 10 000 for children less than 15 years old for the same period. The male to female ratio was 6.58:1. This is the first reported epidemiologic study of autism spectrum disorder in Chinese children. The incidence rate is similar to those reported in Australia and North America and is lower than Europeans. PMID- 18160560 TI - Bleeding management of uterine conization using a pasteurized FVIII/VWF Concentrate in a woman affected by type 2A von willebrand disease. PMID- 18160561 TI - PIA1/A2 polymorphism of the platelet glycoprotein receptor IIb/IIIIa and its correlation to cerebrovascular diseases: an appraisal. AB - Platelet glycoprotein (GP) IIb/IIIa is a membrane receptor for fibrinogen and von Willebrand factor. There is considerable controversy regarding the clinical role of the GP IIb/IIIa PIA1/A2 as a risk factor of cerebrovascular diseases. Here, the author performs a summative analysis on the recent previous reports on the GP IIb/IIIa PIA1/A2 and its correlation to cerebrovascular diseases. The meta analysis was performed to assess the correlation between the pattern of GP IIb/IIIa PIA1/A2 polymorphism and cerebrovascular diseases. From available 4 case control studies, 553 patients and 1059 controls are evaluated. From overall risk estimation, the subjects with PIA2 alleles have 1.18 times higher risk to develop cerebrovascular diseases. According to this analysis, the author proposes that the pattern of GP IIb/IIIa PIA1/A2 polymorphism does not represent a useful marker of increased risk for cerebrovascular diseases. The lack of association between pattern of GP IIb/IIIa PIA1/A2 polymorphism and ethnicity can also be showed in this study. PMID- 18160562 TI - Use of recombinant activated factor VII to arrest uncontrolled bleeding: a case series. AB - A retrospective analysis is described to assess the effects of using recombinant activated factor VII to control bleeding in a series of patients who had failed to respond to conventional hemostatic measures. In all, 18 patients (aged 16-65 years) with a range of conditions resulting in bleeding refractory to conventional methods of control were treated with recombinant activated factor VII (60-120 Amicrog/kg; 1-4 doses). The effects of recombinant activated factor VII on bleeding were noted together with the patients' transfusion requirements and hematological parameters. Administration of recombinant activated factor VII successfully stopped bleeding in 17 of the 18 patients. Therapy with recombinant activated factor VII significantly decreased transfusion requirements for packed red blood cells, fresh frozen plasma, platelets, and cryoprecipitate compared with pretreatment values along with significant improvement in hemostasis. In various serious bleeding situations, treatment with recombinant activated factor VII may effectively arrest bleeding, which has remained refractory to conventional methods of control. PMID- 18160563 TI - Bilateral subdural hematomas in an adult with hereditary factor VII deficiency: A complication of sit-ups and inversion? AB - A 57-year-old man of northern Italian and Polish descent with a late-onset, mild form of hereditary factor VII deficiency developed bilateral intracranial subdural hematomas after performing sit-ups while he was in a head-down position on an inversion table. The pertinence of the Valsalva maneuver and increased intracranial venous and intraocular pressure associated with sit-ups and other exercises in the causation of subdural hematomas and other types of intracranial and ocular hemorrhage are discussed in this study. PMID- 18160564 TI - Thrombophilia in human immunodeficiency virus-infected patients with osteonecrosis: Is there a real connection? The first case-control study. AB - Several reports have described an increased incidence of osteonecrosis in human immunodeficiency virus-infected patients (HIV+), but the cause has not been established. The association between thrombophilia and osteonecrosis in HIV+ was studied. A case-control study in HIV+, 19 cases and 38 controls, was designed. Magnetic resonance imaging was made in both groups to confirm or exclude hip osteonecrosis. The extensive tests of thrombophilia were measured, and the clinical data were recorded, nadir of CD4(+) cell count and well-known risk factors for osteonecrosis. Thrombophilia has been frequently found both in patients with and without osteonecrosis (thrombophilia, 68.4% vs 60.5%), but no specific thrombophilia tests were significantly associated with osteonecrosis. A low nadir of CD4(+) (<60 cells/microL) and corticoid use were significantly (P < .05) associated with osteonecrosis. In multivariate analysis, only nadir of CD4(+) <60 cells/microL remained a predictor of osteonecrosis (odds ratio = 7.33; 95% confidence interval, 1.80-29.82, P = .005). Thrombophilia might have a limited role in the development of osteonecrosis in HIV+. Nadir of CD4(+) <60 cells/microL and corticoid use were main factors. PMID- 18160565 TI - The impact of an aging population on the diagnosis of pulmonary embolism: comparison of young and elderly patients. AB - The influence of age on predisposing factors, diagnostic tests, and clinical presentation of pulmonary embolism was evaluated in 582 subjects with suspected pulmonary embolism (180 aged <65 years; 402 aged > or =65 years) consecutively enrolled at the Emergency Department. Pulmonary embolism was confirmed in 40% of patients, 75% of those were aged >65 years. Age was directly related to the diagnosis, and the observed probability was higher than the expected probability in the 70 to 79 year subgroup. Score at the Cumulative Illness Rating Scale significantly increased as a function of both age and pulmonary embolism. Dyspnea, syncope, jugular distension, and history of previous venous thromboembolism were more frequently observed in elderly patients. In-hospital mortality rate among the elderly and younger patients was 2% and 0.2%, respectively. The authors conclude that age > or =65 years and high comorbidity are risk factors for pulmonary embolism. PMID- 18160566 TI - Association between the Thr325Ile and Ala147Thr polymorphisms of the TAFI gene and the risk of venous thromboembolic disease. PMID- 18160567 TI - Is there any effect of tumor burden on hemostatic parameters in cancer patients? A case-control study of hemostatic abnormalities and anticardiolipin antibodies in solid tumors. AB - Hemostatic complications are one of the leading causes of mortality in cancer patients. In the present study, we assessed the hemostatic parameters and anticardiolipin antibodies in patients with solid tumors (n = 104) and healthy controls (n = 25) and also find out whether the abnormalities in these hemostatic parameters vary related to tumor burden. Prothrombin time, activated partial thromboplastine time, D-dimer, and fibrinogen as hemostatic parameters were determined by photo-optometric clot detection system, and serum anticardiolipin levels were measured by enzyme-linked immunosorbent assay. The plasma levels of fibrinogen, D-dimer, and serum anticardiolipin IgM levels in cancer patients were significantly higher compared with those in controls (P < .001, P = .001, and P = .01, respectively). Only fibrinogen levels were significantly higher in metastatic group than nonmetastatic group (P < .001 ). Hemostatic abnormalities that are detected in asymptomatic cancer patients may not give any clue about tumor burden or stage of the cancer patients. PMID- 18160568 TI - Fatal bleeding due to a heparin-like anticoagulant in a 37-year-old woman suffering from systemic mastocytosis. AB - A 37-year-old female patient with systemic mastocytosis who was admitted with severe unexplained bleeding symptoms is studied. Laboratory procedures established the diagnosis of a patient-derived-heparin-like anticoagulant as a very rare hemostatic abnormality predisposing to bleeding. The patient died from refractory disease despite therapy with protamine, initiation of chemotherapy, and supportive measures. The case illustrates the clinical presentation and diagnosis of heparin-like anticoagulants. Etiology, pathophysiology, and therapeutic options are discussed. PMID- 18160569 TI - Inflammatory and hemostatic activation in patients undergoing off-pump coronary artery bypass grafting. AB - To characterize hemostatic differences imposed by 2 common cardiac surgeries, the authors studied patients undergoing coronary artery revascularization by off-pump (n = 13) or cardiopulmonary bypass on-pump (n = 26) technique. Blood samples collected to 4 days post-surgery were evaluated by flow cytometry and enzyme linked immunosorbent assay. A significant inflammatory response occurred in both the groups after surgery shown by increased interleukin cytokines and C-reactive protein; however, levels peaked lower and hours later in the off-pump group. Platelets (P-selectin; platelet-leukocyte complexes) and leukocytes (CD11b) were activated only in on-pump patients. Thrombin generation was enhanced in both groups after surgery. Only in the on-pump patients, the thrombin-antithrombin complex, pro-thrombin fragment 1.2, and thrombomodulin (vascular integrity) decreased intraoperatively. Tissue plasminogen activator and plasminogen activator inhibitor-1 were greater in the on-pump patients. Off-pump surgery may place patients at higher risk of postoperative hypercoagulability because of normal platelet function, intraoperative thrombin generation, less fibrinolytic activity, and lack of vascular protection. PMID- 18160570 TI - Coagulation activation by lipopolysaccharides. AB - Lipopolysaccharides at approximate plasma reactivities >3 ng/mL or beta-glucans at >0.5-1 Amicrog/mL are toxic for human blood; lipopolysaccharide interacts with membrane components of susceptible cells (eg, monocytes) activating phospholipase A(2) that destroys the cell membrane. Cell fragments (microparticles or DNA) possess polynegative niches that activate intrinsic hemostasis. Pathologic disseminated intravascular coagulation arises. Blood vessels are obstructed by disseminated thrombi, and vital organ areas become ischemic. Multiorgan failure threatens life of the patient. Diagnosis and therapy of pathologic disseminated intravascular coagulation is of extreme clinical importance. For early diagnosis of pathologic disseminated intravascular coagulation, specific activation markers of coagulation (eg, plasmatic amidolytic thrombin activity) or the plasmatic lipopolysaccharide or glucan reactivity can be measured. A new treatment target might be kallikrein or factor XIIa; 10 to 20 mM arginine is the approximate 50% inhibitory concentration against the contact phase of coagulation. The complex interaction between cell fragments and hemostasis causes pathologic disseminated intravascular coagulation in sepsis. PMID- 18160571 TI - Management of uremic patients with heparin-induced thrombocytopenia requiring hemodialysis. AB - Medical records of 122 patients with suspected heparin-induced thrombocytopenia on dialysis were reviewed. Method of dialysis in heparin-induced thrombocytopenia patients with bleeding from various causes (including surgical interventions) and how to cope with blood access occlusion induced by heparin-induced thrombocytopenia were studied. Of 122 patients, 17 who met the criteria of >30% thrombocytopenia, clots in the extracorporeal circulation, positive for heparin/PF4 complex antibodies, and improvement from heparin-induced thrombocytopenia with the use of an alternative anticoagulant or another strategy for heparin-induced thrombocytopenia were chosen. Argatroban was uneventfully introduced in 12 patients having neither bleeding nor blood access failure. In all, 2 of 5 patients were treated with peritoneal dialysis. The others requiring a regional anticoagulant were given nafamostat mesilate. Argatroban as an alternative provides effectively anticoagulation in patients with heparin-induced thrombocytopenia on dialysis. In patients with heparin-induced thrombocytopenia with bleeding and its associated risk, nafamostat mesilate was an alternative. Peritoneal dialysis also was applied in cases of blood access failure due to heparin-induced thrombocytopenia. PMID- 18160572 TI - Fatal giant aortic thrombus presenting with pulmonary edema in a patient with chronic obstructive pulmonary disease. AB - Thrombus formation in a morphologically normal a aorta is a very rare event. A 50 year-old man with a history of chronic obstructive pulmonary disease, presented to the emergency department with pulmonary edema. Transthoracic and transesophageal echocardiography revealed a highly mobile, pedunculated floating thrombus in the descending thoracic aorta 3-4 cm distal to the origin of the left subclavian artery. The original lumen of the aorta was almost obliterated by the thrombus. The aortic wall was free of any atheroma. Thrombolytic treatment was administered, but 3 hours after starting streptokinase, he developed sudden and severe low-back pain accompanied by loss of lower-extremity pulses which were patent on admission. Cardiopulmonary arrest developed within an hour and the patient died despite resuscitation. The potential causes of aortic thrombus, the clinical spectrum that the patients may present, diagnostic methods, and therapeutic options are discussed. PMID- 18160573 TI - Further removal of factor v related antigen from bovine thrombin by utilizing a membrane-filtration step. AB - Topical bovine thrombin is commonly used during surgery to maintain hemostasis and is rarely associated with abnormalities in hemostasis, including coagulopathies and bleeding. Coagulopathies may be related to the formation of cross-reactive antibodies to bovine factor V. Effectiveness of a new filtration step to remove factor V/Va from bovine thrombin was evaluated. A highly sensitive and specific Western blot capable of detecting minute amounts of factor V/Va and/or its fragments was developed. Samples were evaluated for bovine factor V related antigens using the Western blot method and a competitive enzyme-linked immunosorbent assay. Factor Va light chain fragment levels were detectable in crude thrombin and chromatographically purified thrombin but not in chromatographically purified and virally filtered preparations. Therefore, inclusion of the viral-filtration step during purification of thrombin is effective in reducing factor V or its fragments to undetectable levels, thus enhancing product purity. PMID- 18160574 TI - The recalcified coagulation activity. AB - Thrombin activity generated after plasma recalcification is of analytical and clinical interest. Fifty microliters of citrated plasma was recalcified with 5 microL of 250 mM CaCl( 2). After 0 to 90 minutes (37 degrees C) 50 microl 2.5 M arginine, pH 8.6, was added. After 20 minutes, thrombin was chromogenically quantified. In normal recalcified plasma, the generated thrombin activity is about 0.1-0.2 IU/ml (37 degrees C) when fibrin generation starts. Pooling of normal plasmas increases the generated thrombin activity about 3-fold. Plasmas of patients on heparin or coumarin generate about 10-fold less thrombin activity. Freezing of pooled plasma at -20 degrees C and thawing at room temperature or 37 degrees C increases thrombin generation approximately 1.5- or 2-fold, respectively. Only thrombin activities in the ascending part of the thrombin generation curve (RECA-t(2)/RECAt( 1)>1) are valid. So a prothrombotic state in blood or plasma can be diagnosed. PMID- 18160575 TI - Elevated levels of prothrombin fragment 1 + 2 indicate high risk of thrombosis. AB - Prothrombin fragment 1 + 2 (F1 + 2) is considered to be useful for diagnosis of thrombosis. However, the evidence for a diagnosis of thrombosis by F1 + 2 is still not well established. The plasma concentrations of F1 + 2, soluble fibrin, D-dimer, and thrombin-antithrombin complex were measured in 694 patients suspected of having thrombosis and then were correlated with thrombosis. Plasma concentrations of F1 + 2, soluble fibrin, D-dimer, and thrombin-antithrombin complex were significantly higher in patients with thrombosis, compared with patients without thrombosis. When cutoff values of more than 300 pmol/L for F1 + 2 were used for the diagnosis, more than 50% of the patients were thus found to have thrombosis. The findings showed that F1 + 2, soluble fibrin, D-dimer, and thrombin-antithrombin complex have similar diagnostic ability. The plasma concentration of F1 + 2 closely was well correlated with thrombin-antithrombin complex, soluble fibrin, and D-dimer. Finally, F1 + 2 is one of the most useful parameters for the diagnosis of thrombosis. PMID- 18160576 TI - Kallikrein activates prothrombin. AB - Kallikrein is a multitalented enzyme in hemostasis and inflammation. Normally, kallikrein is formed in intrinsic hemostasis and activates factor XII. A total of 10 microL of 0 to 100 microg/mL human plasma kallikrein in 6% human albumin-PBS were incubated with 90 microL 111.1 microg/mL prothrombin in 6% human albumin in absence and presence of 23 mM Ca(++). After 0 to 64 minutes (37 degrees C), 100 microL of 2.5 M arginine, pH 9, were added. Fifty microliters of 0.72 mM HD-CHG Ala-Arg-pNA in 1.36 M arginine were added and increase in absorbance at 405 nm was determined. Within 8 minutes (37 degrees C), 1 microg/mL kallikrein, ie, 2.5% of the normal plasmatic prekallikrein concentration, generates approximately 3 mIU/mL thrombin in absence and 27 mIU/mL thrombin in presence of Ca(++). Kallikrein can directly activate prothrombin; there is a shortcut in the intrinsic hemostasis system that generates catalytic amounts of thrombin without following the known intrinsic clotting pathway. PMID- 18160577 TI - Factor VIII gene haplotypes and linkage disequilibrium for the indirect genetic analysis of hemophilia A in India. AB - Genomic consequences of factor VIII gene haplotypes for the indirect genetic analysis of haemophilia A has not been done in India hitherto. Consequently, BclI/intron18, HindIII/intron 19, and XbaI/intron 22 restriction sites were investigated in 159 individuals from 42 families with hemophilia A. The frequencies of haplotype II, IV, VI, that is, BclI (+)-HindIII (-)- XbaI (+), BclI (+)HindIII (+)-XbaI (-), and BclI (-)-HindIII (-)-XbaI (+) were 0.312, 0.198, and 0.164 respectively. The high heterogeneity of haplotype II highlighted its potential for indirect genetic diagnosis of factor VIII. Analysis revealed strong but incomplete linkage disequilibrium (D' = 0.76, 0.68, and 0.51) between BclI/HindIII, HindIII/XbaI, and BclI/XbaI, respectively. The overall cumulative polymorphism information content (PIC) of these three markers increased from 0.36 to 0.80. Escalation of PIC up to 80% in the present study suggests that haplotyping of factor VIII gene determines better prognosis in the direction of indirect genetic analysis of hemophilia A. PMID- 18160579 TI - Pericardial hemorrhage due to acetylsalicylic acid in a patient with essential thrombocythemia. AB - Essential thrombocythemia is a clonal myeloproliferative disorder that causes thrombocytosis. Essential thrombocythemia is characterized by increased incidence of thrombosis with arterial event more than venous events and hemorrhagic complications. Acetylsalicylic acid enhances both minor and major bleedings. The authors describe pericardial hemorrhage, which is related to the use of low-dose acetylsalicylic acid in a patient with essential thrombocythemia. The patient was successfully managed with clopidogrel therapy during the 16 months follow-up without recurrent thrombotic or hemorrhagic events. PMID- 18160581 TI - Monitoring of functional plasminogen in the blood of patients on fibrinolytics. AB - There are no reliable data on functional plasminogen in the blood of patients receiving fibrinolytic treatment. Here, artifactual in vitro changes of functional plasminogen were prevented by arginine stabilization blood samples of myocardial infarction patients: 12 received 36.4 mg reteplase in bolus, and 1 patient received 100 mg tissue plasminogen activator in continuous infusion. Arginine (1.5 M, 1.3 mL, pH 8.7) was used to stabilize 2.6 mL ethylenediaminetetraacetic acid-blood. The arginine-stabilized plasma was analyzed with a functional oxidative assay for plasminogen. Functional plasminogen decreased within 2 minutes of reteplase treatment by about 40% and by about 80% after 60 minutes. Lowest plasminogen concentrations were found in plasmas with highest plasmin activities. Chloramine oxidation of purified Glu plasminogen increased its activation by urokinase up to 3-fold. Arginine stabilization allows reliable determinations of functional plasminogen in the blood of patients receiving fibrinolytics, enabling the rapid diagnosis of prothrombotic plasminogen consumption. The present findings support the profibrinolytic action of chloramines. PMID- 18160582 TI - Soluble fibrin inhibits lymphocyte adherence and cytotoxicity against tumor cells: implications for cancer metastasis and immunotherapy. AB - Circulating soluble fibrin (sFn) is elevated in many cancer patients. It is a marker for ongoing disseminated intravascular coagulation and may have prognostic significance. We have demonstrated that sFn inhibited monocyte adherence and cytotoxicity by a mechanism involving blockade of monocyte alphaMbeta2 and tumor cell CD54. It was, therefore, hypothesized that sFn also inhibits lymphocyte and interleukin-2-activated lymphocyte (LAK) adherence and cytotoxicity against tumor cells. This study sought to identify the lymphocyte subset responsible for adherence and killing of A375 melanoma cells and whether sFn inhibited these parameters. Lymphocyte and LAK cell adherence and cytotoxicity, which was adherence dependent, were inhibited by preincubation with purified or plasma derived sFn. The lymphocyte and LAK cell activities were primarily a result of CD8(+) MHC (major histocompatibility complex) unrestricted cytotoxic T cells. These results suggest that elevated levels of circulating sFn may be immunosuppressive and may reduce the efficacy of adoptive immunotherapies. PMID- 18160583 TI - The success of warfarin treatment for coronary thrombosis in a young patient with acute coronary syndrome. PMID- 18160584 TI - In vitro simulation of thrombolysis inhibition. AB - Hyperfibrinolysis is a serious clinical complication. The inhibitory concentrations 50% of antifibrinolytics were analyzed in the microtiter plate clot lysis assay, using 50 microL of plasma clots, 10 microL of antifibrinolytic drug, 10 microL of 354 IU/mL (final) urokinase, 4.46 microg/mL (final) tissue type plasminogen activator or 0.6 mg/mL plasmin, and 50 microL of pooled normal plasma as clot supernatant. The inhibitory concentrations 50% of lysine against urokinase or tissue-type plasminogen activator is 2.0 or 4.2 mM, against epsilon amino-caproic acid 0.7 or 1.5 mM, against tranexamic acid 0.03 or 0.17 mM, respectively. The inhibitory concentrations 50% of lysine, epsilon-amino-caproic acid, or tranexamic acid against plasmin is 7.4, 0.4, or 0.04 mM. The inhibitory concentrations 50% of aprotinin against urokinase or tissue-type plasminogen activator is about 60 KIU/mL, against plasmin 19 KIU/mL. Lysine might be a new antifibrinolytic drug with a clinically interesting rapid pharmacokinetic. This data help correct dosing of antifibrinolytics to patients with hyperfibrinolysis. PMID- 18160585 TI - In vitro simulation of extremely activated thrombolysis. AB - A life-threatening thrombus in massive pulmonary embolism has to be eliminated within minutes. Extremely activated plasmatic fibrinolysis destroys such thrombi in time: 50 microL plasma clots were incubated with urokinase or tissue-type plasminogen activator and 50 microL pooled normal plasma. The microtiter plate clot lysis assay was performed. The time point at which 50% of the clot has been lysed is 4 minutes for 8333 IU/mL urokinase or an equimolar concentration of tissue-type plasminogen activator (52498 IU/mL = 105 microg/mL). The effective dose 50% at 5 minutes lysis time is about 800 nM (4320 IU/mL) urokinase or (27220 IU/mL = 54 microg/mL) tissue-type plasminogen activator. Addition of plasminogen to the plasmatic clot supernatant improves thrombolysis if 65 IU/mL of urokinase acts for 10 minutes. The risk for severe intracranial hemorrhage in massive thrombolysis might be much lower than the lethality of a massive pulmonary embolism. Extremely activated plasmatic thrombolysis could be clinically indicated. PMID- 18160586 TI - Adipokines, linking adipocytes and vascular function in hemodialyzed patients, may also be possibly related to CD146, a novel adhesion molecule. AB - Possible correlations between adiponectin, leptin, CD146, a novel adhesion molecule localized at the endothelial junction, and other markers of endothelial cell injury, von Willebrand factor, thrombomodulin, vascular cell adhesion molecule, and intracellular adhesion molecule, and markers of inflammation, tumor necrosis factor-alpha, interleukin-6, and high-sensitivity C-reactive protein in nondiabetic hemodialyzed patients with and without coronary artery disease were studied. Markers of endothelial dysfunction were elevated in hemodialyzed patients, predominantly with coronary artery disease. In multivariate analysis, kinetic urea modeling and plasminogen activator inhibitor-1 remained the only positive predictors of adiponectin. In multivariate analysis, predictors of leptin were triglycerides, tissue plasminogen activator, CD146, and coronary artery disease. In multivariate analysis, predictors of CD146 were age, hemoglobin, and adiponectin. Elevated adiponectin correlated to CD146 may be the expression of a counterregulatory response aimed at mitigating the consequences in endothelial damage and increased cardiovascular risk in renal failure. The data provide further support for a link between adipocytokines, endothelial dysfunction, cardiovascular risk, and renal failure. PMID- 18160587 TI - The effect of plasminogen activator inhibitor-1 -675 4G/5G polymorphism on PAI-1 gene expression and adipocyte differentiation. AB - Obesity is a complex, multifactorial chronic disease frequently associated with cardiovascular risks, hypertriglyceridemia, low high-density lipoprotein cholesterol, high blood pressure, and the insulin resistance that appears to be central to the pathogenesis of Type II diabetes. Plasminogen activator inhibitor 1 expression induced in differentiating adipose tissue, but its role in adipogenesis and obesity is poorly understood. Circulating plasminogen activator inhibitor-1 levels are elevated at an early stage of impaired glucose tolerance, resulting in diabetes and metabolic syndrome. Plasminogen activator inhibitor-1 levels are also significantly elevated in the plasma of obese individuals and in adipose tissues of obese mice and humans. Some investigators proposed that the 675 4G/5G polymorphism in plasminogen activator inhibitor-1 promoter caused overexpression of this gene and predisposed carriers to obesity. In this study, we investigated the role of -675 4G/5G polymorphism in plasminogen activator inhibitor-1 promoter in the expression of this gene and the contribution of plasminogen activator inhibitor-1 to adipogenesis. Using a dual-luciferase promoter assay, we determined that the -675 4G/5G polymorphism contributes significantly to overexpression of plasminogen activator inhibitor-1 in the course of adipogenesis. The antidiabetic agents troglitazone and ciglitazone inhibited reporter gene expression driven by wild-type and -675 4G/5G mutant promoter, as well as the expression of endogenous plasminogen activator inhibitor 1, indicating that suppression of plasminogen activator inhibitor-1 expression may contribute to antidiabetic effects of these agents. The results indicate that absence of plasminogen activator inhibitor-1 in adipocytes may protect the cells against insulin resistance by promoting glucose uptake and adipocyte differentiation via a decrease in the peroxisome proliferator activated receptor gamma expression that modulates the adipocyte differentiation. PMID- 18160588 TI - The role of plasminogen activator inhibitor-1 polymorphism, factor-V-Leiden, and prothrombin-20210 mutations in pulmonary thromboembolism. AB - Polymorphism in plasminogen activator inhibitor-1 gene is suggested to be associated with an increased risk of venous thromboembolism. The aim of this study was to investigate the association of plasminogen activator inhibitor-1 gene polymorphism and its coexistence with factor-V-Leiden and prothrombin-20210 mutations in pulmonary thromboembolism. The authors investigated plasminogen activator inhibitor-1 4G/5G polymorphism, factor-V-Leiden, and prothrombin-20210 mutations in 143 pulmonary thromboembolism patients and 181 controls. Plasminogen activator inhibitor-1 4G/4G, 4G/5G, and 5G/5G gene polymorphisms and prothrombin 20210 mutations were not different between cases and controls. Factor-V-Leiden mutation was present in 21.0% and 7.7% of the cases and controls, respectively, P = .001. Neither different plasminogen activator inhibitor-1 genotypes and 4G allele nor coexistence of the allele with factor-V-Leiden or prothrombin-20210 was associated with the risk of recurrence. As a result, plasminogen activator inhibitor-1 gene polymorphism or its concomitant presence with mentioned mutations was not found to be associated with increased risk for pulmonary thromboembolism or recurrent disease in this study. PMID- 18160589 TI - Ocular vascular thrombotic events: central retinal vein and central retinal artery occlusions. AB - We prospectively assessed associations of thrombophilia- hypofibrinolysis with central retinal vein occlusion (CRVO) (40 patients) and central retinal artery occlusion (CRAO) (9 patients). We used polymerase chain reaction measures for thrombophilia (factor V Leiden, prothrombin, C677T MTHFR, platelet glycoprotein PlA1/A2) and hypofibrinolysis (plasminogen activator inhibitor-1 4G4G). Serologic thrombophilia measures included protein C, protein S (total and free) and antithrombin III, homocysteine, lupus anticoagulant, anticardiolipin antibodies IgG-IgM, and factors VIII and XI. Serologic hypofibrinolysis measures included Lp(a) and plasminogen activator inhibitor activity. For comparison with 40 CRVO and 9 CRAO patients, 80 and 45 race-gender matched controls were studied. The factor V mutation was more common in CRVO (3/40, 8%) than controls (0/79, 0%), P = .036, as was high (>150%) factor VIII (12/40, 30%) versus (4/77, 5%), P = .0002. Low antithrombin III (<80%) was more common in CRVO (5/39, 13%) than in controls (2/73, 3%), P = .049. Homocysteine was high (> or =13.5 micromol/L) in 5/39 (13%) CRVO patients versus 2/78 controls (3%), P = .04. Three of 9 CRAO patients (33%) had low (<73%) protein C versus 2/37 controls (5%), P = .044. Two of 9 CRAO patients (22%) had high (> or =13.5 micromol/L) homocysteine versus 0/42 controls (0%), P =. 028. Four of 9 CRAO patients had the lupus anticoagulant (44%) versus 4/33 (12%) controls (P = .050). CRVO is associated with familial thrombophilia (factor V Leiden, factor VIII, low antithrombin III, homocysteinemia), and CRAO is associated with familial and acquired thrombophilia (low protein C, homocysteinemia, lupus anticoagulant), providing avenues for thromboprophylaxis, and triggering family screening. PMID- 18160590 TI - Recombinant factor VIIa for the prophylaxis of perioperative hemorrhage in a patient with congenital factor XI deficiency undergoing brain tumor neurosurgery. AB - The authors report on the first successful use of recombinant activated factor VII for the prophylaxis of bleeding during brain tumor neurosurgery in a patient suffering from inherited factor XI deficiency. Using the agent, surgery was performed without any bleeding complications. In this setting, off-label use of recombinant activated factor VII appears to be a promising alternative for patients suffering from this rare hemostatic defect with hitherto limited therapeutic options. PMID- 18160591 TI - Severe arterial thrombophilia associated with a homozygous MTHFR gene mutation (A1298C) in a young man with Klinefelter syndrome. AB - Klinefelter syndrome (KS) is the most common sex chromosome disorder in men. It may be associated with an increased risk for venous thrombosis and thromboembolism, which is partially explained by hypofibrinolysis due to androgen deficiency. Additional genetic or acquired thrombophilic states have been shown in KS patients complicated with venous thrombosis as isolated case reports. Arterial thrombotic events had not been previously reported in KS. In this study, a young man with KS who developed acute arterial thrombosis during testosterone replacement therapy is presented. He was homozygous for the A1298C mutation of the methylenetetrahydrofolate reductase (MTHFR) gene. PMID- 18160592 TI - Platelet aggregation and activation in thalassemia major patients in Indonesia. AB - Thromboembolic events and hypercoagulable state have been reported in patients with thalassemia. As platelets play an important role in the pathogenesis of thrombosis, the authors aimed to find the pattern of changes in platelet count, function and activation, and evidence of coagulation activation in patients with thalassemia major in Indonesia. A total of 31 patients with splenectomized and 35 patients with nonsplenectomized thalassemia major were enrolled in this study. Platelet count, platelet aggregation, beta-thromboglobulin, and D-dimer levels were measured. All measured parameters were significantly higher in splenectomized than in nonsplenectomized patients. beta-thromboglobulin level was increased, but D-dimer level was within normal range. The authors concluded that there was an increase in platelet activation in patients with beta-thalassemia major. Platelet activation was higher in splenectomized than in nonsplenectomized patients. PMID- 18160593 TI - Homocysteine activates platelets in vitro. AB - The mechanism of thrombogenicity in hyperhomocysteinemia remains controversial. The authors investigated the association between elevated plasma homocysteine levels, platelet function, and blood coagulation. Blood was collected from healthy subjects and patients with critical limb ischemia. Basal platelet counts and platelet aggregation as well as flow cytometry were performed to assess spontaneous- and agonist-induced platelet aggregation as well as P-selectin and Glycoprotein IIb/IIIa expression at different homocysteine concentrations. Thromboelastography was performed, and platelet shape change was assessed, using a channelyzer, by measuring median platelet volume. Lactate dehydrogenase was measured, to indirectly assess red blood cell membrane integrity, after homocysteine exposure. The study results suggest that platelet activation and hypercoagulability occur after exposure to homocysteine, especially in patients with critical limb ischemia. Homocysteine concentrations of approximately 50 micromol/L appear to be the level at which these changes occur in vitro, and this effect on platelets appears to be indirect. PMID- 18160594 TI - The fibrinogen functional turbidimetric assay. AB - Hitherto, clinical fibrinogen methods were based on coagulation seconds, with assay conditions not similar to a plasma milieu. The fibrinogen functional turbidimetric assay included 50 microL citrated plasma + 100 microL 300 mIU/mL thrombin, 400 microg/mL polybrene, and 6% albumin-phosphate-buffered saline; an increase in absorbance at 405 nm/5 min at room temperature (or 2 minutes at 37 degrees C) was observed. In all, 6% albumin in the fibrinogen functional turbidimetric assay reagent abolishes falsely elevated fibrinogen to fibrin turbidity in hypoproteinemic plasma samples. This assay can detect fibrinogen activity of 250% to 300% of normal, the lower detection limit being 7% of normal (0.2 g/L). The normal range of this assay is 100% +/- 20% (mean value +/- 1 SD; coefficient of variations <4%). This assay imitates fibrinogen to fibrin conversion in clotting blood plasma; it is independent of plasmatic albumin or heparin and can be performed everywhere. This assay has a diagnostic value in pathology-disseminated intravascular coagulation and in assessing risk for atherothrombosis. PMID- 18160595 TI - The effect of ABO blood group on von Willebrand response to exercise. AB - Individuals of O blood group have significantly lower plasma levels of either Factor VIII (FVIII) or the von Willebrand factor (vWF). Conversely, there is accumulating evidence that elevated FVIII-vWF levels may represent an important risk factor for ischemic heart and venous thromboembolic disease. In this study, individuals exercised for 20 minutes at 10% below the first ventilatory threshold (aerobic threshold), which corresponds to 48% of maximum oxygen uptake. People with non-O blood group show higher resting and postexercise vWF levels compared with those of O blood group, as evidenced by a lower maximal heart rate. The groups were compared using the ANOVA one-way test, and a P < 0.05 was considered statistically significant. These results could change the way in which exercise training is designed for both healthy and sick individuals because O group individuals could have a more thrombogenic response to exercise. PMID- 18160596 TI - Pulmonary embolism in medical patients: an autopsy-based study. AB - Pulmonary embolism, though treatable, is a devastating disease and an important cause of morbidity and mortality among hospitalized patients. In all, 1000 autopsies were reviewed in adult medical patients. The overall incidence of pulmonary embolism in adult medical autopsies was 15.9% (159/1000). The incidence of pulmonary embolism contributing significantly to the death of the patients (groups 1 and 2) is 126/1000 (12.6%). Thus, pulmonary embolism very significantly contributed to death in 126/159 (79.24%) of group 1 and 2 patients. Pulmonary embolism affected a younger population as 79.87% of the overall patients, 66.67% of the fatal cases (group 1) and 73% of combined group 1 and 2 cases were below the age of 50 years. Sepsis was the primary diagnosis in 32% of total and in 42% of fatal cases. Hence, pulmonary embolism is considered as an important cause of death in patients admitted to the medical wards. It affects a younger population in India and needs to be tackled appropriately. PMID- 18160597 TI - Anti-factor Xa Assay: an effective method to determine the appropriate dose of heparin in pregnant women. PMID- 18160598 TI - MEF2A sequence variants in Turkish population. AB - Myocyte enhancer factor 2 (MEF2) is present in skeletal, cardiac, and smooth muscles and in neurons. MEF2A gene encodes a transcription factor which was on 15q26. The objective was to study the MEF2A gene in patients with premature MI. The control group consisted of 87 subjects who were older than 45 years with no history of cardiovascular disease or MI and no family history of CAD. The premature MI group consisted of 69 patients with documented MI younger than 45 years. No abnormal bands with single strand conformation polymorphism were detected after screening exon 1 through exon 8. This is the first study that detected 145408: T>C polymorphism in intron 10. In both study groups, the rare polymorphism P279L in exon 7, T>C polymorphism in intron 10, and 21-bp deletion in exon 11 of the gene were not found. The data supported the previous studies indicating no association between MEF2A gene and premature MI. PMID- 18160599 TI - Recurrent abortions in Asian Indians: no role of factor V Leiden Hong Kong/Cambridge mutation and MTHFR polymorphism. AB - Recurrent fetal loss is a frequent health problem. Data accumulated over the past few years have suggested a possible correlation between thrombophilia and fetal loss. Although a clear association has been established between fetal loss and certain thrombophilic states, such as antiphospholipid antibody syndromes, antithrombin deficiency, and combined defects, reports on the prevalence of inherited prothrombotic defects such as factor V Leiden mutation and methylene tetrahydrofolate reductase C677T polymorphism in fetal loss are contradictory. The prevalence of these 2 mutations in Asian Indians with recurrent fetal loss has not yet been studied. In light of this, the present study looked at the prevalence of these mutations in 85 patients with spontaneous recurrent abortion and 31 controls. The authors did not find any significant role of these mutations in the development of recurrent abortion. PMID- 18160600 TI - Plasma plasminogen activator inhibitor-1 levels and nonalcoholic fatty liver in individuals with features of metabolic syndrome. AB - Fatty liver represents the liver component of metabolic syndrome and may be involved in plasminogen activator inhibitor-1 (PAI-1) synthesis. We studied plasma PAI-1 levels and relationships with risk factors for metabolic syndrome, including fatty liver, in 170 patients. Liver ultrasound scan was performed on all patients, and a liver biopsy was performed on those patients with chronically elevated transaminase levels. Plasma PAI-1 levels correlated significantly (P < .05) with body mass index, degree of steatosis, insulin resistance, insulin level, waist circumference, triglycerides, and high-density lipoprotein (HDL) cholesterol. However, only body mass index (beta = .455) and HDL-cholesterol (beta = .293) remained predictors of PAI-1 levels. Liver biopsy revealed a significant correlation (P < .05) between insulin resistance (r = 0.381) or insulin level (r = 0.519) and liver fibrosis. In patients presenting features of metabolic syndrome, plasma PAI-1 levels were mainly conditioned by the whole-body fat content. PMID- 18160601 TI - Factor V Leiden, prothrombin G20210A, and protein C mutation frequency in Turkish venous thrombosis patients. AB - Several inherited polymorphisms are associated with risk of venous thrombosis, including mutation at codon 506 of the factor V gene, mutation at position 20210 of the prothrombin gene, and mutations in the protein C gene. In this study, genotyping for factor V, prothrombin, and protein C mutations was performed in 50 patients and 25 control subjects by polymerase chain reaction-based analysis. The prevalence of factor V and prothrombin mutations was not significantly different from that in the general population. Nine of the patients had heterozygous protein C mutation. There was a high prevalence of the mutated protein C allele in the pulmonary emboli group (42.8%). Protein C mutation incidence was higher in the pulmonary emboli group than in the deep vein thrombosis (8.33%) and cerebral vein thrombosis (16.1%) groups. These results indicate that patients with protein C deficiency have a greater risk of thrombosis than patients with factor V or prothrombin G20210A mutation. PMID- 18160602 TI - The importance of thrombotic risk factors in the development of idiopathic sudden hearing loss. AB - Impaired cochlear blood circulation has been suggested to cause sudden hearing loss. In this study, the role of factor V 1691 G-A (FV 1691 G-A), prothrombin 20210 G-A (PT 20210 G-A), methylene tetrahydrofolate reductase 677 C-T (MTHFR 677 C-T), factor V 4070 A-G (FV 4070 A-G), endothelial cell protein C receptor (EPCR) gene 23-bp insertion, and plasminogen activator inhibitor-1 (PAI-1) 4G/5G mutation was assessed. Fifty-three patients with idiopathic sudden sensorineural hearing loss and 80 individuals comprising the control group were included in this study. The frequency for FV 1691 A was 6.2% in the patient group and 3.7% in the control group, PT 20210 G-A was 1.2% in the patient group and 1.9% in the control group, and FV 4070 A-G was 7.5% in the patient group and 11.3% in the control group. The frequency of MTHFR 677 C-T was significantly higher in the patient group than in the control group, with a P value of .03. PAI-1-675 4G/5G polymorphism was found to be 71.2% and 69.8%, in the control group and the patient group, respectively. The EPCR 23-bp insertion was 0% in the control group and was found in 3 patients (3.7%), which needs further study. PMID- 18160603 TI - Successful pregnancy outcome in women with bad obstetric history and recurrent fetal loss due to thrombophilia: effect of unfractionated heparin and low molecular weight heparin. AB - Acquired and inherited thrombophilias are known to be associated with unfavorable pregnancy outcome including recurrent fetal loss. There are differences of opinion whether these patients need to be treated with aspirin, unfractionated heparin, low-molecular weight heparin, corticosteroids, or intravenous immunoglobulins. In all, 25 consecutive patients with a history of fetal loss and 7 patients who presented in early pregnancy with deep-vein thrombosis were treated, and their pregnancy outcome was noted. All the women were positive either for a solitary or for a combination of acquired and heritable thrombophilia markers. In all, 23 patients were treated with unfractionated heparin and 9 with low-molecular weight heparin. In all, 16 out of 23 patients (69.6%) treated with unfractionated heparin and 9 out of 9 (100%) treated with low-molecular weight heparin had successful pregnancy outcome. There was a complete resolution of thrombus in all the cases. None of the patients had any adverse reactions such as heparin-induced thrombocytopenia, thrombosis, or fracture. Both unfractionated heparin and low-molecular weight heparin were effective in cases of bad obstetric history and recurrent pregnancy loss due to thrombophilia. However, low-molecular weight heparin was found to be more effective than unfractionated heparin along with other advantages of not requiring laboratory monitoring and easy administration. None of the patients in either group had to interrupt the therapy for any adverse treatment-related complications. PMID- 18160604 TI - Correction of the bleeding time with lyophilized platelet infusions in dogs on cardiopulmonary bypass. AB - Lyophilized canine platelets were infused in a single large bolus dose into splenectomized dogs after 2 hours' perfusion on cardiopulmonary bypass to test their possible efficacy in restoring hemostasis after compromise of platelet function. The vessel bleeding time (VBT) was monitored by venipuncture of the exposed jugular vein. During cardiopulmonary bypass, platelet counts fell quickly and the VBTs became prolonged over baseline. Infusion of lyophilized platelets reconstituted in normal saline occurred just before or immediately after weaning from the cardiopulmonary bypass pump. The results showed consistent and persistent lowering of the VBTs by the infused lyophilized platelets. Controls showed continuously prolonged VBTs. The weighted average VBT in infused subjects was significantly lower than the average in controls: 3 minutes 10 seconds versus 6 minutes 59 seconds, respectively (t test, P= .01). These results in this setting indicate the possible effectiveness of similar human lyophilized platelet preparations in reducing postoperative bleeding in open heart surgery. PMID- 18160605 TI - Alterations of von Willebrand factor and ristocetin cofactor activity during atrial fibrillation. AB - The aim of this study was to assess the plasma levels of von Willebrand factor antigen and ristocetin cofactor activity, which are well-known markers of endothelial function, in atrial fibrillation (AF) with or without mitral stenosis (MS). Forty-two patients (16 patients with MS and AF [MS(+)AF(+)], 13 patients with nonvalvular AF [MS(-)AF(+)], and 13 patients with MS and sinus rhythm [MS(+)AF(-)]) were included. Von Willebrand factor antigen levels and ristocetin cofactor activities in all participants were assessed. Overall, von Willebrand factor antigen levels and ristocetin cofactor activities in the AF(+) patients were higher than in the AF(-) patients (P = .003 and P = .002, respectively). Von Willebrand factor antigen levels and ristocetin cofactor activities in the 3 groups were found to be different (P = .012 and P = .01, respectively). Von Willebrand factor antigen levels were similar between the MS(+)AF(+) and MS( )AF(+) groups and were higher than that of the MS(+)AF(-) group. Ristocetin cofactor activity in the MS(-)AF(+) group was significantly higher than in the other 2 groups. The ristocetin cofactor activity and von Willebrand factor antigen levels were significantly higher in diabetic or hypertensive patients than in nondiabetic or normotensive patients. According to the results of this study, circulating von Willebrand factor antigen levels and plasma ristocetin cofactor activities are affected by the presence of AF, MS, and associated comorbidities including type 2 diabetes mellitus and systemic hypertension. Further studies are needed to assess the role of von Willebrand factor antigen and ristocetin cofactor activity in predicting vascular thrombotic events in AF, MS, systemic hypertension, and diabetes mellitus. PMID- 18160606 TI - Unexpected pulseless disease associated with recurrent venous thromboembolisms. AB - Venous thromboembolic disease is a well-documented complication of Klinefelter's syndrome, even if mechanisms underlying this prothrombotic state have not been conclusively established. On the contrary, arterial thrombosis is far less frequent, and a case of a patient with Klinefelter's syndrome presenting with simultaneous venous thrombtoembolic disease and a complete thrombosis of the left subclavian artery is presented. Elevated levels of type 1 plasminogen activator inhibitor, in the absence of other usual thrombophilic abnormalities, raise the question of the role played by this inhibitor of the fibrinolysis in the arterial and venous thromboses presented by this patient. PMID- 18160607 TI - Oxidative stress in young subjects with acute myocardial infarction: evaluation at the initial stage and after 12 months. AB - In 105 subjects (97 men and 8 women) aged <46 years (mean age 39.6 +/- 5.5 years), with recent acute myocardial infarction (T1), thiobarbituric acid reactive substances and total antioxidant status were determined; NO production was evaluated by measuring the nitrite and nitrate (NOx) concentration. The patients with acute myocardial infarction were subdivided according to the main risk factors, number of risk factors, and extent of coronary lesions. The evaluation was repeated after 12 months (T2). In these subjects, thiobarbituric acid reactive substances and NOx were significantly increased and total antioxidant status was significantly decreased at T1. In single risk factor, only NO metabolites were significantly lower in acute myocardial infarction subjects who smoke than in subjects who do not. Subdividing the subjects according to the number of risk factors and number of stenosed coronary vessels, there were no significant differences between the subgroups. At T2, thiobarbituric acid reactive substances and NOx were decreased and total antioxidant status was increased, but all parameters were still altered. PMID- 18160608 TI - Heparin-induced thrombocytopenia complicating hemodialysis. AB - Hemodialysis complicated by heparin-induced thrombocytopenia (HIT) is a rare event requiring anticoagulation with direct-thrombin inhibitors. Contaminant calcific uremic arteriolopathy (calciphylaxis) further complicates this situation due to the possibility that warfarin anticoagulation may exacerbate skin necrosis. The authors report a patient with renal failure and calciphylaxis who developed HIT after starting hemodialysis. She was successfully treated with Argatroban. PMID- 18160609 TI - Pleiotropic effects of heparin and heparinoids in peritoneal dialysis. AB - Unfractionated heparin, low-molecular-weight heparins, and sulodexide belong to the family of glycosaminoglycans. Recent studies report on properties other than anticoagulant activities of these medications. They include modulation of cell growth and proliferation via actions on numerous growth factors affecting the immune system and matrix molecules production and degradation. Long-term peritoneal dialysis remarkably influences peritoneal cavity homeostasis by mechanisms mediated by growth factors. They initiate progression of pathological processes and further account for morphological and functional alterations of the peritoneal membrane. The best-recognized pathologies in peritoneal cavity under these conditions encompass inflammation, fibrosis, and vasculopathy, often leading to fatal encapsulating peritoneal sclerosis. Intraperitoneal heparin and its derivatives, by their pleiotropic actions, may influence these crucial processes and improve the peritoneal dialysis technique survival in a complex and so far understudied way. These issues, novel medical approaches, and their likely mechanisms have been reviewed. PMID- 18160610 TI - Aspirin-resistance frequency: a prospective study in 280 healthy Turkish volunteers. AB - This study reports the frequency of aspirin resistance and its correlation with clinical and biochemical parameters among 280 healthy Turkish volunteers (179 men, 101 women) who were taking 100 mg of aspirin 7 days or more. Aspirin resistance was detected by optical platelet aggregometry, using adenosine diphosphate and arachidonic acid, and defined as a mean aggregation of 64% or more with 5AmicroM adenosine diphosphate and a mean aggregation of 20% or more with 0.5-mg/mL arachidonic acid. Of the study population, 27.5% (26 women [25.5 %] and 51 men [28.5 %]) were aspirin resistant. The current findings indicate that aspirin resistance is an important and real laboratory diagnosis given its frequency of 27.5% in the study population. These results of this large trial evaluating the frequency of aspirin resistance in healthy subjects indicate that aspirin resistance should be diagnosed so that individuals with no response can receive alternative or additional antiplatelet therapy. PMID- 18160611 TI - Further insight into the heparin-releasable and glycosylphosphatidylinositol lipid--anchored forms of tissue factor pathway inhibitor. AB - The release of tissue factor pathway inhibitor (TFPI) from human umbilical vein endothelial cells (HUVECs) was investigated using heparin and phospholipase C. The experiment included incubating HUVECs with 0, 1, or 10 U/mL heparin diluted in Dulbecco Modified Eagle's Medium plus 5% fetal calf serum for 1 or 24 hours. A statistically significant increase in TFPI activity levels was seen at 1 hour, but not at 24 hours. A 20-fold increase in the release of TFPI after phospholipase C treatment of HUVECs was demonstrated, confirming that it is glycosylphosphatidylinositol-lipid (GPI) anchored. Sequential treatment of HUVECs with phospholipase C and heparin was performed, and a trend was observed where GPI-anchored TFPI levels were increased after 1 hour of pretreatment with heparin but were decreased after 24 hours. Serum is a requirement for the heparin dependent release of TFPI from HUVECs. Heparin pretreatment of HUVECs may affect levels of GPI anchored TFPI in a time and dose-dependent manner. PMID- 18160612 TI - The success of clopidogrel treatment for recurrent coronary thrombosis in a patient with aortic and mitral valve replacement. PMID- 18160613 TI - Portal and mesenteric vein thromboses in a patient with prothrombin G20210 mutation, elevated lipoprotein (a), and high factor VIII. AB - A 65-year-old man was examined for abdominal pain. Portal and mesenteric vein thromboses were described by ultrasound and computed tomography. No local cause was found. The patient had a positive history of venous thromboembolism. Thrombophilia workup revealed prothrombin G20210A mutation (heterozygous), C677T mutation of methylenetetrahydrofolate reductase gene (homozygous), elevated level of lipoprotein (a), and high level of coagulation factor VIII. Anticoagulation was started and planned for a long-term duration. The etiology of portal vein thrombosis is often multifactorial, with various combinations of systemic factors (inherited or acquired prothrombotic conditions) and local precipitating factors (inflammation, injury to the portal venous system, cancer of the abdominal organs, cirrhosis). The reported prevalence of hypercoagulable states in patients with portal vein thrombosis has been very heterogeneous so far. Some authors support a role of the prothrombin G20210A mutation. In the reported patient, this mutation was revealed in a combination with other hypercoagulable states. PMID- 18160614 TI - Platelet adhesion testing may predict early hemodialysis arteriovenous graft and fistula failure in end-stage renal disease patients. AB - Vascular access thrombosis (VAT) is the most morbid and costly complication in end-stage renal disease (ESRD) patients. Although hypercoagulability is a major risk factor for VAT, in most patients, the cause of hypercoagulability cannot be identified despite clinical suspicion. In this study, platelet hyperreactivity was investigated for a possible role in the hypercoagulability of ESRD and VAT in 42 patients with arteriovenous (AV) grafts or fistulas. Platelet adhesion, platelet aggregation, and the history of VAT were assessed. The statistics included a nonparametric 2-factor ANOVA, a Mann-Whitney analysis, and a Kaplan Meier analysis of hemodialysis angioaccess survival to examine platelet hyperadhesiveness as a predictor of access survival. The study showed a significant correlation between increased platelet adhesiveness and shortened survival of the primary hemodialysis angioaccess. Collagen-induced platelet aggregation reflected a significantly higher response in those with shortened access survival. These findings may have significant clinical implications for risk assessment and prevention of VAT. PMID- 18160615 TI - Optimal management of an aneurysmal subarachnoid hemorrhage in a patient with known factor XI deficiency: a case report. AB - The authors report a rare case of an acute cerebral aneurysm rupture in a patient with a known factor XI deficiency. Aneurysmal subarachnoid hemorrhage (SAH) accounts for a high mortality and morbidity rate. When SAH is associated with an inherited coagulation disorder such as hemophilia C, an unexpected and possible increase in hemorrhagic stroke and increase in bleeding during surgery and in the postoperative period could lead to an extremely bad outcome. Clinical management consists of rapid correction of the coagulation disorder before undergoing any invasive intracranial procedure. Such an optimal therapeutic strategy must be under the care of a multidisciplinary medical and surgical team. Human factor XI concentrate (Hemoleven, Laboratoire Francais du Fractionnement et des Biotechnologies [LFB], Les Ulis, France) was used successfully in this case report. New treatment using recombinant factor VIIa is discussed. PMID- 18160616 TI - Low-density lipoprotein specifically binds glycoprotein IIb/IIIa: a flow cytometric method for ligand-receptor interaction. AB - Primary platelet aggregation requires agonist-mediated activation of membrane receptor glycoprotein (GP) IIb/IIIa, binding of fibrinogen to GpIIb/IIIa, and cellular events after fibrinogen binding. This study investigated whether fibrinogen receptor GpIIb/IIIa is also the binding site for low-density lipoprotein (LDL) in platelets by using GpIIb/IIIa-coated polystyrene microbeads incubated with various concentrations of fluorescein isothiocyanate (FITC) labeled ligands. Binding was assayed by flow cytometry. Binding of fibrinogen (Fg)-FITC and LDL-FITC to GpIIb/IIIa coated microbeads was concentration dependent, reaching saturation. Binding of LDL-FITC to GpIIb/IIIa coated microbeads was inhibited by fibrinogen. Binding of LDL-FITC or Fg-FITC to freshly isolated platelets gave similar results as those of GpIIb/IIIa coated microbeads. Glycoprotein IIb/IIIa, the fibrinogen receptor on platelets is also one of the binding sites of LDL on platelets. This rapid and reliable flow cytometric technique using coated microbeads may be used as a first step for the study of all ligand receptor interactions. PMID- 18160617 TI - Activated protein C resistance and factor V Leiden in Mexico. AB - A common cause of hereditary thrombophilia is activated protein C resistance (APCR), and most cases result from factor V Leiden mutation. An APCR phenotype without association with factor V Leiden has been described. This transversal, observational, nonrandomized study evaluated these 2 phenomena in healthy indigenous and mestizo Mexican subjects (n = 4345), including 600 Mexican natives. No indigenous subjects had APCR, but 82 mestizo subjects did. After retesting, 50 subjects had a negative test. The remaining 32 subjects had factor V Leiden, giving a 0.85% prevalence of factor V Leiden in the mestizo Mexican population. Only 31% of APCR carriers had factor V Leiden. These results show a very low prevalence of APCR and factor V Leiden in Mexico. Except for factor V Leiden, there are no other mutations in the factor V gene responsible for the APCR phenotype. Acquired APCR is nearly twice as prevalent as the inherited variant. PMID- 18160618 TI - Reduction of chemokine secretion in response to mycobacteria in infliximab treated patients. AB - The use of anti-tumor necrosis factor (TNF) agents as a treatment for chronic inflammatory conditions has been shown to be associated with an increased risk of developing tuberculosis. We studied the effect of the anti-TNF antibody infliximab on antimycobacterial immunity in 26 patients with rheumatoid arthritis or ankylosing spondylitis by use of an in vitro whole-blood model employing a reporter mycobacterium. Blood samples taken before and 30 min and 7 days after a 2-hour infliximab infusion were compared in terms of their abilities both to suppress luminescence of Mycobacterium bovis bacillus Calmette-Guerin lux and to secrete chemokines and cytokines 24 and 96 h after infection. No immediate effect of infliximab on mycobacterial luminescence was detected using this bioassay, irrespective of whether patients were receiving their first (n = 14) or maintenance (n = 12) doses of infliximab. Moreover, no effect on mycobacterial luminescence was detected when blood was taken 7 days after infliximab treatment (n = 7). By contrast, there was a significant reduction in the chemokines implicated in cellular trafficking, namely, interleukin-8, macrophage-inhibitory protein-1alpha (MIP-1alpha), MIP-1beta (24 h and 96 h), and monocyte chemoattractant protein-1 (MCP-1) (24 h) following BCG lux strain infection in the 30-minute post-infliximab-infusion blood samples (P < 0.05). This effect was sustained by MIP-1beta and MCP-1 (24 h; P < 0.05) at 7 days after infusion. Our results suggest that the development of tuberculosis in infliximab-treated patients is not directly related to the mycobactericidal effects of TNF but may be due to inhibition of TNF-dependent chemokine gradients disrupting cellular migration necessary to maintain the integrity of the granuloma. PMID- 18160619 TI - Evaluation of the antigenic relationships among canine parvovirus type 2 variants. AB - The antigenic relationships among the original canine parvovirus type 2 (CPV-2) and the variants CPV-2a, -2b, and -2c were evaluated. Cross-antigenic evaluation revealed clear differences among the CPV variants, which were more appreciable by serum neutralization (SN) than by hemagglutination inhibition. Antigenic differences were found mostly between the original CPV-2 and the variants, but they were also observed among the variants CPV-2a, -2b, and -2c. The variant CPV 2c exhibited a unique antigenic pattern, since it was poorly recognized by the sera of animals immunized with CPV-2, CPV-2a, and CPV-2b. However, animals immunized with CPV-2c exhibited higher SN titers to CPV-2b than to the homologous virus CPV-2c. The observed antigenic differences might drive selection of CPV strains by generating differential immune pressure in the canine population, which raises concerns about vaccine efficacy. PMID- 18160620 TI - Borrelia burgdorferi complement regulator-acquiring surface protein 2 (CspZ) as a serological marker of human Lyme disease. AB - Serological diagnosis of Lyme disease may be complicated by antigenic differences between infecting organisms and those used as test references. Accordingly, it would be helpful to include antigens whose sequences are well conserved by a broad range of Lyme disease spirochetes. In the present study, line blot analyses were performed using recombinant complement regulator-acquiring surface protein 2 (BbCRASP-2) from Borrelia burgdorferi sensu stricto strain B31 and serum samples from human Lyme disease patients from throughout the United States and Germany. The results indicated that a large proportion of the patients had produced antibodies recognizing recombinant BbCRASP-2. In addition, Lyme disease spirochetes isolated from across North America and Europe were found to contain genes encoding proteins with high degrees of similarity to the B. burgdorferi type strain B31 BbCRASP-2, consistent with the high percentage of serologically positive patients. These data indicate that BbCRASP-2 may be valuable for use in a widely effective serological assay. PMID- 18160621 TI - Monoclonal antibodies that bind to common epitopes on the dengue virus type 2 nonstructural-1 and envelope glycoproteins display weak neutralizing activity and differentiated responses to virulent strains: implications for pathogenesis and vaccines. AB - The abilities of monoclonal antibodies (MAbs) that bind to defined sequential epitopes on the dengue virus (DENV) nonstructural-1 (NS1) glycoproteins to cross react with epitopes on the DENV envelope (E) glycoproteins were investigated. In this study, some of these MAbs cross-reacted with the DENV type 2 (DENV-2) E glycoprotein and with synthetic peptides representing X-ray crystallographically confirmed surface-exposed regions on this glycoprotein. MAb 1G5.3 cross-reacted with the flavivirus-conserved 101-WGNGCGLFG-109 fusion sequence, the 273-SSGNL 277 DENV-2 hinge region sequence, and the 156-GKHGKEIKIT-165 sequence of virulent DENV-2 strains. MAb 1G5.4-A1-C3 cross-reacted with the 67-NTTTESRCPT-76 and 156 GKHGKEIKIT-165 sequences of virulent DENV-2 strains, the 338-EIMDLDNRHV-347 sequence from a highly virulent DENV-2 (M2) strain, and two epitopes on a virulent DENV-3 strain (288-KMDKLELKG-296 and 323-RVEYRGEDAP-332), which all contained target ELK/KLE-type motifs (underlined). These MAbs showed reduced cross-reactions with the corresponding sequences from weakly pathogenic strains of all four DENV serotypes and had either no (MAb 1G5.4-A1-C3) or weak (MAb 1G5.3) neutralizing activity against them. MAb 1G5.3 more strongly neutralized DENV-2 strains with higher pathogenic capacities, while MAb 1G5.4-A1-C3 showed increasing neutralizing titers against the virulent DENV-3 strain and the moderately virulent and highly virulent (M2) DENV-2 strains. These cross reactions with the E glycoprotein accord with the observation that MAb 1G5.3 caused dramatic and lethal antibody-enhanced replication (AER) of a DENV-2 strain in vivo. Together with in vivo AER studies of these DENV strains using MAb 1G5.4 A1-C3, these results may account for the increased pathogenic capacities of such strains, which is likely to have important implications for pathogenesis and vaccines. PMID- 18160622 TI - Amelioration of oxidative mitochondrial DNA damage and deletion after renal ischemic injury by the KATP channel opener diazoxide. AB - Renal ischemia was induced in the rat by constriction of the renal artery for 45 min, and the ability of the ATP-sensitive K(+) (K(ATP)) channel opener diazoxide (DZ) to ameliorate renal ischemia-reperfusion (I/R) injury was evaluated. In this model, blood urea nitrogen and creatinine were elevated 2 days after I/R injury but returned closer to normal levels by 7 days after reperfusion. Histological staining for reactive oxygen species (ROS) was clearly positive and oxidized DNA, detected by the presence of the stable adduct 8-hydroxy-2'-deoxyguanosine, was clearly present in the cytoplasm of tubular cells after 1 h of reperfusion and declined 7 days after reperfusion. This finding was confirmed by ELISA, which detected 8-hydroxy-2'-deoxyguanosine in the mitochondrial fraction of kidney homogenates. Despite evidence of improved function measured by blood urea nitrogen and creatinine 7 days after reperfusion, the early changes in tubules were alarming. Mitochondrial DNA showed the common deletion, and the number of TdT-mediated dUTP nick-end label-positive tubular cells increased. Activation of caspase-3 continued, and abnormal levels of ROS were found in the mitochondrial fraction of cellular homogenates. Treatment with DZ before ischemia reduced or prevented the acute and subacute deleterious effects associated with renal I/R injury. We conclude that excess production of ROS by mitochondria on reperfusion is a major upstream event in renal reperfusion injury and that DZ functioned by preventing ROS accumulation in the mitochondria after I/R injury, thereby reducing oxidative stress as measured by the presence of oxidized mitochondrial DNA and features of apoptosis. PMID- 18160623 TI - Arterial stiffness and interdialytic weight gain influence ambulatory blood pressure patterns in hemodialysis patients. AB - Besides an overall increase in blood pressure, hemodialysis patients have marked disturbance in interdialytic ambulatory blood pressure pattern that is characterized by blunted circadian amplitude and a steady rise in blood pressure between dialysis treatments. The pathophysiology of this abnormal blood pressure profile is poorly understood. We hypothesized that the circadian amplitude, the interdialytic increase (linear trend), and the average level of blood pressure (the intercept) are related to the extent of arterial stiffening and the degree of accumulation of salt and water between dialysis treatments. Using a generalized cosinor model, we simultaneously compared the impact of interdialytic weight gain and echo-Doppler-measured aortic pulse wave velocity on the mean level of blood pressure, linear changes over the interdialytic interval, and oscillatory changes in blood pressure. In a cross-sectional analysis of 11,833 blood pressure measurements from 125 long-term hemodialysis patients, we found that aortic pulse wave velocity and interdialytic weight gain had a substantial impact on interdialytic ambulatory blood pressure level, trends, and rhythms. Arterial stiffness was associated with an overall increase in the level (intercept) of systolic, diastolic, and pulse pressure. Interdialytic weight gain, on the other hand, was associated with interdialytic increase (linear trend) in blood pressure. The circadian amplitude was blunted by increments in either arterial stiffness or interdialytic weight gain. Since patterns of ambulatory arterial blood pressure are related to cardiovascular risk factors such as interdialytic weight gain and increased arterial stiffness, the pattern of ambulatory blood pressure recordings may also be of prognostic significance in hemodialysis patients. PMID- 18160624 TI - Inhibitory and excitatory perigenital-to-bladder spinal reflexes in the cat. AB - This study revealed that in awake chronic spinal cord-injured (SCI) cats reflexes from perigenital skin area to the bladder can be either inhibitory or excitatory. Electrical perigenital stimulation at frequencies between 5 and 7 Hz significantly inhibited large-amplitude rhythmic reflex bladder activity, whereas frequencies between 20 and 40 Hz induced large-amplitude bladder contractions even at low bladder volumes when reflex bladder activity was absent. Both inhibitory and excitatory effects were enhanced as the stimulation intensity increased (5-30 V, 0.2-ms pulse width). During cystometrograms, the inhibitory stimulation (7 Hz) significantly increased the micturition volume threshold 35 +/ 13% above the control volume, while the excitatory stimulation (30 Hz) significantly reduced the threshold 21 +/- 3%. Mechanical perigenital stimulation applied by repeated light stroking of the perigenital skin with a cotton swab only induced an excitatory effect on the bladder. Both electrical and mechanical perigenital stimuli induced large-amplitude (>30 cm H(2)O) bladder contractions that were relatively consistent over a range of bladder volumes (10-90% of the capacity). However, the excitatory electrical stimulation only induced bladder contractions lasting on average 42.2 +/- 3.9 s, but the mechanical stimulation induced bladder contractions that lasted as long as the stimulation continued (2 3 min). Excitatory electrical or mechanical perigenital stimulation also induced poststimulus voiding. The ability to either inhibit or excite the bladder by noninvasive methods could significantly transform the current clinical management of bladder function after SCI. PMID- 18160625 TI - Effects of targeted Bcl-2 expression in mitochondria or endoplasmic reticulum on renal tubular cell apoptosis. AB - Bcl-2 family proteins are central regulators of apoptosis. As the prototypic member, Bcl-2 protects various types of cells against apoptotic insults. In mammalian cells, Bcl-2 has a dual subcellular localization, in mitochondria and endoplasmic reticulum (ER). The respective roles played by mitochondrial and ER localized Bcl-2 in apoptotic inhibition are unclear. Using Bcl-2 constructs for targeted subcellular expression, we have now determined the contributions of mitochondrial and ER-localized Bcl-2 to the antiapoptotic effects of Bcl-2 in renal tubular cells. Wild-type Bcl-2, when expressed in renal proximal tubular cells, showed partial colocalizations with both cytochrome c and disulfide isomerase, indicating dual localizations of Bcl-2 in mitochondria and ER. In contrast, Bcl-2 constructs with mitochondria-targeting or ER-targeting sequences led to relatively restricted Bcl-2 expression in mitochondria and ER, respectively. Expression of wild-type and mitochondrial Bcl-2 showed significant inhibitory effects on tubular cell apoptosis that was induced by cisplatin or ATP depletion; however, ER-Bcl-2 was much less effective. During ATP depletion, cytochrome c was released from mitochondria into the cytosol. This release was suppressed by wild-type and mitochondrial Bcl-2, but not by ER-Bcl-2. Consistently, wild-type and mitochondrial Bcl-2, but not ER-Bcl-2, blocked Bax activation during ATP depletion, a critical event for mitochondrial outer membrane permeabilization and cytochrome c release. In contrast, ER-Bcl-2 protected against apoptosis during tunicamycin-induced ER stress. Collectively, the results suggest that the cytoprotective effects of Bcl-2 in different renal injury models are largely determined by its subcellular localizations. PMID- 18160626 TI - Albumin-stimulated DNA synthesis is mediated by Ca2+/PKC as well as EGF receptor dependent p44/42 MAPK and NF-kappaB signal pathways in renal proximal tubule cells. AB - It is now recognized that significant tubular reabsorption of albumin occurs under physiological conditions that may play an important role in maintaining proximal tubular integrity and function. Therefore, this study examined the effect of bovine serum albumin (BSA) on DNA synthesis and its related signal molecules in primary cultured rabbit renal proximal tubule cells (PTCs). BSA increased the level of [(3)H]thymidine incorporation in a dose (> or =3 mg/ml)- and time (> or =3 h)-dependent manner, intracellular Ca(2+) concentration, and the level of protein kinase C (PKC) phosphorylation and stimulated the phosphorylation of the epidermal growth factor receptor (EGFR), which was inhibited by EGTA (extracellular Ca(2+) chelator), 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM, intracellular Ca(2+) chelator), or PKC inhibitors (staurosporine or bisindolylmaleimide I). In addition, the PKC inhibitors or an EGFR inhibitor (AG-1478) blocked the BSA induced phosphorylation of p44/42 mitogen-activated protein kinases (MAPKs). BSA also increased the level of nuclear factor-kappaB (NF-kappaB) and inhibitor of NF kappaB (IkappaB) phosphorylation, which was blocked by staurosporine, AG-1478, or PD-98059 (p44/42 MAPK inhibitor). Inhibition of Ca(2+), PKC, EGFR, p44/42 MAPK, or NF-kappaB signal pathways blocked the BSA-induced incorporation of [(3)H]thymidine. Consequently, the inhibition of Ca(2+), PKC, EGFR, p44/42 MAPKs, or NF-kappaB blocked the BSA-induced increases in cyclin D1, cyclin-dependent kinase (CDK)4, cyclin E, or CDK2 and restored the BSA-induced inhibition of p21(WAF/Cip1) and p27(Kip1) expression. In conclusion, BSA stimulates DNA synthesis that is mediated by Ca(2+)/PKC as well as the EGFR-dependent p44/42 MAPK and NF-kappaB signal pathways in PTCs. PMID- 18160627 TI - Inhibition of proinflammatory genes in anti-GBM glomerulonephritis by targeted dexamethasone-loaded AbEsel liposomes. AB - E-selectin-directed targeted drug delivery was analyzed in anti-glomerular basement membrane glomerulonephritis. Liposomes conjugated with anti-E-selectin antibodies (Ab(Esel) liposomes) were internalized by activated endothelial cells in vitro through E-selectin-mediated endocytosis. At the onset of glomerulonephritis in mice, E-selectin was expressed on glomerular endothelial cells, which resulted in homing of Ab(Esel) liposomes to glomeruli after intravenous administration. Accumulation of Ab(Esel) liposomes in the kidney was 3.6 times higher than nontargeted IgG liposomes, whereas the accumulation of both liposomes in the clearance organs liver and spleen and in heart and lungs was comparable. In glomeruli, the Ab(Esel) liposomes colocalized with the endothelial cell marker CD31. Quantitative RT-PCR analysis of laser-microdissected arterioles, glomeruli, and postcapillary venules demonstrated that targeted delivery of dexamethasone by Ab(Esel) liposomes reduced glomerular endothelial expression of P-selectin, E-selectin, and vascular cell adhesion molecule-1 by 60 70%. The expression of these genes was not modulated in endothelial cells in nontargeted renal microvasculatures. Decrease of glomerular endothelial activation at disease onset was followed by reduced albuminuria at day 7. This study demonstrates the potential of vascular bed-specific drug delivery aimed at disease-induced epitopes on the microvascular endothelial cells as a therapeutic strategy for glomerulonephritis. PMID- 18160628 TI - Bradykinin modulates focal adhesions and induces stress fiber remodeling in renal papillary collecting duct cells. AB - Focal adhesions (FAs) are specialized regions of cell attachment to the extracellular matrix. Previous works have suggested that bradykinin (BK) can modulate cell-matrix interaction. In the present study, we used a physiological cellular model to evaluate the potential role of BK in modulating FAs and stress fibers. We performed a quantitative morphometric analysis of FAs in primary cultured rat renal papillary collecting duct cells, which included size, axial ratio (shape), and average length. After 1, 5, or 10 min of incubation with BK, cultured cells were immunostained and analyzed by confocal microscopy. Although the shape of FAs was not altered, BK induced a decrease in the number of vinculin stained FAs per cell, and a decrease in both their size and their average length, but not in talin-containing FAs, thus suggesting that BK could be inducing a restructuring of FAs. BK also induced a remodeling of the actin filament assemblies rather than their dissipation. Since we have previously demonstrated that BK stimulates activation of PLCbeta in rat renal papillae, we attempted to determine whether BK can modulate FA restructuring by this mechanism, by pretreating cultured cells with the PLCbeta inhibitor U73122. The present study, performed under physiological conditions with cells that were not genetically manipulated, provides new experimental evidence supporting the notion that the intrarenal hormone BK modulates FAs and actin cytoskeleton organization through a mechanism that involves the activation of PLCbeta. We propose this finding as a novel mechanism for BK modulation of tubular collecting duct function. PMID- 18160630 TI - Targeting and readout strategies for fast optical neural control in vitro and in vivo. PMID- 18160631 TI - Systemic and nasal delivery of orexin-A (Hypocretin-1) reduces the effects of sleep deprivation on cognitive performance in nonhuman primates. AB - Hypocretin-1 (orexin-A) was administered to sleep-deprived (30-36 h) rhesus monkeys immediately preceding testing on a multi-image delayed match-to-sample (DMS) short-term memory task. The DMS task used multiple delays and stimulus images and effectively measures cognitive defects produced by sleep deprivation (Porrino et al., 2005). Two methods of administration of orexin-A were tested, intravenous injections (2.5-10.0 microg/kg, i.v.) and a novel method developed for nasal delivery via an atomizer spray mist to the nostrils (dose estimated 1.0 microg/kg). Results showed that orexin-A delivered via the intravenous and nasal routes significantly improved performance in sleep-deprived monkeys; however, the nasal delivery method was significantly more effective than the highest dose (10 microg/kg) of intravenous orexin-A tested. The improvement in performance by orexin-A was specific to trials classified as high versus low cognitive load as determined by performance difficulty under normal testing conditions. Except for the maximum intravenous dose (10 microg/kg), neither delivery method affected task performance in alert non-sleep-deprived animals. The improved performance in sleep-deprived animals was accompanied by orexin-A related alterations in local cerebral glucose metabolism (CMRglc) in specific brain regions shown previously to be engaged by the task and impaired by sleep deprivation (Porrino et al., 2005). Consistent with the differential effects on performance, nasal delivered orexin-A produced a more pronounced reversal of sleep deprivation induced changes in brain metabolic activity (CMRglc) than intravenous orexin-A. These findings provide strong evidence for the effectiveness of intranasal orexin-A in alleviating cognitive deficits produced by loss of sleep. PMID- 18160632 TI - Mosaic removal of hedgehog signaling in the adult SVZ reveals that the residual wild-type stem cells have a limited capacity for self-renewal. AB - The Smoothened gene is necessary for cells to transduce hedgehog signaling. Although we and others have previously shown that embryonic removal of Smoothened in the neural tube results in a loss of stem cells from the postnatal subventricular zone, it was unclear whether this reflected a requirement for hedgehog signaling in the establishment or maintenance of the adult niche. Here, we have examined the consequences of conditional removal of Smoothened gene function within the subventricular zone of the adult neural stem cell niche. We observe that both proliferation and neurogenesis are compromised when hedgehog signaling is removed from subventricular zone stem cells. Moreover, even after a 10 month survival period, the stem cell niche fails to recover. It has been reported that the adult subventricular zone quickly rebounds from an antimitotic insult by increasing proliferation and replenishing the niche. During this recovery, it has been reported that hedgehog signaling appears to be upregulated. When mice in which hedgehog signaling in the subventricular zone has been strongly attenuated are given a similar antimitotic treatment, recovery is limited to the reduced level of proliferation and neurogenesis observed before the mitotic insult. Furthermore, the limited recovery that is observed appears to be largely restricted to the minority of neural stem cells that escape the conditional inactivation of Smoothened gene function. These results demonstrate that ongoing hedgehog signaling is required to maintain adult neural stem cells and that their ability to self-renew is limited. PMID- 18160633 TI - Human neuroma contains increased levels of semaphorin 3A, which surrounds nerve fibers and reduces neurite extension in vitro. AB - Neuroma formation after peripheral nerve injury is detrimental to functional recovery and is therefore a significant clinical problem. The molecular basis for this phenomenon is not fully understood. Here, we show that the expression of the chemorepulsive protein semaphorin 3A (sema3A), but not semaphorin 3F, is increased in human neuroma tissue that has formed in severe obstetric brachial plexus lesions. Sema3A is produced by fibroblasts in the epineurial space and appears to be secreted into the extracellular matrix. It surrounds fascicles, minifascicles, or single axons, suggesting a role in fasciculation and inhibition of neurite outgrowth. Lentiviral vector-mediated knock-down of Neuropilin 1, the receptor for sema3A, leads to increased neurite outgrowth of F11 cells cultured on neuroma tissue, but not of F11 cells cultured on normal nerve tissue. These findings demonstrate the putative inhibitory role of sema3A in human neuroma tissue. Our observations are the first demonstration of the expression of sema3A in human neural scar tissue and support a role for this protein in the inhibition of axonal regeneration in injured human peripheral nerves. These findings contribute to the understanding of the outgrowth inhibitory properties of neuroma tissue. PMID- 18160634 TI - Selective deletion of Bdnf in the ventromedial and dorsomedial hypothalamus of adult mice results in hyperphagic behavior and obesity. AB - Brain-derived neurotrophic factor (BDNF) and its receptor TrkB are expressed in several hypothalamic and hindbrain nuclei involved in regulating energy homeostasis, developmentally and in the adult animal. Their depletion during the fetal or early postnatal periods when developmental processes are still ongoing elicits hyperphagic behavior and obesity in mice. Whether BDNF is a chief element in appetite control in the mature brain remains controversial. The required sources of this neurotrophin are also unknown. We show that glucose administration rapidly induced BDNF mRNA expression, mediated by Bdnf promoter 1, and TrkB transcription in the ventromedial hypothalamus (VMH) of adult mice, consistent with a role of this pathway in satiety. Using viral-mediated selective knock-down of BDNF in the VMH and dorsomedial hypothalamus (DMH) of adult mice, we were able to elucidate the physiological relevance of BDNF in energy balance regulation. Site-specific mutants exhibited hyperphagic behavior and obesity but normal energy expenditure. Furthermore, intracerebroventricular administration of BDNF triggered an immediate neuronal response in multiple hypothalamic nuclei in wild-type mice, suggesting that its anorexigenic actions involve short-term mechanisms. Locomotor, aggressive, and depressive-like behaviors, all of which are associated with neural circuits involving the VMH, were not altered in VMH/DMH-specific BDNF mutants. These findings demonstrate that BDNF is an integral component of central mechanisms mediating satiety in the adult mouse and, moreover, that its synthesis in the VMH and/or DMH is required for the suppression of appetite. PMID- 18160635 TI - Dopamine alters AMPA receptor synaptic expression and subunit composition in dopamine neurons of the ventral tegmental area cultured with prefrontal cortex neurons. AB - Excitatory synapses onto dopamine (DA) neurons of the ventral tegmental area (VTA) represent a critical site of psychostimulant-induced synaptic plasticity. This plasticity involves alterations in synaptic strength through AMPA receptor (AMPAR) redistribution. Here, we report an in vitro model for studying regulation of AMPAR trafficking in DA neurons under control conditions and after elevation of DA levels, mimicking cocaine exposure. We used cocultures containing rat VTA neurons and prefrontal cortex (PFC) neurons from enhanced cyan fluorescent protein-expressing mice. In VTA-PFC cocultures, D1 receptor activation (10 min) increased synaptic and nonsynaptic glutamate receptor subunit 1 (GluR1) and GluR2 surface expression on DA neurons. NMDA or AMPA receptor antagonists blocked this effect, and it was not observed in pure VTA cultures, suggesting that DA agonists acted on D1 receptors on PFC neurons, altering their excitatory transmission onto VTA DA neurons and, thus, influencing AMPARs. To mimic the longer elevation in extracellular DA levels produced by systemic cocaine, cocultures were incubated with DA for 1 h. Synaptic GluR1 was increased 24 h later, reminiscent of the increased AMPA/NMDA ratio at excitatory synapses onto VTA DA neurons 24 h after cocaine injection (Ungless et al., 2001). In contrast, GluR2 was unchanged. Analysis of colocalization of surface GluR1-3 labeling suggested that control DA neurons express a substantial number of GluR1/2, GluR2/3, and homomeric GluR1 receptors and that the increase in surface AMPARs 24 h after DA exposure may in part reflect increased GluR1/3-containing receptors. These results help define the cellular basis for plasticity underlying the development of behavioral sensitization. PMID- 18160636 TI - Quantitative analysis of calcium-dependent vesicle recruitment and its functional role at the calyx of Held synapse. AB - Recruitment of release-ready vesicles at synapses is one of the important factors, which determine dynamic properties of signaling between neurons in the brain. It has been shown that the rate of vesicle recruitment is accelerated by strong synaptic activity. An elevated concentration of calcium ions in the presynaptic terminal ([Ca2+]i) has been proposed to be responsible for this effect. However, the precise relationship between [Ca2+]i and recruitment has not been established yet, and the functional consequences of accelerated recruitment during synaptic activity have not been quantified experimentally. To probe the intracellular Ca2+ dependence of vesicle recruitment and to examine its functional role during trains of action potential (AP)-like stimuli, we monitored [Ca2+]i and synaptic responses simultaneously with paired recordings at the calyx of Held synapse. We found that a distinct, rapidly releasing vesicle pool is replenished with a rate that increases linearly with [Ca2+]i, without any apparent cooperativity. The slope factor for this increase is approximately 1 pool/(microM x s). Blocking Ca2+-dependent recruitment specifically with a calmodulin binding peptide revealed that the steady-state EPSCs during 100 Hz AP like trains were maintained through this Ca2+-dependent recruitment mechanism. Using a simple model of vesicle dynamics, we estimated that the recruitment rate accelerated 10-fold during the steady-state compared with the rate at resting [Ca2+]i. We could also demonstrate an approximate sixfold increase in release probability (facilitation) during the initial 5-15 AP-like stimuli of such trains in our experimental condition, regardless of EPSC depression. PMID- 18160637 TI - Omega-3 fatty acid docosahexaenoic acid increases SorLA/LR11, a sorting protein with reduced expression in sporadic Alzheimer's disease (AD): relevance to AD prevention. AB - Environmental and genetic factors, notably ApoE4, contribute to the etiology of late-onset Alzheimer's disease (LOAD). Reduced mRNA and protein for an apolipoprotein E (ApoE) receptor family member, SorLA (LR11) has been found in LOAD but not early-onset AD, suggesting that LR11 loss is not secondary to pathology. LR11 is a neuronal sorting protein that reduces amyloid precursor protein (APP) trafficking to secretases that generate beta-amyloid (Abeta). Genetic polymorphisms that reduce LR11 expression are associated with increased AD risk. However these polymorphisms account for only a fraction of cases with LR11 deficits, suggesting involvement of environmental factors. Because lipoprotein receptors are typically lipid-regulated, we postulated that LR11 is regulated by docosahexaenoic acid (DHA), an essential omega-3 fatty acid related to reduced AD risk and reduced Abeta accumulation. In this study, we report that DHA significantly increases LR11 in multiple systems, including primary rat neurons, aged non-Tg mice and an aged DHA-depleted APPsw AD mouse model. DHA also increased LR11 in a human neuronal line. In vivo elevation of LR11 was also observed with dietary fish oil in young rats with insulin resistance, a model for type II diabetes, another AD risk factor. These data argue that DHA induction of LR11 does not require DHA-depleting diets and is not age dependent. Because reduced LR11 is known to increase Abeta production and may be a significant genetic cause of LOAD, our results indicate that DHA increases in SorLA/LR11 levels may play an important role in preventing LOAD. PMID- 18160638 TI - Mechanosensory gating of proprioceptor input to modulatory projection neurons. AB - Sensorimotor gating commonly occurs at sensory neuron synapses onto motor circuit neurons and motor neurons. Here, using the crab stomatogastric nervous system, we show that sensorimotor gating also occurs at the level of the projection neurons that activate motor circuits. We compared the influence of the gastro-pyloric receptor (GPR) muscle stretch-sensitive neuron on two projection neurons, modulatory commissural neuron 1 (MCN1) and commissural projection neuron 2 (CPN2), with and without a preceding activation of the mechanosensory ventral cardiac neurons (VCNs). MCN1 and CPN2 project from the paired commissural ganglia (CoGs) to the stomatogastric ganglion (STG), where they activate the gastric mill (chewing) motor circuit. When stimulated separately, the GPR and VCN neurons each elicit the gastric mill rhythm by coactivating MCN1 and CPN2. When GPR is instead stimulated during the VCN-gastric mill rhythm, it slows this rhythm. This effect results from a second GPR synapse onto MCN1 that presynaptically inhibits its STG terminals. Here, we show that, during the VCN-triggered rhythm, the GPR excitation of MCN1 and CPN2 in the CoGs is gated out, leaving only its influence in the STG. This gating effect appears to occur within the CoG and does not result from a ceiling effect on projection neuron firing frequency. Additionally, this gating action enables GPR to either activate rhythmic motor activity or act as a phasic sensorimotor feedback system. These results also indicate that the site of sensorimotor gating can occur at the level of the projection neurons that activate a motor circuit. PMID- 18160639 TI - Conditional knock-out of beta-catenin in postnatal-born dentate gyrus granule neurons results in dendritic malformation. AB - Neurons are continuously added to the brain throughout life, and these neurons must develop dendritic arbors and functional connections with existing neurons to be integrated into neuronal circuitry. The molecular mechanisms that regulate dendritic development of newborn neurons in the hippocampal dentate gyrus are still unclear. Here, we show that beta-catenin is expressed in newborn granule neurons and in neural progenitor cells in the hippocampal dentate gyrus. Specific knock-out of beta-catenin in newborn neurons, without affecting beta-catenin expression in neural progenitor cells, led to defects in dendritic morphology of these newborn neurons in vivo. Majority of newborn neurons that cannot extend dendrites survive <1 month after they were born. Our results indicate that beta catenin plays an important role in dendritic development of postnatal-born neurons in vivo, and is therefore essential for the neurogenesis in the postnatal brain. PMID- 18160640 TI - NMDA receptor activation potentiates inhibitory transmission through GABA receptor-associated protein-dependent exocytosis of GABA(A) receptors. AB - The trafficking of postsynaptic AMPA receptors (AMPARs) is a powerful mechanism for regulating the strength of excitatory synapses. It has become clear that the surface levels of inhibitory GABA(A) receptors (GABA(A)Rs) are also subject to regulation and that GABA(A)R trafficking may contribute to inhibitory plasticity, although the underlying mechanisms are not fully understood. Here, we report that NMDA receptor activation, which has been shown to drive excitatory long-term depression through AMPAR endocytosis, simultaneously increases expression of GABA(A)Rs at the dendritic surface of hippocampal neurons. This NMDA stimulus increases miniature IPSC amplitudes and requires the activity of Ca2+ calmodulin dependent kinase II and the trafficking proteins N-ethylmaleimide-sensitive factor, GABA receptor-associated protein (GABARAP), and glutamate receptor interacting protein (GRIP). These data demonstrate for the first time that endogenous GABARAP and GRIP contribute to the regulated trafficking of GABA(A)Rs. In addition, they reveal that the bidirectional trafficking of AMPA and GABA(A) receptors can be driven by a single glutamatergic stimulus, providing a potent postsynaptic mechanism for modulating neuronal excitability. PMID- 18160641 TI - RGS9-2 negatively modulates L-3,4-dihydroxyphenylalanine-induced dyskinesia in experimental Parkinson's disease. AB - Chronic L-dopa treatment of Parkinson's disease (PD) often leads to debilitating involuntary movements, termed L-dopa-induced dyskinesia (LID), mediated by dopamine (DA) receptors. RGS9-2 is a GTPase accelerating protein that inhibits DA D2 receptor-activated G proteins. Herein, we assess the functional role of RGS9-2 on LID. In monkeys, Western blot analysis of striatal extracts shows that RGS9-2 levels are not altered by MPTP-induced DA denervation and/or chronic L-dopa administration. In MPTP monkeys with LID, striatal RGS9-2 overexpression- achieved by viral vector injection into the striatum--diminishes the involuntary movement intensity without lessening the anti-parkinsonian effects of the D1/D2 receptor agonist L-dopa. In contrasts, in these animals, striatal RGS9-2 overexpression diminishes both the involuntary movement intensity and the anti parkinsonian effects of the D2/D3 receptor agonist ropinirole. In unilaterally 6 OHDA-lesioned rats with LID, we show that the time course of viral vector mediated striatal RGS9-2 overexpression parallels the time course of improvement of L-dopa-induced involuntary movements. We also find that unilateral 6-OHDA lesioned RGS9-/- mice are more susceptible to L-dopa-induced involuntary movements than unilateral 6-OHDA-lesioned RGS9+/+ mice, albeit the rotational behavior--taken as an index of the anti-parkinsonian response--is similar between the two groups of mice. Together, these findings suggest that RGS9-2 plays a pivotal role in LID pathophysiology. However, the findings also suggest that increasing RGS9-2 expression and/or function in PD patients may only be a suitable therapeutic strategy to control involuntary movements induced by nonselective DA agonist such as L-dopa. PMID- 18160642 TI - FMRP phosphorylation reveals an immediate-early signaling pathway triggered by group I mGluR and mediated by PP2A. AB - Fragile X syndrome is a common form of inherited mental retardation and is caused by loss of fragile X mental retardation protein (FMRP), a selective RNA-binding protein that influences the translation of target messages. Here, we identify protein phosphatase 2A (PP2A) as an FMRP phosphatase and report rapid FMRP dephosphorylation after immediate group I metabotropic glutamate receptor (mGluR) stimulation (<1 min) in neurons caused by enhanced PP2A enzymatic activity. In contrast, extended mGluR activation (1-5 min) resulted in mammalian target of rapamycin (mTOR)-mediated PP2A suppression and FMRP rephosphorylation. These activity-dependent changes in FMRP phosphorylation were also observed in dendrites and showed a temporal correlation with the translational profile of select FMRP target transcripts. Collectively, these data reveal an immediate early signaling pathway linking group I mGluR activity to rapid FMRP phosphorylation dynamics mediated by mTOR and PP2A. PMID- 18160643 TI - Dimensions of impulsivity are associated with poor spatial working memory performance in monkeys. AB - Impulsive behavior and novelty seeking are dimensions of temperament that are behavioral determinants of risk for attention deficit/hyperactivity disorder and its neurocognitive endophenotypes, and variation in the dopamine D4 receptor gene (DRD4) explains at least a portion of the variance in the traits. To further characterize the dimensional phenotype associated with impulsiveness, adolescent male monkeys were evaluated using ecologically valid tests of impulsive approach and aggression in response to social or nonsocial stimuli; subsequently, a delayed response task was implemented to assess spatial working memory performance. Subjects were selected into this study based on their response to the social challenge task or by DRD4 genotype, resulting in three groups: low impulsivity/common DRD4 allele, high-impulsivity/common DRD4 allele, or rare DRD4 allele. All animals acquired the delayed response task and could perform at near ceiling levels when a approximately 0 s delay version was imposed, but as delays were lengthened, high-impulsive animals, regardless of DRD4 genotype, made fewer correct responses than did low-impulsive subjects; an inverse relationship existed for working memory and impulsivity. Notably, impulsive behavior evoked by social and nonsocial stimuli explained overlapping and independent portions of the variance in working memory performance. CSF levels of monoamine metabolites did not significantly differentiate the high- and low-impulsive animals, although monkeys carrying the DRD4 rare allele tended to exhibit higher monoamine turnover. These data indicate that dimensions of impulsivity may impact on working memory performance in qualitatively similar ways but through different mechanisms. PMID- 18160644 TI - Using imagination to understand the neural basis of episodic memory. AB - Functional MRI (fMRI) studies investigating the neural basis of episodic memory recall, and the related task of thinking about plausible personal future events, have revealed a consistent network of associated brain regions. Surprisingly little, however, is understood about the contributions individual brain areas make to the overall recollective experience. To examine this, we used a novel fMRI paradigm in which subjects had to imagine fictitious experiences. In contrast to future thinking, this results in experiences that are not explicitly temporal in nature or as reliant on self-processing. By using previously imagined fictitious experiences as a comparison for episodic memories, we identified the neural basis of a key process engaged in common, namely scene construction, involving the generation, maintenance and visualization of complex spatial contexts. This was associated with activations in a distributed network, including hippocampus, parahippocampal gyrus, and retrosplenial cortex. Importantly, we disambiguated these common effects from episodic memory-specific responses in anterior medial prefrontal cortex, posterior cingulate cortex and precuneus. These latter regions may support self-schema and familiarity processes, and contribute to the brain's ability to distinguish real from imaginary memories. We conclude that scene construction constitutes a common process underlying episodic memory and imagination of fictitious experiences, and suggest it may partially account for the similar brain networks implicated in navigation, episodic future thinking, and the default mode. We suggest that additional brain regions are co-opted into this core network in a task-specific manner to support functions such as episodic memory that may have additional requirements. PMID- 18160645 TI - Olig1 and Sox10 interact synergistically to drive myelin basic protein transcription in oligodendrocytes. AB - The oligodendrocyte lineage genes (Olig1/2), encoding basic helix-loop-helix transcription factors, were first identified in screens for master regulators of oligodendrocyte development. OLIG1 is important for differentiation of oligodendrocyte precursors into myelin-forming oligodendrocytes during development and is thought to play a crucial role in remyelination during multiple sclerosis. However, it is still unclear how OLIG1 interacts with its transcriptional cofactors and DNA targets. OLIG1 was reportedly restricted to mammals, but we demonstrate here that zebrafish and other teleosts also possess an OLIG1 homolog. In zebrafish, as in mammals, Olig1 is expressed in the oligodendrocyte lineage. Olig1 associates physically with another myelin associated transcription factor, Sox10, and the Olig1/Sox10 complex activates mbp (myelin basic protein) transcription via conserved DNA sequence motifs in the mbp promoter region. In contrast, Olig2 does not bind to Sox10 in zebrafish, although both OLIG1 and OLIG2 bind SOX10 in mouse. PMID- 18160646 TI - Immediate reward bias in humans: fronto-parietal networks and a role for the catechol-O-methyltransferase 158(Val/Val) genotype. AB - The tendency to choose lesser immediate benefits over greater long-term benefits characterizes alcoholism and other addictive disorders. However, despite its medical and socioeconomic importance, little is known about its neurobiological mechanisms. Brain regions that are activated when deciding between immediate or delayed rewards have been identified (McClure et al., 2004, 2007), as have areas in which responses to reward stimuli predict a paper-and-pencil measure of temporal discounting (Hariri et al., 2006). These studies assume "hot" and "cool" response selection systems, with the hot system proposed to generate impulsive choices in the presence of a proximate reward. However, to date, brain regions in which the magnitude of activity during decision making reliably predicts intertemporal choice behavior have not been identified. Here we address this question in sober alcoholics and non-substance-abusing control subjects and show that immediate reward bias directly scales with the magnitude of functional magnetic resonance imaging bold oxygen level-dependent (BOLD) signal during decision making at sites within the posterior parietal cortex (PPC), dorsal prefrontal cortex (dPFC), and rostral parahippocampal gyrus regions. Conversely, the tendency of an individual to wait for a larger, delayed reward correlates directly with BOLD signal in the lateral orbitofrontal cortex. In addition, genotype at the Val158Met polymorphism of the catechol-O-methyltransferase gene predicts both impulsive choice behavior and activity levels in the dPFC and PPC during decision making. These genotype effects remained significant after controlling for alcohol abuse history. These results shed new light on the neurobiological underpinnings of temporal discounting behavior and identify novel behavioral and neural consequences of genetic variation in dopamine metabolism. PMID- 18160647 TI - Impaired migration in the rostral migratory stream but spared olfactory function after the elimination of programmed cell death in Bax knock-out mice. AB - Rats and mice exhibit neurogenesis of olfactory bulb (OB) interneurons throughout adulthood. To homeostatically maintain stable neuron numbers, it is necessary to continuously remove a subset of OB neurons by programmed cell death (PCD). Here we demonstrate that Bax is critical for the elimination of OB neurons by showing that Bax-KO mice exhibit greatly reduced PCD in the OB. Despite the reduction of PCD, however, proliferation of progenitors and the size of the OB were virtually unaffected in Bax-knock-out (KO) mice. However, reducing PCD by Bax deletion affected the migration of a subset of adult-produced neurons by the disruption of glial tube formation as well as by premature detachment of neuroblasts from the migratory chain. Rescued cells aberrantly remained in the subventricular zone (SVZ)-rostral migratory stream (RMS), in which they differentiated into calretinin+ or GABA-expressing interneurons. Because of the migratory deficit, OB cell homeostasis involving new cell entry and PCD (neuronal turnover) was virtually absent in adult Bax-KO mice. Despite this, Bax-KO mice exhibited normal olfactory behaviors such as odor discrimination and olfactory memory which are thought to be influenced by adult neurogenesis. These results demonstrate that PCD is involved in the regulation of RMS migration and differentiation after OB neurogenesis, but that animals maintain normal olfactory function in the absence of PCD. PMID- 18160648 TI - Analyzing somatosensory axon projections with the sensory neuron-specific Advillin gene. AB - Peripheral sensory neurons detect diverse physical stimuli and transmit the information into the CNS. At present, the genetic tools for specifically studying the development, plasticity, and regeneration of the sensory axon projections are limited. We found that the gene encoding Advillin, an actin binding protein that belongs to the gelsolin superfamily, is expressed almost exclusively in peripheral sensory neurons. We next generated a line of knock-in mice in which the start codon of the Advillin is replaced by the gene encoding human placenta alkaline phosphatase (Avil-hPLAP mice). In heterozygous Avil-hPLAP mice, sensory axons, the exquisite sensory endings, as well as the fine central axonal collaterals can be clearly visualized with a simple alkaline phosphatase staining. Using this mouse line, we found that the development of peripheral target innervation and sensory ending formation is an ordered process with specific timing depending on sensory modalities. This is also true for the in growth of central axonal collaterals into the brainstem and the spinal cord. Our results demonstrate that Avil-hPLAP mouse is a valuable tool for specifically studying peripheral sensory neurons. Functionally, we found that the regenerative axon growth of Advillin-null sensory neurons is significantly shortened and that deletion of Advillin reduces the plasticity of whisker-related barrelettes patterns in the hindbrain. PMID- 18160649 TI - Parietal lobe and episodic memory: bilateral damage causes impaired free recall of autobiographical memory. AB - Does the parietal lobe have a critical role in memory? The neuroimaging literature indicates that it has an important role, especially in episodic memory. However, the neuropsychological literature suggests that its role is more limited to attentional, spatial, or imagery aspects of memory. Here, we present data to adjudicate this disagreement. Two patients with bilateral parietal lobe damage received detailed assessments of their autobiographical memories. The results show that although both patients easily recalled various memories, their freely recalled memories were relatively impoverished, lacking in detail. This deficit was ubiquitous, and not limited to spatial or perceptual aspects of memory. The memory deficit disappeared when memory was specifically probed by asking pointed questions. Additional tests show that it is unlikely that their free recall deficit can be explained by general mental imagery problems. In sum, the parietal lobe appears to have a critical role in recollection aspects of episodic memory. PMID- 18160650 TI - Asymmetry of anticipatory activity in visual cortex predicts the locus of attention and perception. AB - Humans can use advance information to direct spatial attention before stimulus presentation and respond more accurately to stimuli at the attended location compared with unattended locations. Likewise, spatially directed attention is associated with anticipatory activity in the portion of visual cortex representing the attended location. It is unknown, however, whether and how anticipatory signals predict the locus of spatial attention and perception. Here, we show that prestimulus, preparatory activity is highly correlated across regions representing attended and unattended locations. Comparing activity representing attended versus unattended locations, rather than measuring activity for only one location, dramatically improves the accuracy with which preparatory signals predict the locus of attention, largely by removing this positive correlation common across locations. In V3A, moreover, only the difference in activity between attended and unattended locations predicts whether upcoming visual stimuli will be accurately perceived. These results suggest that the locus of attention is coded in visual cortex by an asymmetry of anticipatory activity between attended and unattended locations and that this asymmetry predicts the accuracy of perception. This coding strategy may bias activity in downstream brain regions to represent the stimulus at the attended location. PMID- 18160651 TI - Task-related interaction between basal ganglia and cortical dopamine release. AB - Dopamine (DA) is a powerful neuromodulator for a wide variety of behaviors. Considerable evidence accumulated from rodent and monkey experiments over the last two decades suggests that DA activity in the frontal cortex is reciprocally linked to that in functionally related basal ganglia (BG) structures. However, the functional importance of this in humans is still unknown. To address this issue, we measured endogenous DA release using positron emission tomography in 15 healthy subjects as they practiced the first training session of a finger sequence learning task. Significant results were observed not only in striatal areas but also in extrastriatal "motor" regions, bilaterally. Faster learning was specifically coupled to lower DA release in the sensorimotor part of the globus pallidus pars interna (GPi) contralateral to the moving hand, which was paralleled by a higher increase in DA levels in the pre-supplementary motor area (pre-SMA). This finding provides original evidence supporting a motor-learning related interaction between DA release in left GPi and pre-SMA, a mechanism that may also apply to other anatomically and functionally interconnected BG and frontal cortical areas as a function of behavior. PMID- 18160652 TI - Focusing effect of acetylcholine on neuroplasticity in the human motor cortex. AB - Cholinergic neuromodulation is pivotal for arousal, attention, and cognitive processes. Loss or dysregulation of cholinergic inputs leads to cognitive impairments like those manifested in Alzheimer's disease. Such dysfunction can be at least partially restored by an increase of acetylcholine (ACh). In animal studies, ACh selectively facilitates long-term excitability changes induced by feed-forward afferent input. Consequently, it has been hypothesized that ACh enhances the signal-to-noise ratio of input processing. However, the neurophysiological foundation for its ability to enhance cognition in humans is not well documented. In this study we explore the effects of rivastigmine, a cholinesterase inhibitor, on global and synapse-specific forms of cortical plasticity induced by transcranial direct current stimulation (tDCS) and paired associative stimulation (PAS) on 10-12 healthy subjects, respectively. Rivastigmine essentially blocked the induction of the global excitability enhancement elicited by anodal tDCS and revealed a tendency to first reduce and then stabilize cathodal tDCS-induced inhibitory aftereffects. However, ACh enhanced the synapse-specific excitability enhancement produced by facilitatory PAS and consolidated the inhibitory PAS-induced excitability diminution. These findings are in line with a cholinergic focusing effect that optimizes the detection of relevant signals during information processing in humans. PMID- 18160653 TI - A TAT-DEF-Elk-1 peptide regulates the cytonuclear trafficking of Elk-1 and controls cytoskeleton dynamics. AB - The transcription factor Elk-1 plays a key role in cell differentiation, proliferation and apoptosis. This role is thought to arise from its phosphorylation by activated extracellular signal-regulated kinases (ERKs), a critical posttranslational event for the transcriptional activity of the ternary complex composed of Elk-1 and a dimer of serum response factor (SRF) at the serum response element (SRE) regulatory site of transcription. In addition to its nuclear localization, Elk-1 is found in the dendrites and soma of neuronal cells and recent evidence implicate a cytoplasmic proapoptotic function of Elk-1, via its association with the mitochondrial permeability transition pore complex. Thus, the nuclear versus cytoplasmic localization of Elk-1 seems to be crucial for its biological function. In this study we show that the excitatory neurotransmitter, glutamate, induces an ERK-dependent Elk-1 activation and nuclear relocalization. We demonstrate that Elk-1 phosphorylation on Ser383/389 has a dual function and triggers both Elk-1 nuclear translocation and SRE dependent gene expression. Mutating these sites into inactive residues or using a synthetic penetrating peptide (TAT-DEF-Elk-1), which specifically interferes with the DEF docking domain of Elk-1, prevents Elk-1 nuclear translocation without interfering with ERK nor MSK1 (mitogen- and stress-activated protein kinase 1), a CREB kinase downstream from ERK- activation. This results in a differential regulation of glutamate-induced IEG regulation when compared with classical inhibitors of the ERK pathway. Using the TAT-DEF-Elk-1 peptide or the dominant negative version of Elk-1, we show that Elk-1 phosphorylation controls dendritic elongation, SRF and Actin expression levels as well as cytoskeleton dynamics. PMID- 18160654 TI - A critical function for beta-amyloid precursor protein in neuronal migration revealed by in utero RNA interference. AB - Physiological processing of the beta-amyloid precursor protein (APP) generates amyloid beta-protein, which can assemble into oligomers that mediate synaptic failure in Alzheimer's disease. Two decades of research have led to human trials of compounds that chronically target this processing, and yet the normal function of APP in vivo remains unclear. We used the method of in utero electroporation of shRNA constructs into the developing cortex to acutely knock down APP in rodents. This approach revealed that neuronal precursor cells in embryonic cortex require APP to migrate correctly into the nascent cortical plate. cDNAs encoding human APP or its homologues, amyloid precursor-like protein 1 (APLP1) or APLP2, fully rescued the shRNA-mediated migration defect. Analysis of an array of mutations and deletions in APP revealed that both the extracellular and cytoplasmic domains of APP are required for efficient rescue. Whereas knock-down of APP inhibited cortical plate entry, overexpression of APP caused accelerated migration of cells past the cortical plate boundary, confirming that normal APP levels are required for correct neuronal migration. In addition, we found that Disabled-1 (Dab1), an adaptor protein with a well established role in cortical cell migration, acts downstream of APP for this function in cortical plate entry. We conclude that full-length APP functions as an important factor for proper migration of neuronal precursors into the cortical plate during the development of the mammalian brain. PMID- 18160655 TI - Liver X receptor activation enhances cholesterol loss from the brain, decreases neuroinflammation, and increases survival of the NPC1 mouse. AB - Although cholesterol is a major component of the CNS, there is little information on how or whether a change in sterol flux across the blood-brain barrier might alter neurodegeneration. In Niemann-Pick type C (NPC) disease, a mutation in NPC1 protein causes unesterified cholesterol to accumulate in the lysosomal compartment of every cell, including neurons and glia. Using the murine model of this disease, we used genetic and pharmacologic approaches in an attempt to alter cholesterol homeostasis across the CNS. Genetic deletion of the sterol transporters ATP-binding cassette transporter A1 (ABCA1) and low-density lipoprotein receptor in the NPC1 mouse did not affect sterol balance or longevity. However, deletion of the nuclear receptor, liver X receptor beta (LXRbeta), had an adverse effect on progression of the disease. We therefore tested the effects of increasing LXR activity by oral administration of a synthetic ligand for this transcription factor. Treatment with this LXR agonist increased cholesterol excretion out of brain from 17 to 49 microg per day, slowed neurodegeneration, and prolonged life. This agonist did not alter synthesis of cholesterol or expression of genes associated with the formation of 24(S) hydroxycholesterol or neurosteroids such as CYP46A1, 3alphaHSD, and CYP11A1. However, levels of the sterol transporters ABCA1 and ATP-binding cassette transporter G1 were increased. Concomitantly, markers of neuroinflammation, CD14, MAC1, CD11c, and inducible nitric oxide synthase, were reduced, and microglia reverted from their amoeboid, active form to a ramified, resting configuration. Thus, LXR activation resulted in increased cholesterol excretion from the brain, decreased neuroinflammation, and deactivation of microglia to slow neurodegeneration and extend the lifespan of the NPC1 mouse. PMID- 18160656 TI - Selective targeting of different neural cell adhesion molecule isoforms during motoneuron myotube synapse formation in culture and the switch from an immature to mature form of synaptic vesicle cycling. AB - Characterization of neuromuscular junction formation and function in mice lacking all neural cell adhesion molecule (NCAM) isoforms or only the 180 isoform demonstrated that the 180 isoform was required at adult synapses to maintain effective transmission with repetitive stimulation whereas the 140 and/or 120 isoform(s) were sufficient to mediate the downregulation of synaptic vesicle cycling along the axon after synapse formation. However, the expression and targeting of each isoform and its relationship to distinct forms of synaptic vesicle cycling before and after synapse formation was previously unknown. By transfecting chick motoneurons with fluorescently tagged mouse 180, 140 and 120 isoforms, we show that before myotube contact the 180 and 140 isoforms are expressed in distinct puncta along the axon which are sites of an immature form (Brefeldin A sensitive, L-type Ca2+ channel mediated) of vesicle cycling. After myotube contact the 140 and 180 isoforms are downregulated from the axon and selectively targeted to the presynaptic terminal. This coincided with the downregulation of vesicle cycling along the axon and the expression of the mature form (BFA insensitive, P/Q type Ca2+ channel mediated) of vesicle cycling at the terminal. The synaptic targeting of exogenously expressed 180 and 140 isoforms also occurred when chick motoneurons contacted +/+ mouse myotubes; however only the 180 but not the 140 isoform was targeted on contact with NCAM-/- myotubes. These observations indicate that postsynaptic NCAM is required for the synaptic targeting of presynaptic 140 NCAM but that the localization of presynaptic 180 NCAM occurs via a different mechanism. PMID- 18160657 TI - Inhibiting glycosaminoglycan chain polymerization decreases the inhibitory activity of astrocyte-derived chondroitin sulfate proteoglycans. AB - Chondroitin sulfate proteoglycans (CSPGs) are upregulated in the CNS after injury and participate in the inhibition of axon regeneration mainly through their glycosaminoglycan (GAG) side chains. In the present study, we have identified a new way to alleviate the inhibition of axonal regeneration by CSPG GAGs. We have successfully decreased the amount of CSPG GAG produced by astrocytes by targeting chondroitin polymerizing factor (ChPF), a key enzyme in the CSPG biosynthetic pathway. Using short interfering RNA (siRNA), we reduced ChPF mRNA levels by 70% in both the Neu7 astrocyte cell line and primary rat astrocytes. This reduction leads to a decrease in ChPF protein levels and a reduced amount of CSPG GAG chains in the conditioned media (CM) of these cells. Secretion of neurocan by primary astrocytes and NG2 core protein by Neu7 cells transfected with ChPF siRNA is not decreased, suggesting that inhibiting GAG chain synthesis does not affect core protein trafficking from these cells. CM from siRNA-treated Neu7 cells is a less repulsive substrate for axons than CM from control cells. In addition, axonal outgrowth from cerebellar granule neurons is increased on or in CM from ChPF siRNA-treated Neu7 cells. These data indicate that targeting the biosynthesis of CSPG GAG is a potentially new therapeutic avenue for decreasing CSPG GAG produced by astrocytes after CNS injury. PMID- 18160658 TI - Action and outcome encoding in the primate caudate nucleus. AB - The basal ganglia appear to have a central role in reinforcement learning. Previous experiments, focusing on activity preceding movement execution, support the idea that dorsal striatal neurons bias action selection according to the expected values of actions. However, many phasically active striatal neurons respond at a time too late to initiate or select movements. Given the data suggesting a role for the basal ganglia in reinforcement learning, postmovement activity may therefore reflect evaluative processing important for learning the values of actions. To better understand these postmovement neurons, we determined whether individual striatal neurons encode information about saccade direction, whether a reward had been received, or both. We recorded from phasically active neurons in the caudate nucleus while monkeys performed a probabilistically rewarded delayed saccade task. Many neurons exhibited peak responses after saccade execution (77 of 149) that were often tuned for the direction of the preceding saccade (61 of 77). Of those neurons responding during the reward epoch, one subset showed direction tuning for the immediately preceding saccade (43 of 60), whereas another subset responded differentially on rewarded versus unrewarded trials (35 of 60). We found that there was relatively little overlap of these properties in individual neurons. The encoding of action and outcome was performed by largely separate populations of caudate neurons that were active after movement execution. Thus, striatal neurons active primarily after a movement appear to be segregated into two distinct groups that provide complimentary information about the outcomes of actions. PMID- 18160659 TI - Proprioceptive sensory neuropathy in mice with a mutation in the cytoplasmic Dynein heavy chain 1 gene. AB - Mice heterozygous for the radiation-induced Sprawling (Swl) mutation display an early-onset sensory neuropathy with muscle spindle deficiency. The lack of an H reflex despite normal motor nerve function in the hindlimbs of these mutants strongly suggests defective proprioception. Immunohistochemical analyses reveal that proprioceptive sensory neurons are severely compromised in the lumbar dorsal root ganglia of newborn Swl/+ mice, whereas motor neuron numbers remain unaltered even in aged animals. We have used positional cloning to identify a nine base pair deletion in the cytoplasmic dynein heavy chain 1 gene (Dync1h1) in this mutant. Furthermore, we demonstrate that Loa/+ mice, which have previously been shown to carry a missense point mutation in Dync1h1 that results in late-onset motor neuron loss, also present with a severe, early-onset proprioceptive sensory neuropathy. Interestingly, in contrast to the Loa mutation, the Swl mutation does not delay disease progression in a motor neuron disease mouse model overexpressing a human mutant superoxide dismutase (SOD1(G93A)) transgene. Together, we provide in vivo evidence that distinct mutations in cytoplasmic dynein can either result in a pure sensory neuropathy or in a sensory neuropathy with motor neuron involvement. PMID- 18160660 TI - A rabbit ventricular action potential model replicating cardiac dynamics at rapid heart rates. AB - Mathematical modeling of the cardiac action potential has proven to be a powerful tool for illuminating various aspects of cardiac function, including cardiac arrhythmias. However, no currently available detailed action potential model accurately reproduces the dynamics of the cardiac action potential and intracellular calcium (Ca(i)) cycling at rapid heart rates relevant to ventricular tachycardia and fibrillation. The aim of this study was to develop such a model. Using an existing rabbit ventricular action potential model, we modified the L-type calcium (Ca) current (I(Ca,L)) and Ca(i) cycling formulations based on new experimental patch-clamp data obtained in isolated rabbit ventricular myocytes, using the perforated patch configuration at 35-37 degrees C. Incorporating a minimal seven-state Markovian model of I(Ca,L) that reproduced Ca- and voltage-dependent kinetics in combination with our previously published dynamic Ca(i) cycling model, the new model replicates experimentally observed action potential duration and Ca(i) transient alternans at rapid heart rates, and accurately reproduces experimental action potential duration restitution curves obtained by either dynamic or S1S2 pacing. PMID- 18160661 TI - Modifying L-type calcium current kinetics: consequences for cardiac excitation and arrhythmia dynamics. AB - The L-type Ca current (I(Ca,L)), essential for normal cardiac function, also regulates dynamic action potential (AP) properties that promote ventricular fibrillation. Blocking I(Ca,L) can prevent ventricular fibrillation, but only at levels suppressing contractility. We speculated that, instead of blocking I(Ca,L), modifying its shape by altering kinetic features could produce equivalent anti-fibrillatory effects without depressing contractility. To test this concept experimentally, we overexpressed a mutant Ca-insensitive calmodulin (CaM(1234)) in rabbit ventricular myocytes to inhibit Ca-dependent I(Ca,L) inactivation, combined with the ATP-sensitive K current agonist pinacidil or I(Ca,L) blocker verapamil to maintain AP duration (APD) near control levels. Cell shortening was enhanced in pinacidil-treated myocytes, but depressed in verapamil treated myocytes. Both combinations flattened APD restitution slope and prevented APD alternans, similar to I(Ca,L) blockade. To predict the arrhythmogenic consequences, we simulated the cellular effects using a new AP model, which reproduced flattening of APD restitution slope and prevention of APD/Ca(i) transient alternans but maintained a normal Ca(i) transient. In simulated two dimensional cardiac tissue, these changes prevented the arrhythmogenic spatially discordant APD/Ca(i) transient alternans and spiral wave breakup. These findings provide a proof-of-concept test that I(Ca,L) can be targeted to increase dynamic wave stability without depressing contractility, which may have promise as an antifibrillatory strategy. PMID- 18160662 TI - Sliding of alkylating anticancer drugs along the minor groove of DNA: new insights on sequence selectivity. AB - Currently, little is known about the molecular recognition pathways between DNA alkylating anticancer drugs and their targets despite their pharmacological relevance. In the framework of classical molecular dynamics simulations, here we use umbrella sampling to map the potential of mean force (PMF) associated with sliding along the DNA minor groove of two of these compounds. These are an indole derivative of duocarmycin (DSI) and the putative reactive form of anthramycin (anhydro-anthramycin, IMI). Twenty-three configurations were considered for each drug/DNA complex, corresponding to a movement along approximately 3 basepairs. The alkylation site turns out to be the most favorable for DSI, while a barrier of approximately 6 kcal/mol separates the reactive configuration of IMI.DNA from the absolute minimum. An analysis of various contributions to the PMF reveals that solvent effects play an important role for the largest and more flexible drug DSI. Instead, the PMF of IMI.DNA overall correlates with changes in the binding enthalpy. Implications of these results on the sequence selectivity of the two drugs are discussed. PMID- 18160663 TI - Protein phase behavior in aqueous solutions: crystallization, liquid-liquid phase separation, gels, and aggregates. AB - The aggregates and gels commonly observed during protein crystallization have generally been considered disordered phases without further characterization. Here their physical nature is addressed by investigating protein salting-out in ammonium sulfate and sodium chloride for six proteins (ovalbumin, ribonuclease A, soybean trypsin inhibitor, lysozyme, and beta-lactoglobulin A and B) at 4 degrees C, 23 degrees C, and 37 degrees C. When interpreted within the framework of a theoretical phase diagram obtained for colloidal particles displaying short-range attractive interactions, the results show that the formation of aggregates can be interpreted theoretically in terms of a gas-liquid phase separation for aggregates that are amorphous or gel-like. A notable additional feature is the existence of a second aggregation line observed for both ovalbumin and ribonuclease A in ammonium sulfate, interpreted theoretically as the spinodal. Further investigation of ovalbumin and lysozyme reveals that the formation of aggregates can be interpreted, in light of theoretical results from mode-coupling theory, as a kinetically trapped state or a gel phase that occurs through the intermediate of a gas-liquid phase separation. Despite the limitations of simple theoretical models of short-range attractive interactions, such as their inability to reproduce the effect of temperature, they provide a framework useful to describe the main features of protein phase behavior. PMID- 18160664 TI - Aurora kinase inhibitory VX-680 increases Bax/Bcl-2 ratio and induces apoptosis in Aurora-A-high acute myeloid leukemia. AB - Previously, we and others showed that mitotic Aurora-A kinase (Aur-A) was required for accurate mitotic entry and proper spindle assembly. In this study, we found that expression of Aur-A was markedly elevated in bone marrow mononuclear cells (BMMCs) obtained from a significant portion of de novo acute myeloid leukemia (AML) patients. Targeting human primary AML cells with Aur-A kinase inhibitory VX-680 led to apoptotic cell death in a dose-dependent manner. Importantly, VX-680-induced cell death was preferentially higher in Aur-A-high primary leukemic blasts compared with Aur-A-low AML (P < .001) or normal BMMCs (P < .001), suggesting the possible pharmacologic window in targeting Aurora kinase among Aur-A-high VX-680-sensitive leukemia patients. VX-680-induced cell death in AML cell lines was accompanied by formation of monopolar mitotic spindles, G(2)/M phase arrest, decreased phosphorylated(p)-Akt-1, and increased proteolytic cleavage of procaspase-3 and poly(ADP)ribose polymerase. Notably, VX-680 increased Bax/Bcl-2 expression ratio, a favorable proapoptotic predictor for drug response and survival in AML. Lastly, VX-680 enhanced the cytotoxic effect of the chemotherapeutic agent etoposide (VP16) on AML cells. Together, we concluded that Aurora kinases were potentially therapeutic targets for AML and that Aur-A-high expression may serve as a differential marker for selective treatment. PMID- 18160665 TI - Cooperative signaling through the signal transducer and activator of transcription 3 and nuclear factor-{kappa}B pathways in subtypes of diffuse large B-cell lymphoma. AB - The activated B cell-like (ABC) subgroup of diffuse large B-cell lymphoma (DLBCL) is characterized by constitutive activation of the nuclear factor-kappaB (NF kappaB) pathway. In this study, we showed that the NF-kappaB pathway induced the expression of the cytokines interleukin (IL)-6 and IL-10 in ABC DLBCL cell lines, which also have high levels of total and phosphorylated signal transducer and activator of transcription (STAT) 3 protein, suggesting autocrine signaling. Using RNA interference for STAT3, we defined a gene expression signature of IL-6 and IL-10 signaling through STAT3. Based on this signature, we constructed a molecular predictor of STAT3 signaling that defined a subset of ABC DLBCL tumors with high expression of STAT3, IL-6, and/or IL-10 and their downstream targets. Although the STAT3-high and STAT3-low subsets had equivalent expression of genes that distinguish ABC DLBCL from germinal center B cell-like DLBCL, STAT3-high ABC DLBCLs had higher expression of signatures that reflected NF-kappaB activity, proliferation, and glycolysis. A small-molecule inhibitor of Janus kinase signaling, which blocked STAT3 signature expression, was toxic only for ABC DLBCL lines and synergized with an inhibitor of NF-kappaB signaling. These findings suggest that the biological interplay between the STAT3 and NF-kappaB pathways may be exploited for the treatments of a subset of ABC DLBCLs. PMID- 18160666 TI - Increased proteasomal degradation of Bax is a common feature of poor prognosis chronic lymphocytic leukemia. AB - Many biologic markers are associated with poor prognosis in chronic lymphocytic leukemia (CLL), but their mechanistic role remains unclear. Bax is an essential proapoptotic protein and decreased levels in malignant cells lead to resistance to apoptosis. Using a Bax degradation activity (BDA) assay, CLL cells were found to show variable Bax instability. However, BDA did not correlate with Bax protein levels: BDA positive and negative cases had high and low baseline Bax levels. BDA positive cases showed a marked accumulation of poor prognostic markers-unmutated immunoglobulin heavy chain variable genes, ZAP-70/CD38 positivity, 11q22/17p13 deletion, and short lymphocyte doubling time. Patients with BDA positive cells had a shorter median overall survival (OS; 126 months vs not reached, P = .011) and time to first treatment (16 vs 156 months, P = .029) than BDA negative cases. Dual BDA and ZAP-70 positivity had a median OS of 84 months (P = .012). The BDA assay measures the intrinsic ubiquitin/proteasome activity of CLL cells and dynamic changes in Bax protein levels over time. Mechanistically, Bax instability may represent a final common pathway for disparate prognostic markers, as well as being itself an indicator of poor prognosis. PMID- 18160667 TI - A 2-gene classifier for predicting response to the farnesyltransferase inhibitor tipifarnib in acute myeloid leukemia. AB - At present, there is no method available to predict response to farnesyltransferase inhibitors (FTIs). We analyzed gene expression profiles from the bone marrow of patients from a phase 2 study of the FTI tipifarnib in older adults with previously untreated acute myeloid leukemia (AML). The RASGRP1/APTX gene expression ratio was found to predict response to tipifarnib with the greatest accuracy using a "leave one out" cross validation (LOOCV; 96%). RASGRP1 is a guanine nucleotide exchange factor that activates RAS, while APTX (aprataxin) is involved in DNA excision repair. The utility of this classifier for predicting response to tipifarnib was validated in an independent set of 58 samples from relapsed or refractory AML, with a negative predictive value (NPV) and positive predictive value (PPV) of 92% and 28%, respectively (odds ratio of 4.4). The classifier also predicted for improved overall survival (154 vs 56 days; P < .001), which was independent of other covariates, including a previously described prognostic gene expression classifier. Therefore, these data indicate that a 2-gene expression assay may have utility in categorizing a population of patients with AML who are more likely to respond to tipifarnib. PMID- 18160668 TI - Re-evaluation of the role of the protein S-C4b binding protein complex in activated protein C-catalyzed factor Va-inactivation. AB - Protein S expresses cofactor activity for activated protein C (APC) by enhancing the APC-catalyzed proteolysis at R(306) in factor Va. It is generally accepted that only free protein S is active and that complex formation with C4b-binding protein (C4BP) inhibits the APC-cofactor activity of protein S. However, the present study shows that protein S-C4BP expresses APC-cofactor activity and stimulates APC-catalyzed proteolysis at R(306) more than 10-fold, but instead inhibits proteolysis at R(506) by APC 3- to 4-fold. Free protein S stimulates APC catalyzed cleavage at R(306) approximately 20-fold and has no effect on cleavage at R(506). The resulting net effect of protein S-C4BP complex formation on APC catalyzed factor Va inactivation is a 6- to 8-fold reduction in factor Va inactivation when compared with free protein S, which is not explained by inhibition of APC-cofactor activity of protein S at R(306), but by generation of a specific inhibitor for APCcatalyzed proteolysis at R(506) of factor Va. These results are of interest for carriers of the factor V(Leiden) mutation (R(506)Q), as protein S-C4BP effectively enhances APC-catalyzed factor Va (R(306)) inactivation in plasma containing factor V(Leiden). PMID- 18160669 TI - Bortezomib blocks Bax degradation in malignant B cells during treatment with TRAIL. AB - Proapoptotic Bcl-2 family member Bax is a crucial protein in the induction of apoptosis, and its activation is required for this process. Here we report that Bax is a short-lived protein in malignant B cells and Bax protein levels decreased rapidly when protein synthesis was blocked. Malignant B cells were relatively resistant to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis, and this correlated with low basal Bax protein levels. Furthermore, during treatment with TRAIL, the resistant cell lines showed prominent Bax degradation activity. This degradation activity was localized to mitochondrial Bax and could be prevented by truncated Bid, a BH3-only protein; in contrast, cytosolic Bax was relatively stable. The proteasome inhibitor bortezomib is a potent drug in inducing apoptosis in vitro in malignant B-cell lines and primary chronic lymphocytic leukemic (CLL) cells. In CLL cells, bortezomib induced Bax accumulation, translocation to mitochondria, conformational change, and oligomerization. Accumulation and stabilization of Bax protein by bortezomib-sensitized malignant B cells to TRAIL-induced apoptosis. This study reveals that Bax instability confers resistance to TRAIL, which can be reversed by Bax stabilization with a proteasome inhibitor. PMID- 18160670 TI - Ratio of mutant JAK2-V617F to wild-type Jak2 determines the MPD phenotypes in transgenic mice. AB - An acquired somatic mutation in the JAK2 gene (JAK2-V617F) is present in the majority of patients with myeloproliferative disorders (MPDs). Several phenotypic manifestations (polycythemia vera [PV], essential thrombocythemia [ET], and primary myelofibrosis) can be associated with the same mutation. We generated JAK2-V617F transgenic mice using a human JAK2 gene with the sequences encoding the kinase domain placed in the inverse orientation and flanked by antiparallel loxP sites. Crossing mice of one transgenic line (FF1) with transgenic mice expressing Cre-recombinase under the control of the hematopoiesis specific Vav promoter led to expression of JAK2-V617F that was lower than the endogenous wild type Jak2. These mice developed a phenotype resembling ET with strongly elevated platelet counts and moderate neutrophilia. Induction of the JAK2-V617F transgene with the interferon-inducible MxCre resulted in expression of JAK2-V617F approximately equal to wild-type Jak2 and a PV-like phenotype with increased hemoglobin, thrombocytosis, and neutrophilia. Higher levels of JAK2-V617F in mouse bone marrow by retroviral transduction caused a PV-like phenotype without thrombocytosis. These data are consistent with the hypothesis that the ratio of mutant to wild-type JAK2 is critical for the phenotypic manifestation. A similar correlation was also found in patients with MPD. PMID- 18160671 TI - Identification of somatic JAK1 mutations in patients with acute myeloid leukemia. AB - Somatic mutations in JAK2 are frequently found in myeloproliferative diseases, and gain-of-function JAK3 alleles have been identified in M7 acute myeloid leukemia (AML), but a role for JAK1 in AML has not been described. We screened the entire coding region of JAK1 by total exonic resequencing of bone marrow DNA samples from 94 patients with de novo AML. We identified 2 novel somatic mutations in highly conserved residues of the JAK1 gene (T478S, V623A), in 2 separate patients and confirmed these by resequencing germ line DNA samples from the same patients. Overexpression of mutant JAK1 did not transform primary murine cells in standard assays, but compared with wild-type JAK1, JAK1(T478S), and JAK1(V623A) expression was associated with increased STAT1 activation in response to type I interferon and activation of multiple downstream signaling pathways. This is the first report to demonstrate somatic JAK1 mutations in AML and suggests that JAK1 mutations may function as disease-modifying mutations in AML pathogenesis. PMID- 18160672 TI - Sensorimotor reorganization by proprioceptive training in musician's dystonia and writer's cramp. AB - OBJECTIVE: The sensorimotor organization (SMO) of the motor hand area is abnormal in focal hand dystonia and likely contributes to symptom manifestation. In healthy subjects SMO is changed by training with proprioceptive stimulation. Here we test whether similar interventions reverse the abnormal SMO in musician's dystonia and writer's cramp. If so, they could be developed for therapeutic application. METHODS: In six non-musicians, six professional musicians, six patients with musician's dystonia, and six patients with writer's cramp, SMO was explored by measuring changes in short-interval-intracortical-inhibition (SICI) during short periods of hand muscle vibration before and after two training types: AttVIB, involving attention to 15 minutes vibration of the abductor pollicis brevis (APB); and AttIndex, involving attention to subtle cutaneous stimulation of the index finger. RESULTS: In healthy non-musicians, baseline SMO is spatially differentiated: SICI is reduced in projections to the vibrated, but enhanced to the non-vibrated muscles. Here AttVIB increased and AttIndex reduced the effect of subsequent APB-vibration on SMO. In healthy musicians, baseline SMO is less differentiated. AttVIB reinstated a more differential SMO pattern while AttIndex attenuated the effect of APB vibration. In focal hand dystonia, SMO is completely dedifferentiated. AttVIB tended to restore a more differential SMO in musician's dystonia but not in writer's cramp while AttIndex failed to induce any changes in both groups. CONCLUSION: The intervention effect depends on the pre interventional sensorimotor organization (SMO). In focal hand dystonia, particularly in musician's dystonia, it is possible to retrain an abnormal SMO toward a more differential pattern, which has potential implications for therapy. PMID- 18160673 TI - Association of serum lipid indices with large artery atherosclerotic stroke. AB - BACKGROUND: Low-density lipoprotein cholesterol (LDL) is the primary lipid target for vascular risk reduction in stroke patients, but emerging data suggest that other lipid indices may better predict vascular hazard. We evaluated the relationship between several measures of the classically obtained serum lipid panel and the occurrence of large artery atherosclerotic stroke. METHODS: Data prospectively collected over a 4-year period on subjects admitted with ischemic stroke or TIA to a university medical center were analyzed. Independent associations of fasting serum lipid indices with large artery atherosclerotic (LAA) stroke mechanism were evaluated. RESULTS: Of 1,049 patients, 247 (23.5%) were classified with LAA, 224 (21.4%) were classified with small vessel disease (SVD), and 578 (55%) were non-LAA, non-SVD subtype. Lipid levels were similar between LAA and SVD patients. Total cholesterol, triglycerides, LDL, non-high density lipoprotein cholesterol (HDL), and triglyceride:HDL ratio were significantly higher in LAA vs non-LAA, non-SVD patients. After adjustment for age, hypertension, diabetes, smoking, body mass index, and premorbid statin use, significant odds ratios (ORs) for LAA compared with all other ischemic stroke subtypes for patients in the uppermost lipid quartiles (vs lowest) were triglycerides (OR 2.69, 95% CI 1.44 to 5.02) and non-HDL (OR 2.39, 95% CI 1.40 to 4.11). LDL was not associated with LAA. CONCLUSIONS: Compared with all other ischemic stroke subtypes, elevated levels of serum triglycerides and non-high density lipoprotein, but not low-density lipoprotein (LDL), are associated with large artery atherosclerotic stroke. These non-LDL lipid measures may have utility in delineating atherosclerotic stroke risk. PMID- 18160674 TI - Diagnosis and etiology of congenital muscular dystrophy. AB - OBJECTIVE: We aimed to determine the frequency of all known forms of congenital muscular dystrophy (CMD) in a large Australasian cohort. METHODS: We screened 101 patients with CMD with a combination of immunofluorescence, Western blotting, and DNA sequencing to identify disease-associated abnormalities in glycosylated alpha dystroglycan, collagen VI, laminin alpha2, alpha7-integrin, and selenoprotein. RESULTS: A total of 45% of the CMD cohort were assigned to an immunofluorescent subgroup based on their abnormal staining pattern. Abnormal staining for glycosylated alpha-dystroglycan was present in 25% of patients, and approximately half of these had reduced glycosylated alpha-dystroglycan by Western blot. Sequencing of the FKRP, fukutin, POMGnT1, and POMT1 genes in all patients with abnormal alpha-dystroglycan immunofluorescence identified mutations in one patient for each of these genes and two patients had mutations in POMT2. Twelve percent of patients had abnormalities in collagen VI immunofluorescence, and we identified disease-causing COL6 mutations in eight of nine patients in whom the genes were sequenced. Laminin alpha2 deficiency accounted for only 8% of CMD. alpha7-Integrin staining was absent in 12 of 45 patients studied, and ITGA7 gene mutations were excluded in all of these patients. CONCLUSIONS: We define the distribution of different forms of congenital muscular dystrophy in a large cohort of mixed ethnicity and demonstrate the utility and limitations of current diagnostic techniques. PMID- 18160675 TI - Education, cognitive function, and severity of neuropathology in Alzheimer disease. AB - BACKGROUND: Education may modulate the degree to which the neuropathology of Alzheimer disease (AD) is expressed as impaired cognitive performance. METHODS: We studied 2,051 participants age 65+ years at 27 AD Centers who died and underwent autopsy. All took the Mini-Mental State Examination (MMSE) within 2 years before death. Braak & Braak stage, neuritic plaque density, and Consortium to Establish a Registry for Alzheimer's Disease and National Institute on Aging (NIA)/Reagan diagnostic classifications quantified AD neuropathologic severity. Multivariate analyses modeled MMSE in relation to education and neuropathologic severity, adjusting for age at death, Lewy body pathology, and vascular dementia. RESULTS: Higher education was associated with higher MMSE scores when AD neuropathology was absent or mild. But with more advanced neuropathology, differences in MMSE scores among education levels were attenuated. For example, among patients without AD by NIA/Reagan criteria, fitted MMSE scores ranged from 19.6 for patients with less than high school education to 25.9 with education beyond high school. But among patients with neuropathologically advanced AD, the range of scores by education was only 7.1 to 8.6. CONCLUSIONS: We found no evidence of larger education-related differences in cognitive function when Alzheimer disease (AD) neuropathology was more advanced. Higher Mini-Mental State Examination scores among more educated persons with mild or no AD may reflect better test-taking skills or cognitive reserve, but these advantages may ultimately be overwhelmed by AD neuropathology. PMID- 18160676 TI - Report of the intergovernmental conference of far-Eastern countries on rural hygiene. 1937. PMID- 18160677 TI - Differences in young people's reports of sexual behaviors according to interview methodology: a randomized trial in India. AB - OBJECTIVES: We compared reports of sexual behaviors given in standard face-to face interviews with reports given in audio computer-assisted self-interviews (ACASIs) and culturally specific interactive interviews among adolescents in India. We sought to determine which of the interview methods leads to higher reporting of sexual behaviors among economically disadvantaged 15-19-year-olds in urban India. METHODS: We conducted a randomized trial in which each participant (583 boys and 475 girls) was assigned to 2 interview methods: face-to-face interview and ACASI or interactive interview. We used matched case-control analyses to assess differences in the individual's reporting on the 2 methods. RESULTS: Female participants consistently reported fewer sexual behaviors in ACASIs than in face-to-face interviews, whereas male participants' reports differed according to type of sexual behavior and interview mode. Both male and female participants reported more sexual behaviors during interactive interviews than during face-to-face interviews. Twenty-eight percent of male participants reported having engaged in heterosexual intercourse in interactive interviews, as compared with 20% in face-to-face interviews (P< .01); the corresponding percentages for female participants were 7% and 2% (P<.01). CONCLUSIONS: Our results showed that young people were more likely to report sexual behaviors in culturally specific interactive interviews than in face-to-face interviews. By contrast, ACASIs did not uniformly lead to higher reporting levels than did face to-face interviews. PMID- 18160678 TI - Glial cell missing 1 regulates placental growth factor (PGF) gene transcription in human trophoblast. AB - Placental growth factor (PGF, previously known as PlGF) is prominently expressed by trophoblasts in human placenta, whereas most nontrophoblast cells express low levels of PGF mRNA under normal physiological conditions. We have shown that hypoxia decreases PGF expression in the trophoblast, but little is known about transcriptional regulation of PGF gene expression. We sought to determine promoter regions of the human PGF gene that contribute to its restricted high constitutive expression in the trophoblast. Overlapping putative promoter regions of human PGF gene encompassing 2-1.5 kb were cloned into reporter vectors and co transfected into trophoblast and nontrophoblast cell lines. Promoter activity generated by a 2-1.5-kb clone was significantly higher in trophoblasts than in nontrophoblasts. Selective deletion mutants showed that a clone encompassing the PGF (2-828/++34) region generated promoter activity similar to the 2-1.5-kb region in the trophoblast. However, deletion of another 131 bp from this subclone (2-698/++34) resulted in significantly less promoter activity in the trophoblast. The (2-828/2-698) region significantly enhanced activity of a minimal promoter construct in trophoblast but not in nontrophoblast cells, suggesting that this region contributes to regulating PGF transcription in the trophoblast. Site directed mutagenesis of a glial cell missing 1 (GCM1) motif in the 131-bp region significantly decreased enhancer activity in the trophoblast. Furthermore, overexpression of GCM1 significantly increased PGF 2-1.5-kb promoter activity and PGF mRNA expression in trophoblast and nontrophoblast cells. Forced overexpression of GCM1 restored PGF expression in the hypoxic trophoblast. These data support a functional role for GCM1 contributing to constitutively high trophoblast PGF expression and is the first direct evidence of an oxygen responsive, trophoblast-specific transcription factor contributing to the regulation of PGF expression. PMID- 18160679 TI - Gene-environment interactions, not neonatal growth hormone deficiency, time puberty in female rhesus monkeys. AB - The factors that influence the timing of puberty and the onset of adult fertility are poorly understood. While focus on the juvenile period has provided insights into how growth-related cues affect pubertal timing, growth velocity during infancy that is sustained into the juvenile period may be important. On the other hand, social factors, specifically exposure to psychosocial stressors, can delay sexual maturation, possibly by altering growth velocities during development. Using female rhesus monkeys, the present study used a prospective analysis to determine how neonatal growth hormone (GH) inhibition with a sandostatin analog or suppression of the pituitary-gonadal axis with a GnRH analog affected growth and sexual maturation. A separate retrospective analysis was done assessing the effects of social dominance status during development on pubertal timing. Because a specific polymorphism in the gene encoding the serotonin (5HT) reuptake transporter increases vulnerability to psychosocial stressors, females were also genotyped and were then classified as socially dominant, having both alleles for the long promoter variant or having at least one allele for the short promoter variant, or as socially subordinate, having the long variant or having the short variant. Neonatal treatments were not balanced for social status or genotype, so analyses were performed separately. Although the neonatal treatments reduced GH secretion postnatally and through the juvenile period, neither growth nor sexual maturation was affected. In contrast, the retrospective analysis showed sexual maturation was delayed significantly in subordinate females carrying at least one allele of the short promoter variant in the gene encoding the 5HT reuptake transporter, and this delay was associated with reduced GH and leptin secretion during the juvenile phase but not with differences in growth velocities from birth. These data suggest that decreased neonatal GH secretion does not adversely affect sexual maturation, but that polymorphisms in the gene encoding the 5HT transporter modulate the adverse consequences of social subordination on the timing of puberty in female rhesus monkeys. PMID- 18160680 TI - Weaning and the developmental changes in follicle-stimulating hormone, pituitary adenylate cyclase-activating polypeptide, and inhibin B in the male rat. AB - Pituitary Fshb concentrations increase markedly and selectively beginning on Postnatal Day 20 in the male rat. To evaluate the factors potentially responsible for this rise in FSH, we adjusted the time of weaning, which is generally also on Day 20. Male rat pups were provided nutrients by suckling only and were weaned to laboratory chow earlier (Day 17) or later (Day 23) than normally performed in animal facilities (Day 20). Between ages 17 and 29 days, significant increases were seen in serum LH (1.4-fold) and FSH (2.4-fold) levels; pituitary expression of Lhb (5.4-fold), Fshb (21.3-fold), and inhibin beta B (Inhbb, 2.26-fold) mRNAs; and testicular expression of Inhbb (10-fold) mRNA. Concurrently, significant decreases occurred in serum inhibin B levels (1.8-fold); pituitary adenylate cyclase-activating polypeptide (Adcyap1, 4.2-fold), total follistatin (Fst, 3.5 fold), and Fst isoform 288 (5.6-fold) mRNAs; and testicular expression of inhibin beta A (8.2-fold) mRNA. Early weaning significantly increased serum FSH but not LH and increased pituitary expression of Fshb and GnRH receptor (Gnrhr) mRNAs but not Lhb. Early weaning also significantly decreased serum inhibin B but increased testicular expression of the Inhbb subunit. Early weaning also caused pituitary expression of Fst and Adcyap1 to decline earlier than in the control group. Immediately after weaning, growth accelerated substantially, and the time of weaning produced significant and differential effects on circulating leptin levels that were not related to indices of FSH production. From these observations, we propose the novel hypothesis that the increase in growth rate subsequent to weaning signals circulating inhibin B levels to fall and pituitary Adcyap1 and consequently Fst expression to decrease, and that these events together facilitate the rise in Fshb and Gnrhr expression by increasing pituitary activin signaling. PMID- 18160681 TI - Images in clinical medicine. Small-bowel intussusception. PMID- 18160682 TI - Comparing physicians on efficiency. PMID- 18160683 TI - Is quality improvement improving quality? A view from the doctor's office. PMID- 18160684 TI - One step forward, two steps back--will there ever be an AIDS vaccine? PMID- 18160685 TI - Prophylactic catheter ablation for the prevention of defibrillator therapy. AB - BACKGROUND: For patients who have a ventricular tachyarrhythmic event, implantable cardioverter-defibrillators (ICDs) are a mainstay of therapy to prevent sudden death. However, ICD shocks are painful, can result in clinical depression, and do not offer complete protection against death from arrhythmia. We designed this randomized trial to examine whether prophylactic radiofrequency catheter ablation of arrhythmogenic ventricular tissue would reduce the incidence of ICD therapy. METHODS: Eligible patients with a history of a myocardial infarction underwent defibrillator implantation for spontaneous ventricular tachycardia or fibrillation. The patients did not receive antiarrhythmic drugs. Patients were randomly assigned to defibrillator implantation alone or defibrillator implantation with adjunctive catheter ablation (64 patients in each group). Ablation was performed with the use of a substrate-based approach in which the myocardial scar is mapped and ablated while the heart remains predominantly in sinus rhythm. The primary end point was survival free from any appropriate ICD therapy. RESULTS: The mortality rate 30 days after ablation was zero, and there were no significant changes in ventricular function or functional class during the mean (+/-SD) follow-up period of 22.5+/-5.5 months. Twenty-one patients assigned to defibrillator implantation alone (33%) and eight patients assigned to defibrillator implantation plus ablation (12%) received appropriate ICD therapy (antitachycardia pacing or shocks) (hazard ratio in the ablation group, 0.35; 95% confidence interval, 0.15 to 0.78, P=0.007). Among these patients, 20 in the control group (31%) and 6 in the ablation group (9%) received shocks (P=0.003). Mortality was not increased in the group assigned to ablation as compared with the control group (9% vs. 17%, P=0.29). CONCLUSIONS: In this randomized trial, prophylactic substrate-based catheter ablation reduced the incidence of ICD therapy in patients with a history of myocardial infarction who received ICDs for the secondary prevention of sudden death. (Current Controlled Trials number, ISRCTN62488166 [controlled-trials.com].). PMID- 18160686 TI - Paclitaxel plus bevacizumab versus paclitaxel alone for metastatic breast cancer. AB - BACKGROUND: In an open-label, randomized, phase 3 trial, we compared the efficacy and safety of paclitaxel with that of paclitaxel plus bevacizumab, a monoclonal antibody against vascular endothelial growth factor, as initial treatment for metastatic breast cancer. METHODS: We randomly assigned patients to receive 90 mg of paclitaxel per square meter of body-surface area on days 1, 8, and 15 every 4 weeks, either alone or with 10 mg of bevacizumab per kilogram of body weight on days 1 and 15. The primary end point was progression-free survival; overall survival was a secondary end point. RESULTS: From December 2001 through May 2004, a total of 722 patients were enrolled. Paclitaxel plus bevacizumab significantly prolonged progression-free survival as compared with paclitaxel alone (median, 11.8 vs. 5.9 months; hazard ratio for progression, 0.60; P<0.001) and increased the objective response rate (36.9% vs. 21.2%, P<0.001). The overall survival rate, however, was similar in the two groups (median, 26.7 vs. 25.2 months; hazard ratio, 0.88; P=0.16). Grade 3 or 4 hypertension (14.8% vs. 0.0%, P<0.001), proteinuria (3.6% vs. 0.0%, P<0.001), headache (2.2% vs. 0.0%, P=0.008), and cerebrovascular ischemia (1.9% vs. 0.0%, P=0.02) were more frequent in patients receiving paclitaxel plus bevacizumab. Infection was more common in patients receiving paclitaxel plus bevacizumab (9.3% vs. 2.9%, P<0.001), but febrile neutropenia was uncommon (<1% overall). CONCLUSIONS: Initial therapy of metastatic breast cancer with paclitaxel plus bevacizumab prolongs progression free survival, but not overall survival, as compared with paclitaxel alone. (ClinicalTrials.gov number, NCT00028990 [ClinicalTrials.gov].). PMID- 18160687 TI - Local dystrophin restoration with antisense oligonucleotide PRO051. AB - BACKGROUND: Duchenne's muscular dystrophy is associated with severe, progressive muscle weakness and typically leads to death between the ages of 20 and 35 years. By inducing specific exon skipping during messenger RNA (mRNA) splicing, antisense compounds were recently shown to correct the open reading frame of the DMD gene and thus to restore dystrophin expression in vitro and in animal models in vivo. We explored the safety, adverse-event profile, and local dystrophin restoring effect of a single, intramuscular dose of an antisense oligonucleotide, PRO051, in patients with this disease. METHODS: Four patients, who were selected on the basis of their mutational status, muscle condition, and positive exon skipping response to PRO051 in vitro, received a dose of 0.8 mg of PRO051 injected into the tibialis anterior muscle. A biopsy was performed 28 days later. Safety measures, composition of mRNA, and dystrophin expression were assessed. RESULTS: PRO051 injection was not associated with clinically apparent adverse events. Each patient showed specific skipping of exon 51 and sarcolemmal dystrophin in 64 to 97% of myofibers. The amount of dystrophin in total protein extracts ranged from 3 to 12% of that found in the control specimen and from 17 to 35% of that of the control specimen in the quantitative ratio of dystrophin to laminin alpha2. CONCLUSIONS: Intramuscular injection of antisense oligonucleotide PRO051 induced dystrophin synthesis in four patients with Duchenne's muscular dystrophy who had suitable mutations, suggesting that further studies might be feasible. PMID- 18160688 TI - COL4A1 mutations and hereditary angiopathy, nephropathy, aneurysms, and muscle cramps. AB - BACKGROUND: COL4A3, COL4A4, and COL4A5 are the only collagen genes that have been implicated in inherited nephropathies in humans. However, the causative genes for a number of hereditary multicystic kidney diseases, myopathies with cramps, and heritable intracranial aneurysms remain unknown. METHODS: We characterized the renal and extrarenal phenotypes of subjects from three families who had an autosomal dominant hereditary angiopathy with nephropathy, aneurysms, and muscle cramps (HANAC), which we propose is a syndrome. Linkage studies involving microsatellite markers flanking the COL4A1-COL4A2 locus were performed, followed by sequence analysis of COL4A1 complementary DNA extracted from skin-fibroblast specimens from the subjects. RESULTS: We identified three closely located glycine mutations in exons 24 and 25 of the gene COL4A1, which encodes procollagen type IV alpha1. The clinical renal manifestations of the HANAC syndrome in these families include hematuria and bilateral, large cysts. Histologic analysis revealed complex basement-membrane defects in kidney and skin. The systemic angiopathy of the HANAC syndrome appears to affect both small vessels and large arteries. CONCLUSIONS: COL4A1 may be a candidate gene in unexplained familial syndromes with autosomal dominant hematuria, cystic kidney disease, intracranial aneurysms, and muscle cramps. PMID- 18160689 TI - Clinical practice. Localized prostate cancer. PMID- 18160690 TI - Images in clinical medicine. Mapping the atrioventricular node. PMID- 18160691 TI - Case records of the Massachusetts General Hospital. Case 40-2007. A 38-year-old man with weakness in the hands. PMID- 18160692 TI - Ablation after ICD implantation--bridging the gap between promise and practice. PMID- 18160693 TI - Skipping toward personalized molecular medicine. PMID- 18160694 TI - Military-civilian collaboration in trauma care and the senior visiting surgeon program. PMID- 18160695 TI - Effectiveness of influenza vaccination. PMID- 18160696 TI - Sexuality and health among older adults in the United States. PMID- 18160697 TI - Minimizing ventricular pacing in sinus-node disease. PMID- 18160698 TI - Multiple autoimmune diseases after autologous stem-cell transplantation. PMID- 18160699 TI - Aspirin and hormone therapy for prostate cancer. PMID- 18160700 TI - A brief history of great discoveries in pharmacology: in celebration of the centennial anniversary of the founding of the American Society of Pharmacology and Experimental Therapeutics. PMID- 18160701 TI - 5-HT receptor regulation of neurotransmitter release. AB - Serotoninergic neurons in the central nervous system impinge on many other neurons and modulate their neurotransmitter release. This review focuses on 1) the function of presynaptic 5-hydroxytryptamine (5-HT) heteroreceptors on axon terminals of central cholinergic, dopaminergic, noradrenergic, or GABAergic neurons and 2) the role of GABAergic interneurons expressing 5-HT heteroreceptors in the regulation of acetylcholine, dopamine, or noradrenaline release. In vitro studies on slices or synaptosomes and in vivo microdialysis experiments have shown that 5-HT(1A), 5-HT(1B), 5-HT(2A), 5-HT(2C), 5-HT(3), and/or 5-HT(4) heteroreceptors mediate this modulation. 5-HT(1B) receptors on neocortical cholinergic, striatal dopaminergic, or hippocampal GABAergic axon terminals are examples for release-inhibiting 5-HT heteroreceptors; 5-HT(3) receptors on hippocampal GABAergic or 5-HT(4) receptors on hippocampal cholinergic axon terminals are examples for release-facilitating 5-HT heteroreceptors. GABA released from GABAergic interneurons upon activation of facilitatory 5-HT receptors, e.g., 5-HT(2A) or 5-HT(3) receptors, mediates inhibition of the release of other neurotransmitters such as prefrontal neocortical dopamine or neocortical acetylcholine release, respectively. Conversely, attenuated GABA release in response to activation of inhibitory 5-HT heteroreceptors, e.g., 5 HT(1A) or 5-HT(1B) receptors on GABAergic interneurons is involved in paradoxical facilitation of hippocampal acetylcholine and striatal dopamine release, respectively. Such 5-HT heteroreceptors are considered potential targets for appropriate 5-HT receptor ligands which, by enhancing the release of a relevant neurotransmitter, can compensate for its hypothesized deficiency in distinct brain areas. Examples for such deficiencies are the impaired release of hippocampal or neocortical acetylcholine, striatal dopamine, and hippocampal or neocortical noradrenaline in disorders such as Alzheimer's disease, Parkinson's disease, and major depression, respectively. PMID- 18160702 TI - Basic helix-loop-helix transcription factors cooperate to specify a cortical projection neuron identity. AB - Several transcription factors are essential determinants of a cortical projection neuron identity, but their mode of action (instructive versus permissive) and downstream genetic cascades remain poorly defined. Here, we demonstrate that the proneural basic helix-loop-helix (bHLH) gene Ngn2 instructs a partial cortical identity when misexpressed in ventral telencephalic progenitors, inducing ectopic marker expression in a defined temporal sequence, including early (24 h; Nscl2), intermediate (48 h; BhlhB5), and late (72 h; NeuroD, NeuroD2, Math2, and Tbr1) target genes. Strikingly, cortical gene expression was much more rapidly induced by Ngn2 in the dorsal telencephalon (within 12 to 24 h). We identify the bHLH gene Math3 as a dorsally restricted Ngn2 transcriptional target and cofactor, which synergizes with Ngn2 to accelerate target gene transcription in the cortex. Using a novel in vivo luciferase assay, we show that Ngn2 generates only approximately 60% of the transcriptional drive in ventral versus dorsal telencephalic domains, an activity that is augmented by Math3, providing a mechanistic basis for regional differences in Ngn2 function. Cortical bHLH genes thus cooperate to control transcriptional strength, thereby temporally coordinating downstream gene expression. PMID- 18160703 TI - Contribution of annexin 2 to the architecture of mature endothelial adherens junctions. AB - The vascular endothelial cadherin (VE-cad)-based complex is involved in the maintenance of vascular endothelium integrity. Using immunoprecipitation experiments, we have demonstrated that, in confluent human umbilical vein endothelial cells, the VE-cad-based complex interacts with annexin 2 and that annexin 2 translocates from the cytoplasm to the cell-cell contact sites as cell confluence is established. Annexin 2, located in cholesterol rafts, binds to both the actin cytoskeleton and the VE-cad-based complex so the complex is docked to cholesterol rafts. These multiple connections prevent the lateral diffusion of the VE-cad-based complex, thus strengthening adherens junctions in the ultimate steps of maturation. Moreover, we observed that the down-regulation of annexin 2 by small interfering RNA induces a delocalization of VE-cad from adherens junctions and consequently a destabilization of these junctions. Furthermore, our data indicate that the decoupling of the annexin 2/p11 complex from the VE-cad based junction, triggered by vascular endothelial growth factor treatment, facilitates the switch from a quiescent to an immature state. PMID- 18160704 TI - Wingless signaling induces widespread chromatin remodeling of target loci. AB - How signaling cascades influence gene regulation at the level of chromatin modification is not well understood. We studied this process using the Wingless/Wnt pathway in Drosophila. When cells sense Wingless ligand, Armadillo (the fly beta-catenin) becomes stabilized and translocates to the nucleus, where it binds to the sequence-specific DNA binding protein TCF to activate transcription of target genes. Here, we show that Wingless signaling induces TCF and Armadillo recruitment to a select subset of TCF binding site clusters that act as Wingless response elements. Despite this localized TCF/Armadillo recruitment, histones are acetylated over a wide region (up to 30 kb) surrounding the Wingless response elements in response to pathway activation. This widespread histone acetylation occurs independently of transcription. In contrast to Wingless targets, other active genes not regulated by the pathway display sharp acetylation peaks centered on their core promoters. Widespread acetylation of Wingless targets is dependent upon CBP, a histone acetyltransferase known to bind to Armadillo and is correlated with activation of target gene expression. These data suggest that pathway activation induces localized recruitment of TCF/Armadillo/CBP to Wingless response elements, leading to widespread histone acetylation of target loci prior to transcriptional activation. PMID- 18160705 TI - An evolutionarily conserved nuclear export signal facilitates cytoplasmic localization of the Tbx5 transcription factor. AB - During cardiac development, the T-box transcription factor Tbx5 displays dynamic changes in localization from strictly nuclear to both nuclear and cytoplasmic to exclusively cytoplasmic along the actin cytoskeleton in cells coexpressing its binding protein LMP4. Although nuclear localization signals (NLSs) have been described, the mechanism by which Tbx5 exits the nucleus remained elusive. Here, we describe for Tbx5 a nuclear export signal (NES) that is recognized by the CRM1 export protein. Site-directed mutagenesis of a critical amino acid(s) within this sequence determined the functionality of this NES. Confocal localization studies and luciferase transcriptional reporter assays with NES mutant Tbx5 forms demonstrated retention in the nucleus, regardless of the presence of LMP4. Coimmunoprecipitation and pharmacological interference studies demonstrated a direct interaction between Tbx5 and CRM1, revealing that Tbx5 is using the CRM1 pathway for nuclear export. In addition to Tbx5, we identified NESs in all T-box proteins and demonstrated interaction of the family members Tbx3 and Brachyury with the CRM1 exporter, suggesting general significance. This first demonstration of evolutionarily conserved NESs in all T-box proteins in conjunction with NLSs indicates a primordial function of T-box proteins to dynamically shuttle between nuclear and cytoplasmic compartments of the cell. PMID- 18160706 TI - Sizn1 is a novel protein that functions as a transcriptional coactivator of bone morphogenic protein signaling. AB - Bone morphogenic proteins (BMPs) play pleotrophic roles in nervous system development, and their signaling is highly regulated at virtually every step in the pathway. We have cloned a novel gene, Sizn1 (Smad-interacting zinc finger protein), which functions as a transcriptional coactivator of BMP signaling. It positively modulates BMP signaling by interacting with Smad family members and associating with CBP in the transcription complex. Sizn1 is expressed in the ventral embryonic forebrain, where, as we will show, it contributes to BMP dependent, cholinergic-neuron-specific gene expression. These data indicate that Sizn1 is a positive modulator of BMP signaling and provide further insight into how BMP signaling can be modulated in neuronal progenitor subsets to influence cell-type-specific gene expression and development. PMID- 18160707 TI - Integration of transforming growth factor beta and RAS signaling silences a RAB5 guanine nucleotide exchange factor and enhances growth factor-directed cell migration. AB - Transforming growth factor beta (TGF-beta) receptor (TbetaR) signaling contributes to normal development as well as tumorigenesis. Here we report that RIN1, a RAB5 guanine nucleotide exchange factor (GEF) and down regulator of receptor tyrosine kinases (RTKs), promotes TbetaR signaling through enhanced endocytosis. TbetaR activation induces SNAI1 (Snail), a transcription repressor that reduces RIN1 expression, providing a negative feedback mechanism to control TbetaR trafficking and downstream signaling. Persistent RAS signaling disrupts this equilibrium by stabilizing SNAI1 protein, resulting in strong silencing of RIN1 and stabilization of RTKs. TGF-beta-induced RIN1 silencing in breast cancer cells prolonged sensitivity to hepatocyte growth factor, a ligand for the MET type RTK, and enhanced growth factor-directed cell motility. We conclude that in some tumor cells TbetaR and RAS signals are integrated through the silencing of RIN1, leading to a reduction in RAB5-mediated endocytosis. These findings shed new light on the basis for distinct interpretations of TGF-beta signaling by normal versus transformed cells. PMID- 18160708 TI - Loss of singleminded-2s in the mouse mammary gland induces an epithelial mesenchymal transition associated with up-regulation of slug and matrix metalloprotease 2. AB - The short splice variant of the basic helix-loop-helix Per-Arnt-Sim transcription factor Singleminded-2, SIM2s, has been implicated in development and is frequently lost or reduced in primary breast tumors. Here, we show that loss of Sim2s causes aberrant mouse mammary gland ductal development with features suggestive of malignant transformation, including increased proliferation, loss of polarity, down-regulation of E-cadherin, and invasion of the surrounding stroma. Additionally, knockdown of SIM2s in MCF-7 breast cancer cells contributed to an epithelial-mesenchymal transition (EMT) and increased tumorigenesis. In both Sim2(-/-) mammary glands and SIM2s-depleted MCF7 cells, these changes were associated with increased SLUG and MMP2 levels. SIM2s protein was detectable on the SLUG promoter, and overexpression of SIM2s repressed expression from a SLUG controlled reporter in a dose-dependent manner. To our knowledge, SIM2s is the first protein shown to bind and repress the SLUG promoter, providing a plausible explanation for the development role and breast tumor-suppressive activity of SIM2s. Together, our results suggest that SIM2s is a key regulator of mammary ductal development and that loss of SIM2s expression is associated with an invasive, EMT-like phenotype. PMID- 18160709 TI - Myocyte enhancer factor 2 and chorion factor 2 collaborate in activation of the myogenic program in Drosophila. AB - The process of myogenesis requires the coordinated activation of many structural genes whose products are required for myofibril assembly, function, and regulation. Although numerous reports have documented the importance of the myogenic regulator myocyte enhancer factor 2 (MEF2) in muscle differentiation, the interaction of MEF2 with cofactors is critical to the realization of muscle fate. We identify here a genomic region required for full MEF2-mediated activation of actin gene expression in Drosophila, and we identify the zinc finger transcriptional regulator chorion factor 2 (CF2) as a factor functioning alongside MEF2 via this region. Furthermore, although both MEF2 and CF2 can individually activate actin gene expression, we demonstrate that these two factors collaborate in regulating the Actin57B target gene in vitro and in vivo. More globally, MEF2 and CF2 synergistically activate the enhancers of a number of muscle-specific genes, and loss of CF2 function in vivo results in reductions in the levels of several muscle structural gene transcripts. These findings validate a general importance of CF2 alongside MEF2 as a critical regulator of the myogenic program, identify a new regulator functioning with MEF2 to control cell fate, and provide insight into the network of regulatory events that shape the developing musculature. PMID- 18160710 TI - ATP binding by monarch-1/NLRP12 is critical for its inhibitory function. AB - The recently discovered nucleotide binding domain-leucine rich repeat (NLR) gene family is conserved from plants to mammals, and several members are associated with human autoinflammatory or immunodeficiency disorders. This family is defined by a central nucleotide binding domain that contains the highly conserved Walker A and Walker B motifs. Although the nucleotide binding domain is a defining feature of this family, it has not been extensively studied in its purified form. In this report, we show that purified Monarch-1/NLRP12, an NLR protein that negatively regulates NF-kappaB signaling, specifically binds ATP and exhibits ATP hydrolysis activity. Intact Walker A/B motifs are required for this activity. These motifs are also required for Monarch-1 to undergo self-oligomerization, Toll-like receptor- or CD40L-activated association with NF-kappaB-inducing kinase (NIK) and interleukin-1 receptor-associated kinase 1 (IRAK-1), degradation of NIK, and inhibition of IRAK-1 phosphorylation. The stable expression of a Walker A/B mutant in THP-1 monocytes results in increased production of proinflammatory cytokines and chemokines to an extent comparable to that in cells in which Monarch-1 is silenced via short hairpin RNA. The results of this study are consistent with a model wherein ATP binding regulates the anti-inflammatory activity of Monarch-1. PMID- 18160711 TI - Recombination-based telomere maintenance is dependent on Tel1-MRN and Rap1 and inhibited by telomerase, Taz1, and Ku in fission yeast. AB - Fission yeast cells survive loss of the telomerase catalytic subunit Trt1 (TERT) through recombination-based telomere maintenance or through chromosome circularization. Although trt1Delta survivors with linear chromosomes can be obtained, they often spontaneously circularize their chromosomes. Therefore, it was difficult to establish genetic requirements for telomerase-independent telomere maintenance. In contrast, when the telomere-binding protein Taz1 is also deleted, taz1Delta trt1Delta cells are able to stably maintain telomeres. Thus, taz1Delta trt1Delta cells can serve as a valuable tool in understanding the regulation of telomerase-independent telomere maintenance. In this study, we show that the checkpoint kinase Tel1 (ATM) and the DNA repair complex Rad32-Rad50-Nbs1 (MRN) are required for telomere maintenance in taz1Delta trt1Delta cells. Surprisingly, Rap1 is also essential for telomere maintenance in taz1Delta trt1Delta cells, even though recruitment of Rap1 to telomeres depends on Taz1. Expression of catalytically inactive Trt1 can efficiently inhibit recombination based telomere maintenance, but the inhibition requires both Est1 and Ku70. While Est1 is essential for recruitment of Trt1 to telomeres, Ku70 is dispensable. Thus, we conclude that Taz1, TERT-Est1, and Ku70-Ku80 prevent telomere recombination, whereas MRN-Tel1 and Rap1 promote recombination-based telomere maintenance. Evolutionarily conserved proteins in higher eukaryotic cells might similarly contribute to telomere recombination. PMID- 18160712 TI - Doubling the size of the glucocorticoid receptor ligand binding pocket by deacylcortivazol. AB - A common feature of nuclear receptor ligand binding domains (LBD) is a helical sandwich fold that nests a ligand binding pocket within the bottom half of the domain. Here we report that the ligand pocket of glucocorticoid receptor (GR) can be continuously extended into the top half of the LBD by binding to deacylcortivazol (DAC), an extremely potent glucocorticoid. It has been puzzling for decades why DAC, which contains a phenylpyrazole replacement at the conserved 3-ketone of steroid hormones that are normally required for activation of their cognate receptors, is a potent GR activator. The crystal structure of the GR LBD bound to DAC and the fourth LXXLL motif of steroid receptor coactivator 1 reveals that the GR ligand binding pocket is expanded to a size of 1,070 A(3), effectively doubling the size of the GR dexamethasone-binding pocket of 540 A(3) and yet leaving the structure of the coactivator binding site intact. DAC occupies only approximately 50% of the space of the pocket but makes intricate interactions with the receptor around the phenylpyrazole group that accounts for the high-affinity binding of DAC. The dramatic expansion of the DAC-binding pocket thus highlights the conformational adaptability of GR to ligand binding. The new structure also allows docking of various nonsteroidal ligands that cannot be fitted into the previous structures, thus providing a new rational template for drug discovery of steroidal and nonsteroidal glucocorticoids that can be specifically designed to reach the unoccupied space of the expanded pocket. PMID- 18160713 TI - CARD6 is interferon inducible but not involved in nucleotide-binding oligomerization domain protein signaling leading to NF-kappaB activation. AB - We have previously reported the cloning and characterization of CARD6, a caspase recruitment domain (CARD)-containing protein that is structurally related to the interferon (IFN)-inducible GTPases. CARD6 associates with microtubules and with receptor-interacting protein 2 (RIP2). RIP2 mediates NF-kappaB activation induced by the intracellular nucleotide-binding oligomerization domain (NOD) receptors that sense bacterial peptidoglycan. Here we report that the expression of CARD6 and RIP2 in bone marrow-derived macrophages is rapidly induced by beta IFN and gamma IFN. This IFN-induced upregulation of CARD6 is suppressed by lipopolysaccharide (LPS), in contrast to LPS's enhancement of IFN-induced RIP2 upregulation. We generated CARD6-deficient (CARD6(-/-)) mice and carried out extensive analyses of signaling pathways mediating innate and adaptive immune responses, including the NOD pathways, but did not detect any abnormalities. Moreover, CARD6(-/-) mice were just as susceptible as wild-type mice to infection by Salmonella enterica serovar Typhimurium, Listeria monocytogenes, Candida albicans, lymphocytic choriomeningitis virus, or mouse adenovirus type 1. Thus, although structural and in vitro analyses strongly suggest an important role for CARD6 in immune defense, the physiological function of CARD6 remains obscure. PMID- 18160714 TI - Mosaic complementation demonstrates a regulatory role for myosin VIIa in actin dynamics of stereocilia. AB - We have developed a bacterial artificial chromosome transgenesis approach that allowed the expression of myosin VIIa from the mouse X chromosome. We demonstrated the complementation of the Myo7a null mutant phenotype producing a fine mosaic of two types of sensory hair cells within inner ear epithelia of hemizygous transgenic females due to X inactivation. Direct comparisons between neighboring auditory hair cells that were different only with respect to myosin VIIa expression revealed that mutant stereocilia are significantly longer than those of their complemented counterparts. Myosin VIIa-deficient hair cells showed an abnormally persistent tip localization of whirlin, a protein directly linked to elongation of stereocilia, in stereocilia. Furthermore, myosin VIIa localized at the tips of all abnormally short stereocilia of mice deficient for either myosin XVa or whirlin. Our results strongly suggest that myosin VIIa regulates the establishment of a setpoint for stereocilium heights, and this novel role may influence their normal staircase-like arrangement within a bundle. PMID- 18160715 TI - Deubiquitylating enzyme UBP64 controls cell fate through stabilization of the transcriptional repressor tramtrack. AB - Protein ubiquitylation plays a central role in multiple signal transduction pathways. However, the substrate specificity and potential developmental roles of deubiquitylating enzymes remain poorly understood. Here, we show that the Drosophila ubiquitin protease UBP64 controls cell fate in the developing eye. UBP64 represses neuronal cell fate but promotes the formation of nonneuronal cone cells. Using a proteomics approach, we identified the transcriptional repressor Tramtrack (TTK) as a primary UBP64 substrate. In common with TTK, reduced UBP64 levels lead to a loss of cone cells, supernumerary photoreceptors, and mechanosensory bristle cells. Previously, it was demonstrated that the blockade of neuronal cell fate was relieved by SINA-dependent ubiquitylation and degradation of TTK. We found that UBP64 counteracts SINA function by deubiquitylating TTK, leading to its stabilization and thereby promoting a nonneuronal cell fate. Mass spectrometric mapping revealed that SINA ubiquitylates multiple sites dispersed throughout TTK, which are duly deubiquitylated by UBP64. This observation suggests that both E3 SINA and UBP64 use a scanning mechanism to (de)ubiquitylate TTK. We conclude that the balance of TTK ubiquitylation by SINA and deubiquitylation by UBP64 constitutes a specific posttranslational switch controlling cell fate. PMID- 18160716 TI - A novel mechanism for the control of translation initiation by amino acids, mediated by phosphorylation of eukaryotic initiation factor 2B. AB - Eukaryotic initiation factor 2B (eIF2B) plays a key role in controlling the initiation of mRNA translation. eIF2B is heteropentamer whose catalytic (epsilon) subunit promotes GDP/GTP exchange on eIF2. We show here that depriving human cells of amino acids rapidly results in the inhibition of eIF2B, independently of changes in eIF2 phosphorylation. Although amino acid deprivation also inhibits signaling through the mammalian target of rapamycin complex 1 (mTORC1), the inhibition of eIF2B activity by amino acid starvation is independent of mTORC1. Instead, amino acids repress the phosphorylation of a novel site in eIF2Bepsilon. We identify this site as Ser525, located adjacent to the known phosphoregulatory region in eIF2Bepsilon. Mutation of Ser525 to Ala abolishes the regulation of eIF2B and protein synthesis by amino acids. This indicates that phosphorylation of this site is crucial for the control of eIF2B and protein synthesis by amino acids. These findings identify a new way in which amino acids regulate a key step in translation initiation and indicate that this involves a novel amino acid sensitive signaling mechanism. PMID- 18160717 TI - Beta-catenin expression results in p53-independent DNA damage and oncogene induced senescence in prelymphomagenic thymocytes in vivo. AB - The expression of beta-catenin, a potent oncogene, is causally linked to tumorigenesis. Therefore, it was surprising that the transgenic expression of oncogenic beta-catenin in thymocytes resulted in thymic involution instead of lymphomagenesis. In this report, we demonstrate that this is because the expression of oncogenic beta-catenin induces DNA damage, growth arrest, oncogene induced senescence (OIS), and apoptosis of immature thymocytes. In p53-deficient mice, the expression of oncogenic beta-catenin still results in DNA damage and OIS, but the thymocytes survive and eventually progress to thymic lymphoma. beta Catenin-induced thymic lymphomas are distinct from lymphomas that arise in p53(-/ ) mice. They are CD4(-) CD8(-), while p53-dependent lymphomas are largely CD4(+) CD8(+), and they develop at an earlier age and in the absence of c-Myc expression or Notch1 signaling. Thus, we report that oncogenic beta-catenin-induced, p53 independent growth arrest and OIS and p53-dependent apoptosis protect developing thymocytes from transformation by oncogenic beta-catenin. PMID- 18160718 TI - Centrin/Cdc31 is a novel regulator of protein degradation. AB - Rad23 is required for efficient protein degradation and performs an important role in nucleotide excision repair. Saccharomyces cerevisiae Rad23, and its human counterpart (hHR23), are present in a complex containing the DNA repair factor Rad4 (termed XPC, for xeroderma pigmentosum group C, in humans). XPC/hHR23 was also reported to bind centrin-2, a member of the superfamily of calcium-binding EF-hand proteins. We report here that yeast centrin, which is encoded by CDC31, is similarly present in a complex with Rad4/Rad23 (called NEF2). The interaction between Cdc31 and Rad23/Rad4 varied by growth phase and reflected oscillations in Cdc31 levels. Strikingly, a cdc31 mutant that formed a weaker interaction with Rad4 showed sensitivity to UV light. Based on the dual function of Rad23, in both DNA repair and protein degradation, we questioned if Cdc31 also participated in protein degradation. We report here that Cdc31 binds the proteasome and multiubiquitinated proteins through its carboxy-terminal EF-hand motifs. Moreover, cdc31 mutants were highly sensitive to drugs that cause protein damage, failed to efficiently degrade proteolytic substrates, and formed altered interactions with the proteasome. These findings reveal for the first time a new role for centrin/Cdc31 in protein degradation. PMID- 18160719 TI - Protein kinase A-dependent phosphorylation modulates beta1Pix guanine nucleotide exchange factor activity through 14-3-3beta binding. AB - beta(1)Pix is a guanine nucleotide exchange factor (GEF) for the small GTPases Rac and Cdc42 which has been shown to mediate signaling pathways leading to cytoskeletal reorganization. In the present study, we show that the basal association between endogenous betaPix and endogenous 14-3-3beta was increased after forskolin stimulation and significantly inhibited by protein kinase A inhibitor. However, forskolin stimulation failed to increase the interaction between 14-3-3beta and a beta(1)Pix mutant that is insensitive to protein kinase A phosphorylation, beta(1)Pix(S516A, T526A). We present evidence indicating that forskolin-induced binding of 14-3-3beta to beta(1)Pix results in inhibition of Rac1 GTP loading in 293 cells and in vitro. Furthermore, we show that deletion of 10 amino acid residues within the leucine zipper domain is sufficient to block beta(1)Pix homodimerization and 14-3-3beta binding and modulates beta(1)Pix-GEF activity. These residues also play a crucial role in beta(1)Pix intracellular localization. These results indicate that 14-3-3beta negatively affects the GEF activity of dimeric beta(1)Pix only. Altogether, these results provide a mechanistic insight into the role of 14-3-3beta in modulating beta(1)Pix-GEF activity. PMID- 18160720 TI - Jak2 FERM domain interaction with the erythropoietin receptor regulates Jak2 kinase activity. AB - Janus kinases are essential for signal transduction by a variety of cytokine receptors and when inappropriately activated can cause hematopoietic disorders and oncogenesis. Consequently, it can be predicted that the interaction of the kinases with receptors and the events required for activation are highly controlled. In a screen to identify phosphorylation events regulating Jak2 activity in EpoR signaling, we identified a mutant (Jak2-Y613E) which has the property of being constitutively activated, as well as an inactivating mutation (Y766E). Although no evidence was obtained to indicate that either site is phosphorylated in signaling, the consequences of the Y613E mutation are similar to those observed with recently described activating mutations in Jak2 (Jak2 V617F and Jak2-L611S). However, unlike the V617F or L611S mutant, the Y613E mutant requires the presence of the receptor but not Epo stimulation for activation and downstream signaling. The properties of the Jak2-Y613E mutant suggest that under normal conditions, Jak2 that is not associated with a receptor is locked into an inactive state and receptor binding through the FERM domain relieves steric constraints, allowing the potential to be activated with receptor engagement. PMID- 18160722 TI - Putting the "bio" back into biomarkers: orienting proteomic discovery toward biology and away from the measurement platform. PMID- 18160723 TI - Preanalytical influences in clinical proteomic studies: raising awareness of fundamental issues in sample banking. PMID- 18160724 TI - Fifty-eight years of hemoglobin analysis. PMID- 18160725 TI - Inflammation in atherosclerosis: from vascular biology to biomarker discovery and risk prediction. AB - Recent investigations of atherosclerosis have focused on inflammation, providing new insight into mechanisms of disease. Inflammatory cytokines involved in vascular inflammation stimulate the generation of endothelial adhesion molecules, proteases, and other mediators, which may enter the circulation in soluble form. These primary cytokines also induce production of the messenger cytokine interleukin-6, which stimulates the liver to increase production of acute-phase reactants such as C-reactive protein. In addition, platelets and adipose tissue can generate inflammatory mediators relevant to atherothrombosis. Despite the irreplaceable utility of plasma lipid profiles in assessment of atherosclerotic risk, these profiles provide an incomplete picture. Indeed, many cardiovascular events occur in individuals with plasma cholesterol concentrations below the National Cholesterol Education Program thresholds of 200 mg/dL for total cholesterol and 130 mg/dL for low-density lipoprotein (LDL) cholesterol. The concept of the involvement of inflammation in atherosclerosis has spurred the discovery and adoption of inflammatory biomarkers for cardiovascular risk prediction. C-reactive protein is currently the best validated inflammatory biomarker; in addition, soluble CD40 ligand, adiponectin, interleukin 18, and matrix metalloproteinase 9 may provide additional information for cardiovascular risk stratification and prediction. This review retraces the biology of atherothrombosis and the evidence supporting the role of inflammatory biomarkers in predicting primary cardiovascular events in this biologic context. PMID- 18160726 TI - Influence of 5,10-methylenetetrahydrofolate reductase polymorphism on whole-blood folate concentrations measured by LC-MS/MS, microbiologic assay, and bio-rad radioassay. AB - BACKGROUND: The 5,10-methylenetetrahydrofolate reductase (NADPH) (MTHFR) C677T polymorphism may affect whole-blood folate pattern measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and total folate measured by LC-MS/MS, microbiologic assay, and Bio-Rad radioassay (BR). METHODS: We analyzed 171 whole blood hemolysates from 2 blood banks for folate pattern and total folate concentrations using these 3 methods and determined MTHFR genotype. RESULTS: The median (range) total folate concentration by LC-MS/MS was higher in the US set [378 (228-820) nmol/L; n = 96] than in the European set [250 (122-582) nmol/L; n = 75]. The whole-blood folate pattern [median (range)] was similar for individuals with C/C (n = 73) and C/T (n = 66) genotype: 88% (71%-91%) and 86% (50%-91%), respectively, for 5-methyltetrahydrofolic acid (5CH(3)THF) vs 12% (9% 29%) and 14% (9%-51%) for forms other than 5-methyltetrahydrofolic acid (non 5CH(3)THF). Individuals with T/T (n = 32) genotype had 58% (22%-87%) 5CH(3)THF vs 42% (13%-78%) non-5CH(3)THF. Compared with microbiologic assay results, LC-MS/MS (r = 0.94) and BR (r = 0.87) results were significantly lower (-10% and -45%, respectively); however, these differences were concentration dependent and also genotype dependent for the BR assay (-48% for C/C+C/T and -31% for T/T). The microbiologic assay completely recovered [mean (SD)] folates added to a whole blood hemolysate, except for tetrahydrofolic acid (THF) [46.4% (8.1%)]. The BR assay under-recovered 5CH(3)THF [51% (4.1%)] and 5-formyltetrahydrofolic acid [18% (0.1%)], and over-recovered THF [152% (19%)]. CONCLUSION: MTHFR C677T polymorphism influences the folate pattern in whole blood. The agreement between total folate by LC-MS/MS and microbiologic assay, independent of the MTHFR genotype, allows the use of one regression equation. Because BR results are genotype dependent, different regression equations should be used. PMID- 18160727 TI - Unbound bilirubin: a better predictor of kernicterus? PMID- 18160730 TI - Use of single-value protein compensation of the Jaffe creatinine assay contributes to clinically significant inaccuracy in results. PMID- 18160731 TI - Stability of thiopurine metabolites: a potential analytical bias. PMID- 18160732 TI - Drug monitoring and toxicology (DMT). PMID- 18160733 TI - False-positive rates for the qualitative analysis of urine benzodiazepines and metabolites with the reformulated Abbott Multigent reagents. PMID- 18160734 TI - Are clinical laboratories prepared for accurate testing of 25-hydroxy vitamin D? PMID- 18160735 TI - Biological variation of myeloperoxidase. PMID- 18160736 TI - Mismeasure of C-type natriuretic peptide. PMID- 18160737 TI - Hexaprimer amplification refractory mutation system PCR for simultaneous single tube genotyping of 2 close polymorphisms. PMID- 18160739 TI - Human apolipoprotein C-I expression in mice impairs learning and memory functions. AB - The H2 allele of APOC1, giving rise to increased gene expression of apolipoprotein C-I (apoC-I), is in genetic disequilibrium with the APOE4 allele and may provide a major risk factor for Alzheimer's disease (AD). We found that apoC-I protein is present in astrocytes and endothelial cells within hippocampal regions in both human control and AD brains. Interestingly, apoC-I colocalized with beta-amyloid (Abeta) in plaques in AD brains, and in vitro experiments revealed that aggregation of Abeta was delayed in the presence of apoC-I. Moreover, apoC-I was found to exacerbate the soluble Abeta oligomer-induced neuronal death. To establish a potential role for apoC-I in cognitive functions, we used human (h) APOC1(+/0) transgenic mice that express APOC1 mRNA throughout their brains and apoC-I protein in astrocytes and endothelial cells. The hAPOC1(+/0) mice displayed impaired hippocampal-dependent learning and memory functions compared with their wild-type littermates, as judged from their performance in the object recognition task (P = 0.012) and in the Morris water maze task (P = 0.010). ApoC-I may affect learning as a result of its inhibitory properties toward apoE-dependent lipid metabolism. However, no differences in brain mRNA or protein levels of endogenous apoE were detected between transgenic and wild-type mice. In conclusion, human apoC-I expression impairs cognitive functions in mice independent of apoE expression, which supports the potential of a modulatory role for apoC-I during the development of AD. PMID- 18160740 TI - Cryopreservation of sperm after chemotherapy. PMID- 18160741 TI - Molecular profiling of the human testis reveals stringent pathway-specific regulation of RNA expression following gonadotropin suppression and progestogen treatment. AB - Gonadotropin withdrawal induces changes in gene expression in all 3 major cell types of the testis. Knowledge of the genes affected, in both the presence and absence of additional progestogen, will give insight into the regulation of human testicular function and aid development of novel contraceptive methods. We have undertaken a whole-genome analysis of RNA expression in testicular biopsies from normal men and after 4 weeks of gonadotropin suppression induced by gonadotropin releasing hormone antagonist plus testosterone administration sufficient to cause marked suppression of spermatogenesis. Microarray analysis shows that interindividual variability is markedly low, and the response to treatment is focused on a small subset of genes particularly related to pathways in steroidogenesis and cholesterol biosynthesis or metabolism, the Leydig cell gene INSL3, and genes involved in early meiosis or Sertoli-germ cell junctions. These changes in expression were confirmed by quantitative reverse transcriptase polymerase chain reaction. No major changes in gene expression were identified in men additionally treated with a progestogen, although FLJ35767, an expressed sequence tag that is expressed in the germ cell compartment, did show a small but significant additional effect of progestogen. Overall, the results of this investigation disclose a remarkably stringent regulation of testicular gene expression, revealing the genes most sensitive to gonadotropin withdrawal, and might reflect the most labile pathways in the regulation of testicular function. PMID- 18160742 TI - A health perspective of indoor air pollution. PMID- 18160743 TI - External proficiency testing programme & quality of haematology tests in India. PMID- 18160744 TI - Cognitive functioning in schizophrenia : its relevance to rehabilitation. PMID- 18160745 TI - An update on newer beta-lactamases. AB - The resistance to beta-lactam antibiotics is an increasing problem worldwide and beta lactamases production is the most common mechanism of drug resistance. Both global and Indian figures showed a marked increase in the number of beta lactamases producing organisms. These enzymes extended spectrum beta-lactamases (ESBLs) are numerous and continuous mutation has led to the development of enzymes having expanded substrate profile. To date, there are more than 130 TEM type and more than 50 sulphydryl variable (SHV) type beta-lactamases found in Gram negative bacilli. ESBL producing Enterobacteriaceae are, as a rule, resistant to all cephalosporins and extended spectrum penicillins including the monobactam, aztreonam, while resistance to trimethoprim - sulphamethaxazole and aminoglycosides is frequently co-transferred on the same plasmid. Many ESBL producing organisms also express Amp C beta-lactamases. Amp C- beta-lactamases are clinically significant, as these confer resistance to cephalosporins in the oxyimino group, 7 alpha-methoxy cephalosporins, and are poorly inhibited by clavulanic acid. Carbepenems are the drugs of choice for the treatment of infections caused by ESBL producing organisms but carbapenemases (MBLs) have emerged and have spread from Pseudomonas aeruginosa to Enterobacteriaceae. The routine clinical microbiology laboratories should employ simple methods to recognize these enzymes using various substrates and inhibitors. These organisms may lead to therapeutic dead ends. Presently, the therapy relies on beta-lactam/ beta-lactamases inhibitor combinations, carbepenems and piperacillin - tazobactam plus aminoglycoside combination. Proper infection control practices and barrier precautions are essential to contain the organisms producing beta-lactamases. PMID- 18160746 TI - Impact of external haematology proficiency testing programme on quality of laboratories. AB - BACKGROUND & OBJECTIVE: A reliable and reproducible report from a laboratory needs internal quality control within the laboratory and participation in external proficiency testing programmes (EPTP). This study conducted at the Department of Haematology, All India Institute of Medical Sciences (AIIMS), New Delhi, which has been conducting an EPTP since 1992, was undertaken to assess the efficacy of this programme in improving the performance of participating laboratories in reporting test samples sent for Hb, total leucocyte count (TLC), reticulocyte count and assessment of peripheral blood smear (PBS). METHODS: The samples were prepared in our laboratory according to the International Standards Organization (ISO) guidelines. The performance of individual laboratories was assessed using robust Z score, which is an indicator of acceptability of the test result. RESULTS: An improvement in the overall percentage of laboratories with acceptable reports was seen during the study period. It has increased from 38,40,40 per cent in 1992 to 85, 90,94.7 per cent in 2006 for Hb, TLC, reticulocyte count, respectively. However, the results for peripheral smear assessment improved only marginally. INTERPRETATION & CONCLUSION: The external haematology proficiency testing programme run by our department for Hb, TLC, reticulocyte count, and peripheral blood smear assessment, has helped in improving the reporting standards of these parameters in Indian laboratories. PMID- 18160747 TI - Cognitive functions in stable schizophrenia & euthymic state of bipolar disorder. AB - BACKGROUND & OBJECTIVES: Some cognitive deficits in schizophrenia and bipolar disorders persist after the subsidence of active symptoms. We carried out this study to assess and compare the cognitive functioning of patients with stable schizophrenia and bipolar disorder. METHODS: Fifteen each of stable maintained schizophrenic patients and euthymic bipolar-I patients attending outpatient clinic in a tertiary care psychiatric hospital in north India were included in the study as also equal number of age and education matched control subjects. Cognitive assessments were done using Wisconsin's Card Sorting Test (WCST), Spatial Working Memory Test (SWMT) and Continuous Performance Test (CPT). RESULTS: Stable schizophrenia patients performed poorly on all the neurocognitive parameters as compared to both controls and bipolar euthymic patients. Euthymic bipolar patients showed significant difference on executive functions with normal controls. Patterns of cognitive disturbances in tasks of executive function are similar in both groups but are quantitatively more marked in schizophrenia. INTERPRETATION & CONCLUSIONS: Our results showed that stable schizophrenia patients performed significantly worse on cognitive measures than patients of euthymic bipolar disorder which was consistent with their poorer functional outcome. The results further indicated that stable schizophrenia and euthymic bipolar disorders may be distinguished qualitatively in neuropsychological terms with different profiles of cognitive impairment. PMID- 18160748 TI - Effect of pretreatment with chromium picolinate on haematological parameters during dengue virus infection in mice. AB - BACKGROUND & OBJECTIVE: Dengue virus (DV) has caused severe epidemics of dengue fever (DF) and dengue haemorrhagic fever (DHF) and is endemic all over India. We have earlier reported that exposure of mice to hexavalent chromium [Cr(VI)] compounds increased the severity of dengue virus infection. Trivalent chromium picolinate (CrP) is used worldwide as micronutrient and nutritional supplement. The present study was therefore, carried out to investigate the effects of CrP on various haematological parameters during DV infection of mice. METHODS: The Swiss Albino smice were inoculated with dengue virus (1000 LD50, intracerebrally) and fed with chromium picolinate (CrP) in drinking water (100 and 250 mg/l) for 24 wk. Peripheral blood leucocytes and other haematological parameters, and spleens were studied on days 4 and 8 after virus inoculations and the findings were compared with those given only CrP and the normal control age matched mice. RESULTS: CrP in drinking water for 24 wk had no significant effects on peripheral blood cells of mice. On the other hand, there was significant decrease in different haematological parameters following inoculation of normal mice with DV. In CrP fed mice the effects of DV infection were abolished on most of the haematological parameters. INTERPRETATION & CONCLUSION: The findings of present study showed that the adverse effects of DV infection, specially on platelets and leucocytes, were abrogated by pretreatment of mice with CrP. The therapeutic utility of CrP in viral infections including dengue needs to be studied in depth. PMID- 18160749 TI - A new strategy for elimination of kala-azar from rural Bihar. AB - BACKGROUND & OBJECTIVE: Bihar State of India has been an endemic State for kala azar. There has been many phases of DDT sprays for vector control. An outbreak of kala-azar occurred in Goanpura, 6 km from Patna, Bihar, in 2003. We undertook this study with a new approach of kala-azar elimination in this village with priority to treatment of cases followed by supplementation with supervised DDT spray for vector control. METHODS: This study included a camp approach to collect patients at in the camp, screening of patients with rK-39, transporting the patients to the hospital of Balaji Utthan Sansthan, Patna, Bihar, confirming the diagnosis by demonstration of Leishman-Donovan (LD) bodies in splenic aspirates after proper clinical and pathological investigations, and treatment with amphotericin B (Fungizone) at a dose of 1mg/kg body wt for 20 days. If parasites persisted after 20 days, five more infusions were given. The State Government was persuaded to do supervised DDT spray as per the guidelines. All patients were followed up for 6 months for any relapse, and the village of Goanpura for 3 years for occurrence of any new case. RESULTS: A camp was held in that village on September 11, 2003 after due publicity. A total of 368 patients having different ailments, attended the camp; 25 patients were screened with rk-39 for kala-azar and 21 patients gave positive results. All 21 patients were shifted to Balaji Utthan Sansthan for treatment. After thorough clinical examination and pathological tests splenic aspiration was done. All 21 patients were positive for LD bodies. One patient died and the remaining 20 completed full course of treatment and were cured. No patients relapsed within six months of follow up. Two rounds of supervised DDT spray were done. No new case occurred in the village during three years of follow up. No sandfly was detected in the village during the years of follow up. INTERPRETATION & CONCLUSION: Camp strategy to collect patients at one place, screening of patients with rk-39, transporting rk-39 positive patients to the hospital, treatment with an effective drug amphotericin B with no incidence of unresponsiveness and relapsed minimized transmission of the disease; only two rounds of supervised intensive spray of DDT eliminated sandflies from the village. This new cost effective approach in which treatment of patients was done with an effective drug followed by supervised DDT spray may be adopted for elimination of kala-azar from Bihar. PMID- 18160750 TI - Sputum conversion at the end of intensive phase of Category-1 regimen in the treatment of pulmonary tuberculosis patients with diabetes mellitus or HIV infection: An analysis of risk factors. AB - BACKGROUND & OBJECTIVE: New smear-positive pulmonary tuberculosis (PTB) patients in the Revised National Tuberculosis Control Programme (RNTCP) are treated with a 6-month short-course chemotherapy (SCC) regimen irrespective of co-morbid conditions. We undertook this retrospective analysis to compare sputum conversion rates (smear, culture) at the end of intensive phase (IP) of Category-1 regimen among patients admitted to concurrent controlled clinical trials: pulmonary tuberculosis alone (PTB) or with type 2 diabetes mellitus (DM-TB) or HIV infection (HIV-TB), and to identify the risk factors influencing sputum conversion. METHODS: In this retrospective analysis sputum conversion rates at the end of intensive phase (IP) in three concurrent studies undertaken among PTB, DM-TB and HIV-TB patients, during 1998 - 2002 at the Tuberculosis Research Centre (TRC), Chennai, were compared. Sputum smears were examined by fluorescent microscopy. HIV infected patients did not receive anti-retroviral treatment (ART). Patients with DM were treated with oral hypoglycaemic drugs or insulin (sc). RESULTS: The study population included 98, 92 and 88 patients in the PTB, DM-TB and HIV-TB studies. At the end of IP the smear conversion (58, 61, and 62%) and culture conversion (86, 88 and 92%) rates were similar in the three groups respectively. The variables associated with lack of sputum smear or culture conversion were age >45 yr, higher pre-treatment smear and culture grading, and extent of the radiographic involvement. INTERPRETATION & CONCLUSION: Our findings confirm that the current policy of the control programme to treat all pulmonary TB patients with or with out co-morbid conditions with Category-I regimen appears to be appropriate. PMID- 18160751 TI - Production & purification of a fibrinolytic enzyme (thrombinase) from Bacillus sphaericus. AB - BACKGROUND & OBJECTIVE: Treatment of thromboembolic vascular disease has relied on anticoagulants. However, recognition that lysis of preformed fibrin could be accomplished in vivo by a process involving the conversion of inactive plasminogen to active plasmin enzyme led to an alternative enzyme-based approach. The drugs used for this therapy are called the fibrinolytic enzymes. In this study we attempted the production, purification and characterization of fibrinolytic enzyme from Bacillus sphaericus. METHODS: The seed was prepared in nutrient yeast salt medium (NYSM) in shake flask and organism was produced in 100 l pilot fermentor. Biomass was separated by centrifugation and crude protein was prepared by ammonium sulphate precipitation. Purification was done by ion exchange chromatography using Q sepharose followed by gel filtration chromatography using Sephacryl S- 300. Molecular weight was determined through HPLC. Fibrinolytic activity was assayed by fibrin plate method. RESULTS: The production method yielded 64 mg/l of the crude enzyme and after purification it was 6.3 mg/l. The molecular weight of the compound was 18.6 kDa. INTERPRETATION & CONCLUSION: The enzyme exhibited similar fibrinolytic activity as that of streptokinase, on fibrin plates that were devoid of plasminogen, suggesting that its fibrinolytic action is independent of plasminogen and it is not a plasminogen activator. PMID- 18160752 TI - SCA 1, SCA 2 & SCA 3/MJD mutations in ataxia syndromes in southern India. AB - BACKGROUND & OBJECTIVE: Spinocerebellar ataxias (SCAs) are often caused by expansions of CTG/ CAG trinucleotide repeat in the genome. Expansions at the SCA1, 2 and 3 loci are the most frequent, but differences in their relative proportion in regions occur across the world. We carried out this study to assess the occurrence of SCA1, 2 and 3, at a tertiary neuro-psychiatric center in Bangalore, Karnataka. METHODS: Probands (N=318) who were diagnosed to have an ataxia syndrome (progressive degenerative ataxia of unknown cause) attending the clinical services of the National Institute of Mental Health and Neuro Sciences (NIMHANS), Bangalore, were evaluated over a period of three years. Standard protocols were used for both clinical and molecular diagnosis. RESULTS: Genotyping established that SCA1, 2 and 3 accounted for more than one third of the ataxia cases seen in the clinic. In the cases with established family history and autosomal dominant inheritance SCA1 was most prevalent followed by SCA2 and SCA3. INTERPRETATION & CONCLUSION: Our findings suggested SCA1 rather than SCA2 to be the more common mutation in southern India. Large numbers of SCA3 probands were also identified. Differences in prevalence of these syndromes within India need to be explored further for founder effects, correlations with phenotype, and patterns of outcome. Family history was not apparent in almost a fifth of those tested positive, highlighting the value of testing even in the absence of family history. Molecular testing should be extended to cover the other forms of ataxia, of which a large number are now known. Combined efforts to confirm the presence of these less common forms, as well as family studies to detect novel mutations, are necessary in this context in India. PMID- 18160753 TI - Effect of vitamin C on blood glucose, serum lipids & serum insulin in type 2 diabetes patients. AB - BACKGROUND & OBJECTIVE: Diabetes mellitus is one of the most common metabolic disorders that causes micro- and macro-vascular complications. Because of additive effects of hyperglycaemia and hyperlipidaemia for cardiovascular diseases, lipid abnormalities should be evaluated in diabetes. As vitamin C is known for its beneficial effects on serum lipids and glycated haemoglobin (HbA1c), we evaluated the effect of different doses of vitamin C on blood glucose, serum lipids and serum insulin in individuals with type 2 diabetes mellitus. METHODS: A total of 84 patients with type 2 diabetes referred to Yazd Diabetes Research Center, Iran, were included in the study. They received randomly either 500 mg or 1000 mg daily of vitamin C for six weeks. Fasting blood sugar (FBS), triglyceride (TG), total cholesterol (TC), low and high density lipoprotein (LDL, HDL), glycated haemoglobin HbA(Ic) and serum insulin were measured before and after vitamin C consumption and the results were analyzed. RESULTS: A significant decrease in FBS, TG, LDL, HbA1c and serum insulin was seen in the group supplemented with 1000 mg vitamin C. The dose of 500 mg vitamin C, however, did not produce any significant change in any of the parameters studied. INTERPRETATION & CONCLUSION: Our results indicate that daily consumption of 1000 mg supplementary vitamin C may be beneficial in decreasing blood glucose and lipids in patients with type 2 diabetes and thus reducing the risk of complications. PMID- 18160754 TI - Prevalence of goitre in 6 -12 years school-going children of Panchmahal district in Gujarat, India. AB - BACKGROUND & OBJECTIVE: Iodine deficiency disorders (IDDs) are one of the worldwide public health problems of today. IDDs are endemic in various districts of Gujarat. We undertook this survey in Panchmahal district of Gujarat to study the prevalence of goiter in a sample of school children, to asses the level of iodine in salt samples, and to determine median urinary iodine concentration in a sample of children. METHODS: In a cross-sectional study, 15,900 school-going children in the age group of 6-12 yr were studied from 10 talukas. A total of 15 clusters were selected by cluster sampling method. Goitre was assessed in the children and urine and salt samples were studied for iodine concentration. RESULTS: The overall prevalence of goitre was 20.5 per cent among the children examined. The median iodine concentration was 70 mg/l indicating mild iodine deficiency; also 61 per cent of the urine samples examined showed iodine insufficiency. Iodine content was found to be adequate in 54.3 per cent of salt samples. INTERPRETATION & CONCLUSION: Our study showed that goitre is still an important public health problem in the district. This calls for identification of factors to strengthen NIDDCP and the need to reimpose ban on sale of non-iodized salt in Gujarat. PMID- 18160755 TI - Effect of NR-ANX-C (a polyherbal formulation) on haloperidol induced catalepsy in albino mice. AB - BACKGROUND & OBJECTIVE: Use of typical antipsychotics like haloperidol in treatment of schizophrenia is associated with a high incidence of extrapyramidal side effects. In rodents, administration of haloperidol leads to the development of a behavioural state called catalepsy, in which the animal is not able to correct an externally imposed posture. In the present study we evaluated the anticataleptic efficacy of NR-ANX-C, a polyherbal formulation containing bioactives of Withania somnifera, Ocimum sanctum, Camellia sinensis, triphala and shilajit in haloperidol induced catalepsy in mice. METHODS: Five groups (n = 6) of male albino mice were used in the study. Catalepsy was induced by ip administration of haloperidol (1mg/kg). The degree of catalepsy (cataleptic score) was measured as the time the animal maintained an imposed posture. We compared the anticataleptic efficacy of NR-ANX-C (10, 25 and 50 mg/kg) with scopolamine (1 mg/kg). The superoxide dismutase (SOD) level in brain tissue was also estimated to correlate the levels of oxidative stress and degree of catalepsy in the animal. RESULTS: Significant (P<0.01) reduction in the cataleptic scores was observed in all NR-ANX-C treated groups and maximum reduction was observed in the NR-ANX-C (25 mg/kg) treated group. Significant (P<0.05) reduction in SOD activity was observed in NR-ANX-C (25 and 50 mg/kg) treated groups and maximum reduction was observed in NR-ANX-C (25mg/kg) treated group. INTERPRETATION & CONCLUSION: In our study, maximum reduction in cataleptic score was observed in NR-ANX-C (25 mg/kg) treated group. The maximum reduction in SOD activity was also observed in the same group. These findings suggest a possible involvement of the antioxidant potential of NRANX- C in alleviating haloperidol induced catalepsy. PMID- 18160756 TI - Chronic kidney disease in India. PMID- 18160757 TI - Detection of ESBLs using third & fourth generation cephalosporins in double disc synergy test. PMID- 18160758 TI - Agar gel disk as an alternative to paper disk diffusion antimicrobial susceptibility test. PMID- 18160759 TI - High incidence of actinobacillus actinomycetemcomitans infection in acute coronary syndrome. AB - Recent epidemiological studies suggest that periodontitis is an important risk factor for coronary heart disease (CHD). The aim of this study was to evaluate the association between periodontitis and CHD, particularly acute coronary syndrome (ACS), focusing on microbiological and immunological features. Twenty eight CHD patients, 15 with ACS and 13 with chronic CHD, were included in this study. Coronary angiography, periodontal examination, and dental radiography were performed in all patients. Subgingival plaque, saliva, and blood samples were analyzed for the periodontopathogens Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythensis, Treponema denticola, and Prevotella intermedia using polymerase chain reaction. Specific serum antibody titers to the 5 periodontal pathogens were determined by enzyme-linked immunosorbent assay. It was found that 33% of the ACS patients (5/15) harbored A. actinomycetemcomitans in oral samples, whereas no A. actinomycetemcomitans (0/13) was found in the chronic CHD patients (P < 0.05). Furthermore, ACS patients showed significantly higher serum IgG titers to A. actinomycetemcomitans (P < 0.05) compared with chronic CHD. More tooth loss and alveolar bone loss were noted in ACS patients than in chronic CHD patients, although the differences were not statistically significant. Periodontal pathogens, particularly A. actinomycetemcomitans, may play a role in the development of ACS. PMID- 18160760 TI - Effect of early versus late in-hospital initiation of statin therapy on the clinical outcomes of patients with acute coronary syndrome. AB - Statins provide effective secondary prevention of cardiovascular events after acute coronary syndrome (ACS). Current guidelines recommend that statins be initiated in ACS patients before hospital discharge. In this retrospective study, we investigated the influence of early compared with late in-hospital initiation of statin therapy on the clinical outcomes of ACS patients. Two hundred and ten ACS patients who had no history of statin treatment before hospitalization were enrolled. The patients were divided into early (statin treatment initiated < or = 2 days after admission) and late (statin treatment initiated > 2 days after admission and before discharge) statin groups. We examined the association between early statin use and clinical outcomes in these patients using Cox proportional hazards models. Four months after discharge, event-free survival was 85% in the early statin group and 79% in the late statin group. In multivariate analyses, Killip IV classification and abnormal renal function were independent predictors of the composite endpoint of cardiovascular death, recurrent myocardial infarction, angina requiring rehospitalization, revascularization, and stroke. Early statin treatment had no significant influence (hazard ratio: 1.11; 95% confidence interval: 0.54 to 2.25; P = 0.78) on the composite endpoint 4 months after discharge. At 12 months after discharge, early statin treatment still had no significant influence on the composite endpoint (hazard ratio: 1.53; 95% confidence interval: 0.83 to 2.81; P = 0.17). We concluded that ensuring all eligible ACS patients begin statin therapy during their hospital stay is more important than emphasizing immediate statin use after admission. PMID- 18160761 TI - Comparison of long-term clinical outcome after sirolimus-eluting stent implantation in patients with and without hemodialysis. AB - Poor clinical outcomes for hemodialysis (HD) patients compared to non-HD patients after coronary intervention have been reported. Although coronary intervention using sirolimus-eluting stents (SESs) might be expected to reduce restenosis in HD patients, little is known about the efficacy of the SESs. The purpose of the present study was to compare the clinical and angiographic outcomes of HD patients with non-HD patients after SES implantation. The study population consisted of 170 consecutive patients (234 lesions) who had undergone successful coronary SES implantation. The patients were classified into 2 groups, an HD group (18 patients, 27 lesions) and a non-HD group (152 patients, 207 lesions). The incidence of any clinical event was significantly higher in the HD group than in the non-HD group (50.0% versus 12.5%, P < 0.0001). Target lesion revascularization was necessary in 6 patients (33.3%) in the HD group and in 7 patients (4.6%) in the non-HD group (P < 0.0001). The Cox proportional-hazards regression model on cardiac events identified HD patients (P = 0.0301, hazard ratio = 2.704) as an explanatory factor. Moreover, the Cox proportional-hazards regression model on target lesion revascularization identified HD (P = 0.0004, hazard ratio = 6.921) and in-stent re-stenosis lesion (P = 0.0293, hazard ratio = 3.323) as explanatory factors. The present study suggests that compared with non HD patients, HD patients with coronary artery disease treated by SESs have a poorer clinical outcome. PMID- 18160762 TI - Development of a signal-averaged vector-projected 187-channel high-resolution electrocardiogram for the evaluation of the spatial location of high-frequency potentials and abnormal ventricular repolarization. AB - Noninvasive risk stratification is important for screening for lethal arrhythmia. We developed a 187-channel signal-averaged vector-projected high-resolution electrocardiograph (187-ch SAVP-ECG) for detecting abnormalities in the spatial location of ventricular high-frequency late potentials (HFLPs) and ventricular repolarization. The subjects consisted of 30 normal controls (CONTROL) and 13 patients with HFLPs (6 with myocardial infarction [MI], 6 with cardiomyopathy, and 1 with Brugada syndrome). The modified X, Y, Z-lead ECG and the synthesized signals from vector-projected 187-channel ECGs were amplified and passed through a digital filter. We calculated the integration of the HFLPs area between QRS(end) and 30 ms before QRS(end). The integrated HFLPs map was superimposed on the corrected recovery time (RTc) and Tpeak-end dispersion maps composed by 187 ch SAVP-ECG. All patients received an examination by 64-channel magnetocardiography (64-ch MCG) on the same day. The spatial distribution of HFLPs by the 187-ch SAVP-ECG map was in agreement with the location of increased RT dispersion in MI. The spatial distribution of HFLPs in DCM demonstrated a wide variety of patterns. Interestingly, the spatial distribution of HFLPs in cases with ARVC was located at around a right ventricular outflow region. The spatial distribution of HFLPs by 187-ch SAVP-ECG was in agreement with those determined by 64-ch MCG. The 187-ch SAVP-ECG might be useful for evaluating the spatial distribution of nonuniform conduction and ventricular repolarization heterogeneity. PMID- 18160763 TI - A new therapeutic strategy for hypertrophic nonobstructive cardiomyopathy in humans. A randomized and prospective study with an Angiotensin II receptor blocker. AB - A randomized and prospective study was designed to prove the efficacy of angiotensin II receptor blockers (ARB) in the amelioration of myocardial impairment in hypertrophic nonobstructive cardiomyopathy (HNCM). Nineteen consecutive patients with HNCM were randomly assigned to two groups and then underwent cine magnetic resonance evaluation of left ventricular mass (LVM) twice just before and after one year of observation. In the ARB group, 50 mg of losartan potassium was administered once daily during the observation period. The ratio of LVM after the observation period over that before the period was blindly compared between the two groups to estimate morphologically the ameliorative effect of ARB. In the ARB group, LVM was 203 +/- 47 cm(3) before the treatment period and 190 +/- 55 cm(3) after the period and the ratio of the final LVM over the initial LVM was 0.93 +/- 0.10. In the non-ARB group the initial and final LVM values were 177 +/- 48 cm(3) and 179 +/- 45 cm(3), and the ratio of the final LVM over the initial LVM was 1.02 +/- 0.07. The ratio of the final LVM over the initial LVM in the ARB group was significantly smaller (P = 0.03) than that in the non-ARB group. The smaller ratio in the ARB group strongly indicates that ARB ameliorated the natural course of HNCM during the one year observation period. Thus, this is the first demonstration of the therapeutic efficacy of ARB in human HNCM. PMID- 18160764 TI - Tea catechin consumption reduces circulating oxidized low-density lipoprotein. AB - It has been reported that green tea consumption reduces the risk of coronary artery disease and cardiac events. Catechin is a major constituent of Japanese green tea and an antioxidant. Lipids and oxidization of low-density lipoprotein cholesterol (LDL-C) play important roles in atherosclerosis. Therefore, we evaluated the effect of catechin intake on the lipid profile and plasma oxidized LDL. The study population consisted of 40 healthy adult volunteers (10 men, 30 women). Catechin was extracted from green tea leaves. The subjects were randomly divided into two groups, a catechin group (n = 29) and a control group (n = 11). In the catechin group, catechin (500 mg: equivalent to 6 or 7 cups of green tea) was administered orally. Venous blood samples were obtained before eating a meal at the start and after 4 weeks without any lifestyle modification. Plasma oxidized LDL assay was performed with a sandwich-type enzyme immunoassay using anti-oxidized phosphatidylcholine monoclonal antibody. The baseline lipid profiles and tea consumptions were similar between the two groups. Plasma oxidized LDL was significantly decreased after catechin administration (from 9.56 +/- 9.2 to 7.76 +/- 7.7 U/mL, P = 0.005), while plasma LDL-C, triglyceride, and HDL-C concentrations did not change. Catechin decreased the plasma oxidized LDL concentration without significant change in plasma LDL concentration. The mechanism of the beneficial effects of green tea on coronary artery disease might result from a decrease in plasma oxidized LDL. PMID- 18160765 TI - The relationship between obesity and transforming growth factor beta on renal damage in essential hypertension. AB - The aim of this study was to evaluate the effect of obesity on renal functions and the possible relationship between TGF-beta1 and obesity in hypertensive patients. Seventy newly diagnosed, hypertensive patients (male/female 36/34, aged 45.0 +/- 8.0 years) and 30 (male/female 17/13, aged 41.8 +/- 7.7 years) normotensive controls were included. Patients in both groups were analyzed for serum levels of glucose, creatinine, uric acid, lipids, and TGF-beta1. A 24-hour urine sample was also obtained; creatinine clearance rate and urinary albumin excretion (UEA) were investigated. TGF-beta1 levels were significantly higher (40.7 +/- 13.6 versus 34.2 +/- 12.1 pg/mL, P = 0.02), and creatinine clearance was significantly lower in patients compared with controls (98.9 +/- 25.5 versus 124.5 +/- 23.1 mL/min. per. 1.73 m(2), P = 0.001). Serum TGF-beta1 levels (45.2 +/- 14 versis 38.0 +/- 12.8 pg/mL, P = 0.03), creatinine clearance rates (109.8 29.9 versus 93.0 +/- 20.8 mL/min. per. 1.73 m(2), P = 0.001), and urinary albumin excretion (55.7 +/- 62.0 versus 12.7 +/- 12.6 mg/24 h, P = 0.002) were higher in obese hypertensive patients than in nonobese patients. In hypertensive patients, TGF-beta1 levels correlated with body mass index (r = 0.296, P = 0.01) and creatinine clearance (r = 0.238, P = 0.04). The results suggest that increased body mass index is associated with increased creatinine clearance, urinary albumin excretion, and TGF-beta1 levels in essential hypertension. In addition, TGF-beta1 is positively correlated with body mass index and creatinine clearance in patients with essential hypertension. PMID- 18160766 TI - Factors for successful weaning from a percutaneous cardiopulmonary support system (PCPS) in patients with low cardiac output syndrome after cardiovascular surgery. AB - The objective of this study was to analyze the early predictive factors for successful weaning from a percutaneous cardiopulmonary support system (PCPS) in patients with low cardiac output syndrome after cardiovascular surgery. A total of 938 patients underwent cardiovascular surgery with cardiopulmonary bypass (CPB) from January 1991 to September 2006 at Gunma University Hospital. Of these 938 patients, 13 (1.4%) required PCPS to maintain hemodynamics within 48 hours after surgery. The mean age of the 13 patients was 66 years (range, 45 to 86 years). Nine patients underwent open-heart surgery, 3 repair of a thoracic aortic aneurysm, and 1 a pericardiectomy. The patients were divided into 2 groups; group A (n = 4) who were removed from PCPS and group B (n = 9) who were not removed from PCPS. The conditions during the operation and after PCPS support were compared between the 2 groups. The mean age was higher, and operation time, CPB time, and aortic cross-clamping time were significantly (P < 0.05) longer in patients with PCPS than in those without PCPS. The mean PCPS time in all 13 patients was 190 +/- 122 hours. The mean age was higher, and CPB time and the aortic cross-clamping time were longer in group B than in group A (NS). The mean duration of PCPS support was significantly (P < 0.05) shorter in group A than in group B (117 +/- 42 hours versus 235 +/- 136 hours). PCPS flow in group A could be reduced from 48 hours after PCPS induction. However, PCPC flow in group B could not be reduced, and there were significant (P < 0.05) differences in PCPS flow at 72 and 96 hours after starting PCPS. Significant (P < 0.05) differences in the absolute values of the APACHE II score, serum lactate levels, administered epinephrine dose, and levels of total bilirubin (T-Bil), serum creatinine (sCr), and lactate dehydrogenase (LDH) were found between the 2 groups within 96 hours after PCPS induction. In addition, there were significant (P < 0.05) differences in the rate of change compared with the baseline control value obtained prior to PCPS use in PCPS flow, APACHE II score, and levels of T-Bil, sCr, and LDH within 96 hours after PCPS induction. Significant differences in the rate of change of sCr and LDH were found, especially from the early phase after PCPS use, compared with other parameters. In the patients removed from PCPS, PCPS flow could be reduced within 48 hours after commencement of PCPS. Improvements in the APACHE II score and biochemical variables within 96 hours appear to be reliable prognostic factors for PCPS patients. PMID- 18160767 TI - Increased functional importance of the Na,Ca-exchanger in contracting failing human myocardium but unchanged activity in isolated vesicles. AB - The present study aimed to investigate the hypothesis that the function of the Na,Ca-exchanger (NCX) is of higher importance for contractility and Ca(2+) homeostasis in left ventricle from terminally failing than from nonfailing human hearts. The effect of decreasing extracellular [Na](e) (140 to 25 mmol/L) on force of contraction in isolated left ventricular papillary muscle strips was studied as a reflection of NCX function in multicellular preparations (terminally failing, DCM, dilated cardiomyopathy, NYHA IV, n = 13; nonfailing, NF, donor hearts, n = 10). Decreasing [Na](e) has previously been shown to increase contractility in vitro secondary to a decreased Ca(2+)-extrusion by the NCX. In addition, the NCX activity was measured as Na(+)-dependent (45)Ca(2+)-uptake into isolated myocardial vesicles as a function of time and Ca(2+)-concentration (DCM n = 8, NF n = 8). Decreasing [Na](e) enhanced the contractility of papillary muscle strips in both DCM and NF, but the contractility of DCM was increased at smaller reductions of [Na](e) than NF. The NCX activity in isolated myocardial vesicles was unchanged as a function of time (T(1/2): DCM 2.4 +/- 0.3 s versus NF 2.5 +/- 0.3 s) and as a function of Ca(2+) (DCM 0.99 +/- 0.08 versus NF 0.96 +/- 0.07 nmol/mg protein x 3 s, K(1/2): DCM 39.2 microM versus NF 38.3 microM). These results demonstrate a higher sensitivity of the failing human myocardium towards Na,Ca-exchanger mediated positive inotropic effects, suggesting a higher significance of the Na,Ca-exchanger for the extrusion of Ca(2+)-ions in intact failing versus nonfailing human myocardium. Since the activity and the Ca (2+) affinity of the Na,Ca-exchanger in isolated vesicles was unchanged, we propose that alterations in Ca(2+)-and Na(+)-homeostasis (due to impaired function of the sarcoplasmic reticulum and the Na(+), K(+)-ATPase) or the prolonged action potential are the reason for this observation. PMID- 18160768 TI - Late multiple stent fractures following deployment of sirolimus-eluting stents for diffuse right coronary artery stenosis. AB - We report a case of late multiple stent fractures following the deployment of sirolimus-eluting stents (SESs) for diffuse right coronary artery (RCA) stenosis. A 44-year-old male with hyperlipidemia was referred to our hospital for acute myocardial infarction (AMI). Percutaneous coronary intervention (PCI) was performed for total occlusion of the proximal segment of the RCA, and 5 SESs were consecutively implanted for long, diffuse stenotic lesions. A follow-up coronary angiography (CAG) performed 8 months later revealed strut fractures in the middle of all the stents, except the most proximal one. Multislice computed tomography confirmed the locations of the fractured struts inside of the stents. PMID- 18160769 TI - Noninvasive detection of Takotsubo [corrected] cardiomyopathy using multi detector row computed tomography. AB - We present a 60-year-old woman with chest pain preceded by emotional stress. Trans-thoracic echocardiography demonstrated localized left ventricular hypokinesis around the apical area. Multi-detector row computed tomography (MDCT) revealed no significant stenosis in the coronary arteries, which comfirmed Takotsubo-like cardiomyopathy. We show here the usefulness of MDCT for the noninvasive differentiation of Takotsubo cardiomyopathy from acute coronary syndrome. PMID- 18160770 TI - Coumestrol decreases intestinal alkaline phosphatase activity in post-delivery mice but does not affect vitamin D receptor and calcium channels in post-delivery and neonatal mice. AB - In this study, we investigated the effects of administration of coumestrol during pregnancy on calcium (Ca) metabolism in post-delivery maternal and neonatal mice. From 6.5 to 16.5 days post coitus (dpc), pregnant females were administered daily doses of coumestrol (200 microg/kg body weight/day). One day after parturition, blood samples and the kidneys, liver, jejunum and duodenum were obtained from each of maternal mouse, and blood samples and the kidneys and liver were obtained from neonatal mice. Coumestrol did not have any significant effect on the Ca and inorganic phosphorus concentrations in the sera of the maternal and neonatal mice. No notable effects of coumestrol were observed in relation to Vitamin D receptor expression in the maternal and neonatal mice by immunohistochemical analysis. Coumestrol did not affect the Vitamin D receptor and epithelial calcium channel and 2 mRNA levels in any of the organs investigated. Enzyme histochemical analysis showed that coumestrol decreased intestinal alkaline phosphatase activity in the maternal jejunum and duodenum. In the duodenum, coumestrol decreased expression of intestinal alkaline phosphatase, c-fos and vascular endothelial growth factor at the mRNA level. However, we did not observe any significant effects of coumestrol on the expression of these genes. In conclusion, coumestrol decreased intestinal alkaline phosphatase activity in the small intestines of maternal mice at the level used in the present study, and the mechanisms underlying this effect are different for the jejunum and duodenum. PMID- 18160771 TI - Determination of the estrogenic activity of wild phytoestrogen-rich Pueraria mirifica by MCF-7 proliferation assay. AB - The aim of this study was to evaluate the estrogenic activity of tuberous samples of wild, phytoestrogen-rich Pueraria mirifica collected from 28 out of 76 provinces of Thailand by MCF-7 proliferation assay. The plant extracts were administered to MCF-7, ER alpha positive human mammary adenocarcinoma cell cultures, for 3 days at dosages of 0.1, 1, 10, 100 and 1,000 microg/ml and were compared with 17 beta-estradiol at concentrations of 10(-12)-10(-6) M. The mean P. mirifica population at 1 mug/ml exhibited significant proliferation. Two plant samples exhibited levels of proliferation in MCF-7 that were similar to 17beta estradiol. The mean P. mirifica populations at 100 and 1,000 microg/ml exhibited significant cytotoxicity in MCF-7. Analysis of the estrogenic activity of puerarin, representative of major isoflavonoids in P. mirifica tubers, revealed proliferation in MCF-7 only at the highest dose (10(-6) M) that was 10(2)-10(5) times less active than 17 beta-estradiol. Puerarin and 17 beta-estradiol at concentration of 10(-12)-10(-6) M exhibited no cytotoxicity in MCF-7. PMID- 18160772 TI - hTERT-immortalized cells useful for analyzing effects of low-dose-rate radiation on human cells. AB - To establish immortal human cells, we introduced the cDNA of the human telomere reverse transcriptase (hTERT) gene into skin fibroblast cells obtained from normal and ataxia telangiectasia (AT) individuals of Japanese origin. hTERT immortalized cells retained their original characteristics and radiosensitivity except for immortalization, suggesting that these cells might be useful for analyzing the effects of radiation on human cells.hTERT-immortalized cells from a normal individual showed a greater resistance after low-dose-rate irradiation than after high-dose-rate irradiation. In contrast, cells from AT patients irradiated at a low-dose rate showed virtually the same survival as those irradiated at a high-dose rate. In hTERT-immortalized normal cells, the genetic effects of low-dose-rate radiation were quantitatively and qualitatively less severe than those of high-dose-rate radiation. In hTERT-immortalized AT cells, some fraction of DNA damage such as DNA double-strand breaks might not be repaired, and AT cells were severely affected by low-dose-rate radiation. The activation of ataxia telangiectasia mutated (ATM) protein was confirmed during low-dose-rate radiation, and may play an important role in repair of DNA damage induced by low-dose-rate radiation. This paper reviews briefly the current research at our laboratory. The hTERT-immortalized cells may be useful in determining the effects of low-dose and low-dose-rate radiation on human cells. PMID- 18160774 TI - DNA degradation during maturation of erythrocytes - molecular cytogenetic characterization of Howell-Jolly bodies. AB - Howell-Jolly bodies (HJBs) are small DNA-containing inclusions of erythrocytes and are often present after splenectomy. The genetic composition of HJBs is unknown at present. We isolated individual erythrocytes that had inclusion bodies from five splenectomized patients and performed DNA amplification using degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) with subsequent reverse painting on normal male metaphase spreads. We also measured the sizes of HJBs in erythrocytes from a splenectomized patient using an inverted microscope. Two-dimensional positions of HJBs were projected onto a virtual erythrocyte. The average size of HJBs was 0.73 +/- 0.17 microm (range 0.4-1.1 microm). Inside the erythrocyte the HJBs were found to be equally distributed. Small HJBs contained DNA from one or two centromeres and larger HJBs contained DNA from up to eight different centromeres. Centromeric DNA from chromosomes 1/5, 7, 8, and 18 was most frequently observed. Signals from the centromeric regions of chromosomes 3, 4, 9, and 10 were not observed. Signals from euchromatic regions were detected in a few cases. We hypothesize that in addition to enucleation and nucleus fragmentation DNA degradation during maturation of erythrocytes preferentially eliminates euchromatic DNA. Similarities between these processes and those described for embryonic stem cells suggest that most stem cells are able to degrade DNA in a genetically controlled manner. PMID- 18160775 TI - The molecular basis of the folate-sensitive fragile site FRA11A at 11q13. AB - We report on the molecular basis of the rare, folate-sensitive fragile site FRA11A in chromosome band 11q13 in a family with cytogenetic expression. Five individuals express the fragile site and one was mentally retarded. Expansion of a polymorphic CGG-repeat located at the 5' end of the C11orf80 gene causes FRA11A. The CGG-repeat elongation coincides with hypermethylation of the adjacent CpG island and subsequent transcriptional silencing of the C11orf80 gene. This gene has no homology with known genes. A relationship between cytogenetic expression of the fragile site and the mental handicap seems unlikely, as FRA11A was found in a mentally retarded patient as well as in phenotypically normal carriers from the same family. However, incomplete penetrance cannot be entirely excluded. PMID- 18160776 TI - Molecular studies of segmental aneusomy: FISHing for the atypical cry in del(5)(p15.3). AB - We report a newborn male with multiple congenital anomalies including growth retardation, hypotonia, dysmorphic facies, widely-spaced nipples, micropenis, cryptorchidism, optic nerve hypoplasia, heart disease, and a striking, high pitched cry. Chromosome analysis revealed de novo partial trisomy 11q due to a der(5)t(5;11)(p15.3;q22). Fluorescence in situ hybridization (FISH) showed loss of the 5p telomere signal on the der(5) chromosome, indicating the infant has partial monosomy 5p in addition to partial trisomy 11q. Among cases involving trisomy 11q, an unusual cry has only been documented in the presence of a der(5)t(5p;11q). This apparent dependence of the abnormal cry on monosomy 5p suggested the same genetic mechanism that occurs in Cri du chat syndrome (CDCS) may be responsible for the atypical cry in der(5)t(5p;11q) individuals. Neither a commercial CDCS probe (LSI D5S23, D5S721) nor a series of BAC clones encompassing distal regions implicated in the CDCS-associated cat-cry were deleted in our patient. These results suggest a second cry-modifying locus maps telomeric to BAC RP11-94J21 in band 5p15.33. This locus may not only cause the abnormal cry in individuals with a der(5)t(5p;11q) but could also contribute to the phenotypic variability and discordant mapping studies observed for CDCS. PMID- 18160777 TI - Microdeletions in 9p21.3 induce false negative results in CDKN2A FISH analysis of Ewing sarcoma. AB - Deletion of the CDKN2A locus at 9p21.3 has been reported to be a poor prognostic sign in the Ewing sarcoma family of tumours. In clinical applications CDKN2A deletion is primarily detected using fluorescent in situ hybridisation (FISH) with a commercial probe, size approximately 190 kb. Due to limitations in resolution, FISH analysis may fail to detect microdeletions smaller than 190 kb. In the present study, we performed 44K array comparative genomic hybridisation (CGH) on eleven Ewing sarcoma cell lines and 26 tissue samples in order to define the sizes of 9p21.3 deletions. Microarray CGH analysis revealed 9p21.3 deletions encompassing the CDKN2A locus in eight cell lines (73%) and in six tumours (23%). In four cases (two cell lines and two tissue samples) the deletion was less than 190 kb in size. In one cell line sample, we detected a microdeletion of approximately 58 kb in 9p21.3 harbouring the CDKN2A locus. We confirmed this result using 244K microarray CGH and TaqMan quantitative RT-PCR analysis and further performed FISH analysis on this cell line sample. Here, we show that CDKN2A FISH analysis can give false negative results in cases with small microdeletions. Our results suggest that new and more accurate FISH methods should be developed for detection of deletions in the CDKN2A locus. PMID- 18160778 TI - Discontinuities and unsynapsed regions in meiotic chromosomes have a trans effect on meiotic recombination of some chromosomes in human males. AB - During meiosis, homologous chromosome pairing and synapsis are essential for subsequent meiotic recombination (crossing-over). Discontinuous regions (gaps) and unsynapsed regions (splits) were most frequently observed in the heterochromatic regions of bivalent synaptonemal complex (SC) 9, and we have previously demonstrated that gaps and splits significantly altered the distribution of MLH1 recombination foci on SC 9. Here, immunofluorescence techniques (using antibodies against SC proteins and the crossover-associated MLH1 protein) were combined with a centromere-specific fluorescence in situ hybridization technique that allows identification of every individual chromosome. The effect of gaps/splits on meiotic recombination patterns in autosomes other than chromosome 9 during the pachytene stage of meiotic prophase was then examined in 6,026 bivalents from 262 pachytene cells from three human males. In 64 analyzed cells with a gapped SC 9, the frequency of MLH1 foci in SCs 5 and 10 and in SC arms 10q, 11p and 16q was decreased compared to 168 analyzed cells with a normally-synapsed SC 9 (controls). In 24 analyzed cells with splits in SC 9, there was a significant reduction in MLH1 focus frequency for SC 5q and the whole SC5 bivalent. The positioning of MLH1 foci on other SCs in cells with gapped/split SC 9 was not altered. These studies suggest that gaps and splits not only have a cis effect, but may also have a trans effect on meiotic recombination in humans. PMID- 18160779 TI - PLAG1 activation in lipoblastoma coinciding with low-level amplification of a derivative chromosome 8 with a deletion del(8)(q13q21.2). AB - Lipoblastoma is a benign uncommon soft-tissue-tumor resembling fetal adipose tissue affecting mainly children under three years of age. In lipoblastoma, the typical cytogenetic changes are clonal rearrangements involving chromosomal region 8q11-->q13. The oncogene PLAG1 (pleomorphic adenoma gene 1) is located within this chromosomal region on band 8q12. Recent reports have demonstrated that in lipoblastoma, the PLAG1 gene is activated by 'promoter-swapping'. Herein, we demonstrate that in lipoblastoma, the PLAG1 gene may also be activated by low level amplification. We report on a lipoblastoma with the karyotype 48 approximately 50,XX,del(8)(q13q21.2),+del(8)(q13q21.2)x4[cp12]. Subsequent FISH analysis on uncultured tumor cells confirmed this result and demonstrated a low level amplification of the chromosomal region 8pter-->8q13 and 8q21.2-->8qter. A partial monosomy was seen for the chromosomal region 8q13-->8q21.2. No other gains or losses were observed by CGH analysis. RT-PCR analysis showed that the PLAG1 gene is activated in the tumor sample of the lipoblastoma analyzed, in contrast to normal fatty tissue without PLAG1 expression. In conclusion, our results demonstrate that low-level amplification is a further mechanism of PLAG1 activation in lipoblastomas. PMID- 18160780 TI - Oligonucleotide array comparative genomic hybridization refines the structure of 8p23.1, 17q12 and 20q13.2 amplifications in gastric carcinomas. AB - Oligonucleotide array comparative genomic hybridization (aCGH) was applied on fifteen gastric cancer (GCA) samples to reveal information of DNA copy number changes at an exon-level resolution. Twelve of the samples represented the intestinal (IGCA) and three the diffuse (DGCA) type of GCA. The samples had previously been assessed for genetic stability by microsatellite analysis and categorized into microsatellite phenotypes according to the type of alterations. As compared to our previous results obtained using cDNA platforms, the oligonucleotide platforms revealed more aberrations per sample (0-45 vs. 0-22). A total of 22 amplifications were detected by the oligonucleotide arrays. Ten of the amplicons had also been detected on the cDNA platform, but five of them spanned only one or a few cDNA clones, thus resembling apparent outliers. Two tumors showed five or more amplifications by oligonucleotide aCGH, suggesting the presence of an amplifier phenotype. The amplifications occurred irrespective of the microsatellite phenotypes. None of the DGCA tumors showed more than one aberration, whereas the IGCA tumors showed several aberrations. The increased resolution of the oligonucleotide arrays enabled the detection of amplicon boundaries at gene level, allowing, e.g., the determination of the 17q12 core amplicon and interstitial losses within the 8p23.1-->p22 and 20q13.2-->q13.1 amplifications. Previously no losses have been reported within amplified regions in GCA. In addition to novel amplified regions, the oligonucleotide array results describe novel targets for amplicons at 8p11 (SFRP1), 11p12 (LRRC4C), and 19q13.2 (CEACAM6). PMID- 18160781 TI - Gene copy number analysis in malignant pleural mesothelioma using oligonucleotide array CGH. AB - Conventional cytogenetic analyses and comparative genomic hybridization have revealed a complex and even chaotic nature of chromosomal aberrations in pleural malignant mesothelioma (MM). We set out to describe the complex gene copy number changes and screen for novel genetic aberrations using a high-density oligonucleotide microarray platform for comparative genomic hybridization (aCGH) of a series of 26 well-characterized MM tumor samples. The number of copy number changes varied from zero to 40 per sample. Gene copy number losses predominated over gains, and the most frequent region of loss was 9p21.3 (17/26 cases), the locus of CDKN2A and CDKN2B, both known to be commonly lost in MM. The most recurrent minimal regions of losses were 1p31.1--> p13.2, 3p22.1-->p14.2, 6q22.1, 9p21.3, 13cen-->q14.12, 14q22.1-->qter, and 22qcen-->q12.3. Previously unreported gains included 9p13.3, 7p22.3-->p22.2, 12q13.3, and 17q21.32-->qter. The results suggest that gene copy number losses are a major mechanism of MM carcinogenesis and reveal a recurrent pattern of copy number changes in MM. PMID- 18160782 TI - Double minutes (dmin) and homogeneously staining regions (hsr) in myeloid disorders: a new case suggesting that dmin form hsr in vivo. AB - We describe a case of an 87-year-old woman with myelodysplastic syndrome (MDS) in transformation. At presentation, the patient suffered from refractory anemia with excess of blasts (RAEB). The karyotype showed a clone with 3 to >20 double minutes (dmin) that were MYC-positive and, in addition, a cryptic deletion of MYC on one of the chromosome 8 homologs. The dmin varied both in size and in fluorescence intensity for the MYC probe following FISH. After six months, the disease had progressed to refractory anemia with excess of blasts in transformation (RAEBT). The corresponding karyotype showed a second cell line with two markers that appeared like rings and, again, a cryptic MYC deletion on one of the chromosome 8 homologs. The markers contained centromeres and amplified MYC sequences. The deletion of MYC on one of the chromosome 8 homologs and the variable sizes and intensities of the dmin supports an episome model for dmin formation. Chronologically, the MYC-positive dmin appeared first, followed by the appearance of MYC-positive marker chromosomes, suggesting that the dmin may have evolved to form homogeneously staining regions (hsr) in vivo. PMID- 18160783 TI - Telomere dynamics and genome stability in the human pancreatic tumor cell line MIAPaCa-2. AB - Telomeres are specialized structures found at the ends of eukaryotic chromosomes serving as guardians of genome stability. In normal cells telomeres shorten with each cell division, but immortal cells undergoing multiple divisions constantly have to maintain telomere lengths above a critical level. This is accomplished either through expression of telomerase or the alternative recombination pathway (ALT). In the present study, we analyzed telomere dynamics of the telomerase positive human pancreatic tumor cell line MIAPaCa-2. The cells demonstrated genomic instability with a high frequency of chromosomal aberrations resulting in differences between individual karyotypes within the same cell population. The telomeres were short when compared with normal human fibroblasts, and about 39% of the chromosome ends did not have detectable telomere repeats as demonstrated by PNA-FISH. In many cases telomere signals were missing even when sister chromatids were strongly labeled. In addition, we used an internal PNA probe specific for the X chromosome, present in a single copy in these cells, in order to follow telomere dynamics on individual chromatids. High heterogeneity in telomere signals among individual X chromosomes as well as between their sister chromatids suggested sudden and stochastic loss or gain of telomere repeats. Such constant genomic instability often results in apoptosis and death of a fraction of cells present in the culture at all times. We discuss possible molecular mechanisms that may explain this observed telomere heterogeneity and possible adaptive repair mechanisms by which these cells maintain their chromosomes in order to survive such extreme and permanent genomic instability. PMID- 18160784 TI - Alternative splicing of exon 3b gives rise to ODF2 and Cenexin. AB - ODF2 was first identified as the major component of the sperm tail outer dense fibers. Additionally, ODF2 is a critical component of the mature centriole of the animal centrosome where it locates to the distal appendages. Moreover, generation of primary cilia strictly depends on ODF2. The mature centriole is characterized further by recruitment of Cenexin. Albeit highly similar in sequence the relationship between ODF2 and Cenexin has not been investigated. We demonstrate here that ODF2 and Cenexin are alternative splice products by identifying a novel exon 3b encoding Cenexin specific amino acids. Even though ODF2 is the main isoform in testicular tissue RT-PCR analyses revealed that isoforms are not restricted to specific tissues. PMID- 18160785 TI - The putative peroxisomal gene Pxt1 is exclusively expressed in the testis. AB - Genes reported to be crucial for spermatogenesis are often exclusively expressed in the testis. We have identified a novel male germ cell-specific expressed gene named peroxisomal testis specific 1 (Pxt1) with expression starting at the spermatocyte stage during mouse spermatogenesis. The putative amino acid sequence encoded by the cDNA of the Pxt1 gene contains a conserved Asn-His-Leu (NHL)-motif at its C-terminal end, which is characteristic for peroxisomal proteins. Pxt1 EGFP fusion protein is co-localized with known peroxisomal marker proteins in transfected NIH3T3 cells. In addition, we could demonstrate that the peroxisomal targeting signal NHL is functional and responsible for the correct subcellular localization of the Pxt1-EGFP fusion protein. In male germ cells peroxisomes were reported only in spermatogonia. The Pxt1 gene is so far the first gene coding for a putative peroxisomal protein which is expressed in later steps of spermatogenesis, namely in pachytene spermatocytes. PMID- 18160786 TI - Emergence of complex rearrangements at translocation breakpoints in a transgenic mouse; implications for mechanisms involved in the formation of chromosome rearrangements. AB - Cryptic complex rearrangements as a result of a reciprocal chromosome translocation have been characterised in a transgenic mouse strain. Analysis of the breakpoint junctions in our previous studies showed that the ada transgene was integrated at the breakpoint forming a fusion gene with Golga3 (Mea2). In this study, further detailed analysis around the translocation junctions revealed that the surrounding regions were composed of 13 fragments of defined transgenic chromosome origins over approximately 1.9-Mb areas. Exactly the same cluster structure of these 13 breakpoint fragments already existed in the second generation of the transgenic mice. Our results show that this highly complex rearrangement has been conserved as the incipient form without any additional changes for 18 years up to the present generation, suggesting simultaneous occurrence of multiple events in the founder mouse. PMID- 18160787 TI - Similarities in the chromosomal distribution of AG and AC repeats within and between Drosophila, human and barley chromosomes. AB - Two simple sequence repeats (SSRs), AG and AC, were mapped directly in the metaphase chromosomes of man and barley (Hordeum vulgare L.), and in the metaphase and polytene chromosomes of Drosophila melanogaster. To this end, synthetic oligonucleotides corresponding to (AG)(12) and (AC)(8) were labelled by the random primer technique and used as probes in fluorescent in situ hybridisation (FISH) under high stringency and strict washing conditions. The distribution and intensity of the signals for the repeat sequences were found to be characteristic of the chromosomes and genomes of the three species analysed. The AC repeat sites were uniformly dispersed along the euchromatic segments of all three genomes; in fact, they were largely excluded from the heterochromatin. The Drosophila genome showed a high density of AC sequences on the X chromosome in both mitotic and polytene nuclei. In contrast, the AG repeats were associated with the euchromatic regions of the polytene chromosomes (and in high density on the X chromosome), but were only seen in specific heterochromatic regions in the mitotic chromosomes of all three species. In Drosophila, the AG repeats were exclusively distributed on the tips of the Y chromosome and near the centromere on both arms of chromosome 2. In barley and man, AG repeats were associated with the centromeres (of all chromosomes) and nucleolar organizer regions, respectively. The conserved chromosome distribution of AC within and between these three phylogenetically distant species, and the association of AG in specific chromosome regions with structural or functional properties, suggests that long clusters of these repeats may have some, as yet unknown, role. PMID- 18160788 TI - A radiation hybrid map of river buffalo (Bubalus bubalis) chromosome 1 (BBU1). AB - The largest chromosome in the river buffalo karyotype, BBU1, is a submetacentric chromosome with reported homology between BBU1q and bovine chromosome 1 and between BBU1p and BTA27. We present the first radiation hybrid map of this chromosome containing 69 cattle derived markers including 48 coding genes, 17 microsatellites and four ESTs distributed in two linkage groups spanning a total length of 1330.1 cR(5000). The RH map was constructed based on analysis of a recently developed river buffalo-hamster whole genome radiation hybrid (BBURH(5000)) panel. The retention frequency of individual markers across the panel ranged from 17.8 to 52.2%. With few exceptions, the order of markers within linkage groups is identical to the order established for corresponding cattle RH maps. The BBU1 map provides a starting point for comparison of gene order rearrangements between river buffalo chromosome 1 and its bovine homologs. PMID- 18160789 TI - Localization of single-copy sequences on chicken synaptonemal complex spreads using fluorescence in situ hybridization (FISH). AB - Synaptonemal complex (SC) spreads from bird oocytes and spermatocytes show the complete chromosome complement and can be observed at the light microscope using immunostaining of the proteins that compose the lateral elements. To investigate the use of avian SC spreads as substrates for fluorescent in situ hybridization (FISH) in combination with immunostaining, we applied two single-copy sequences to chicken oocyte spreads. Signals for both target sequences were consistently observed on the short arm of bivalent 1 in a large number of nuclei. Based on previous data about the size of chromosome 1 and from measurements on probed SC spreads, an estimate of the physical distance in Mb between each sequence and the telomere was calculated. The crossover frequencies along SC 1 obtained by immunolocalization of MLH1 foci during pachytene were used to calculate the distances in cM to the target sequences and to compare this cytogenetic SC map with the consensus linkage map for GGA1. The combination of SC-FISH and immunostaining could be generally applied to obtain high-resolution mapping of single-copy sequences in birds and, coupled with MLH1 crossover maps, it could be a reliable approach to obtain genetic distances between markers to test the genetic linkage maps generated from molecular markers. PMID- 18160790 TI - An integrated and comparative genetic map of the turkey genome. AB - An integrated genetic linkage map was developed for the turkey (Meleagris gallopavo) that combines the genetic markers from the three previous mapping efforts. The UMN integrated map includes 613 loci arranged into 41 linkage groups. An additional 105 markers are tentatively placed within linkage groups based on two-point LOD scores and 19 markers remain unlinked. A total of 210 previously unmapped markers has been added to the UMN turkey genetic map. Markers from each of the 20 linkage groups identified in the Roslin map and the 22 linkage groups of the Nte map are incorporated into the new integrated map. Overall map distance contained within the 41 linkage groups is 3,365 cM (sex averaged) with the largest linkage group (94 loci) measuring 533.1 cM. Average marker interval for the map was 7.86 cM. Sequences of markers included in the new map were compared to the chicken genome sequence by 'BLASTN'. Significant similarity scores were obtained for 95.6% of the turkey sequences encompassing an estimated 91% of the chicken genome. A physical map of the chicken genome based on positions of the turkey sequences was built and 36 of the 41 turkey linkage groups were aligned with the physical map, five linkage groups remain unassigned. Given the close similarities between the turkey and chicken genomes, the chicken genome sequence could serve as a scaffold for a genome sequencing effort in the turkey. PMID- 18160791 TI - Two rare aneutriploids in the unisexual Ambystoma (Amphibia, Caudata) identified by GISH indicating two different types of meiotic errors. AB - We report two types of aneutriploids in unisexual salamanders Ambystomalaterale 2jeffersonianum (LJJ) and Ambystoma 2 laterale-jeffersonianum (LLJ). One karyotype has 3n = 42: L27 (L8-); J15 (J8p+), and we suggest that it was induced by homoeologous pairing after premeiotic endomitosis followed by an unequal L8;J8 segregation. The second karyotype has 3n = 43: L14 (L10q); J29 (J12+), which can be explained by meiotic nondisjunction followed by unbalanced segregation. These two rare aneutriploids demonstrate two different types of meiotic errors that might help to explain the high mortality observed in this complex. Case one also indicates that contemporary intergenomic exchanges and homoeologous recombinations may occur after a premeiotic chromosome doubling event. Our study provides additional evidence for the extremely flexible reproduction of unisexual Ambystoma. PMID- 18160792 TI - A haploid-diploid-triploid mosaic of the Amazon molly, Poecilia formosa. AB - We report the finding of the first haploid-diploid-triploid mosaic fish from the family Poeciliidae. The animal was derived from a laboratory cross of a female F1 hybrid of Poecilia mexicana and P. latipinna with a male from an ornamental strain derived from P. mexicana and P. sphenops (Black molly). It was identified because of its unusual pigmentation pattern and molecular methods (flow cytometry, NOR staining) confirmed its mosaic genotype. The mode of mosaic formation and the possible importance for poeciliid fish evolution are discussed. PMID- 18160793 TI - Histone H3 lysine 9 acetylation pattern suggests that X and B chromosomes are silenced during entire male meiosis in a grasshopper. AB - The facultative heterochromatic X chromosome in leptotene spermatocytes of the grasshopper Eyprepocnemis plorans showed marked hypoacetylation for lysine 9 in the H3 histone (H3-K9) with no sign of histone H2AX phosphorylation. Since H3-K9 hypoacetylation precedes the meiotic appearance of phosphorylated H2AX (gamma H2AX), which marks the beginning of recombinational DNA double-strand breaks (DSBs), it seems that meiotic sex-chromosome inactivation (MSCI) in this grasshopper occurs prior to the beginning of recombination and hence synapsis (which in this species begins later than recombination). In addition, all constitutively heterochromatic chromosome regions harbouring a 180-bp tandem repeat DNA and rDNA (B chromosomes and pericentromeric regions of A chromosomes) were H3-K9 hypoacetylated at early leptotene even though they will synapse at subsequent stages. This also suggests that meiotic silencing in this grasshopper might be independent of synapsis. The H3-K9 hypoacetylated state of facultative and constitutive heterochromatin persisted during subsequent meiotic stages and was even apparent in round spermatids. Finally, the fact that B chromosomes are differentially hypoacetylated in testis and embryo interphase cells suggests that they might be silenced early in development and remain this way for most (or all) life-cycle stages. PMID- 18160794 TI - Physical mapping of rDNA and satDNA in A and B chromosomes of the grasshopper Eyprepocnemis plorans from a Greek population. AB - Adult males and females of the grasshopper Eyprepocnemis plorans from a Greek population were analysed by C-banding, silver impregnation and double FISH for two DNA probes, i.e. ribosomal DNA (rDNA) and a 180-bp tandem repeat DNA (satDNA). This population shows characteristics of rDNA location in A chromosomes that are intermediate between those previously reported for eastern (Caucasus) and western (Spain and Morocco) populations. The four rDNA clusters revealed by FISH in chromosomes X, 9, 10 and 11 in Greek specimens imply two more than the two observed in chromosomes 9 and 11 in the Caucasus, but less than the 12 observed in all chromosomes in Morocco. Remarkably, the X chromosome bears one of the new rDNA locations in Greece with respect to the Caucasus, but it appears to be inactive, in contrast to X chromosomes in western populations, which are usually active. B chromosomes were very frequent in the Greek population, and three variants differing in size were observed, all of these being largely composed of rDNA, with the exception of a small pericentromeric satDNA cluster. The high B frequency suggests that B chromosomes in this population might behave parasitically, in resemblance to Bs in western populations. PMID- 18160795 TI - Molecular characterisation and chromosomal localisation of a telomere-like repetitive DNA sequence highly enriched in the C genome of Brassica. AB - The aim of this work was to find C genome specific repetitive DNA sequences able to differentiate the homeologous A (B. rapa) and C (B. oleracea) genomes of Brassica, in order to assist in the physical identification of B. napus chromosomes. A repetitive sequence (pBo1.6) highly enriched in the C genome of Brassica was cloned from B. oleracea and its chromosomal organisation was investigated through fluorescent in situ hybridisation (FISH) in B. oleracea (2n = 18, CC), B. rapa (2n = 20, AA) and B. napus (2n = 38, AACC) genomes. The sequence was 203 bp long with a GC content of 48.3%. It showed up to 89% sequence identity with telomere-like DNA from many plant species. This repeat was clearly underrepresented in the A genome and the in situ hybridisation showed its B. oleracea specificity at the chromosomal level. Sequence pBo1.6 was localised at interstitial and/or telomeric/subtelomeric regions of all chromosomes from B. oleracea, whereas in B. rapa no signal was detected in most of the cells. In B. napus 18 to 24 chromosomes hybridised with pBo1.6. The discovery of a sequence highly enriched in the C genome of Brassica opens the opportunity for detailed studies regarding the subsequent evolution of DNA sequences in polyploid genomes. Moreover, pBo1.6 may be useful for the determination of the chromosomal location of transgenic DNA in genetically modified oilseed rape. PMID- 18160796 TI - A neocentromere derived from a supernumerary marker deleted from the long arm of chromosome 6. AB - Parental chromosome studies were referred to us after initial finding of a balanced translocation involving chromosomes 4 and 15 in their phenotypically abnormal male child (cytogenetic analysis was done at another laboratory). In addition to the same 4;15 translocation, the father also had an interstitial deletion of the long arm of one chromosome 6 and a marker chromosome. In this article, we report a neocentromere on this marker, which was determined to be composed of chromosome 6 material by FISH. The child's karyotype was re interpreted to be unbalanced due to the presence of the abnormal chromosome 6, but without the marker. The clinical phenotype associated with the interstitial deletion of chromosome 6 is also reported. PMID- 18160797 TI - Two independent chromosomal rearrangements, a very small (550 kb) duplication of the 7q subtelomeric region and an atypical 17q11.2 (NF1) microdeletion, in a girl with neurofibromatosis. AB - Most patients with neurofibromatosis (NF1) are endowed with heterozygous mutations in the NF1 gene. Approximately 5% show an interstitial deletion of chromosome 17q11.2 (including NF1) and in most cases also a more severe phenotype. Here we report on a 7-year-old girl with classical NF1 signs, and in addition mild overgrowth (97th percentile), relatively low OFC (10th-25th percentile), facial dysmorphy, hoarse voice, and developmental delay. FISH analysis revealed a 17q11.2 microdeletion as well as an unbalanced 7p;13q translocation leading to trisomy of the 7q36.3 subtelomeric region. The patient's mother and grandmother who were phenotypically normal carried the same unbalanced translocation. The 17q11.2 microdeletion had arisen de novo. Array comparative genomic hybridization (CGH) demonstrated gain of a 550-kb segment from 7qter and loss of 2.5 Mb from 17q11.2 (an atypical NF1 microdeletion). We conclude that the patient's phenotype is caused by the atypical NF1 deletion, whereas 7q36.3 trisomy represents a subtelomeric copy number variation without phenotypic consequences. To our knowledge this is the first report that a duplication of the subtelomeric region of chromosome 7q containing functional genes (FAM62B, WDR60, and VIPR2) can be tolerated without phenotypic consequences. The 17q11.2 microdeletion (containing nine more genes than the common NF1 microdeletions) and the 7qter duplication were not accompanied by unexpected clinical features. Most likely the 7qter trisomy and the 17q11.2 microdeletion coincide by chance in our patient. PMID- 18160798 TI - Trisomy of medial 15q as result of an analphoid supernumerary ring chromosome detected by CGH and FISH. AB - We report a 21-year-old patient with a de novo mosaic, analphoid ring of chromosome 15q22.2-->q24.1. The clinical features of this patient are mild and include tall stature, obesity, striae distensae in the hypogastrium, malocclusion and bilateral gynecomastia with scarce glandular tissue. M-FISH and FISH using a chromosome 15 painting probe indicated that the ring is of chromosome 15 origin. Further CGH analysis and FISH with the PML locus-specific probe demonstrated that the extra material derived from the medial part of the long arm of chromosome 15, including two bands, q22 and q23. Additionally, FISH with BAC probes specific for 15q allowed for a localization of the breakpoints at 15q22.2 and 15q24.1, distal to clones RP11-30M4 and RP11-500O23 respectively. We discuss the relationship between the patient's genotype and phenotype comparing it to reported cases of trisomy of medial 15q. PMID- 18160799 TI - Molecular determinants of bat wing development. AB - The specialization of the forelimb into a wing allowed bats to become the only mammals to achieve powered flight. Recent studies in developmental biology have begun to elucidate the molecular mechanisms behind elements of this important morphological transformation. Specifically, researchers have identified molecular changes contributing to: the formation of the bat wing membrane, the elongation of skeletal elements of the bat wing and the reduction of the bat ulna. The general picture emerging from this research is that small changes in the expression of genes critical to many aspects of development have driven large changes in bat wing morphology. Thus, bats can be added to the growing list of groups in which expression changes in key developmental genes have been linked to the evolution of morphological innovations (e.g. early bilaterians, cetaceans, insects). PMID- 18160800 TI - Growth and development of two species of bats in a shared maternity roost. AB - Skeletogenesis was studied in two species of bats, Myotis austroriparius (southeastern brown bat) and Tadarida brasiliensis (Brazilian free-tailed bat), occupying a maternity roost in central Florida. These bats often use distinct maternity roost environments, so this provided an opportunity to examine differential patterns of long bone growth while fetuses and newborn developed under similar environmental conditions. Some differences in the timing of onset of osteogenesis were revealed in the bats, indicating that some elements of the hindlimb develop relatively more rapidly in T. brasiliensis than in M. austroriparius. Some variance was also noted, with similarity to other species previously studied by others, in the exact timing and elongation of both long bones, as well as carpal and tarsal bones. In contrast to many elements of the long appendicular skeleton of developing Mus musculus, the bats all exhibit relatively precocial patterns of osteogenesis during which cartilaginous precursors are replaced by bone tissue. The relative advanced timing of osteogenesis in select hindlimb bones of T. brasiliensis may account for its relatively low neonatal mortality compared to M. austroriparius newborn in the same roost. PMID- 18160801 TI - Forelimb versus hindlimb skeletal development in the big brown bat, Eptesicus fuscus: functional divergence is reflected in chondrocytic performance in Autopodial growth plates. AB - The morphology of the chiropteran forelimb demonstrates musculoskeletal specializations for powered flight essentially unique among mammals, including extreme elongation of the distal skeletal elements. Recent studies have focused primarily on the relative timing and levels of gene expression during early stages of endochondral ossification in the chiropteran embryo for clues to the molecular basis of the evolutionary origins of flight in these species. The goal of the current study was to examine how elongation of skeletal elements of the forelimb autopod is achieved through a combination of cellular proliferation, cellular enlargement and matrix synthesis during a period of rapid postnatal growth in Eptesicus fuscus. Quantitative analyses were done of multiple performance parameters of growth plate chondrocytes during all phases of the differentiation cascade. Fourteen autopodial growth plates from the forelimb and hindlimb of one individual, as well as the proximal tibial growth plate, were collected and analyzed. Significant differences were seen in all performance parameters examined. Particularly striking were the differences between growth plates of the manus and pes in the size of the pool of chondrocytes in all cellular zones and rates of turnover of terminal cells. The magnitude of hypertrophy of chondrocytes in growth plates of the manus in E. fuscus far exceeded what has been reported previously in any species, even in rapidly elongating rodent long bones. Volume changes approaching x70 and height changes of 50-60 mum/cell (paralleling the direction of growth) occurred after proliferation in the most rapidly growing growth plates. The data demonstrate that final differences in lengths of homologous skeletal elements in the autopod of the forelimb and hindlimb of this species result not just from an initiating factor early in development, but from continued quantitative differences in chondrocytic performance during postnatal bone elongation as measured by multiple kinetic-based parameters. PMID- 18160802 TI - Postnatal bone elongation of the manus versus pes: analysis of the chondrocytic differentiation cascade in Mus musculus and Eptesicus fuscus. AB - Bones elongate postnatally by endochondral ossification as cells of the cartilaginous growth plate undergo a differentiation cascade of proliferation, cellular hypertrophy and matrix synthesis. Interspecific comparisons of homologous bones elongating at different rates has been a useful approach for studying the dynamics of this process. The purpose of this study was to measure quantitative stereological parameters of growth plates of the third digit of the manus and pes of the laboratory mouse, and make comparisons to chondrocytic performance parameters in the homologous bones of the big brown bat, Eptesicus fuscus, where extremely rapid postnatal elongation of bones of the manus is associated with skeletal modifications for powered flight. Measurements were made across all zones of forelimb and hindlimb autopod growth plates by dividing each growth plate into strata of equal height (from thirteen 200-mum-high strata in the metacarpus to five 40-mum-high strata in phalangeal bones of the pes). Results indicate that all chondrocytic performance parameters known to quantitatively contribute to the elongation potential of a growth plate change together. A significant finding was that in growth plates of the chiropteran manus, final hypertrophic cell size and shape were achieved early in the zone of hypertrophy, indicating that interstitial expansion of the growth plate resulting from the incremental chondrocytic height increase in the direction of elongation was completed soon after the transition from the cessation of proliferation to the initiation of hypertrophy. This is unlike what has been reported in most mammalian growth plates previously analyzed, but is the situation in the proximal tibial growth plate of rapidly growing frogs and precocial birds. This suggests that a similar adaptation for stabilization of a rapidly elongating bone has evolved independently in three widely separated groups that have in common rapid growth in limbs to be used for early active, powered locomotion. PMID- 18160803 TI - Biomechanics of the bat limb skeleton: scaling, material properties and mechanics. AB - BACKGROUND/AIMS: Wing skeletons of bats are uniquely specialized for flight, reflecting both evolutionary history and the need to maintain structural integrity while generating aerodynamic forces. METHODS: We analyzed the anatomical structure of bat wing skeletons in the context of scaling patterns relative to other mammals, material properties and the mechanical function of the wing bones during flight. RESULTS: Compared with nonvolant mammals, the bones of the bat forelimb are elongated, even after correcting for shared phylogenetic history. Bats have consistently larger-diameter bones in the forelimb than do nonvolant mammals but significantly narrower hindlimb bones. Mineralization in the cortical bone of wings is lower than in the long bones of other adult mammals, with a proximodistal gradient of decreasing mineralization. The distal phalanges have only a small amount of mineralized tissue underlying the articular cartilage. Loads required to elicit a 10% length deflection in the wing bones of Glossophaga soricina varied approximately 50-fold along the wing and flexural stiffness nearly 200-fold. Commensurate with low mineralization and flexural stiffness, bat bones experience extraordinarily high bending strains during flight. CONCLUSION: Bat limb skeletons share features with other mammals and possess specialized characteristics, mostly related to the mechanical demands of flight. PMID- 18160805 TI - Impact and management of skin toxicity associated with anti-epidermal growth factor receptor therapy: survey results. AB - Biologic agents targeting the epidermal growth factor receptor (EGFR) have emerged as a robust treatment option for various solid tumors. Despite lower systemic side effects than conventional chemotherapy, the majority of patients treated with these agents experience cutaneous toxicities, including papulopustular rashes, hair and nail changes, xerosis and pruritus, which have a significant impact on health and quality of life. Currently no consensus or management guidelines exist for these untoward events. Therefore, a retrospective survey was carried out across 110 oncology practioners in the US that were administering EGFR inhibitors. Providers were queried on the impact and management of these untoward events in their practices. Responses suggest that combination therapies (topical and oral) were more effective than either therapy alone, and also lead to a more rapid resolution of the papulopustular rash. Providers also reported that patients frequently complained of physical symptoms associated with the rash (itching and pain), and that they had a positive perception when being treated for their cutaneous side effects. The survey results support that attentive cutaneous care is important in patients treated with EGFR inhibitors, and that proactive/combined interventions may enhance quality of life and optimize consistent drug administration. PMID- 18160806 TI - Targeting the epidermal growth factor receptor in colorectal cancer: a potential therapeutic role for gastrin-releasing peptide receptor antagonists. PMID- 18160807 TI - Testicular mixed germ cell tumor in an adolescent with cowden disease. AB - Cowden disease (also known as Cowden syndrome) is characterized by multiple organ hamartomatous tumors and an increased risk of malignancy, in particular of the breast, thyroid and endometrium. Testicular tumors including seminoma have previously been reported in adult patients. We are reporting, for the first time, a case of testicular mixed germ cell tumor in an adolescent with Cowden disease. An association of testicular malignancy in Cowden disease could be explained by the previous observation of strong PTEN gene expression in the basal cell layer around seminiferous tubules and increased frequency of PTEN mutations in cultured testicular cancer cell lines. Surveillance for breast, thyroid, endometrial and renal cancer has been recommended for individuals with Cowden disease. The association of Cowden disease and testicular malignancy in our case suggests the need for additional screening of testes. PMID- 18160808 TI - High incidence of extracellular matrix metalloproteinase inducer expression in non-small cell lung cancers. Association with clinicopathological parameters. AB - OBJECTIVE: Extracellular matrix metalloproteinase inducer (EMMPRIN) is a highly glycosylated transmembrane protein that is widely present on the surface of various tumor cells, assisting in tumor progression by stimulating the production of several matrix metalloproteinases in adjacent stromal cells. However, its clinical relevance remains to be evaluated in lung cancers. Therefore, we aimed to investigate the relationship between EMMPRIN expression in non-small cell lung cancer (NSCLC) and clinicopathological characteristics and prognosis. METHODS: EMMPRIN expression was semiquantified by immunohistochemistry with anti-human EMMPRIN monoclonal antibody in 208 surgically resected NSCLCs and was analyzed statistically in relation to various characteristics. RESULTS: EMMPRIN expression was seen in most NSCLC samples (92%). High levels of EMMPRIN expression were significantly associated with differentiation and pT(1) stage in adenocarcinomas. There were no significant differences in overall survival between patients with tumors having high and low levels of EMMPRIN expression in pathological stage I NSCLCs (5-year survival rates, 69 vs. 60%). CONCLUSIONS: EMMPRIN was preferentially expressed in most NSCLCs. High levels of expression were associated with early T stage and well-differentiated adenocarcinoma, and were not a prognostic factor in NSCLC. PMID- 18160809 TI - Impact of plasma tissue inhibitor of matrix metalloproteinase-1 on long-term survival in patients with colorectal cancer. AB - Tissue inhibitor of metalloproteinase-1 (TIMP-1) not only inhibits matrix metalloproteinases but also stimulates tumor growth. In this study, long-term follow-up results were analyzed to clarify the prognostic value of plasma TIMP-1. Preoperative plasma TIMP-1 was measured from peripheral blood samples of 87 Japanese patients with colorectal carcinoma. All the patients underwent surgical resection and were followed for 5 years prospectively. The median follow-up period was 70 months (60-79 months). The cutoff value of plasma TIMP-1 was set at 170 ng/ml based on the ROC curve. Sensitivity and specificity to predict 5-year survival was 66.7 and 55.0% with this cutoff value. In univariate analyses for overall survival, lymph node metastasis, serosal invasion, peritoneal metastasis, liver metastasis, metastasis to other distant organs and TIMP-1 were significant. In multivariate analyses, lymph node and liver metastases, metastasis to other distant organs and plasma TIMP-1 were independent prognosticators, but p values of TIMP-1 did not reach statistical significance. Our results suggested that the preoperative plasma TIMP-1 concentration could be a useful prognosticator of long term survival in patients with colorectal carcinoma. PMID- 18160810 TI - Feasibility of using cetuximab and bevacizumab in a patient with colorectal cancer and terminal renal failure. PMID- 18160811 TI - N-terminal-pro-B type natriuretic peptide as a useful tool to evaluate pulmonary hypertension and cardiac function in CDH infants. AB - OBJECTIVE: In congenital diaphragmatic hernia (CDH) the severity of pulmonary hypertension (PH) is considered, by several authors, determinant of clinical outcome. Plasmatic N-terminal-pro-B type natriuretic peptide (NT-proBNP) might be useful in diagnosis and management of PH in newborns, although its interest in CDH infants remains to be defined. Early NT-proBNP levels were assessed in CDH infants and correlated with cardiovascular echocardiographic parameters. PATIENTS AND METHODS: 28 newborns, CDH and age-matched controls were enrolled in a prospective study. Clinical condition, NT-proBNP plasmatic levels, echo parameters of PH and biventricular function were assessed at 24 h after delivery as well as survival outcome. RESULTS: Estimated mean pulmonary pressure and NT proBNP were significantly higher in CDH than control infants. NT-proBNP significantly correlated with estimated pulmonary artery pressure, right ventricular Tei index, and tricuspid E/A ratio. Additionally, we found that CDH infants with NT-proBNP >11,500 pg/ml experienced a worse prognosis. CONCLUSIONS: We demonstrated that PH is associated with NT-proBNP elevation and diastolic impairment in CDH infants. Early elevations in NT-proBNP levels seem to alert for a subset of CDH infants with worse prognosis. PMID- 18160812 TI - The protective effect of TiF4, SnF2 and NaF against erosion-like lesions in situ. AB - The aim of this in situ study was to compare the protective effect of TiF4, SnF2 and NaF on the development of erosion-like lesions in human enamel. Fourteen human molars were each divided into 4 specimens, mounted on acrylic mouth appliances and worn by 7 volunteers for 9 days. In order to mimic a feasible treatment procedure for patients with voluntary or involuntary gastric reflux, the specimens were etched for 2 min twice a day (0.01 M HCl) and fluoride applications were performed every third day (2 min). The controls were treated as the other specimens except for the fluoride applications. Etch depths and surface roughness changes (R(q)) were measured by white light interferometry. Compared with the control, TiF4 and SnF2 reduced the etching depth by 100% (p < 0.001) and 91% (p < 0.001), respectively, and both treatments resulted in an observable surface layer. NaF had no significant protective effect (p = 0.46). It can be concluded that although SnF2 provided significant protection for the enamel surfaces, TiF4 showed the best protection against acid attack, while NaF had no significant protective effect in this study. PMID- 18160813 TI - Approximal fluoride concentration using different fluoridated products alone or in combination. AB - The aim of the present investigation was to measure the approximal fluoride (F) concentration after using different F-containing products, either alone or in combination. Ten subjects participated on a total of 17 occasions, which were randomized, distributed and divided into two trials (called trial I and trial II). The F concentration was studied after treatment with toothpicks, dental flosses, an interdental brush dipped in 0.2 or 0.32% F gels, brushing with toothpaste containing 0.32% F and a mouthrinse with 0.2% NaF solution. The results of trial I revealed that the rinsing solution resulted in the highest approximal F values, followed by toothpicks and brushing. When brushing was combined with either flossing or toothpick, the sequence order of the two methods had less influence. However, when combined with rinsing it appeared to be better to use the floss or the toothpicks first, prior to the mouthrinse. In trial II, treatment with the gels alone, and in combination with brushing, resulted in higher approximal F concentrations compared with brushing alone. The data indicate that the order of usage when combining products may be of importance when it comes to obtaining high F levels at the approximal sites and that an interdental brush with F gel after toothbrushing may be a suitable method, called 'Inter Dental Brush Gel Method', to produce elevated F concentrations at approximal sites. PMID- 18160814 TI - Indoxyl sulfate reduces superoxide scavenging activity in the kidneys of normal and uremic rats. AB - BACKGROUND/AIM: Indoxyl sulfate (IS) is a uremic toxin that accelerates the progression of chronic renal failure (CRF). This study aimed at determining whether IS impairs antioxidative systems (redox status) in the kidney. METHODS: IS was orally administered to normal and subtotally nephrectomized (three fourths and five sixths) rats (CRF rats) for 2 weeks. By use of in vivo and ex vivo electron spin resonance spectroscopy, the kidney redox status was evaluated using carbamoyl-PROXYL as a radical spin probe in living rats, and the kidney superoxide scavenging activity was measured. Immunohistochemistry of superoxide dismutase (SOD) in the kidney was performed. RESULTS: Administration of IS increased serum and kidney levels of IS and serum creatinine and decreased creatinine clearance. CRF rats showed reduced spin reduction rate, prolonged half life of the spin probe, and reduced superoxide scavenging activity and SOD positive areas in the kidney as compared with normal rats. Administration of IS further reduced radical spin reduction rate, prolonged half-life of the spin probe, and reduced superoxide scavenging activity and SOD-positive areas in the kidneys. CONCLUSIONS: Administration of IS reduced superoxide scavenging activity in the kidneys of normal and CRF rats. Thus, the nephrotoxicity of IS may be induced by impairing the antioxidative systems in the kidney. PMID- 18160815 TI - alpha-Klotho: a regulator that integrates calcium homeostasis. AB - Intensive study on calcium homeostasis regulation over the past several decades has established a systematized construal of its role in living phenomena, leaving us with the impression that this field is fairly defined and understood. However, the unveiling of the molecular function of alpha-Klotho has recently given new insight into this field. alpha-Klotho is a unique molecule that plays pivotal roles in: (i) the rapid tuning of extracellular Ca(2+) concentration through transepithelial Ca(2+) transport; (ii) parathyroid hormone secretion and subsequent Ca(2+) increase in the serum, and (iii) the signal transduction of FGF23 that adjusts the calcium concentration by downregulating the production of 1,25(OH)(2)D(3). Through these pathways, alpha-Klotho participates in the regulation of calcium homeostasis of the CSF and blood/body fluids by its actions in the choroid plexus, parathyroid glands and DCT nephrons. In this regard, alpha Klotho is a key player that integrates 'a multi-step regulatory system of calcium homeostasis' that rapidly adjusts the extracellular calcium concentration and continuously maintains its concentration within a narrow physiological range. PMID- 18160816 TI - SLC40A1 c.1402G-->a results in aberrant splicing, ferroportin truncation after glycine 330, and an autosomal dominant hemochromatosis phenotype. AB - BACKGROUND/AIMS: To determine the molecular basis of a mild hemochromatosis phenotype in a man of Scottish-Irish descent. METHODS: We sequenced genomic DNA to detect mutations of HFE, SLC40A1, TFR2, HAMP, and HFE2. RNA isolated from blood mononuclear cells was used to make cDNA. RT-PCR was performed to amplify ferroportin from cDNA, and amplified products were visualized by electrophoresis and sequenced. RESULTS: The proband was heterozygous for the novel mutation c.1402G-->A (predicted G468S) in exon 7 of the ferroportin gene (SLC40A1). Located in the last nucleotide before the splice junction, this mutation results in aberrant splicing to a cryptic upstream splice site located at nt 990 within the same exon. This causes truncation of ferroportin after glycine 330 and the addition of 4 irrelevant amino acids before terminating. The truncated ferroportin protein, missing its C-terminal 241 amino acids, would lack all structural motifs beyond transmembrane region 7. The patient was also heterozygous for the common HFE H63D polymorphism, but did not have coding region mutations in TFR2, HAMP, or HFE2. CONCLUSIONS: We conclude that this patient represents a unique example of hemochromatosis due to a single base-pair mutation of SLC40A1 that results in aberrant splicing and truncation of ferroportin. PMID- 18160817 TI - Pulmonary hypertension associated with myeloproliferative disorders: a retrospective study of ten cases. AB - BACKGROUND: Pulmonary hypertension (PH) is a severe hemodynamic disorder in which the pulmonary artery pressure is persistently elevated, leading to right-sided heart failure. Some studies have suggested an association between PH and myeloproliferative diseases (MPD). OBJECTIVES: This study describes clinical, hematological and hemodynamic characteristics of PH associated with MPD. METHODS: We retrospectively reviewed 10 cases of PH associated with MPD: polycythemia vera (8 patients) and essential thrombocythemia (2 patients), followed between 1993 and 2002. The baseline evaluation was established by right-sided heart catheterization, ventilation/perfusion lung scan and pulmonary angiography if required. RESULTS: Six patients had confirmed chronic thromboembolic pulmonary hypertension (CTEPH) and 4 had pulmonary arterial hypertension (PAH) associated with MPD without other risk factors for PAH. The hemodynamic characteristics of CTEPH and PAH associated with MPD were similar. The diagnosis of CTEPH was concomitant to that of MPD in all cases (5 polycythemia vera and 1 essential thrombocythemia). The PAH associated with MPD occurred later in the evolution of the MPD (3 polycythemia vera and 1 essential thrombocythemia) with a median of 162 months after the diagnosis of MPD, and it was associated with myeloid metaplasia (p < 0.01). CONCLUSION: We describe 2 distinct forms of PH in the context of MPD: CTEPH, which is diagnosed at an early stage of the MPD, and PAH, which occurs later in the course of the MPD and is associated with myeloid metaplasia. Progressively increasing dyspnea in a patient with an MPD warrants further investigation to rule out PAH and CTEPH, while a diagnosis of CTEPH warrants ruling out MPD. PMID- 18160818 TI - Cellular and molecular events underlying the dysregulated response of the aged brain to stroke: a mini-review. AB - BACKGROUND: Age-related brain injuries, including stroke, are a major cause of physical and mental disabilities. OBJECTIVE: Therefore, studying the basic mechanism underlying functional recovery after brain stroke in aged subjects is of considerable clinical interest. METHODS: This review summarizes the effects of age on recovery after stroke in an animal model, with emphasis on the underlying cellular mechanisms. RESULTS: Data from our laboratory and elsewhere indicate that, behaviorally, aged rats were more severely impaired by stroke than young rats, and they also showed diminished functional recovery. Infarct volume did not differ significantly between young and aged animals, but critical differences were apparent in the cytological response to stroke, most notably an age-related acceleration in the development of the glial scar. Early infarct in older rats is associated with premature accumulation of BrdU-positive microglia and astrocytes, persistence of activated oligodendrocytes, a high incidence of neuronal degeneration and accelerated apoptosis. In aged rats, neuroepithelial-positive cells were rapidly incorporated into the glial scar, but these neuroepithelial like cells did not make a significant contribution to neurogenesis in the infarcted cortex in young or aged animals. The response of plasticity-associated proteins like MAP1B, was delayed in aged rats. Tissue recovery was further delayed by an age-related increase in the amount of the neurotoxic C-terminal fragment of the beta-amyloid precursor protein (A-beta) at 2 weeks poststroke. CONCLUSION: The available evidence indicates that the aged brain has the capability to mount a cytoproliferative response to injury, but the timing of the cellular and genetic response to cerebral insult is dysregulated in aged animals, thereby further compromising functional recovery. Elucidating the molecular basis for this phenomenon in the aging brain could yield novel approaches to neurorestoration in the elderly. PMID- 18160819 TI - Cerebral hypoperfusion increases estrogen receptor abundance in the ovine fetal brain and pituitary. AB - BACKGROUND/AIMS: Estrogen is an important component of fetal neuroendocrine function in late-gestation fetal sheep; however, little is known about the regulation of estrogen receptor abundance in the brain and pituitary of fetuses. The present study was performed to test the hypotheses that estrogen receptor abundance in the fetal brain and pituitary are influenced by circulating estradiol concentrations and that they are acutely regulated after cerebral hypoperfusion. METHODS: We studied 16 time-dated fetal sheep (124-128 days gestation) that were chronically catheterized and instrumented at least 5 days before study. Four groups (n = 4 each) were studied in which fetuses received estradiol (0.25 mg/day, producing physiological increases in fetal plasma estradiol concentrations) or placebo implants, and in which fetuses received a 10 min period of brachiocephalic occlusion (BCO) or sham-BCO. One hour after BCO or sham-BCO, fetuses were euthanized and tissues rapidly removed for analysis of estrogen receptors (ER)-alpha and -beta at the mRNA and protein levels. RESULTS: Both BCO and estradiol treatment were effective in changing ER expression, although the effects were region-specific. BCO dramatically increased ER-alpha in the pituitary and both ER-alpha and ER-beta in the brainstem, while decreasing ER alpha expression in the hypothalamus. Estradiol treatment decreased ER-alpha expression in the hypothalamus, whereas it increased ER-alpha expression in the brainstem, cerebral cortex and hippocampus. CONCLUSIONS: We conclude that the expression of ER-alpha and ER-beta in the brain and pituitary of fetal sheep are influenced by circulating estrogen concentrations and acutely regulated in response to cerebral hypoperfusion. PMID- 18160820 TI - Adhesion of human hematopoietic progenitor cells to mesenchymal stromal cells involves CD44. AB - BACKGROUND: Direct cell-cell contact between hematopoietic progenitor cells (HPC) and their cellular microenvironment is essential for maintenance of 'stemness'. We have previously demonstrated that a feeder layer of human mesenchymal stromal cells (MSC) could provide a surrogate model as a niche for human HPC. Maintenance of long-term culture-initiating cells was significantly lower on fibroblasts. METHODS: Adhesion of HPC to MSC was further analyzed using our recently described adhesion assay based on gravitational force upon inversion and in combination with specific antibodies against CD44. RESULTS: Adhesion of KG1a and CD34+ cells was significantly reduced by administration of a monoclonal CD44 antibody and for KG1a to a greater extent than for CD34+ cells. Interaction of HPC and MSC was further analyzed by laser scanning confocal microscopy. CD44 was located on the uropod of CD34+ cells at the site of contact with MSC and both cell types were interwoven by a network of fibronectin. CONCLUSION: Various adhesion proteins, including CD44, are involved in the contact of human HPC and human MSC and further analysis of the relative significance and interaction of these proteins will be crucial for the understanding of the mechanism of this specific cell-cell interaction. PMID- 18160821 TI - Cell polarity and asymmetric cell division within human hematopoietic stem and progenitor cells. AB - Like other somatic stem cells, hematopoietic stem cells (HSC) contain the capacity to self-renew and to give rise to committed progenitor cells that are able to replenish all hematopoietic cell types. To keep a constant level of HSC, the decision whether their progeny maintain the stem cell fate or become committed to differentiation needs to be highly controlled. In this context it became evident that HSC niches fulfill important functions in keeping the level of HSC more or less constant. Before discovering such niches, it was widely assumed that HSC divide asymmetrically to give birth to a daughter cell maintaining the stem cell fate and to another one which is committed to differentiation. Here, I summarize some of the experimental data being compatible with the model of asymmetric cell division and review some of our latest findings, which demonstrate the occurrence of asymmetric cell divisions within the HSC and hematopoietic progenitor cell compartment. Since cell polarity is an essential prerequisite for asymmetrically dividing as well as for migrating cells, I will also discuss some aspects of cell polarity of primitive hematopoietic cells. PMID- 18160822 TI - The origins of human embryonic stem cells: a biological conundrum. AB - Human inner cell mass (ICM) cells isolated from in vitro fertilized blastocysts are the progenitor cells used to establish in vitro stable human embryonic stem cells (hESCs) which are pluripotent and self-renew indefinitely. This long-term perpetuation of hESCs in the undifferentiated state is thought to be an in vitro adaptation of the ICM cells. To investigate at the molecular level how hESCs acquired their unique properties, transcriptional profiles of isolated ICM cells and undifferentiated hESCs were compared. We identified 33 genes enriched in the ICM compared to the trophectoderm and hESCs. These genes are involved in signaling cascades (SEMA7A and MAP3K10), cell proliferation (CUZD1 and MS4A7) and chromatin remodeling (H1FOO and HRMT1L4). Furthermore, primordial germ cell specific genes (SGCA and TEX11) were detected as expressed in the ICM cells and not hESCs. We propose that the transcriptional differences observed between ICM cells and hESCs might be accounted for by adaptive reprogramming events induced by the in vitro culture conditions which are distinct from that of in vitro fertilized blastocysts. hESCs are a distinct cell type lacking in the human embryo but, nonetheless, resemble the ICM in their ability to differentiate into cells representative of the endodermal, ectodermal and mesodermal cell lineages. PMID- 18160823 TI - Stem cell differentiation: cardiac repair. AB - Cellular transplantation has been employed for several years to deliver donor cardiomyocytes to normal and injured hearts. Recent reports of a variety of stem cells with apparent cardiomyogenic potential have raised the possibility of cell transplantation-based therapeutic interventions for heart disease. Here we review the preclinical studies demonstrating that intracardiac transplantation of skeletal myoblasts, cardiomyocytes and cardiomyogenic stem cells is feasible. In addition, recent clinical studies of skeletal myoblast and adult stem cell transplantation for heart disease are discussed. PMID- 18160824 TI - New insights into the cell biology of hematopoietic progenitors by studying prominin-1 (CD133). AB - Prominin-1 (alias CD133) has received considerable interest because of its expression by several stem and progenitor cells originating from various sources, including the neural and hematopoietic systems. As a cell surface marker, prominin-1 is now used for somatic stem cell isolation. Its expression in cancer stem cells has broadened its clinical value, as it might be useful to outline new prospects for more effective cancer therapies by targeting tumor-initiating cells. Cell biological studies of this molecule have demonstrated that it is specifically concentrated in various membrane structures that protrude from the planar areas of the plasmalemma. Prominin-1 binds to the plasma membrane cholesterol and is associated with a particular membrane microdomain in a cholesterol-dependent manner. Although its physiological function is not yet determined, it is becoming clear that this cell surface protein, as a unique marker of both plasma membrane protrusions and membrane microdomains, might reveal new aspects of the cell biology of rare stem and cancer stem cells. The aim of this review is to outline the recent discoveries regarding the dynamic reorganization of the plasma membrane of rare CD133+ hematopoietic progenitor cells during cell migration and division. PMID- 18160825 TI - Tenascin C in stem cell niches: redundant, permissive or instructive? AB - The stem cell niche provides the specialized environment that is able to sustain the lifelong maintenance of stem cells in their discrete locations within organs. The niche is usually composed of several different cell types and a specialized extracellular matrix consisting of many different constituents. Additionally, a variety of growth factors are secreted into the extracellular space and contribute to the functional organization of the niche. Here, I will concentrate on the multimodular extracellular matrix glycoprotein tenascin C (Tnc) and discuss it as an exemplary molecule that is present in several stem cell niches. In spite of its intuitively suggestive presence, it has been difficult to provide functional evidence for the importance of Tnc in the context of stem cells. In the nervous system, the careful analysis of Tnc-deficient mice has revealed that the developmental program neural stem cell pass-through is delayed due to changes in growth factor responsiveness. To gain further insight, we have employed the gene trap technology in neural stem cells to identify potential Tnc target genes. This approach has surfaced 2 interesting candidates that may contribute to a better understanding of the signal(s) elicited by Tnc in neural stem/progenitor cells in the niche. PMID- 18160826 TI - [Incidence and histologic patterns of bone marrow involvement of malignant lymphoma based on the World Health Organization classification: a single institution study]. AB - BACKGROUND: Working Formulation and Revised European American Classification of Lymphoid Neoplasms (REAL) have mainly been used in the studies for bone marrow involvement of malignant lymphoma in Korea. We investigated the incidence and histologic patterns of malignant lymphoma according to the WHO classification. METHODS: This study included 507 cases of malignant lymphoma that were requested for bone marrow study for the staging during the period January 1999-December 2005 in Korea Cancer Center Hospital. Medical records, peripheral blood smears, bone marrow aspiration smears, biopsy sections, and histopathologic findings were analyzed retrospectively. RESULTS: Of the 507 cases of malignant lymphoma, 473 (93.3%) were non-Hodgkin lymphoma (NHL) and 34 (6.7%) were Hodgkin lymphoma (HL). The overall incidence of bone marrow involvement by NHL and HL was 12.5% (59/473) and 11.8% (4/34), respectively. Among NHL cases, the incidence of bone marrow involvement by B-cell and T-cell neoplasms was 11.4% (43/377) and 16.7% (16/96), respectively. Although the incidences of bone marrow involvement by several B cell neoplasms were more than 30%, diffuse large B cell lymphoma showed a relatively low incidence of bone marrow involvement (4.6%). Of bone marrow involvement patterns, diffuse infiltration pattern was the most common (40.0%). Peripheral blood involvement by lymphoma was observed in 35.6% of cases with bone marrow involvement. CONCLUSIONS: We used WHO classification in the study for the bone marrow involvement of malignant lymphoma, and this single-institution study should give a useful, up-to-date histopathologic information. PMID- 18160827 TI - [Prognostic effect of cytoplasmic CD79a expression in acute myeloid leukemia with t(8;21)]. AB - BACKGROUND: Although cytoplasmic CD79a (cytCD79a) is a highly lineage-specific marker of B lymphoid cells and plays an important role in the diagnosis of acute leukemia, its clinical significance is not fully understood. We aimed to investigate the relationship between cytCD79a positivity and survival probability, and to evaluate the prognostic value of cytCD79a expression in AML with t(8;21) (q22;q22). METHODS: A total of 68 cases of AML with t(8;21)(q22;q22) were diagnosed based on conventional morphology, cytochemistry, flow cytometrty, and cytogenetic and molecular genetic analysis. Immunohistochemistry of cytCD79a was performed retrospectively. Laboratory and clinical findings were reviewed. RESULTS: Five patients among 68 AML with t(8;21)(q22;q22) revealed cytCD79a positive reaction; scores for myeloid lineage/B-lymphoid lineage were 5/3-3.5. Among the five cytCD79a positive patients, only one patient was a child. Three patients were with refractory AML or relapsed, and two patients died within 10 months. Median survival time of cytCD79a positive group was shorter (8.0 months) than that (61.3 months) of cytCD79a negative group. The survival probability of the cytCD79a expression group was significantly lower than classical AML with t(8;21)(q22;q22) (P=0.0001). CONCLUSIONS: These findings emphasize the necessity of investigating cytCD79a, especially in AML with t(8;21)(q22;q22), for a different clinical prognostic value. PMID- 18160828 TI - [New leukocyte counting method of cerebrospinal fluid: using transparent ruler tape]. AB - BACKGROUND: To enumerate leukocyte count in cerebrospinal fluid (CSF) is important for diagnosing bacterial meningitis. Using automated hematology analyzer for enumeration of leukocyte in CSF is below the sensitivity, so microscopic hemocytometric method is standard method. But this requires sufficient practical experience and has limitation of accuracy and stability. So we developed new manual method and evaluated it. METHODS: We designed new method using transparent ruler tape. We performed correlation, accuracy and precision test by counting leukocyte in diluted EDTA blood with three methods: new method, Neubauer and Nageotte hemocytometry. Twenty two CSF were used for stability test, which determines leukocyte count according to time (within one hour and after 2, 4 and 12 hr), by new method and Neubauer hemocytometry at room temperature. RESULTS: There was no clinical significant difference between three methods in correlation test, whereas Neubauer and Nageotte hemocytometry showed a bias to underestimation relative to the results obtained with new method in case with low leukocyte count. The new method showed the lowest CV and most accurate result. In stability test, leukocyte counts decreased being 44.4%, 72.1% of initial values after 2 hr, 14.8%, 31.1%, after 4 hr and 4.2%, 8.7%, after 12 hr, by Nageotte hemocytometry and new method, respectively. CONCLUSIONS: The new method we devised is simple, easy and applicable to use in a laboratory and offers advantages of improved precision and stability. It may be sufficient for replacing standard methods for leukocyte counting in CSF. PMID- 18160829 TI - [Clinical significance of serum osteopontin in patients with multiple myeloma]. AB - BACKGROUND: Angiogenesis and osteoclastogenesis are increased in the bone marrow of multiple myeloma (MM) patients in parallel with the tumor progression. Osteopontin (OPN) is a multifunctional protein that is involved in angiogenesis and bone destruction and, eventually, in tumor progression in MM. OPN is known to increase in MM patients as the disease progresses and bone is destroyed. We studied the clinical usefulness of OPN as a monitoring marker for treatment response in patients with MM. METHODS: We obtained 70 serial sera from 27 MM patients and 14 sera from healthy individuals. OPN was measured by a sandwich ELISA method. The hospital records were reviewed, and the clinically important markers for monitoring the treatment response, such as monoclonal component, immunoglobulin, free light chain, and hemoglobin, etc, were analyzed together with OPN levels. RESULTS: There was no significant difference in OPN levels between MM patients and healthy controls. OPN showed no significant correlations with the markers used for monitoring of treatment response such as M component, immunoglobulin, and free light chain levels. There was no difference in OPN levels between the 3 groups classified by the amount of M component. In addition, OPN levels showed no compatible changes to the treatment response of MM patients. CONCLUSIONS: Although OPN has been known to have an important role in the formation and progression of MM by involving angiogenesis and bone destruction, our results show that OPN is not valuable as a clinical marker for monitoring the treatment response in MM patients because of inconsistency in its levels in MM patients. PMID- 18160830 TI - [Proportions of cells expressing CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, or CD13,33+/CD34+ in the regenerating bone marrows during complete remission of acute leukemia or after bone marrow transplantation]. AB - BACKGROUND: The hemopoietic stem cells increase in number during the regeneration after chemotherapy or bone marrow transplantation (BMT). Although the proportion of hemopoietic stem cells and their differentiation have been studied by immunophenotyping using the flow cytometry, no substantial research efforts have been directed toward the regenerating marrow. We attempted to discover the proportions of undifferentiated stem cells, committed stem cells, B cell precursors, and myeloid precursors in the regenerating bone marrows during complete remission (CR) and after engraftment of BMT. METHODS: Bone marrow samples from 82 patients with acute leukemia in CR and from 25 patients after BMT engraftment, along with 22 control samples, were used to find the numbers of CD38 /CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells in the large lymphocyte gate by flow cytometry. We cross-analyzed our results in terms of groups: CR, BMT, and initial diagnosis groups. We performed significance tests on age, relapse, chromosomal abnormalities, clinical outcomes, and initial immunophenotypes of the leukemic cells. RESULTS: The proportions of CD38-/CD34+, CD38+/CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells are more highly distributed in acute B-lymphoblastic leukemia than the normal group and also in the CR than the BMT group. CD19+/CD34+ cells were increased in the relapse group and CD38+/ CD34+, CD19+/CD34+, and CD13,33+/CD34+ cells were increased in the group with chromosomal abnormality. The results were irrelevant to the initial immunophenotype of the leukemic blasts. CONCLUSIONS: The increases of the markers spanned too widely to apply one specific cutoff value to analyze them. They seemed to be the results of normal regeneration, irrelevant to relapse or initial immunophenotype of leukemic blasts. PMID- 18160831 TI - [Evaluation of two beta 2-microglobulin reagents by turbidimetric immunoassay]. AB - BACKGROUND: An increased level of beta 2-microglobulin (beta 2M) is seen in diseases such as lymphoproliferative diseases, renal diseases, solid tumors, liver diseases, certain viral infection, or chronic inflammatory diseases, etc. In this study, we evaluated a quantitative beta 2M assay for precision and linearity using an automated turbidimetric immunoassay (TIA) by Hitachi 7600-110 (Hitachi High Technologies Co., Japan). The TIA of beta 2M was compared with a chemiluminescent immunoassay (CLIA) by Immulite 2000 (Diagnostic Products Corporation, USA). METHODS: Two TIAs, the Hitachi 7600-110 with Roche reagent (Roche-TIA) and the Hitachi 7600- 110 with HBI reagent (HBI-TIA), were evaluated for within-run precision, within-day precision, betweendays precision, and linearity. With 68 serum samples, two TIAs were compared with Immulite 2000 using DPC reagent (DPC-CLIA). These data were analyzed by Passing-Bablok analysis and Bland- Altman analysis. RESULTS: The coefficients of variation (CVs) of within run precision, within-day precision, and between- days precision were less than 7% in all groups. The linearity tests of the two TIAs were maintained well (Roche TIA: R2=0.9952; HBI-TIA: R2=0.9946). The comparison study indicated good correlations (Roche-TIA/DPC-CLIA: r=0.9738, y=0.9625x-0.0375; HBI-TIA/DPC-CLIA: r=0.9725, y= 1.1000x-0.3100). In Bland-Altman analysis, less than 2SD differences were observed between the two groups. CONCLUSIONS: Both Roche-TIA and HBI-TIA showed a good precision and excellent correlations with DPC-CLIA; therefore, TIA could be used in the routine laboratory to determine a quantitative analysis of beta 2M. PMID- 18160832 TI - Detection of 12 respiratory viruses with two-set multiplex reverse transcriptase PCR assay using a dual priming oligonucleotide system. AB - BACKGROUND: We intended to evaluate the diagnostic usefulness of a multiplex reverse transcriptase- PCR (mRT-PCR) assay kit under dual priming oligonucleotide system (DPO) for the childhood acute respiratory tract infections. METHODS: Two hundred nasopharyngeal aspirates were taken from children < or =5 yr old admitted due to acute respiratory infections in 2004. Direct fluorescent antibody (FA) assays were performed with fresh specimens; then, mRT-PCRs for the detection of 12 respiratory viruses (Seeplex RV detection kit, SeeGene, Seoul, Korea) were tested with frozen specimens. RESULTS: FA assays for five common respiratory viruses showed positive results in 66 patients (33.0%), while mRT-PCR detected causative viruses in 112 patients (56.0%), including 16 co-infected cases (8.0%). A total of 129 viruses were identified: respiratory syncytial virus A/B (38.0%/7.8%), influenza virus A/B (10.1%/5.4%), parainfluenza virus 1/2/3 (7.0%/3.1%/7.8%), coronavirus 229E or NL63 (6.2%), human metapneumovirus (4.7%), adenovirus (4.7%), rhinovirus (3.9%), and coronavirus OC43 (1.6%). CONCLUSIONS: DPO-based mRT-PCR was found as a sensitive tool for the detection of the viruses that cause childhood respiratory infections. Clinical significances of the agents detected by mRTPCR need further evaluations. PMID- 18160833 TI - [Coproduction of qnrB and armA from extended-spectrum beta-lactamase-producing Klebsiella pneumoniae]. AB - BACKGROUND: Multidrug-resistant Enterobacteriaceae is a worldwide problem. Although various resistance mechanisms have been recognized with increasing frequency, only a few cases of triple resistance of extended-spectrum beta lactamase (ESBL)-producing Klebsiella pneumoniae have been reported. This study was designed to evaluate the coexistence of qnr (qnrA, qnrB, and qnrS) and 16S rRNA methylase (armA, rmtA, rmtB, and rmtC) in ESBL-producing K. pneumoniae. METHODS: We tested 44 isolates of ESBL-producing K. pneumoniae at Chungnam National University Hospital from March to September 2006. Antimicrobial susceptibilities were tested by broth microdilution method, and transconjugation test was performed using E. coli J53 with azide resistance. Search for qnr (qnrA, qnrB, and qnrS) and 16S rRNA methylase (armA, rmtA, rmtB, and rmtC) genes was conducted by PCR amplification, and the genotypes were determined by direct nucleotide sequence analysis of the amplified products. Epidemiologic study was performed by Enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). RESULTS: All ESBL-positive strains produced qnrB; however, armA was detected in 68.2%. The coproduction rate of qnrB and armA in ESBL-producing K. pneumoniae was 68.2%. Two types (A and B) were dominant in ERIC-PCR results. CONCLUSIONS: K. pneumoniae producing qnrB, armA, and ESBL are spreading widely. PMID- 18160834 TI - [Comparisons of latex agglutination, immunochromatography and enzyme immunoassay methods for the detection of rotavirus antigen]. AB - BACKGROUND: Rotavirus is the most common cause of childhood gastroenteritis during winter season. Rapid, accurate diagnosis is essential for preventing severe complications of rotaviral gastroenteritis. The sensitivity and specificity of five detection test kits for rotavirus including latex agglutination (LAT), enzyme immunoassay (EIA) and three immunochromatographic methods (ICG) were evaluated in this study. METHODS: A total of 95 stool samples collected from patients with acute gastroenteritis were studied. The test kits were as follows: LAT (Slidex latex, bioMerieux Vitek, France); three kinds of ICG (Dipstick ROTA, Eiken, Japan; SAS Rota Test, SA Scientific, Inc., USA; and ASAN Easy Test Rota strip, ASAN Pharmaceutical., Korea); and EIA (VIDAS Rotavirus, bioMerieux Vitek). The samples showing discordant results were reevaluated by reverse-transcription (RT) PCR and clinical manifestations. RESULTS: Of a total of 95 cases, 56 (58.9%) were positive and 39 (41.1%) were negative. Thirteen cases showed discordant results. Sensitivity and specificity were, respectively, 85.7% and 100% for LAT, 100% and 95% for both of Dipstick ROTA and SAS Rota, 86.7% and 87.5% for ASAN Rota strip and 98.1% and 97.3% for EIA. CONCLUSIONS: LAT was rapid and easy to perform and showed the lowest sensitivity among the five test kits. ICG showed a good agreement with EIA and RT-PCR. EIA was the best in respect of sensitivity and specificity, but difficulty in interpretations of equivocal results and time-consuming procedures were limitations. In conclusion, ICG, which is easy to perform at a low cost, may be an optimal method in place of LAT for the detection of rotavirus. PMID- 18160835 TI - [Human leukocyte antigen typing proficiency surveys in Korea, 2005-2006]. AB - BACKGROUND: To monitor the performance of histocompatibility testing laboratories, HLA proficiency survey in Korea has been conducted biannually since 1996. In this report, we summarized the results of the surveys performed in recent two years (2005-2006). METHODS: A total of four proficiency surveys were performed, in which 59-61 laboratories participated. Each survey included three tests for HLA class I (serology and DNA) and class II (DNA) typing and six tests for HLA crossmatch. RESULTS: The overall concordance of serologic typing was 98.9% (355/359) for HLA-A, 97.5% (350/ 359) for HLA-B, and 94.7% (337/356) for HLA-C. The antigens assigned correctly by less than 95% of the participating laboratories were A26 (93.8%), B38 (94.2%), Cw3/Cw10 (90.9%), Cw6 (94.4%), and Cw8 (74.3%). The overall concordance rates of DNA typing were 99.6% (533/535) for HLA-A, 99.8% (539/540) for HLA-B, and 100% (392/392) for HLA-C. Correct assignment of HLA-DRB1 and -DQB1 was reported by 99.2% (98.1-100%) and 96.7% (88.9-100%) for the generic level and 100% and 95.8% (75-100%) for the allelic level, respectively. On the average 3.8% (0-7.7%) of the total laboratories showed unacceptable results in the crossmatch tests. CONCLUSIONS: The rates of correct antigen identification and of unacceptable crossmatch were similar to those of previous surveys, which were considered satisfactory. The Korean proficiency survey program may have contributed to a high quality of HLA tests today and should be continued for further improvements of the tests tomorrow. PMID- 18160836 TI - Comparison of VERSANT Hepatitis B virus DNA 3.0 Assay with Digene Hybrid Capture II Hepatitis B virus DNA test in relation to clinical status of Hepatitis B virus. AB - BACKGROUND: Some differences exist among various Hepatitis B virus (HBV) DNA quantification assays due to lack of standardization and besides clinical usefulness has not been firmly elucidated in Korean HBV patients. METHODS: We compared Bayer VERSANT HBV DNA 3.0 Assay (VERSANT 3.0) with Digene Hybrid Capture II HBV DNA Test (HC-II) according to HBeAg status and ALT levels in 232 HBV infected Korean patients. One hundred and seventeen sera with undetectable DNA levels by HC-II were further analyzed by Real-Q HBV quantification assay (BioSewoom). RESULTS: Although VERSANT 3.0 and HC-II showed an excellent correlation (r=0.9739), the results (copies/mL) by VERSANT 3.0 were 0.45 log(10) higher than those by HC-II. HBV DNA levels were higher in HBeAg-positive group than in HBeAg-negative group (P=0.002), and in abnormal ALT group than in normal ALT group (P<0.0001). The detection rate of HBV DNA by VERSANT 3.0 was lower in HBeAg-negative and normal ALT group (n=68) than in HBeAg-positive or abnormal ALT group (n=164) (35.3% vs 89.6%, P<0.0001). Fifty two sera out of 61 sera with undetectable DNA by VERSANT 3.0 were measurable by Real-Q with mean value of 3.26 log(10) copies/mL. CONCLUSIONS: VERSANT 3.0 and HC-II showed an excellent correlation, but a little difference (0.45 log10) existed. VERSANT 3.0 effectively measured clinically relevant HBV DNA levels in most HBV-infected patients in Korea. However, more sensitive assays are needed for patients with negative HBeAg and normal ALT to see the low copies of HBV DNA levels. PMID- 18160837 TI - [Development of a web-based program for the identification of human leukocyte antigen antibody specificities]. AB - BACKGROUND: Panel reactive antibody (PRA) test is used to determine whether a patient awaiting transplantation is previously sensitized. Tail analysis algorithm is widely used to identify antibody specificities, but it is very difficult to perform manually. METHODS: To develop a web-based program, PHP (5.1.2), Apache (2.0.55), and MySQL (5.0.22) were used. Tail analysis algorithm was applied to identify specificities, which analyzed statistically 2 x 2 tables representing reactivities to broad antigens, splits and cross reactive groups (CREG). Exploiting two CREG classifications of Rodey (R) and Takemoto (T), antibody specificities were identified by 3 methods (ABC, R-ABC, T-ABC) simultaneously. Performance of the system was evaluated using 159 samples that showed > or =6 PRA% by a lymphocytotoxicity assay. RESULTS: A web-based system that can identify HLA antibody specificities was implemented on www.koreanhla.com. Among 159 samples tested, antibody specificities were identified in 151 (95.0 %), but not in 8 samples with PRA >97%. Among the 151 samples, 110 showed broad or split specificities and 41 CREG specificities. CONCLUSIONS: We developed a web-based computer program for the identification of HLA antibody specificities. Accessible to everyone on the internet, this program should be of help in sharing PRA results among laboratories. PMID- 18160839 TI - Mass spectrometry based proteomic analysis of human stem cells: a brief review. AB - Stem cells can give rise to various cell types and are capable of regenerating themselves over multiple cell divisions. Pluripotency and self-renewal potential of stem cells have drawn vast interest from different disciplines, with studies on the molecular properties of stem cells being one example. Current investigations on the molecular basis of stem cells pluripotency and self-renewal entail traditional techniques from chemistry and molecular biology. In this mini review, we discuss progress in stem cell research that employs proteomics approaches. Specifically, we focus on studies on human stem cells from proteomics perspective. To our best knowledge, only the following types of human stem cells have been examined via proteomics analysis: human neuronal stem cells, human mesenchymal stem cells, and human embryonic stem cells. Protein expression serves as biomarkers of stem cells and identification and expression level of such biomarkers are usually determined using two-dimensional electrophoresis coupled mass spectrometry or non-gel based mass spectrometry. PMID- 18160838 TI - DNA vaccines for cervical cancer: from bench to bedside. AB - More than 99% of cervical cancers have been associated with human papillomaviruses (HPVs), particularly HPV type 16. The clear association between HPV infection and cervical cancer indicates that HPV serves as an ideal target for development of preventive and therapeutic vaccines. Although the recently licensed preventive HPV vaccine, Gardasil, has been shown to be safe and capable of generating significant protection against specific HPV types, it does not have therapeutic effect against established HPV infections and HPV-associated lesions. Two HPV oncogenic proteins, E6 and E7, are consistently co-expressed in HPV expressing cervical cancers and are important in the induction and maintenance of cellular transformation. Therefore, immunotherapy targeting E6 and/or E7 proteins may provide an opportunity to prevent and treat HPV-associated cervical malignancies. It has been established that T cell-mediated immunity is one of the most crucial components to defend against HPV infections and HPV-associated lesions. Therefore, effective therapeutic HPV vaccines should generate strong E6/E7-specific T cell-mediated immune responses. DNA vaccines have emerged as an attractive approach for antigen-specific T cell-mediated immunotherapy to combat cancers. Intradermal administration of DNA vaccines via a gene gun represents an efficient way to deliver DNA vaccines into professional antigen-presenting cells in vivo. Professional antigen-presenting cells, such as dendritic cells, are the most effective cells for priming antigen-specific T cells. Using the gene gun delivery system, we tested several DNA vaccines that employ intracellular targeting strategies for enhancing MHC class I and class II presentation of encoded model antigen HPV-16 E7. Furthermore, we have developed a strategy to prolong the life of DCs to enhance DNA vaccine potency. More recently, we have developed a strategy to generate antigen-specific CD4(+) T cell immune responses to further enhance DNA vaccine potency. The impressive pre- clinical data generated from our studies have led to several HPV DNA vaccine clinical trials. PMID- 18160840 TI - The association of Pro12Ala polymorphism of peroxisome proliferator-activated receptor-gamma gene with serum osteoprotegerin levels in healthy Korean women. AB - Recent evidences suggest that the activation of peroxisome proliferator-activated receptor (PPAR)-gamma, which is an important transcriptional factor in adipocyte differentiation, also plays an important role in the bone microenvironment. The objective of the study was to clarify whether Pro12Ala polymorphism was related to the serum OPG levels and bone mineral metabolism in healthy Korean women. In 239 Korean women (mean age 51 years), who participated in medical check-up program in a health promotion center, anthropometric measurements, lumbar spine and femoral neck bone mineral density (BMD), bone turnover markers, such as serum total alkaline phosphatase (ALP) levels, urine deoxypyridinoline levels, and 24-h urine calcium excretion were measured. Serum levels of OPG were measured with ELISA method. DNAs were extracted from the samples and the genotyping of the Pro12Ala polymorphism (rs1801282) in the PPAR-gamma gene was performed via an allelic discrimination assay using a TaqMan probe. In addition, we examined the haplotype analysis between two polymorphisms of PPAR-gamma gene, Pro12Ala in exon B and C161T in exon 6 (rs3856806). Allelic frequencies were 0.950 for Pro allele and 0.050 for Ala allele, which was in compliance with Hardy- Weinberg equilibrium, and there was no Ala12Ala genotype among the genotyped subjects. Mean serum OPG level was significantly lower (P=0.035), and serum total ALP was significantly higher (P=0.014) in the Pro12Ala genotype group compared with the Pro12Pro genotype group, which were consistently significant even after adjustment for weight, height, and serum follicle stimulating hormone (FSH). In multiple regression analysis with serum OPG as the dependent variable and age, weight, ALP, femoral neck BMD and Pro12Ala genotype included in the model, only Pro12Ala genotype was significant determinant of serum OPG level (b=??0.136, P=0.035). The haplotype analysis with C161T polymorphism revealed that subjects with Ala and T alleles showed significantly lower serum OPG levels compared with those with Pro12Pro/CC genotype, which were consistently significant even after adjustment for age, weight, height and FSH (P=0.010). This result suggests statistically significant association of Pro12Ala polymorphisms with serum OPG levels in Korean females. PMID- 18160841 TI - Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death. AB - Although N-myc downstream regulated gene 2 (NDRG2) has been known to be a tumor suppressor gene, the function of this gene has not been elucidated. In the present study, we investigated the expression and function of NDRG2 in human gastric cancer. Among seven gastric cancer and two non-cancer cell lines, only two gastric cancer cell lines, SNU-16 and SNU-620, expressed NDRG2, which was detected in the cytoplasm. Interestingly, NDRG2 was highly expressed in normal gastric tissues, but gastric cancer patients were divided into NDRG2-positive and -negative groups. The survival rate of NDRG2-negative patients was lower than that of NDRG2-positive patients. We confirmed that the loss of NDRG2 expression was a significant and independent prognostic indicator in gastric carcinomas by multivariate analysis. To investigate the role of NDRG2 in gastric cancer cells, we generated a NDRG2-silenced gastric cancer cell line, which stably expresses NDRG2 siRNA. NDRG2-silenced SNU-620 cells exhibited slightly increased proliferation and cisplatin resistance. In addition, inhibition of NDRG2 decreased Fas expression and Fas-mediated cell death. Taken together, these data suggest that inactivation of NDRG2 may elicit resistance against anticancer drug and Fas-mediated cell death. Furthermore, case studies of gastric cancer patients indicate that NDRG2 expression may be involved in tumor progression and overall survival of the patients. PMID- 18160842 TI - Celastrol inhibits production of nitric oxide and proinflammatory cytokines through MAPK signal transduction and NF-kappaB in LPS-stimulated BV-2 microglial cells. AB - Excessive production of nitric oxide (NO) and proinflammatory cytokines from activated microglia play an important role in human neurodegenerative disorders. Here, we investigated whether celastrol, which has been used as a potent anti inflammatory and anti-oxidative agent in Chinese medicine, attenuates excessive production of NO and proinflammatory cytokines such as TNF-alpha and IL-1betal in LPS-stimulated BV-2 cells, a mouse microglial cell line. We report here that the LPS-elicited excessive production of NO, TNF-alpha, and IL-1beta in BV-2 cells was largely inhibited in the presence of celastrol, and the attenuation of inducible iNOS and these cytokines resulted from the reduced expression of mRNAs of iNOS and these cytokines, respectively. The molecular mechanisms that underlie celastrol-mediated attenuation were the inhibition of LPS-induced phosphorylation of MAPK/ERK1/2 and the DNA binding activity of NF-kappaB in BV-2 cells. The results indicate that celastrol effectively attenuated NO and proinflammatory cytokine production via the inhibition of ERK1/2 phosphorylation and NF-kappaB activation in LPS-activated microglia. Thus, celastrol may be an effective therapeutic candidate for use in the treatment of neurodegenerative human brain disorders. PMID- 18160843 TI - Gene expression responses in vivo by human telomerase reverse transcriptase (hTERT)-targeting trans-splicing ribozyme. AB - A trans-splicing ribozyme which can specifically reprogram human telomerase reverse transcriptase (hTERT) RNA was previously suggested as a useful agent for tumor-targeted gene therapy. In this study, we evaluated in vivo function of the hTERT-targeting trans-splicing ribozymes by employing the molecular analysis of expression level of genes affected by the ribozyme delivery into peritoneal carcinomatosis mice model. To this effect, we constructed adenoviral vector encoding the specific ribozyme. Noticeably, more than four-fold reduction in the level of hTERT RNA was observed in tumor nodules by the systemic infection of the ribozyme-encoding virus. Such hTERT RNA knockdown in vivo induced changes in the global gene expression profile, including the suppression of specific genes associated with anti-apoptosis including bcl2, and genes for angiogenesis and metastasis. In addition, specific trans-splicing reaction with the targeted hTERT RNA took place in the tumors established as peritoneal carcinomatosis in mice by systemic delivery of the ribozyme. In conclusion, this study demonstrates that an hTERT-specific RNA replacement approach using trans-splicing ribozyme represents a potential modality to treat cancer. PMID- 18160844 TI - Blockade of airway inflammation and hyper-responsiveness by an angiopoietin-1 variant, COMP-Ang1. AB - Inflammation of the asthmatic airway is usually accompanied by increased vascular permeability and plasma exudation. Angiopoietin-1 (Ang1) has potential therapeutic applications in preventing vascular leakage. Recently, we developed a soluble, stable, and potent Ang1 variant, COMP-Ang1. COMP-Ang1 is more potent than native Ang1 in phosphorylating the tyrosine kinase with immunoglobulin and epidermal growth factor homology domain 2 receptor in lung endothelial cells. We have used a mouse model for allergic airway disease to determine effects of COMP Ang1 on allergen-induced bronchial inflammation and airway hyper-responsiveness. These mice develop the following typical pathophysiological features of allergic airway disease in the lungs: increased numbers of inflammatory cells of the airways, airway hyper-responsiveness, increased levels of Th2 cell cytokines (IL 4, IL-5, and IL-13), adhesion molecules (intercellular adhesion molecule-1 and vascular cell adhesion molecule-1), and chemokines (eotaxin and RANTES), and increased vascular permeability. Intravenous administration of COMP-Ang1 reduced bronchial inflammation and airway hyper-responsiveness. In addition, the increased plasma extravasation in allergic airway disease was significantly reduced by the administration of COMP-Ang1. These results suggest that COMP-Ang1 attenuates airway inflammation and hyper-responsiveness, prevents vascular leakage, and may be used as a therapeutic agent in allergic airway disease. PMID- 18160845 TI - Detection of homodimer formation of CD99 through extracelluar domain using bimolecular fluorescence complementation analysis. AB - Although various functions of CD99 have been reported, such as apoptosis and homotypic aggregation of thymocyte and transendothelial migration of immune cells, biochemical/molecular natures of CD99 are still elusive. Using mouse CD99 gene, we show that CD99 forms homodimer through its extracellular domain. Expression of mouse CD99 is up-regulated on T cells after CD3-mediated activation, like the case for human CD99. The potential of CD99 to form homodimer was tested with a recently developed bimoleular fluorescence complementation analysis (BiFC). In BiFC analysis, the dimerization-induced fluorescence was strong near the perinuclear region and was faded at the cell membrane. However, surface expression of CD99 was still detected by flow cytometry, suggesting that CD99 either in monomer form or in association with other molecules exists on the cell surface. In BiFC analysis using CD99 mutants with its extracellular, transmembrane, or cytosolic domains changed to corresponding human CD4 domains, the mutant replaced with human CD4-extracellular domain did not produce fluorescence. Purified soluble CD99-Fc fusion proteins bound to CD99-Fc immobilized onto the gold sensor chip in surface plasmon resonance analysis, confirming that the extracellular domain was responsible for dimer formation. Intracytoplasmic staining for CD99 expression in the thymocytes and mature T cells showed that most of the cells, even the cells with low surface level of CD99, contained the molecule inside the cell. Our results suggest that majority of CD99 homodimers may exit in the cell and be exported to the cell surface, dissociating from each other, after a certain regulatory signal is delivered. PMID- 18160846 TI - A novel thiol compound, N-acetylcysteine amide, attenuates allergic airway disease by regulating activation of NF-kappaB and hypoxia-inducible factor 1alpha. AB - Reactive oxygen species (ROS) play an important role in the pathogenesis of airway inflammation and hyperresponsiveness. Recent studies have demonstrated that antioxidants are able to reduce airway inflammation and hyperreactivity in animal models of allergic airway disease. A newly developed antioxidant, small molecular weight thiol compound, N-acetylcysteine amide (AD4) has been shown to increase cellular levels of glutathione and to attenuate oxidative stress related disorders such as Alzheimer's disease, Parkinson's disease, and multiple sclerosis. However, the effects of AD4 on allergic airway disease such as asthma are unknown. We used ovalbumin (OVA)-inhaled mice to evaluate the role of AD4 in allergic airway disease. In this study with OVA-inhaled mice, the increased ROS generation, the increased levels of Th2 cytokines and VEGF, the increased vascular permeability, the increased mucus production, and the increased airway resistance in the lungs were significantly reduced by the administration of AD4. We also found that the administration of AD4 decreased the increases of the NF kappaB and hypoxia-inducible factor-1alpha (HIF-1alpha) levels in nuclear protein extracts of lung tissues after OVA inhalation. These results suggest that AD4 attenuates airway inflammation and hyperresponsiveness by regulating activation of NF-kappaB and HIF-1alpha as well as reducing ROS generation in allergic airway disease. PMID- 18160847 TI - Comparative bioinformatics analysis of prion proteins isolated from reptile, rodent, ruminant, and human species. AB - Prion proteins (PrPs) are infectious pathogens that cause a group of invariably fatal, neurodegenerative diseases, including Creutzfeldt-Jakob disease, by means of an entirely novel mechanism. They are produced by various species, including reptile, rodent, ruminant and mammals, during normal metabolic processes, but they can be slowly changed into pathogenic isoforms upon contact with other infectious PrP isoforms. This transmission can occur across species barriers. In the present study, phylogram for each PrP sequence was generated by PAUP* 4.0 program using Neighbor-Joining method with 1,000 times bootstrapping process for the phylogenetic analysis. The molecular dynamics (MD) simulations were performed by the SANDER module in the AMBER 7 package using Amber 99 force field. All the simulation process was conducted in the IBM p690 Supercomputing System in Korea Institute of Science and Technology Information. To reduce the calculation time, we used the Generalized Born (GB) model. We compared the sequences and structural characteristics of normal and pathogenic (E200K) human PrPs with those of other reptile, rodent, ruminant and mammalian PrPs. Phylogenetic analysis revealed that, although the turtle PrP sequence is the most distinct of the PrPs analyzed, it nonetheless retains five conserved secondary structural elements that are similar to those found in the mammalian PrPs, suggesting that these elements have important functions in vivo. The RMS deviation between the normal and E200K human PrPs was larger than that between the normal human and bovine PrPs, and all of the beta-sheet structures in human E200K PrP were very stable during MD simulations. PMID- 18160848 TI - Suppression of HIV-1 Tat-induced monocyte adhesiveness by a cell-permeable superoxide dismutase in astrocytes. AB - HIV-1 Tat is considered to be one of key players to facilitate monocyte entry into the CNS, which is characteristic feature of AIDS-related encephalitis and dementia. This study was performed to determine the regulatory function of superoxide dismutase (SOD) on the HIV-1 Tat-induced signaling pathways leading to NF-kappaB activation, expression of adhesion molecules, and monocyte adhesion in CRT-MG human astroglioma cells by using cell-permeable SOD. When cell-permeable SOD was added to the culture medium of CRT-MG cells, it rapidly entered the cells in dose- and time-dependent manners. Treatment of astrocytes with cell-permeable SOD led to decrease in Tat-induced ROS generation as well as NF-kappaB activation. Cell-permeable SOD inhibited the activation of MAP kinases including ERK, JNK and p38 by HIV-1 Tat. Treatment of CRT-MG cells with cell-permeable SOD significantly inhibited protein and mRNA levels of ICAM-1 and VCAM-1 up-regulated by HIV-1 Tat, as measured by Western blot analysis and RT-PCR. Furthermore, enhanced adhesiveness of monocyte to astrocyte by HIV-1 Tat was significantly abrogated by pretreatment with cell-permeable SOD fusion proteins. These data indicate that SOD has a regulatory function for HIV-1 Tat-induced NF-kappaB activation in astrocytes and suggest that cell-permeable SOD can be used as a feasible therapeutic agent for regulation of ROS-related neurological diseases. PMID- 18160849 TI - Identification of a kidney-specific mouse organic cation transporter like-1 (mOCTL1). AB - Organic ion transporters are expressed in various tissues that transport endogenous and exogenous compounds including their metabolites. There are organic anion transporter (OAT), organic cation transporter (OCT), organic anion transporter like protein (OATLP) and organic cation transporter like (OCTL). Considering the variety of charged organic ionic compounds, the existence of numerous isoforms of organic ion transporters can be assumed. In the present study, we have searched for a new isoform in the expressed sequence tag (EST) database using human organic anion transporter 4 (hOAT4) amino acid sequence as a "query". We found a candidate clone (BC021449) from the mouse kidney cDNA library. This clone was identified as an ortholog of ORCTL3 or OCTL-1. The mOCTL1 cDNA consists of 2016 base pairs encoding 551 amino acid residues with 12 putative transmembrane domains. The deduced amino acid sequence of mOCTL1 showed 35 to 40% identity to those of the other members of the OATs and OCTs. According to the tissue distribution, examined by Northern blot analysis, about a 2.4-kb transcript of mOCTL1 was observed in the kidney. About a 90-kDa band was detected when Western blot analysis in the mouse kidney was done by using antibody against synthesized oligopeptide of mOCTL1. The immunohistochemical result showed that mOCTL1 was stained at the glomerulus (the parietal epithelial cells and podocytes), pars recta of proximal tubule, distal convoluted tubules, connecting tubules and collecting tubules. From these results, we conclude that mOCTL1 may be a candidate for an organic ion transporter isoform in the mouse kidney. PMID- 18160850 TI - Downregulation of regenerating islet-derived 3 alpha (REG3A) in primary human gastric adenocarcinomas. AB - Gastric carcinoma is considered to be one of the most prevalent cancers worldwide. We have performed differential-display polymerase chain reaction (DD PCR) in order to compare the gene expression profile of gastric carcinoma and that of a normal stomach, in an attempt to identifiy differentially expressed genes associated with primary human gastric cancers. One of the down-regulated genes in gastric cancers was identified as regenerating islet-derived 3 alpha (REG3A), also known as hepatocarcinoma-intestine-pancreas/ pancreatitis associated protein (HIP/PAP). REG3A exhibited relatively high expression levels in normal gastric mucosa. However, REG3A was found to be down-regulated in 67% (20 out of 30 samples) of primary human gastric cancers, as determined by RT-PCR. In addition, REG3A mRNA expression was not detected in stomach cancer cell lines, SNU cells. Immunohistochemical analysis further confirmed the down-regulation of REG3A expression in primary human gastric cancers. Treatment with the demethylating agent, 5-aza-2'-deoxycytidine (5-Aza-dC) resulted in the restoration of REG3A mRNA expression in the gastric cancer cell line, indicating that the transcriptional silencing of REG3A in SNU cell lines was caused by DNA methylation. Taken together, these data indicate that REG3A is down-regulated in most primary human gastric cancer cells, and might be useful in the diagnosis of gastric cancer. Further characterization of the differentially expressed gene, REG3A, should lead to a better understanding of the changes occurring at the molecular level during the development and progression of primary human gastric cancer. PMID- 18160851 TI - Association of FOXJ1 polymorphisms with systemic lupus erythematosus and rheumatoid arthritis in Korean population. AB - The forkhead-box J1 (FOXJ1) transcription factor could suppress a spontaneous activation of T cells and B cells through an induction of IkappaBbeta that results in repression of NF-kappaB activity. In Foxj1 deficiency mice, systemic autoimmune inflammation is quite common symptom. Therefore, deregulated Foxj1 is supposed to be associated with autoimmune diseases and/or other inflammatory diseases. Previously, we identified that polymorphisms of human FOXJ1 gene (g.??460C>T, g.1805G>T and g.3375G>C) are associated with allergic rhinitis in a Korean population. In present study, we compared the genotype and allele frequencies of these SNPs between healthy controls and systemic lupus erythematosus (SLE) or rheumatoid arthritis (RA) patients. We also investigated the relationships between each genotype and the expression levels of anti- nuclear antibodies in SLE patients, and rheumatoid factor and anti-cyclic citrullinated peptide in RA patients. The frequencies of haplotypes constructed by these FOXJ1 SNPs were compared between controls and SLE (or RA) patients. The results of genotype and allele analysis showed that the prevalence of polymorphism g.3375G>C was associated with the susceptibility of SLE (P = 0.0072 and 0.0042, respectively). But no significant association was found with RA. In the haplotype analysis, however, the main CGG showed a weak association between controls and RA patients (P = 0.048). PMID- 18160852 TI - Interleukin-10 endogenously expressed in microglia prevents lipopolysaccharide induced neurodegeneration in the rat cerebral cortex in vivo. AB - A degree of brain inflammation is required for repair of damaged tissue, but excessive inflammation causes neuronal cell death. Here, we observe that IL-10 is expressed in LPS-injected rat cerebral cortex, contributing to neuronal survival. Cells immunopositive for IL-10 were detected as early as 8 h post-injection and persisted for up to 3 d, in parallel with the expression of IL-1beta, TNF-alpha, and iNOS. Double immunofluorescence staining showed that IL-10 expression was localized mainly in activated microglia. Next, we examined the neuroprotective effects of IL-10 using IL-10 neutralizing antibody (IL-10NA). Blockade of IL-10 action caused a significant loss of neurons both 3 d and 7 d after LPS injection. Further, the induction of mRNA species encoding IL-1beta, TNF-alpha, and iNOS, reactive oxygen species (ROS) production, and NADPH oxidase activation, increased after co-injection of LPS and IL-10NA, compared to the levels seen after injection of LPS alone. Taken together, these results clearly suggest that LPS induced endogenous expression of IL-10 in microglia contributes to neuronal survival by inhibiting brain inflammation. PMID- 18160853 TI - ATP released from beta-amyloid-stimulated microglia induces reactive oxygen species production in an autocrine fashion. AB - Present study demonstrated that fibrillar beta-amyloid peptide (fAbeta1-42) induced ATP release, which in turn activated NADPH oxidase via the P2X7 receptor (P2X7R). Reactive oxygen species (ROS) production in fAbeta1-42- treated microglia appeared to require Ca2+ influx from extracellular sources, because ROS generation was abolished to control levels in the absence of extracellular Ca2+. Considering previous observation of superoxide generation by Ca2+ influx through P2X7R in microglia, we hypothesized that ROS production in fAbeta-stimulated microglia might be mediated by ATP released from the microglia. We therefore examined whether fAbeta1-42-induced Ca2+ influx was mediated through P2X7R activation. In serial experiments, we found that microglial pretreatment with the P2X7R antagonists Pyridoxal-phosphate-6-azophenyl-2',4'- disulfonate (100 microM) or oxidized ATP (100 microM) inhibited fAbeta-induced Ca2+ influx and reduced ROS generation to basal levels. Furthermore, ATP efflux from fAbeta1-42- stimulated microglia was observed, and apyrase treatment decreased the generation of ROS. These findings provide conclusive evidence that fAbeta-stimulated ROS generation in microglial cells is regulated by ATP released from the microglia in an autocrine manner. PMID- 18160854 TI - The centrosomal localization of KM-HN-1 (MGC33607) depends on the leucine zipper motif and the C-terminal coiled-coil domain. AB - KM-HN-1 is a C-terminal coiled-coil domain containing protein previously referred to as image clone MGC33607. This protein has been previously identified as a cancer/testis antigen and reported as nuclear and chromatin localizing protein. We raised polyclonal antisera with the GST fusion protein and identified them as a 105 kDa protein. Motif analysis showed that this protein harbors the leucine zipper motif in internal 1/3 region and the coiled-coil domain in the C-terminal region. Using the full length and various deletion mutants, we determined the motif that governs the subcellular localization of KM-HN-1. Immunofluorescence staining of the endogenous KM-HN-1 and various kinds of GFP-tagged KM-HN-1 revealed that KM-HN-1 localizes to the centrosomes as well as nucleus. The centrosomal localization-determining region of this protein is C-terminal coiled coil domain in which the leucine zipper motif and the nuclear export signal (NES) harbor. PMID- 18160855 TI - Caring competencies for a complex healthcare environment: part 1. PMID- 18160856 TI - Take time to care. PMID- 18160857 TI - Leadership in caring. PMID- 18160858 TI - Measuring the emotional intelligence of clinical staff nurses: an approach for improving the clinical care environment. AB - BACKGROUND: Significant correlations have been demonstrated between emotional intelligence (EI) and high levels of performance and other organizational variables related to retention and workplace environment. Although these variables are also important for safe care and healthy work relationships, there is little research on the EI of clinical staff nurses. The purpose of this study was to measure EI as related to performance level of clinical staff nurses, and to collect data on important related career/organizational variables. METHODS: Thirty-six nurses in 3 urban hospitals in Honolulu, Hawaii, participated. EI, nursing performance level, and career/organizational variables were measured. DISCUSSION: Significant positive correlations between clinical performance level and EI scores were observed. However, a large percentage of the clinical staff demonstrated below average total EI scores and subscores (perceiving emotions, experiencing emotions). CONCLUSIONS: Findings from this study offer the first evidence that the performance level of clinical staff nurses correlates positively with EI. A high percentage of nurses in this study scored below average on both total EI score and important subscores. These findings warrant further investigation. EI may offer a framework and specific skills to enhance collaboration, positive conflict behaviors, and healthy relationships in the clinical care environment. PMID- 18160859 TI - Creating a profile of a nurse effective in caring. AB - BACKGROUND: Watson's Theory of Caring and Caritas proposes that caring and love potentiate healing. Creating a profile of the characteristics of a "Caritas nurse" has implications for integrating caring and love into patient care, and measuring the impact of caring on patient and operational outcomes. METHODS: This psychometric study examined the profile of nurses effective in caring. The Caring Factor Survey and the Healthcare Environment Survey were selected to measure caring and work environment conditions as reported by the patient and by the nurse, respectively. RESULTS: The results of this study revealed that nurses of all ages who received high scores in caring were most frustrated with the work environment, were most experienced, worked only the hours scheduled, were most affected by the relationship with the patient, derived the most enjoyment from the relationship with their coworkers, and provided continuity of care most consistently. DISCUSSION AND IMPLICATIONS: Findings from this study indicate that further inquiry into the profile of nurses effective in caring and evaluation of the presence of caring and love on patient outcomes is warranted. PMID- 18160860 TI - The dimensions of leadership: a foundation for caring competency. AB - The Institute of Medicine (as recorded in their publication, Keeping Patients Safe: Transforming the Work Environment of Nurses), the American Nurses Credentialing Center's Magnet Recognition Program, and the American Organization of Nurse Executives have all called for the attention of caring competencies for nurse leaders. These competencies include skills and behaviors such as communication, relationship management, and building and sustaining trust. The Center for Nursing Leadership's 9 dimensions of leadership-holding the truth, [recognizing the] intellectual and emotional self, discovery of potential, [the] quest for the adventure toward knowing, diversity as a vehicle to wholeness, appreciation of ambiguity, knowing something of life, holding multiple perspectives without judgment, and keeping commitments to oneself-are applied to the behavior and performance of nurse leaders across settings and scope of practice. This article illustrates the embodiment of caring competencies by nurse leaders through the use of the dimensions of leadership. PMID- 18160861 TI - Nurses supporting nurses: creating a mentoring program for staff nurses to improve the workforce environment. AB - BACKGROUND: The hospital workforce environment has been recognized as an important factor for nurse retention and patient safety, yet there is ongoing evidence that inadequate communication, intraprofessional oppression, and lack of collaboration and conflict resolution continue to disempower nurses and hinder improvement of workforce conditions. PURPOSE: A 3-year academic-hospital partnership developed and used a registered nurse (RN) mentor and advocacy program to improve the RN work environment and selected patient outcomes. METHOD: The partnership initiated mentor-mentee teams and a Workforce Environment Governance Board, and obtained preliminary data on outcomes related to mentor mentee teamwork, changes in the level of support within each unit, and the impact of improved working conditions on nurse and patient satisfaction, nurse vacancy and turnover rates, and 3 patient safety outcomes related to fall and pressure ulcer prevention and use of restraints. DISCUSSION: Dedicated mentors not only engaged in supporting fellow nurses but also assisted with enhancing the overall work environment for RNs. The partnership enhanced mutual respect between frontline RNs and managers, and allowed frontline RNs to improve the culture of support. CONCLUSIONS: The comprehensive approach to incorporating mentor-mentee teams changes the way fellow nurses and others perceive nurses, augments support by managers and coworkers, and improves patient care outcomes. PMID- 18160862 TI - Interdisciplinary collaboration for healthcare professionals. AB - Interdisciplinary collaboration has the capacity to affect both healthcare providers and patients. Research has shown that the lack of communication and collaboration may be responsible for as much as 70% of the adverse events currently reported. The purpose of this article is 2-fold: to examine factors that may influence interdisciplinary collaboration and consequently patient outcomes and to examine the relationship between interdisciplinary collaboration and King's theory of goal attainment as a theory to support the phenomenon of interdisciplinary collaboration. PMID- 18160863 TI - Organizational culture: the silent political force. AB - Organizational cultures are affected by, and affect, organization politics. Underlying all cultures is the values of the individuals in the organization. These need to be identified and understood by anyone who seeks to influence or change behaviors. This article describes a series of studies done to increase the understanding of values of professional nurses. The results point out differences between specialties, organizations, and communities that can help leaders make better decisions in areas ranging from staffing to leadership styles, while assisting colleagues in the expression of their authentic selves. PMID- 18160864 TI - Exploring a culture of caring. AB - AIM: The delivery of patient-centered care is basic to a large midwestern healthcare institution's mission and highly valued by the department of nursing. Even so, nurses on one medical unit questioned whether caring behaviors were devalued in a technology-oriented environment of providing care. The nursing leadership on the unit responded to the inquiry by conducting a research study. This study explored the state of patient-centered nursing care on a medical unit as perceived by the nursing staff and patients, using Watson's Theory of Human Caring as a framework. SUBJECTS AND METHODS: The study utilized surveys for both nursing staff (n = 31) and patients (n = 62), and included a focus group of nursing staff (n = 8) to explore ideas for innovation. RESULTS AND CONCLUSIONS: Both nurses and patients perceived a high level of caring on the unit. The overall theme from the focus group was that "caring begets caring," with 2 subthemes: "relationships of care" and "the context of caring." Caring for each other was identified as essential to keep staff energized and able to work lovingly with patients. Nursing leadership brought the research findings to all staff on the unit for discussion and implementation of structural support for the unit culture of caring. PMID- 18160865 TI - Evoking the essence of caring: experiences from a Peruvian adventure relevant for leaders in healthcare today. AB - There are times when an experience is so powerful that it evokes caring in its purest form. This is what happened when a group of healthcare providers traveled together on a trip to Peru. This article recounts a harrowing experience that called each person to give caring and receive learning regarding his or her own journey. It is about how caring can simply happen with ease and purpose. Communication and collaboration occurred effortlessly. In this experience, there was a remembering of the passion and purpose that exists when we are called to serve in healthcare settings. The learning gained included 4 key elements of caring: intent, presence, action, and surrender. This learning offers insight, direction for healthcare environments today. Healthcare leaders must be responsive to creating cultures and providing opportunities to support those serving patients to intentionally think about their caring approaches. It is possible to evoke caring through providing opportunities that help healthcare providers transcend the daily barriers to being present and be more effective in their caring. It is imperative that they experience ways to examine their ability to care for themselves and others and to reconnect to the passion of choosing the healthcare profession. PMID- 18160866 TI - Caring communications: how technology enhances interpersonal relations, part I. AB - It can help improve communication, collaboration, conflict resolution in healthcare by creating a foundation of caring capabilities. PMID- 18160867 TI - Complex healthcare regulation: managing the continued challenge of competence. PMID- 18160872 TI - We were Wong. PMID- 18160876 TI - On alert for postpartum C. sordellii infection. PMID- 18160878 TI - Waking up to hospital bed entrapment risks. PMID- 18160879 TI - How chronic pain affects sexuality. PMID- 18160880 TI - Administering an orally disintegrating tablet. PMID- 18160881 TI - Reporting hollow threats. PMID- 18160882 TI - Putting a dent in pressure ulcer rates. PMID- 18160884 TI - Don't let the bedbugs bite. PMID- 18160885 TI - Assessing blood flow with CT angiography. PMID- 18160886 TI - Myths & facts... About obstructive sleep apnea. PMID- 18160888 TI - Why can't I hear this heart murmur? PMID- 18160890 TI - Women & heart disease: What's new? AB - Find out by reading this analysis of the new AHA guidelines for assessing and managing risk. PMID- 18160892 TI - Transferring a patient from bed to stretcher. PMID- 18160893 TI - Wonder woman. Like my childhood heroine, Mrs. Betty also had amazing powers. PMID- 18160894 TI - Keeping an eye on abnormalities. Use these photos to help identify some common abnormalities of the eyelids and eyes. PMID- 18160895 TI - Keep your patient safe during PCA. AB - Go beyond the push-button approach to patient-controlled analgesia and learn what it takes to keep him both safe and pain-free. PMID- 18160901 TI - Choosing a wound dressing, part 1. PMID- 18160903 TI - Frostbite. PMID- 18160905 TI - Anatomy and surgical approaches of the temporal bone and adjacent areas. PMID- 18160922 TI - The natural history of back pain after a randomised controlled trial of acupuncture vs usual care--long term outcomes. AB - INTRODUCTION: There is growing evidence about the effectiveness of acupuncture in the short term treatment of chronic low back pain but little is known about long term outcomes. To address this question we followed up participants of a past randomised controlled trial of acupuncture to assess outcomes after 5.5 to 7 years. METHODS: A postal questionnaire assessing pain, quality of life, disability, experience with back pain and healthcare resource use was sent to all 239 participants of the York Acupuncture for Back Pain trial. RESULTS: Response to the survey was low at 43.9%. Pain measured by the SF-36 Bodily Pain dimension was maintained in the acupuncture group since the last follow up 3.5 to 5 years previously. The usual care group had improved over the intervening years and there was now no evidence of a difference between the groups (difference -0.4 points, 95% confidence interval -10.1 to 9.7). The results were unchanged on sensitivity analysis using multiple imputation. In both groups back pain had not completely resolved and worry about back health was moderate. Physiotherapy and acupuncture were used at similar rates for continuing treatment. DISCUSSION: We theorise that exposure to a short course of acupuncture speeds natural recovery from a back pain episode, but improvements plateau after two years. Acupuncture is often accessed privately for long term management of back pain but is rarely available within the health service. While our study methods were robust, the low response rate means that our findings should be interpreted with caution. PMID- 18160923 TI - Acupuncture needling versus lidocaine injection of trigger points in myofascial pain syndrome in elderly patients--a randomised trial. AB - AIM: To compare the efficacy of acupuncture needling and 0.5% lidocaine injection of trigger points in myofascial pain syndrome of elderly patients. METHODS: Thirty nine participants with myofascial pain syndrome of one or both upper trapezius muscles were randomised to treatment with either acupuncture needling (n=18) or 0.5% lidocaine injection (n=21) at all the trigger points on days 0, 7 and 14, in a single-blinded study. Pain scores, range of neck movement, pressure pain intensity and depression were measured up to four weeks from the first treatment. RESULTS: Local twitch responses were elicited at least once in 94.9% of all subjects. Both groups improved, but there was no significant difference in reduction of pain in the two groups at any time point up to one month. Overall, the range of cervical movement improved in both groups, apart from extension in the acupuncture needling group. Changes in depression showed only trends. CONCLUSION: There was no significant difference between acupuncture needling and 0.5% lidocaine injection of trigger points for treating myofascial pain syndrome in elderly patients. PMID- 18160924 TI - Acupuncture in the treatment of fibromyalgia in tertiary care--a case series. AB - AIMS: Fibromyalgia is a common cause of chronic widespread pain. The benefit of medication is often limited by its side effects, and the improvements obtained with exercise and education are inconsistent. Many patients seek acupuncture treatment, which is reported to be helpful in some cases. This study aimed to explore the acceptability and benefits of acupuncture offered in the setting of a tertiary referral clinic. METHODS: An open, uncontrolled observational study was conducted among patients who met the usual fibromyalgia criteria and who had a pain score of at least 30 on a 100mm Visual Analogue Scale (VAS). Patients were allowed to continue other treatments but not to introduce new ones. Acupuncture was given using a Western approach according to a protocol developed by consensus. Patients were offered eight treatments in eight weeks. Outcome measures included VAS of pain intensity and Fibromyalgia Impact Questionnaire (range 0 - 100), and were taken before and after treatment, and at 14, 20 and 34 weeks from enrolment. RESULTS: Twenty four eligible patients were enrolled in a 12 month period. Baseline mean pain VAS score for these 24 patients was 74 (SD 18) and mean Fibromyalgia Impact Questionnaire score 78 (SD 12.4). Only 14 patients completed the course of treatment within about 10 weeks. Compliance was poor in the remaining patients because of difficulty attending clinic, and in two cases because of exacerbation of pain. Completion of outcome measures was variable and therefore the analysis of data is limited. Five patients scored at least 20% reduction in Fibromyalgia Impact Questionnaire score which is a clinically relevant improvement. Two of these scored at least 50% reduction. CONCLUSION: Acupuncture appears to offer symptomatic improvement to some patients with fibromyalgia in a tertiary clinic who have failed to respond to other treatments. In view of its safety, further acupuncture research is justified in this population. PMID- 18160925 TI - Intramuscular and periosteal acupuncture for anxiety and sleep quality in patients with chronic musculoskeletal pain--an evaluator blind, controlled study. AB - OBJECTIVES: Periosteal acupuncture seems clinically to have a stronger effect on pain than standard intramuscular acupuncture. The aim of the present study was to compare their relative effectiveness on promoting psychological functioning and quality of sleep. METHODS: Consecutive patients (n=144) referred to a physiotherapist for treatment of chronic nociceptive pain in the neck or low back for more than three months, aged 18-70, were alternately allocated to an intramuscular acupuncture group (n=59), to a periosteal acupuncture group (n=55), and, for the latter part of the study, to an information control group (n=30). Eight treatments were administered during a five week period with two optional treatments after one month. All patients were encouraged to stay active. Psychological functioning was estimated with the Hospital Anxiety and Depression Scale (HAD) and quality of sleep with a visual analogue scale. All estimations were performed prior to treatment, one week after the end of treatment, and one, three and six months after end of treatment. Non-parametric statistics were used. RESULTS: There were no significant differences between the acupuncture groups, nor between the acupuncture and control groups during the treatment period. One month after treatment, the level of anxiety was lower in both acupuncture groups compared with the control group. The proportions of non-cases (HAD score 0-7) increased in the intramuscular acupuncture group from 39 to 47 (not significant), in the periosteal acupuncture group from 37 to 49 (P<0.001), and in the control group from 15 to 16. CONCLUSION: No differences between periosteal and intramuscular acupuncture were found. One month after treatment both acupuncture interventions reduced anxiety in patients suffering from chronic nociceptive musculoskeletal pain in the neck or low back when compared with a control intervention. PMID- 18160926 TI - De qi: Chinese acupuncture patients' experiences and beliefs regarding acupuncture needling sensation--an exploratory survey. AB - INTRODUCTION: While de qi, the acupuncture needling sensation, has been considered as an important component of acupuncture, little is known of the acupuncture patient's experience and beliefs about de qi in clinical settings. The aim of this study was to describe Chinese acupuncture patients' perceived sensations of, and beliefs about, acupuncture needling. METHODS: We developed a questionnaire and conducted a survey study at two time periods among 200 subjects at six outpatient acupuncture clinics in Beijing, China. RESULTS: Respondents were 55% female and had a mean age of 41 years. The most common types of needling sensations reported by subjects were the terms 'distended' (94%), 'sore' (81%), 'electric' (81%) and 'numb' (78%). Eighty-nine percent of subjects reported that the needling sensation travelled away from the puncturing points or travelled among the needling points. Eighty-two percent of subjects believed that the needling sensation was very important for acupuncture treatment, and 68% further indicated that the stronger the needling sensation, the more effective the therapy. Eighty-one percent of subjects found the acupuncture process to be very comfortable and relaxing. CONCLUSION: Chinese acupuncture patients described the common characteristics of de qi and its migratory nature. The sensations were believed to be important in producing clinical efficacy by most patients. Measuring the sensations described as de qi in future prospective studies will help us understand the degree to which this phenomenon has an effect on the physiological outcome and clinical response to acupuncture. There appears to be a limit to the number of sensations that can be discriminated by each individual patient, and further development of the questionnaire is planned. PMID- 18160927 TI - Point injection as an alternative acupuncture technique--an exploratory study of responses in healthy subjects. AB - INTRODUCTION: Point injection as a therapeutic technique is well documented, but its physiological effects have not been formally compared with traditional acupuncture. One aim of this study was to compare the effects of the two techniques at one acupuncture point, as a step towards understanding the mode of action of point injection and validating its clinical use. A second aim was to explore whether repeated point injection at the same site might provide a way of increasing the dose of stimulation, in the hope of identifying a dose response curve which could be an alternative strategy to placebo control in demonstrating the biological effects of acupuncture. METHODS: Sixty nine healthy subjects (age range 18-56 years, mean 29.9; 48 females) completed the study, which employed a counterbalanced experimental design with two stimulation sessions of LI4 approximately one week apart. One half of the participants received point injection first, and the other half received traditional acupuncture first. Baseline physiological data were recorded, then measurements were made before, during and after stimulation; each subject also reported needle sensation (de qi). The measures were heart rate, derived pressure rate product and mean arterial pressure. RESULTS: Although stronger sensations of de qi were reported with point injection, no significant differences were found for mean heart rate (HR), pressure rate product (PRP) and mean arterial pressure (MAP) before and after stimulation by the two techniques. No subject gender or age bias was encountered and previous exposure to acupuncture had no effect on outcome. Power spectral analysis of heart rate variability (HRV) made on data from a small subset (n=10) of this cohort also showed no significant differences in autonomic response. CONCLUSION: Point injection and traditional acupuncture seem to provoke similar physiological responses, although the greater needle sensation seen with point injection might indicate it could have more powerful clinical effects. Further studies of repeated point injection are necessary to indicate whether this technique may provide a method of increased strength of point stimulation, as an alternative to traditional needling in acupuncture research. PMID- 18160928 TI - Analgesic effects of indirect moxibustion on an experimental rat model of osteoarthritis in the knee. AB - AIMS: The analgesic effects of moxibustion on an experimental model of osteoarthritis of the knee were investigated. METHODS: Male Wistar rats (n=36, 296-421 g) were used. Intra-articular injection of mono-iodoacetic acid (MIA) was performed to induce knee osteoarthritis. Indirect moxibustion was applied to the lateral aspect of the knee joint every other day for 28 days (14 treatments). Weight bearing of the hind legs was measured directly by the downward pressure applied to footplates, using an Incapacitance Tester. Morphine was injected for testing the validity of weight bearing as a pain measure, and naloxone was used to examine the participation of endogenous opioids in the mechanism of moxibustion analgesia. Data were analysed by calculating the area under the curve. RESULTS: Injection of MIA significantly reduced weight bearing. No analgesic effects of moxibustion were observed during the initial 7 days (unpaired t test, P=0.83). Continued moxibustion treatments increased weight bearing at the 14th day significantly, and this effect continued until the end of the experiment on the 28th day (P<0.05). A single moxibustion treatment had no immediate effect on weight bearing. The analgesia due to the cumulative effect of moxibustion was antagonised by naloxone injection. Morphine injection in control MIA injected rats increased weight bearing to the normal range, confirming the validity of the measurements. CONCLUSIONS: These results highlight the importance of repeated moxibustion treatments for pain relief in experimental knee osteoarthritis and suggest the existence of sustained inhibitory modulation by endogenous opioids in the moxibustion group. PMID- 18160929 TI - Did 'The Princess on the Pea' suffer from fibromyalgia syndrome? The influence on sleep and the effects of acupuncture. AB - Fibromyalgia syndrome (FMS) is a chronic pain syndrome characterised by central sensitisation resulting in hypersentivity of the skin and deeper tissues as well as fatigue. Possibly the princess in Hans Christian Andersen's 'The Princess and the Pea' suffered from FMS since chronic sleep disturbances are typical in FMS. These sleep disturbances have been attributed to a dysfunction in the systems regulating sleep and wakefulness resulting in loss of deep sleep. In addition, many patients with FMS experience cognitive dysfunction, characterised by impaired concentration and short term memory consolidation, a complaint also commonly reported in other sleep disorders. In recent reviews evaluating the efficacy of acupuncture in FMS it has been concluded that acupuncture has no specific effect. A prerequisite for this conclusion is that all the major symptoms in the syndrome have been assessed. However, previous studies have generally focused on the pain alleviating effect of acupuncture in FMS. We have observed that not only pain but also sleep and cognitive dysfunction may be ameliorated in response to acupuncture, suggesting that these variables should be taken into account when evaluating the effects of acupuncture in FMS. Furthermore, the results demonstrated great individual variability apart from the systematic effects related to the group, indicating that individually performed treatment strategies are required. Our suggestion is supported by experimental and clinical studies showing that acupuncture may affect insomnia and alertness, and that there may be neurophysiologic bases for these specific effects. PMID- 18160930 TI - Chronic prostatitis/chronic pelvic pain syndrome and acupuncture--a case report. AB - A 47 year old man had suffered from chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) for eight years, and received conventional treatment from a university clinic without effect during the entire period. The patient presented for acupuncture treatment, complaining of constant pain in the perineal region, varying in severity from 5 to 9 on a 0 to 10 pain scale. After three treatments with acupuncture, the pain was reduced to 3.5; after a further three treatments the pain was reduced to 2.5; and after 10 months, the pain was reduced to either 2 or 3 and has remained stable for two years since then. The acupuncture points used were BL28, BL29 and KI3, all located within the S2-S4 segments. It seems likely that the reported beneficial effect may be due to the acupuncture treatment and further cases should be studied. PMID- 18160931 TI - Treatment of juvenile stroke using Yamamoto New Scalp Acupuncture (YNSA) - a case report. AB - In Germany, stroke affects approximately 300 children each year. Treatment options for stroke patients, particularly for children, are limited. A case is described of an 18 month old girl with hemiparesis, who showed a considerable degree of recovery over the course of 21 months while receiving Yamamoto New Scalp Acupuncture using soft laser. This may represent a new option for the therapy of such patients. PMID- 18160932 TI - Use of di- and tripropionate substrate analogs to probe the active site of human recombinant coproporphyrinogen oxidase. AB - BACKGROUND: Defects in the enzyme coproporphyrinogen oxidase result in accumulation of porphyrins which may affect the severity of a subset of porphyrias. Thus evaluation of this enzyme for substrate selectivity is of value. Kinetic evaluations of recombinant human coproporphyrinogen oxidase have been undertaken using six di- and tripropionate analogs of the natural substrate coproporphyrinogen-III. These substrate analogs were modified by having alkyl groups in place of one or both of the ring 13- or 17-propionate moieties. MATERIAL/METHODS: Cloned human enzyme was incubated with analogs under apparent first order conditions and with various substrate concentrations. The kinetic values, K(m) and V(max), were determined. RESULTS: Relative to the authentic substrate, the K(m) values for the 13-ethyl, dimethyl and diethyl porphyrinogens were very comparable whereas the K(m) values were much higher using dipropyl and dibutyl porphyrinogen and much lower for the 17-ethyl analog. For the dipropionate analogs, the V(max) values were an apparent function of the carbon length of the substituent on the C and D rings, with longer carbon length severely reducing product formation by some 4-5 orders of magnitude. Also, the two isomeric tripropionates that were tested indicated that it was more detrimental to have an ethyl group at the 13-position for both binding and catalysis. CONCLUSIONS: This work extends our understanding of porphyrin ring substituent effects reported by Cooper et al. (2005). The substituents on both the C and D rings have significant effects on both the substrate binding and catalysis by this important enzyme. PMID- 18160933 TI - The protective effect of resveratrol on the intestinal mucosal barrier in rats with severe acute pancreatitis. AB - BACKGROUND: Severe acute pancreatitis (SAP) can result in intestinal mucosal barrier (IMB) dysfunction. The aim of this study is to demonstrate the protective effect of resveratrol (RES) on the IMB in rats with SAP. MATERIAL/METHODS: Fifty four male Sprague-Dawley (SD) rats were randomly divided into three equal groups: a sham-operated group (SO), a severe acute pancreatitis group (SAP), and a resveratrol-treated group (RES), each group containing 6 rats which were evaluated at 3, 6, and 12 h. Serum endotoxin levels were determined by turbidimetry. The apoptosis rate of intestinal mucosal cells was detected by using the TUNEL method and the expressions of the apoptosis-related proteins Bcl 2 and Bax were observed using an immunohistochemical technique. RESULTS: The serum endotoxin levels were significantly higher in the SAP group than in the SO group (P<0.05). The endotoxin levels of the RES group were lower than those of the SAP group at all the time-points (P<0.05). In the SAP group, pancreatic and intestinal mucosal congestion, edema, and inflammatory cell infiltration were apparent. In the RES group, pancreatic and intestinal morphological changes were alleviated at all time-points. The apoptotic cell index of the mucosal cells in the RES group was lower than that of the SAP group (P<0.05). In comparison with the SAP group, the RES group's expression of Bcl-2 increased and that of Bax decreased significantly (P<0.05). CONCLUSIONS: An increase in intestinal mucosal cell apoptosis is involved in the intestinal mucosal dysfunction of SAP rats. Resveratrol could decrease endotoxin translocation and protect IMB function. PMID- 18160934 TI - Effect of short-duration electrically induced tachycardia on regional myocardial contractility. AB - BACKGROUND: Tachycardia is a physiological mechanism for adapting cardiac output to modifications in energy consumption in the organism. The repercussions that short-duration tachycardia has on myocardial contractility have not been sufficiently studied.
To study the effects of short-duration tachycardia on regional myocardial function in the anterior face of the left ventricle depending on the rate, duration, and origin of the electrical stimulation producing the tachycardia. MATERIAL/METHODS: Two series were studied in an in vivo canine experimental model. Stimulation was performed in the right atrium and in the base of the left ventricle at a frequency 20 and 40% above the basal rate. The duration of each pacing episode was 10, 20, and 40 minutes followed by a recovery period of 40 minutes. ECG, left ventricular pressure, and regional function curves of a myocardial segment were studied. RESULTS: During atrial pacing there was a similar decrease in both regional segment lengths (end-systolic and end diastolic lengths) and no variation in the shortening fraction or in hemodynamic parameters. During ventricular pacing, systolic pressure in the left ventricle decreased, end-diastolic pressure increased, end-diastolic length shortened (meaning less ventricular filling), and the end-systolic length increased, leading to a reduced shortening fraction or regional contractility alteration which was temporarily maintained once pacing ceased. CONCLUSIONS: The transient persistence of segment dysfunction after induced ventricular tachycardia ceases indicates the activation of a different, unknown mechanism. PMID- 18160935 TI - Influence of classical and rock music on red blood cell rheological properties in rats. AB - BACKGROUND: A number of studies have reported physiological effects of music. Different types of music have been found to induce different alterations. Although some physiological and psychological parameters have been demonstrated to be influenced by music, the effect of music on hemorheological parameters such as red blood cell (RBC) deformability and aggregation are unknown. This study aimed at investigating the effects of classical and rock music on hemorheological parameters in rats. MATERIAL/METHODS: Twenty-eight rats were divided into four groups: the control, noise-applied, and the classical music- and rock music applied groups. Taped classical or rock music were played repeatedly for 1 hour a day for 2 weeks and 95-dB machine sound was applied to the noise-applied rats during the same period. RBC deformability and aggregation were measured using an ektacytometer. RESULTS: RBC deformability was found to be increased in the classical music group. Exposure to both classical and rock music resulted in a decrement in erythrocyte aggregation, but the decline in RBC aggregation was of a higher degree of significance in the classical music group. Exposure to noise did not have any effect on the parameters studied. CONCLUSIONS: The results of this study indicate that the alterations in hemorheological parameters were more pronounced in the classical music group compared with the rock music group. PMID- 18160936 TI - Differential and opposed transcriptional effects of interleukin-18 gene polymorphisms (-137, +113, and +127) in human HepG-2, HeLa, U937, and THP-1 cells. AB - BACKGROUND: Polymorphisms in the IL-18 promoter region (-137, +113, and +127), with perfect linkage disequilibria (Delta=1, p<0.001) among them, were observed in Singaporean Chinese, Malays, and Indians. These polymorphisms appear to have no association with atopic phenotypes. However, functional studies indicate that IL-18 gene polymorphisms strongly affect its activity in cultured cells. The reasons for such conflicting results remain to be determined. It is likely that they are related to the responding cells or to the varying cytokine milieus in the course of the atopic diseases. In this study the effects of IL-18 gene polymorphisms in various IL-18-producing cells were investigated. MATERIAL/METHODS: Three observed alleles were cloned and transfected into HepG-2, HeLa, U937, and THP-1 cells. The transfectional activities of the cultured cells were analyzed. RESULTS: Inserted fragments had significant, but opposite transfectional activity in HepG-2 and HeLa cells, while there was no difference in transfectional activity in U937 and THP-1 cells. CONCLUSIONS: These data support the authors' previous observation in which they were unable to detect an association between IL-18 gene polymorphisms and atopic phenotypes in this population. This might be due to different regulatory elements that affect the functional assessment of polymorphisms in different IL-18-producing cells. The functional significance of genetic variants in experimental studies may not be consistent due to complex human traits, but it can also be due to variations in the experimental approaches used, such as the use of different cell or tissue types. PMID- 18160937 TI - Physical activity assessed with three different methods and the Framingham Risk Score on 10-year coronary heart disease risk. AB - BACKGROUND: Physical activity (PA) protects against coronary heart disease (CHD) by favorably altering several CHD risk factors. In order to best understand the true nature of the relationship between PA and CHD, the impact different PA assessment methods have on the relationships must first be clarified. The purpose of this study was to examine associations between the Framingham Risk Score on 10 year estimated risk of CHD (10-year CHD risk) and three different PA assessment methods. MATERIAL/METHODS: Female (n=143) and male (n=155) adults wore activity monitors and recorded PA in logs during a seven-day period after which they completed a seven-day PA recall and had CHD risk factors assessed. The 10-year CHD risk was calculated from the CHD risk factors. RESULTS: Multiple regression analyses showed 10-year CHD risk was negatively associated with physical activity measured by the activity monitor (p<0.05) but not the log or recall in women. In men, significant inverse relationships between 10-year CHD risk and physical activity measured by the activity monitor and recall were observed. The log was not significantly associated with 10-year CHD risk in men. CONCLUSIONS: The results of this study concur with previous research in that PA is associated with the risk of developing CHD; however, the findings clearly suggest that it is important to consider the PA assessment method when interpreting this relationship. PMID- 18160938 TI - Idiopathic central precocious puberty in 28 boys. AB - BACKGROUND: Idiopathic central precocious puberty (CPP) is rare in boys. The aim was to to analyze the presentation and evaluate the frequency of familial factor in boys with idiopathic CPP. MATERIAL/METHODS: Data for 28 boys seen consecutively by the same physician for idiopathic CPP were analyzed. RESULTS: Puberty started after seven years in all the boys. The associations were intrauterine growth retardation in two, one of whom had Silver Russell syndrome, bilateral retinal degeneration (one case), epilepsy (one case), cryptorchidism (two cases), and inguinal hernia (two cases). All patients had normal basal plasma concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH). The LH/FSH peaks ratio after a gonadotropin hormone-releasing hormone (GnRH) test was <2 in 8/26 and plasma testosterone <0.5 ng/ml in 5/28. Familial early puberty was found in 12 cases (40%). Familial and non-familial forms had similar characteristics, except that body mass index was greater in the familial form (P<0.04). Plasma testosterone of one patient, whose mother had menstruated at 11 years, remained >1 ng/ml despite shortening the interval between GnRH analogue injections. CONCLUSIONS: Puberty started after seven years in all cases of idiopathic CPP, suggesting that pubertal onset before this age suggests organic CPP. Almost half the cases had familial early puberty. PMID- 18160939 TI - Intraobserver and interobserver variability of measuring waist circumference. AB - BACKGROUND: The measurement of waist circumference for assessing abdominal obesity has become widely accepted. MATERIAL/METHODS: To evaluate the reliability of measuring waist circumference, anthropometric parameters were measured in 150 adult patients by two diabetes educators on two consecutive days. The intraobserver difference was defined as the difference in the measurements made by the same educator on different days and the interobserver difference as the difference in the measurements made by the two educators on the same days. RESULTS: The correlation coefficients (r values) of the measurements were >0.99 for the educators and days. The interobserver difference was statistically significant for waist circumference (96.23 cm vs. 97.08 cm, p<0.0001). Although the %Delta values (percent difference in the two means) for waist circumference proved to be 2.5-6.3 times higher and the% absolute Delta values (percent average difference) 1.5-2.8 times higher than those for body mass index, the absolute values of the differences (Delta, the difference of the two means, and absolute delta, the average difference) were small (waist circumference Delta: 0.17 cm and 0.85 cm, absolute Delta: 1.51 cm and 2.15 cm; body mass index Delta: 0.02 kg/m(2) and 0.04 kg/m(2), absolute Delta: 0.292 kg/m(2) and 0.226 kg/m(2); intraobserver and interobserver differences, respectively). CONCLUSIONS: The intraobserver and interobserver variability for waist circumference were higher than those for body mass index. Nevertheless, the differences in repeated measurements of waist circumference were small when expressed in absolute values. The reliability of waist circumference should be considered in the clinical practice. PMID- 18160940 TI - Comparison of iron and hematological indices in Giardia lamblia infection before and after treatment in 102 children in Ahwaz, Iran. AB - BACKGROUND: Up to now there have been few reports about hematological alterations induced by Giardia lamblia infection. The aim of this study was to evaluate the effects of giardiasis treatment on hematological indices in children in the Khuzestan area by comparing these indices before and after treatment with metronidazole. MATERIAL/METHODS: One hundred two children ranging in age from 5 to 11 years infected with Giardia lamblia between July 2002 and July 2003 and without histories of other diseases were selected from kindergartens and primary schools in the city of Ahwaz. Diagnosis was based on stool examination. The children were treated with metronidazole (15 mg/kg/day for 5 days); hematological values were determined immediately before treatment and repeated 40-45 days after beginning treatment and compared. RESULTS: Iron deficiency anemia was found in 26.5% of the Giardia-infected children. Mean serum hemoglobin, serum hematocrit, serum ferritin, and the ratio of serum iron to total iron-binding capacity were increased (P<0.0001) and mean red-cell distribution width and iron-binding capacity were decreased after treatment (P<0.0001). CONCLUSIONS: Treating giardiasis in children with metronidazole improves iron indices and it appears that in evaluating anemia one should rule out giardiasis and treat it in asymptomatic cases to aid in better health status in pediatrics. PMID- 18160941 TI - Hysterectomy surveillance in the United States, 1997 through 2005. AB - BACKGROUND: To identify patterns and trends in hysterectomy from 1997 through 2005. MATERIAL/METHODS: Analyses are based on hysterectomy prevalence data from the Behavior Risk Factor Surveillance System, hysterectomy incidence data from the National Hospital Discharge Survey, and population estimates from the US Census Bureau. RESULTS: Hysterectomy rates significantly decreased 1.9% per year between 1997 and 2005 (-0.5% for ages 18-44, -3.1% for ages 45-64, and -5.0% for ages 65 years and older). The estimated annual decrease in rates was significant in the Northeast (-2.9%), Northwest (-1.7%), and South (-2.6%), but not in the West. For hysterectomies performed among women ages 18-44 years, the percentage in 1997-98 compared with 2004-05 resulting from leiomyoma (fibroids) decreased (31.4% vs. 26.9%), from uterine bleeding increased (14.6% vs. 25.2%), from endometriosis decreased (17.3% in vs. 16.2%), and from pain increased (10.4% vs. 11.7%); the most common procedure, total abdominal hysterectomy, decreased (65.0% vs. 60.5%), the second most common procedure, vaginal hysterectomy, decreased (32.0% vs. 30.7%), and the third most common procedure, subtotal hysterectomy, increased (1.6% in 1997-98 and 7.5% in 2004-05). Decreases in hysterectomy rates occurred for most of the reproductive health conditions resulting in hysterectomy. Exceptions included pain and bleeding in the age group 18-44 and bleeding in the age group 45-64. An increase occurred in subtotal abdominal hysterectomy rates in each of the age groups. CONCLUSIONS: Continued monitoring of hysterectomy rates provides an indication of female reproductive health and how women are being treated for selected reproductive problems. PMID- 18160942 TI - Lack of changes in the concentration of thiobarbituric acid-reactive substances (TBARS) and in the activities of erythrocyte antioxidant enzymes in alcohol dependent patients after detoxification. AB - BACKGROUND: Chronic alcohol consumption induces an increase in oxidative stress. There are studies that indicate changes in the activity of antioxidant enzymes and the concentration of lipid peroxidation products in the blood of organisms as a result of alcohol consumption, but published results are somewhat conflicting. The aim was to study the effect of alcohol withdrawal and detoxification on the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in erythrocytes and on the concentration of thiobarbituric acid reactive substances (TBARS) in the erythrocytes and blood plasma of alcohol dependent patients. MATERIAL/METHODS: The study group consisted of 42 alcohol dependent men who underwent seven-day detoxification at an addiction treatment department. The control group consisted of 20 healthy male volunteers. Blood for analysis was taken from the antecubital vein before detoxification, after the first detoxification, and after several (> or =3) detoxifications. RESULTS: Before detoxification, the concentration of TBARS in the men's blood plasma was 40% higher (p<0.001) than in the control group. After detoxification, the concentration of TBARS decreased, but remained higher than in the control group. The activities of SOD and GPx in erythrocytes of alcohol-dependent persons were statistically significantly lower than in the control group both before and after detoxification. However, after detoxification, a slight increase in the activity of SOD was observed compared to that before detoxification. CONCLUSIONS: This study confirms that alcohol-dependency leads to oxidative stress in the peripheral blood. Seven-day alcohol withdrawal and detoxification has no effect on TBARS concentration and antioxidant enzyme activity. PMID- 18160943 TI - The correlations between endogenous dehydroepiandrosterone sulfate and some atherosclerosis risk factors in premenopausal women. AB - BACKGROUND: Dehydroepiandrosterone (DHEA) is postulated to have antiatherogenic properties, but the possible mechanism of this action is unclear. The aim of this study was to determine the influence of endogenous DHEA-S on the levels of some factors playing significant roles in atherogenesis. MATERIAL/METHODS: In a group of 40 premenopausal women, relationships between endogenous DHEA-S and serum lipids and the apolipoproteins A1 (apoA1) and B (apoB), serum lipid peroxide (LPO), and total antioxidant system (TAS) concentrations as markers of the serum antioxidant-prooxidant balance were measured as well as clinical and biochemical parameters playing roles in atheromatosis such as the type of obesity and the serum glucose, insulin, insulin-like growth factor (IGF-1) and homocysteine (HCY) concentrations. RESULTS: Statistical analysis revealed significant correlation (p<0.05) between serum DHEA-S level and the serum concentrations of: HDL(2)-C (r=0.53), HDL(2)-C/HDL(3)-C (r=0.58), TG (r=0.35), IGF-1 (r=0.39), and HCY (r= 0.44). There was no statistically significant correlation between DHEA-S level and other biochemical and clinical parameters (age, BMI, WHR) found in this study. CONCLUSIONS: Despite unfavorable correlation between DHEA-S and TG concentration, the results of this study indicate a potential antiatherogenic action of DHEA which may occur through various mechanisms: by increasing HDL(2)-C and the HDL(2)-C/HDL(3)-C ratio, which has an atheroprotective effect, by elevating the serum IGF-1 concentration, or by decreasing the HCY level. These preliminary results, however, require further investigation. PMID- 18160944 TI - Levels of plasma fibrinogen and D-dimer in subjects with subclinical hyperthyroidism. AB - BACKGROUND: During the last 15 years, several risk markers for atherosclerosis, such as fibrinogen and D-dimer, have been identified. The role of elevated fibrinogen levels as an independent risk factor for coronary, cerebral, and peripheral vascular disease is well established on the basis of clinical and epidemiological studies. Increased D-dimer levels are associated with increased risk of future myocardial infarction, stroke, and peripheral vascular disease. The aim of this study was to evaluate the alterations in fibrinogen and D-dimer, which indicates overall thrombotic activity, in subjects with subclinical hyperthyroidism. MATERIAL/METHODS: Thirty-six subclinical hyperthyroidic subjects and 36 euthyroidic control subjects matched for age, gender, and body mass index were selected. The levels of plasma fibrinogen and D-dimer in all subjects were measured. RESULTS: The level of fibrinogen was significantly higher in the subclinical hyperthyroidic group than in the euthyroidic group (296.9+/-74.3 mg/dl vs. 255.0+/-41.7 mg/dl, p<0.001). The level of D-dimer was significantly higher in the subclinical hyperthyroidic group than in the euthyroidic group (261.9+/-47.8 mg/dl vs. 216.4+/-32.1 mg/dl, p<0.000). CONCLUSIONS: The results suggest that subjects with subclinical hyperthyroidism present a relatively hypercoagulable state. This state could contribute to increased thromboembolic risk in subclinical hyperthyroidism. PMID- 18160945 TI - Serum malondialdehyde, erythrocyte glutathione peroxidase, and erythrocyte superoxide dismutase levels in women with early spontaneous abortions accompanied by vaginal bleeding. AB - BACKGROUND: Malondialdehyde (MDA) is a marker of lipid peroxidation. Glutathione peroxidase (GPX) and superoxide dismutase (SOD) are the main enzymes responsible for the detoxification of superoxide anion. The aim was to assess whether serum MDA, erythrocyte GPX, and erythrocyte SOD levels altered during early spontaneous abortions presenting with vaginal bleeding. MATERIAL/METHODS: A group of pregnant women at less than 8 weeks' gestation with spontaneous abortion and vaginal bleeding (n=23) and a control group of healthy pregnancies with similar characteristics (n=25) were included. Serum MDA levels, erythrocyte GPX, and SOD activities were determined and compared among the groups. RESULTS: Characteristics, including maternal age, parity, gestational age, complete blood count values, serum total protein, serum albumin, and serum lipid profile, were similar across the groups. Spontaneous abortion prior to 8 weeks of gestation was associated with increased mean serum MDA levels and decreased mean erythrocyte SOD activity. Erythrocyte GPX values did not differ among the groups. CONCLUSIONS: Increased lipid peroxidation and inhibition of SOD activity might be involved in the termination of spontaneous abortions and expulsion of fetoplacental material out of the uterine cavity. PMID- 18160946 TI - Profile of viral hepatitis A, B, and C in a Saudi Arabian hospital. AB - BACKGROUND: The objective of the study was to investigate the pattern and epidemiology of viral hepatitis among patients of the Saudi Aramco Medical Services Organization (SAMSO) in the Eastern Province of Saudi Arabia. MATERIAL/METHODS: A retrospective analysis of the reported cases of viral hepatitis was conducted from January 2000 to June 2005. RESULTS: A total of 1214 patients with viral hepatitis were identified during the study period. Of hepatitis A, B, and C, HBV was the most predominant type of hepatitis, accounting for 49.3% of the cases, followed by HCV (40.7%) and HAV (10%). Nine (0.7%) patients had both HBV and HCV. The male-to-female ratio was higher in HBV (1.8:1), whereas HAV and HCV showed no significant differences. HAV infection predominates in children (1-20 years), HBV in young adults (31-50 years), and HCV in older adults (51-70 years). CONCLUSIONS: HBV continues to be the most predominant type of viral hepatitis and is a disease of adults, whereas HCV is mainly a disease of older adults. Continued surveillance of hepatitis cases is needed to further delineate the risk factors and to establish effective preventive strategies. PMID- 18160947 TI - Gastric multicentric glomangioma: a case report of this rare cause of abdominal pain. AB - BACKGROUND: A glomus tumor is a rare neoplasm derived from glomus cells, specialized cells that surround small blood vessels which are important in regulating peripheral blood flow. Glomangiomas are a subset of glomus tumors. They are usually localized in the skin and subcutaneous tissue. They have also been reported in viscera, most notably in the gastrointestinal tract. CASE REPORT: A case of gastric multicentric glomangioma is described in an 18-year-old male who presented with chronic continuous abdominal pain over a 4-6 month period. Preoperative diagnosis, operative findings, histology, and immunohistochemistry of the tumor are discussed in detail. CONCLUSIONS: Case reports of this rare tumor are important because of the paucity of studies noted in the gastro-intestinal literature as a result of poor identification prior to the advent of modern immunohistochemistry. The significance of accurately diagnosing a gastrointestinal glomangioma is crucial for appropriate treatment. PMID- 18160948 TI - Wavelet analysis and neural networks for intrapartum fetal monitoring. Can we long for a new technique? Is it doable? PMID- 18160949 TI - Compound Salvia droplet pill, a traditional Chinese medicine, for the treatment of unstable angina pectoris: a systematic review. AB - Compound Salvia droplet pill (CSDP) has been frequently used for patients with angina pectoris in China. However, the efficacy and safety of CSDP for unstable angina pectoris (UA) have not been systematically evaluated. The aim was to assess the efficacy and safety of CSDP for UA and provide information for current practice or future research. An extensive search was performed in Jan 2007 which included MEDLINE, EMBASE, Chinese Biomedical Database (CBM), and the Cochrane Center Controlled Trials Register. Randomized controlled trials (RCTs) about CSDP for UA were included irrespective of language. The main comparisons was CSDP plus current western drugs compared with western drugs alone. The quality of each trial was assessed according to criteria from the Cochrane Reviewers' Handbook. Statistical software (RevMan 4.2) provided by the Cochrane Collaboration was applied. Seventeen RCTs were included in this review. The quality of the trials was low. Statistical analysis of the results showed that CSDP plus western routine drugs compared with routine drugs alone had significant effect on relieving angina symptoms (P<0.00001, pooled RR=1.23, 95%CI: 1.16-1.30) and improving ECG (P<0.00001, pooled RR=1.34, 95%CI: 1.23-1.46). Some trials' data suggest that CSDP can also improve hemorheology and blood lipid level. CSDP had a significant effect for treatment of UA with few adverse events. However, the evidence is not reliable enough because of the low quality of the methodology of the included trials. To produce good evidence, high-quality clinical trials are needed. PMID- 18160950 TI - DNA repair alterations in common pediatric malignancies. AB - DNA repair is an important defense mechanism against DNA damage and includes four distinct pathways: direct, excision, mismatch, and double-strand break repair systems. Recent evidence suggests that alterations in proteins participating in the DNA repair systems may result in cellular senescence, cell death, and neoplastic transformation. Malignancies in adulthood exhibit genomic instability and an increased mutation rate due to underlying defects in DNA repair. However, our knowledge on DNA repair defects, both in germline and somatic mutations, and their relationship with childhood malignancies remains incomplete. Mutations, gene deletions, and inversions in various DNA repair genes have been reported and special attention has recently been focused on the interaction between these abnormalities and malignant transformation in childhood. The purpose of this review is to summarize the existing clinical information concerning components of the DNA repair systems and their influence on the development of the most common pediatric malignancies, including leukemia, tumors of the central nervous system, rhabdomyosarcoma, and retinoblastoma. Such information could possibly explain the response or resistance to chemotherapy and the possible risk of relapse in childhood malignancies presenting specific DNA repair defects. Additionally, these data could be beneficial for the development of novel therapeutic strategies. PMID- 18160952 TI - Neuroscience grows in China. PMID- 18160951 TI - Morbidity, demography, life style, and self-perceived health of Hungarian medical doctors 25 years after graduation. AB - BACKGROUND: This study aimed to chart the health, perceived health, medical career, life style, and sociological factors of Hungarian physicians who graduated in 1979. The results were analyzed for differences between genders and professional specialty groups (primary, surgical, non-surgical, diagnostic). MATERIAL/METHODS: Of 228 questionnaire completed by doctors 25 years after graduation from Semmelweis Medical University, Budapest, Hungary, 186 were analyzable. RESULTS: More men were in surgical professions; a larger proportion of women became primary specialists. Women had to modify their specialty or place of work more often. The average number of children was 2.26 for men and 1.87 for women. Primary specialist men and non-surgical women showed the highest increases in body weight. Hypertension and failure to attend regular screenings were more common in males and they were more often dissatisfied with received treatment. Physical exercise, typically sports, were reduced after graduation and the preferred types of activity also changed. Female physicians considered regular exercise more important. Smokers were mainly male surgeons and women in primary care. Surgeons and women in non-surgical specialties consumed more alcohol. As patients, male physicians followed medical advice more faithfully. Doctors judged their own health status as better than their patients'. Knowledge of foreign languages was higher in the men. Ten percent of the physicians had postgraduate degrees in research. CONCLUSIONS: Physicians give advice and provide care, but they do not always follow such advice themselves. Their social circumstances and scientific careers depend mostly on the social and health system in which they work. PMID- 18160954 TI - The bouncer in the brain. AB - Efficiency variations in the filtering of relevant from irrelevant information could contribute to individual differences in working memory. A new functional imaging study suggests that the basal ganglia act as this filter because activity in this region before stimulus presentation was inversely correlated with unnecessary storage. PMID- 18160955 TI - Dynamin-independent synaptic vesicle retrieval? AB - A new study proposes that synaptic vesicle endocytosis at a large synaptic terminal is partly independent of dynamin and GTP hydrolysis, suggesting a new mechanism leading to vesicle fission and maintenance of neurotransmission. PMID- 18160956 TI - Glia and romance. AB - Drosophila courtship is a complex behavior. A new study shows that glia modulate neurotransmission to influence male preference, but the authors should have resisted the temptation to describe their results in tabloid language. PMID- 18160957 TI - More than a feeling: sensation from cortical stimulation. AB - Changes in neuronal firing underlie sensation, but how many neurons are needed to perceive these activity shifts? Two new studies in Nature suggest that the experimental modulation of only a few neurons can influence perception. PMID- 18160958 TI - Impact of hemodialysis duration on the removal of uremic retention solutes. AB - Several studies have stressed the importance of dialysis time in the removal of uremic retention solutes. To further investigate this, nine stable chronic hemodialysis patients were dialyzed for 4, 6, or 8 h processing the same total blood and dialysate volume by the Genius system and high-flux FX80 dialyzers. Inlet blood and outlet dialysate were analyzed for urea, creatinine, phosphorus, and beta2-microglobulin at various times. Total solute removal, dialyzer extraction ratios, and total cleared volumes were significantly larger during prolonged dialysis for urea, creatinine, phosphorus, and beta2-microglobulin. Reduction ratios increased progressively, except for phosphate and beta2 microglobulin, where the ratios remained constant after 2 h. In contrast, no significant difference was found for the reduction ratios of all solutes and Kt/V(urea) between the three different sessions. With longer dialyses, solutes are efficiently removed from the deeper compartments of the patient's body. Our study shows that care must be taken when using Kt/Vurea or reduction ratios as the only parameters to quantify dialysis adequacy. PMID- 18160959 TI - Achieving blood pressure targets during dialysis improves control but increases intradialytic hypotension. AB - Cardiovascular disease remains the most common cause of mortality in patients with end-stage kidney disease treated by regular hemodialysis. To improve blood pressure control and reduce cardiovascular risk, the United Kingdom Renal Association standards committee introduced pre- and post-dialysis target blood pressures of less than 140/90 and 130/80 mm Hg, respectively. We audited blood pressure control and symptomatic intradialytic hypotension requiring fluid resuscitation in the Greater London area renal centers that serve 2630 patients. The study captured 7890 hemodialysis sessions during a 1-week period where only 36% of the patients achieved the pre-dialysis target and 42% the post-dialysis target, with a wide variation between centers. Different antihypertensive medication prescriptions did not affect achievement of these targets. Fifteen percent of the patients suffered symptomatic hypotension requiring fluid resuscitation associated with significantly greater interdialytic weight gains. Our study found that intradialytic hypotension was significantly greater in centers that achieved better post-dialysis blood pressure targeting. PMID- 18160960 TI - CD14(++)CD16+ monocytes but not total monocyte numbers predict cardiovascular events in dialysis patients. AB - Migration of monocytes into the vessel wall contributes to the onset and progression of atherosclerosis. Because monocytes are a heterogeneous population, we determined potential associations between monocyte subsets and cardiovascular events in a prospective cohort of 94 dialysis patients followed for 35 months. The incidence of cardiovascular events and death measured by Kaplan-Meier plots and flow cytometric analysis of monocyte subsets showed that total leukocyte and monocyte numbers failed to predict event-free survival. Among monocyte subsets, a high CD14(++)CD16(+) monocyte number was associated with higher rates of cardiovascular events and death. In a multivariate proportional hazards model adjusted for classical cardiovascular risk factors, patients with CD14(++)CD16(+) monocyte numbers in the top quartile were at higher risk of cardiovascular events and death compared to patients in the lowest quartile. Our study suggests that the number of CD14(++)CD16(+) monocytes was independently associated with cardiovascular events and death in a high-risk population of dialysis patients. PMID- 18160961 TI - The RIFLE criteria and mortality in acute kidney injury: A systematic review. AB - In 2004, the Acute Dialysis Quality Initiative workgroup proposed a multilevel classification system for acute kidney injury (AKI) identified by the acronym RIFLE (Risk, Injury, Failure, Loss of kidney function, and End-stage kidney disease). Several studies have been published aiming to validate and apply it in clinical practice, verifying whether outcome progressively worsened with the severity of AKI. A literature search from August 2004 to June 2007 was conducted: 24 studies in which the RIFLE classification was used to define AKI were identified. In 13 studies, patient-level data on mortality were available for Risk, Injury, and Failure patients, as well as those without AKI (non-AKI). Death was reported at ICU discharge, hospital discharge, 28, 30, 60, and 90 days. The pooled estimate of relative risk (RR) for mortality for patients with R, I, or F levels compared with non-AKI patients were analyzed. Over 71 000 patients were included in the analysis of published reports. With respect to non-AKI, there appeared to be a stepwise increase in RR for death going from Risk (RR=2.40) to Injury (RR=4.15) to Failure (6.37, P<0.0001 for all). There was significant intertrial heterogeneity as expected with the varying patient populations studied. The RIFLE classification is a simple, readily available clinical tool to classify AKI in different populations. It seems to be a good outcome predictor, with a progressive increase in mortality with worsening RIFLE class. It also suggests that even mild degrees of kidney dysfunction may have a negative impact on outcome. Further refinement of RIFLE nomenclature and classification is ongoing. PMID- 18160962 TI - Antihypertensive therapy with aliskiren. AB - Aliskiren represents the first member in a new class of antihypertensive drugs. Inhibiting the renin-angiotensin system at its rate-limiting step is an idea that has been pursued for >30 years; however, earlier compounds failed because of problems related to efficacy, bioavailability, and/or side effects. Aliskiren, a 610 Da nonpeptide molecule, has exceptional affinity for the human renin enzymatic site and a half-life of about 40 h, which make its 3% bioavailability clinically unimportant with continued administration. The drug is not metabolized by CYP P450 enzymes and is excreted >90% unchanged by the fecal route. No adjustments are necessary for renal function, liver function, age, ethnicity, or other prescribed drugs. Blood pressure reductions are similar to those provided by other monotherapies. Interestingly, aliskiren combined with angiotensin receptor blocker or angiotensin-converting enzyme inhibitor therapy leads to a further blood pressure reduction as does combination with a diuretic or calcium channel blocker. The fact that plasma renin activity is reduced to low levels with aliskiren could provide a theoretical advantage over other treatments, while increases in total renin (prorenin) after the drug poses additional food for thought. Studies with primary cardiovascular and renal end points to address these possibilities are in progress. PMID- 18160963 TI - A new adjuvant improves the immune response to hepatitis B vaccine in hemodialysis patients. AB - Prehemodialysis and hemodialysis patients are at an increased risk of hepatitis B infection and have an impaired immune response to hepatitis B vaccines. We evaluated the immune response to the new adjuvant of hepatitis B vaccine AS04 (HBV-AS04) in this population. We measured antibody persistence for up to 42 months, and the anamnestic response and safety of booster doses in patients who were no longer seroprotected. The primary vaccination study showed that HBV-AS04 elicited an earlier antibody response and higher antibody titers than four double doses of standard hepatitis B vaccine. Seroprotection rates were significantly higher in HBV-AS04 recipients throughout the study. The decline in seroprotection over time was significantly less in the HBV-AS04 group with significantly fewer primed patients requiring a booster dose over the follow-up period. Solicited/unsolicited adverse events were rare following booster administration. Fifty-seven patients experienced a serious adverse event during the follow-up; none of which was vaccine related. When HBV-AS04 was used as the priming immunogen, the need for a booster dose occurred at a longer time compared to double doses of standard hepatitis B vaccine. Hence, in this population, the HBV AS04 was immunogenic, safe, and well-tolerated both as a booster dose after HBV AS04 or standard hepatitis B vaccine priming. PMID- 18160964 TI - Kidney injury molecule-1 expression in transplant biopsies is a sensitive measure of cell injury. AB - Kidney injury molecule-1 (KIM-1) is a specific histological biomarker for diagnosing early tubular injury on renal biopsies. In this study, KIM-1 expression was quantitated in renal transplant biopsies by immunohistochemistry and correlated with renal function. None of the 25 protocol biopsies showed detectable tubular injury on histologic examination, yet 28% had focal positive KIM-1 expression. Proximal tubule KIM-1 expression was present in all biopsies from patients with histological changes showing acute tubular damage and deterioration of kidney function. In this group, higher KIM-1 staining predicted a better outcome with improved blood urea nitrogen (BUN), serum creatinine, and estimated glomerular filtration rate (eGFR) over an ensuing 18 months. KIM-1 was expressed focally in affected tubules in 92% of kidney biopsies from patients with acute cellular rejection. By contrast, there was little positive staining for Ki-67, a cell proliferation marker, in any of the groups. KIM-1 expression significantly correlated with serum creatinine and BUN, and inversely with the eGFR on the biopsy day. Our study shows that KIM-1 staining sensitively and specifically identified proximal tubular injury and correlated with the degree of renal dysfunction. KIM-1 expression is more sensitive than histology for detecting early tubular injury, and its level of expression in transplant biopsies may indicate the potential for recovery of kidney function. PMID- 18160965 TI - Towards a better bowl of rice: assigning function to tens of thousands of rice genes. AB - Rice, one of the most important food crops for humans, is the first crop plant to have its genome sequenced. Rice whole-genome microarrays, genome tiling arrays and genome-wide gene-indexed mutant collections have recently been generated. With the availability of these resources, discovering the function of the estimated 41,000 rice genes is now within reach. Such discoveries have broad practical implications for understanding the biological processes of rice and other economically important grasses such as cereals and bioenergy crops. PMID- 18160966 TI - Understanding the molecular machinery of genetics through 3D structures. AB - Detailed knowledge of the three-dimensional structures of biological molecules has had an enormous impact on all areas of biological science, including genetics, as structure can reveal the fine details of how molecules perform their biological functions. Here we consider how changes in protein sequence affect the corresponding 3D structure, and describe how structural information about proteins, DNA and chromatin has shed light on gene regulatory mechanisms and the storage and transmission of epigenetic information. Finally, we describe how structure determination is benefiting from the high-throughput technologies of the worldwide structural genomics projects. PMID- 18160967 TI - Maintenance of hair follicle immune privilege is linked to prevention of NK cell attack. AB - Hair follicles (HFs) enjoy a relative immune privilege (IP) that is characterized by downregulation of major histocompatibility complex (MHC) class I and local expression of potent immunosuppressants. Normally, natural killer (NK) cells attack cells with absent/low MHC class I expression. However, because few perifollicular NK cells are found around healthy human anagen HFs, we asked how HFs escape from NK cell attack. This study suggests that this happens via an active NK cell suppression. Alopecia areata (AA), an organ-specific autoimmune disease thought to result from a collapse of HF-IP, in contrast, shows striking defects in NK cell inhibition/containment. We show that the NK cell inhibitor macrophage migration inhibitory factor is strongly expressed by the HF epithelium, and very few CD56(+)/NKG2D(+) NK cells are observed in and around normal anagen HFs compared to AA with prominent aggregations of CD56(+)/NKG2D(+) NK around AA-HFs. By flow cytometry, many fewer NK function-activating receptors (NKG2D, NKG2C) and significantly more killer cell Ig-like receptors-2D2/2D3 were found to be expressed on peripheral blood CD56(+) NK cells of healthy controls than on those of AA patients. In addition, only weak immunoreactivity for MHC class I chain-related A gene was observed in normal anagen HFs compared to AA. To our knowledge, this defect is previously unreported and must be taken into account in AA pathogenesis and its management. PMID- 18160968 TI - The Revised Observed Tasks of Daily Living: A Performance-Based Assessment of Everyday Problem Solving in Older Adults. AB - The Revised Observed Tasks of Daily Living (OTDL-R), a performance-based test of everyday problem solving, was administered to a sample of community-dwelling older adults. The OTDL-R included nine tasks, representing medication use, telephone use, and financial management. The OTDL-R had a desirable range of difficulty and satisfactory internal consistency and showed a relatively invariant pattern of relations between measured tasks and the underlying latent dimensions they represent across White and non-White subsamples. The OTDL-R also correlated significantly with age, education, self-rated health, a paper-and pencil measure of everyday problem solving, and measures of basic cognitive functioning. Thus, the OTDL-R is a reliable and valid objective measure of everyday problem solving that has great practical utility for assessing performance in diverse populations. PMID- 18160970 TI - Separately reconstructing the structural and functional parameters of a fluorescent inclusion embedded in a turbid medium. AB - We report a novel imaging technique for fluorescence diffuse optical tomography (FDOT). Unlike conventional FDOT, this technique separates the imaging procedure into two steps to respectively reconstruct the structural information (such as the center position and the radius), and the functional information (such as the fluorophore concentration and/or lifetime) of a fluorescing target embedded in a turbid medium. The structural parameters of the target were estimated from the amplitude ratio and phase difference of fluorescence signals received at different detectors, because the amplitude ratio and phase difference were found independent of, or weakly related to, the functional parameters. Based on the estimated structural parameters, a dual-zone mesh technique was utilized to reconstruct the fluorophore concentration. Results of simulations and phantom experiments showed that the structural parameters could be accurately recovered, without knowing the functional information, and that the reconstruction accuracy of the functional parameter was greater than 80%. PMID- 18160971 TI - On Spin Hamiltonian Fits to Mossbauer Spectra of High-Spin Fe(II) Porphyrinate Systems. AB - Fits to Mossbauer spectra of high-spin iron(II) porphyrinates have been applied to the Fe(II) model compounds octaethylporphyrin(1,2-dimethylimidazole) and tetra paramethoxyporphyrin(1,2-dimethylimidazole). Mossbauer spectra have been measured on these compounds at 4.2 K in large applied fields. Spin Hamiltonians were used for fitting both the electronic and nuclear interactions. The fits are done by adjusting the Hamiltonian parameters to simultaneously minimize the total chi(2) for three different applied fields. In order to get best fits, the EFG tensor need to be rotated relative to the ZFS tensor. A comparative sensitivity analysis of their Spin Hamiltonian parameters has also been done on the ZFS parameters D, and the EFG asymmetry parameter eta. The best fits suggest that both systems definitely have a negative quadrupole splitting, and that largest EFG component is tilted far from the z-axis of the ZFS tensor, which is likely to be near the heme normal. PMID- 18160969 TI - Infectious Disease: Connecting Innate Immunity to Biocidal Polymers. AB - Infectious disease is a critically important global healthcare issue. In the U.S. alone there are 2 million new cases of hospital-acquired infections annually leading to 90,000 deaths and 5 billion dollars of added healthcare costs. Couple these numbers with the appearance of new antibiotic resistant bacterial strains and the increasing occurrences of community-type outbreaks, and clearly this is an important problem. Our review attempts to bridge the research areas of natural host defense peptides (HDPs), a component of the innate immune system, and biocidal cationic polymers. Recently discovered peptidomimetics and other synthetic mimics of HDPs, that can be short oligomers as well as polymeric macromolecules, provide a unique link between these two areas. An emerging class of these mimics are the facially amphiphilic polymers that aim to emulate the physicochemical properties of HDPs but take advantage of the synthetic ease of polymers. These mimics have been designed with antimicrobial activity and, importantly, selectivity that rivals natural HDPs. In addition to providing some perspective on HDPs, selective mimics, and biocidal polymers, focus is given to the arsenal of biophysical techniques available to study their mode of action and interactions with phospholipid membranes. The issue of lipid type is highlighted and the important role of negative curvature lipids is illustrated. Finally, materials applications (for instance, in the development of permanently antibacterial surfaces) are discussed as this is an important part of controlling the spread of infectious disease. PMID- 18160972 TI - Comparison of two detector systems for cone beam CT small animal imaging - a preliminary study. AB - To compare two detector systems - one based on the charge-coupled device (CCD) and image amplifier, the other based on a-Si/CsI flat panel, for cone beam computed-tomography (CT) imaging of small animals.A high resolution, high framing rate detector system for the cone beam CT imaging of small animals was developed. The system consists of a 2048x3072x12 bit CCD optically coupled to an image amplifier and an x-ray phosphor screen. The CCD has an intrinsic pixel size of 12 mum but the effective pixel size can be adjusted through the magnification adjustment of the optical coupling systems. The system is used in conjunction with an x-ray source and a rotating stage for holding and rotating the scanned object in the cone beam CT imaging experiments. The advantages of the system include but are not limited to the ability to adjust the effective pixel size and to achieve extremely high spatial resolution and temporal resolution. However, the need to use optical coupling compromises the detective quanta efficiency (DQE) of the system. In this paper, the imaging characteristics of the system were presented and compared with those of an a-Si/CsI flat-panel detector system. PMID- 18160973 TI - Selective toxicity of zinc oxide nanoparticles to prokaryotic and eukaryotic systems. AB - We report on the toxicity of ZnO nanoparticles (NPs) to gram-negative and gram positive bacterial systems, Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), and primary human immune cells. ZnO NP (~13 nm) showed complete inhibition of E. coli growth at concentrations 3.4 mM, whereas growth of S. aureus was completely inhibited for 1 mM. Parallel experiments using flow cytometry based assays clearly demonstrated that growth inhibitory properties of ZnO NP were accompanied by a corresponding loss of cell viability. Identical ZnO NP had minimal effects on primary human T cell viability at concentrations toxic to both gram-negative and gram-positive bacteria. Collectively, these experiments demonstrate selectivity in the toxic nature of ZnO NP to different bacterial systems and human T lymphocytes. Developing selective toxicity to biological systems and controlling it by NP design could lead to biomedical and antibacterial applications. PMID- 18160974 TI - Effect of ionic liquids on epoxide hydrolase-catalyzed synthesis of chiral 1,2 diols. AB - Ionic liquids (ILs) offer new possibilities for epoxide hydrolase (EH) catalyzed resolution of epoxides and for synthesis of chiral 1,2-diols. Soluble EHs from cress and mouse (csEH and msEH) and microsomal EH from rat (rmEH) were tested in several ILs. For all the enzymes tested, higher enantioselectivities were obtained in [bmim][N(Tf)(2)] and [bmim][PF(6)]. The optimized amount of water for EH activity in these ILs was established. Classical problems arising from low solubility of epoxides in water or from the high tendency of the oxirane ring to undergo chemical hydrolysis were avoided using these new media. PMID- 18160975 TI - Nutrient Intakes of Third Graders: Results from the Child and Adolescent Trial for Cardiovascular Health (CATCH) Baseline Survey. AB - The purpose of this article is to report on baseline intakes of 1874 third-grade children representing a subsample of the Child and Adolescent Trial for Cardiovascular Health (CATCH) cohort. Intakes were assessed using a single, food record-assisted, 24-hour recall. The sample is unique in that it is drawn from four states and includes students from various ethnic backgrounds. Nutrients of interest include total energy, sodium, dietary cholesterol, and percent of energy from total fat and saturated fat. At baseline, third-grade students were consuming above nationally recommended levels of energy from fat, saturated fat, and sodium. The CATCH findings show a mean energy intake of 2031 kcal with significant differences by sex. Significant differences by site were seen for percent of energy from total fat, saturated fat, and dietary cholesterol. Children from Minnesota consumed the lowest proportion of energy from total fat and saturated fat while children from Texas had the highest proportion of energy from total fat and saturated fat. Intake of dietary cholesterol was lowest in Minnesota and highest in Louisiana. Nutrient differences by ethnic group were seen only for energy, with African Americans having the highest energy intake and Hispanics having the lowest energy intake. The number of meals consumed from school food service significantly influenced children's nutrient, intake; children consuming two meals from school food service had significantly greater intakes of energy, saturated fat, and dietary cholesterol compared to students consuming one or no meals from school food-service. The results are compared to other national nutritional surveys of children. PMID- 18160976 TI - Population preference of net texture prior to bed net trial in Kala-Azar-endemic areas. AB - Prior to a community-based efficacy trial of long-lasting insecticidal nets (LLINs) in the prevention of visceral leishmaniasis (VL; also called kala-azar), a pilot study on preference of tools was held in endemic areas of India and Nepal in September 2005.LLINs made of polyester and polyethylene were distributed to 60 participants, who used the nets sequentially for 7 d. Acceptability and preference were evaluated via indirect indicators through questionnaires at three defined time points before and after use of the LLINs and through focus group discussions (FGDs). In the latter, preferences for color and size were also assessed. Untreated bed nets were owned by 87% of the households prior to the study. All users liked textures of both LLIN types after 7 d of use, but had a slight preference for those made of polyester if they were to recommend a LLIN to relatives or friends (p<0.05), mainly because of their relatively greater softness in comparison to polyethylene LLINs. Users reported that both net types reduced mosquito bites and number of insects, including sand fly (bhusana; genus Phlebotomus), inside the house. Side effects were minor and disappeared quickly.The large-scale intervention trial considered the preferences of the study population to decide on the best tool of intervention--light-blue, rectangular, polyester LLINs of different sizes. PMID- 18160977 TI - Risk factors for buruli ulcer: a case control study in Cameroon. AB - BACKGROUND: Buruli ulcer is an infectious disease involving the skin, caused by Mycobacterium ulcerans. This disease is associated with areas where the water is slow-flowing or stagnant. However, the exact mechanism of transmission of the bacillus and the development of the disease through human activities is unknown. METHODOLOGY/PRINCIPAL FINDINGS: A case-control study to identify Buruli ulcer risk factors in Cameroon compared case-patients with community-matched controls on one hand and family-matched controls on the other hand. Risk factors identified by the community-matched study (including 163 pairs) were: having a low level of education, swamp wading, wearing short, lower-body clothing while farming, living near a cocoa plantation or woods, using adhesive bandages when hurt, and using mosquito coils. Protective factors were: using bed nets, washing clothes, and using leaves as traditional treatment or rubbing alcohol when hurt. The family-matched study (including 118 pairs) corroborated the significance of education level, use of bed nets, and treatment with leaves. CONCLUSIONS/SIGNIFICANCE: Covering limbs during farming activities is confirmed as a protective factor guarding against Buruli ulcer disease, but newly identified factors including wound treatment and use of bed nets may provide new insight into the unknown mode of transmission of M. ulcerans or the development of the disease. PMID- 18160978 TI - Treatment of helminth co-infection in individuals with HIV-1: A systematic review of the literature. AB - BACKGROUND AND OBJECTIVES: The HIV-1 pandemic has disproportionately affected individuals in resource-constrained settings. It is important to determine if other prevalent infections affect the progression of HIV-1 in co-infected individuals in these settings. Some observational studies suggest that helminth infection may adversely affect HIV-1 progression. We sought to evaluate existing evidence on whether treatment of helminth infection impacts HIV-1 progression. REVIEW METHODS: This review was conducted using the HIV/AIDS Cochrane Review Group (CRG) search strategy and guidelines. Published and unpublished studies were obtained from The Cochrane Library (Issue 3, 2006), MEDLINE (November 2006), EMBASE (November 2006), CENTRAL (July 2006), and AIDSEARCH (August 2006). Databases listing conference abstracts and scanned reference lists were searched, and authors of included studies were contacted. Data regarding changes in CD4 count, HIV-1 RNA levels, clinical staging and/or mortality were extracted and compared between helminth-treated and helminth-untreated or helminth-uninfected individuals. RESULTS: Of 6,384 abstracts identified, 15 met criteria for potential inclusion, of which 5 were eligible for inclusion. In the single randomized controlled trial (RCT) identified, HIV-1 and schistosomiasis co infected individuals receiving treatment for schistosomiasis had a significantly lower change in plasma HIV-1 RNA over three months (-0.001 log(10) copies/mL) compared to those receiving no treatment (+0.21 log(10) copies/mL), (p = 0.03). Four observational studies met inclusion criteria, and all of these suggested a possible beneficial effect of helminth eradication on plasma HIV-1 RNA levels when compared to plasma HIV-1 RNA changes prior to helminth treatment or to helminth-uninfected or persistently helminth-infected individuals. The follow-up duration in these studies ranged from three to six months. The reported magnitude of effect on HIV-1 RNA was variable, ranging from 0.07-1.05 log(10) copies/mL. None of the included studies showed a significant benefit of helminth treatment on CD4 decline, clinical staging, or mortality. CONCLUSION: There are insufficient data available to determine the potential benefit of helminth eradication in HIV-1 and helminth co-infected adults. Data from a single RCT and multiple observational studies suggest possible benefit in reducing plasma viral load. The impact of de-worming on markers of HIV-1 progression should be addressed in larger randomized studies evaluating species-specific effects and with a sufficient duration of follow-up to document potential differences on clinical outcomes and CD4 decline. PMID- 18160979 TI - Targeted screening strategies to detect Trypanosoma cruzi infection in children. AB - BACKGROUND: Millions of people are infected with Trypanosoma cruzi, the causative agent of Chagas disease in Latin America. Anti-trypanosomal drug therapy can cure infected individuals, but treatment efficacy is highest early in infection. Vector control campaigns disrupt transmission of T. cruzi, but without timely diagnosis, children infected prior to vector control often miss the window of opportunity for effective chemotherapy. METHODS AND FINDINGS: We performed a serological survey in children 2-18 years old living in a peri-urban community of Arequipa, Peru, and linked the results to entomologic, spatial and census data gathered during a vector control campaign. 23 of 433 (5.3% [95% CI 3.4-7.9]) children were confirmed seropositive for T. cruzi infection by two methods. Spatial analysis revealed that households with infected children were very tightly clustered within looser clusters of households with parasite-infected vectors. Bayesian hierarchical mixed models, which controlled for clustering of infection, showed that a child's risk of being seropositive increased by 20% per year of age and 4% per vector captured within the child's house. Receiver operator characteristic (ROC) plots of best-fit models suggest that more than 83% of infected children could be identified while testing only 22% of eligible children. CONCLUSIONS: We found evidence of spatially-focal vector-borne T. cruzi transmission in peri-urban Arequipa. Ongoing vector control campaigns, in addition to preventing further parasite transmission, facilitate the collection of data essential to identifying children at high risk of T. cruzi infection. Targeted screening strategies could make integration of diagnosis and treatment of children into Chagas disease control programs feasible in lower-resource settings. PMID- 18160980 TI - A comparative study of leptospirosis and dengue in Thai children. AB - BACKGROUND: Leptospirosis is an emerging zoonosis that is often under-recognized in children and commonly confused with dengue in tropical settings. An enhanced ability to distinguish leptospirosis from dengue in children would guide clinicians and public health personnel in the appropriate use of limited healthcare resources. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a prospective, hospital-based, study of children with acute febrile illnesses and dengue in Thailand. Among the children without dengue, we identified those with leptospirosis using anti-leptospira IgM and microscopic agglutination titers in paired acute and convalescent blood samples. We then performed a case-control comparison of symptoms, signs, and clinical laboratory values between children with leptospirosis and dengue. In a semi-rural region of Thailand, leptospirosis accounted for 19% of the non-dengue acute febrile illnesses among children presenting during the rainy season. None of the children with leptospirosis were correctly diagnosed at the time of hospital discharge, and one third (33%) were erroneously diagnosed as dengue or scrub typhus. A predictive model to distinguish pediatric leptospirosis from dengue was generated using three variables: the absolute neutrophil count, plasma albumin, and aspartate aminotransferase levels in the first 72 hours of illness. CONCLUSIONS/SIGNIFICANCE: Unrecognized leptospirosis can be a significant cause of "dengue-like" febrile illness in children. Increased awareness of pediatric leptospirosis, and an enhanced ability to discriminate between leptospirosis and dengue early in illness, will help guide the appropriate use of healthcare resources in often resource-limited settings. PMID- 18160981 TI - Community and school-based health education for dengue control in rural Cambodia: a process evaluation. AB - Dengue fever continues to be a major public health problem in Cambodia, with significant impact on children. Health education is a major means for prevention and control of the National Dengue Control Program (NDCP), and is delivered to communities and in schools. Drawing on data collected in 2003-2004 as part of an ethnographic study conducted in eastern Cambodia, we explore the approaches used in health education and their effectiveness to control dengue. Community health education is provided through health centre outreach activities and campaigns of the NDCP, but is not systematically evaluated, is under-funded and delivered irregularly; school-based education is restricted in terms of time and lacks follow-up in terms of practical activities for prevention and control. As a result, adherence is partial. We suggest the need for sustained routine education for dengue prevention and control, and the need for approaches to ensure the translation of knowledge into practice. PMID- 18160982 TI - Neglected diseases and poverty in "The Other America": the greatest health disparity in the United States? PMID- 18160983 TI - Magnetic beads for schistosomiasis diagnosis. PMID- 18160984 TI - The Global Burden of Disease assessments--WHO is responsible? AB - The Global Burden of Disease (GBD) concept has been used by the World Health Organization (WHO) for its reporting on health information for nearly 10 years. The GBD approach results in a single summary measure of morbidity, disability, and mortality, the so-called disability-adjusted life year (DALY). To ensure transparency and objectivity in the derivation of health information, WHO has been urged to use reference groups of external experts to estimate burden of disease. Under the leadership and coordination of WHO, expert groups have been appraising and abstracting burden of disease information. Examples include the Child Health Epidemiology Reference Group (CHERG), the Malaria Monitoring and Evaluation Reference Group (MERG), and the recently established Foodborne Disease Burden Epidemiology Reference Group (FERG). The structure and functioning of and lessons learnt by these groups are described in this paper. External WHO expert groups have provided independent scientific health information while operating under considerable differences in structure and functioning. Although it is not appropriate to devise a single "best practice" model, the common thread described by all groups is the necessity of WHO's leadership and coordination to ensure the provision and dissemination of health information that is to be globally accepted and valued. PMID- 18160985 TI - Combining residual insecticide spraying campaigns with targeted detection and specific chemotherapy for Trypanosoma cruzi infection in children. PMID- 18160986 TI - Risk factors for tungiasis in Nigeria: identification of targets for effective intervention. AB - BACKGROUND: The parasitic skin disease tungiasis (caused by the flea Tunga penetrans) affects resource-poor communities in Latin America, the Caribbean and sub-Saharan Africa. Prevalences in endemic areas are high, and severe pathology occurs commonly. However, risk factors for infestation have never been assessed in Africa. METHODS AND FINDINGS: A cross-sectional study was conducted in Erekiti, a rural community in Lagos State (Nigeria), where tungiasis is endemic. Individuals were examined clinically for the presence of tungiasis, and a questionnaire was applied. Data from 643 individuals (86.6% of the target population) were analyzed; 252 (42.5%) were infested with T. penetrans. In the multivariate logistic regression analysis, presence of pigs on the compounds (adjusted odds ratio = 17.98; 95% confidence interval: 5.55-58.23), sand or clay floor inside houses (9.33; 5.06-17.19), and having the common resting place outside the house (7.14; 4.0-14.29) were the most important risk factors identified. The regular use of closed footwear (0.34; 0.18-0.62) and the use of insecticides indoors (0.2; 0.05-0.83) were protective against infestation. The population attributable fractions associated with tungiasis were: sand or clay floor inside the house (73.7%), resting usually outside the house (65.5%), no regular use of closed footwear (51.1%), and pigs on the compound (37.9%). CONCLUSION: The presence of tungiasis in Erekiti is determined to an important extent by a limited number of modifiable variables. Effective and sustainable intervention measures addressing these factors need to be implemented in this and other West African communities with high disease burden. PMID- 18160987 TI - Filariasis in travelers presenting to the GeoSentinel Surveillance Network. AB - BACKGROUND: As international travel increases, there is rising exposure to many pathogens not traditionally encountered in the resource-rich countries of the world. Filarial infections, a great problem throughout the tropics and subtropics, are relatively rare among travelers even to filaria-endemic regions of the world. The GeoSentinel Surveillance Network, a global network of medicine/travel clinics, was established in 1995 to detect morbidity trends among travelers. PRINCIPAL FINDINGS: We examined data from the GeoSentinel database to determine demographic and travel characteristics associated with filaria acquisition and to understand the differences in clinical presentation between nonendemic visitors and those born in filaria-endemic regions of the world. Filarial infections comprised 0.62% (n = 271) of all medical conditions reported to the GeoSentinel Network from travelers; 37% of patients were diagnosed with Onchocerca volvulus, 25% were infected with Loa loa, and another 25% were diagnosed with Wuchereria bancrofti. Most infections were reported from immigrants and from those immigrants returning to their county of origin (those visiting friends and relatives); the majority of filarial infections were acquired in sub-Saharan Africa. Among the patients who were natives of filaria nonendemic regions, 70.6% acquired their filarial infection with exposure greater than 1 month. Moreover, nonendemic visitors to filaria-endemic regions were more likely to present to GeoSentinel sites with clinically symptomatic conditions compared with those who had lifelong exposure. SIGNIFICANCE: Codifying the filarial infections presenting to the GeoSentinel Surveillance Network has provided insights into the clinical differences seen among filaria-infected expatriates and those from endemic regions and demonstrated that O. volvulus infection can be acquired with short-term travel. PMID- 18160988 TI - Pediatric measles vaccine expressing a dengue antigen induces durable serotype specific neutralizing antibodies to dengue virus. AB - Dengue disease is an increasing global health problem that threatens one-third of the world's population. Despite decades of efforts, no licensed vaccine against dengue is available. With the aim to develop an affordable vaccine that could be used in young populations living in tropical areas, we evaluated a new strategy based on the expression of a minimal dengue antigen by a vector derived from pediatric live-attenuated Schwarz measles vaccine (MV). As a proof-of-concept, we inserted into the MV vector a sequence encoding a minimal combined dengue antigen composed of the envelope domain III (EDIII) fused to the ectodomain of the membrane protein (ectoM) from DV serotype-1. Immunization of mice susceptible to MV resulted in a long-term production of DV1 serotype-specific neutralizing antibodies. The presence of ectoM was critical to the immunogenicity of inserted EDIII. The adjuvant capacity of ectoM correlated with its ability to promote the maturation of dendritic cells and the secretion of proinflammatory and antiviral cytokines and chemokines involved in adaptive immunity. The protective efficacy of this vaccine should be studied in non-human primates. A combined measles dengue vaccine might provide a one-shot approach to immunize children against both diseases where they co-exist. PMID- 18160989 TI - Impact of placental Plasmodium falciparum malaria on pregnancy and perinatal outcome in sub-Saharan Africa: I: introduction to placental malaria. AB - Placental malaria is one of the major features of malaria during pregnancy and has been widely used as a standard indicator to characterize malaria infection in epidemiologic investigations. Although pathogenesis of placental malaria is only partially understood, placental sequestration of Plasmodium falciparum results in the accumulation of parasitized erythrocytes in the intervillous space, infiltration by inflammatory cells, and release of pro-inflammatory mediators, which cause pathologic alterations that could impair materno-fetal exchanges, often resulting in adverse pregnancy outcome. In this report, the impact of placental malaria on pregnancy and perinatal outcome is reviewed using data from studies conducted in sub-Saharan Africa. Generally, placental malaria was associated with increased risk of maternal anemia, HIV infection, and maternal mortality, with younger women and primigravidae more likely to be affected. A variety of adverse perinatal outcomes, including low birth weight, preterm delivery, intrauterine growth retardation, reduced fetal anthropometric parameters, fetal anemia, congenital malaria, increased mother-to-child HIV transmission, and perinatal mortality, were associated with placental malaria. There were, however, conflicting reports on whether the risk of these adverse perinatal outcomes associated with placental malaria were statistically significant. There is a clear need to strengthen the malaria prevention and intervention measures for pregnant women in sub-Saharan Africa. PMID- 18160990 TI - Hypoxia-Inducible Factor (HIF)-1 regulatory pathway and its potential for therapeutic intervention in malignancy and ischemia. AB - Hypoxia-Inducible Factor (HIF)-1 is a dimeric protein complex that plays an integral role in the body's response to low oxygen concentrations, or hypoxia. HIF-1 is among the primary genes involved in the homeostatic process, which can increase vascularization in hypoxic areas such as localized ischemia and tumors. It is a transcription factor for dozens of target genes; HIF-1 is also essential for immunological responses and is a crucial physiological regulator of homeostasis, vascularization, and anaerobic metabolism. Furthermore, HIF-1 is increasingly studied because of its perceived therapeutic potential. As it causes angiogenesis, enhancement of this gene within ischemic patients could promote the vessel proliferation needed for oxygenation. In contrast, as HIF-1 allows for survival and proliferation of cancerous cells due to its angiogenic properties, inhibition potentially could prevent the spread of cancer. With a growing understanding of the HIF-1 pathway, the inhibition and stimulation of its transcriptional activity via small molecules is now an attractive goal. Gene therapy to achieve both vessel proliferation and tumor regression has been demonstrated in animal studies but requires significant improvement and modification before becoming commercially available. This review focuses on the potential of the HIF-1 pathway in therapeutic intervention for the treatment of diseases such as cancer and ischemia. PMID- 18160991 TI - City chaos, contagion, Chadwick, and social justice. PMID- 18160992 TI - Stranger in a strange land: reflections on my first medical relief mission. PMID- 18160993 TI - The apology. AB - This is the second issue featuring a selected piece from the Yale Internal Medicine Residency Program's Writers' Workshop. The annual workshop began in 2003. Abraham Verghese and Richard Selzer, among the best known physician-writers in the United States, have served as workshop leaders, teaching the craft of writing to more than 35 residents. In designing the workshop, Anna B. Reisman, assistant professor in the Department of Internal Medicine at the Yale University School of Medicine and VA Connecticut Health Care System, and Dr. Asghar Rastegar had the goal of making participants better physicians by providing a creative outlet for reflection. The stories and essays written by the Writers' Workshop participants present a range of experiences, real and imagined, and take readers deep into the minds of young doctors trying to make sense of what they do. PMID- 18160994 TI - The 2007 anatomy ceremony: a service of gratitude: I: collected experiences. AB - Yale University medical and PA students, classes of 2010 and 2008 respectively, express their gratitude in a compilation of reflections on learning human anatomy. In coordination with the Section of Anatomy and Experimental Surgery at the School of Medicine, the Yale Journal of Biology and Medicine encourages you to hear the stories of the body as narrated by the student. PMID- 18160996 TI - Diabetic modifier QTLs in F(2) intercrosses carrying homozygous transgene of TGF beta. AB - When the homozygous active form of porcine TGF-beta1 transgene (Tgf/Tgf) (under control of the rat glucagon promoter) is introduced into the nonobese diabetic mouse (NOD) genetic background, the mice develop endocrine and exocrine pancreatic hypoplasia, low serum insulin concentrations, and impaired glucose tolerance. To identify genetic modifiers of the diabetic phenotypes, we crossed hemizygous NOD-Tgf with DBA/2J mice (D2) or C3H/HeJ mice (C3H) and used the "transgenic mice" for quantitative trait loci (QTL) analysis. Genome-wide scans of F(2)-D Tgf/Tgf (D2 x NOD) and F(2)-C Tgf/Tgf (C3H x NOD), homozygous for the TGF-beta1 transgene, identified six statistically significant modifier QTLs: one QTL (Tdn1) in F(2)-D Tgf/Tgf, and five QTLs (Tcn1 to Tcn5) in F(2)-C Tgf/Tgf. Tdn1 (Chr 13, LOD = 4.39), and Tcn3 (Chr 2, LOD = 4.94) showed linkage to body weight at 8 weeks of age. Tcn2 (Chr 7, LOD = 4.38) and Tcn4 (Chr 14, LOD = 3.99 and 3.78) showed linkage to blood glucose (BG) concentrations in ipGTT at 30, 0, and 120 min, respectively. Tcn1 (Chr 1, LOD = 4.41) and Tcn5 (Chr 18, LOD = 4.99) showed linkage to serum insulin concentrations in ipGTT at 30 min. Tcn2 includes the candidate gene, uncoupling protein 2 (Ucp2), and shows linkage to Ucp2 mRNA levels in the soleus muscle (LOD = 4.90). Identification of six QTLs for diabetes related traits in F(2)-D Tgf/Tgf and F(2)-C Tgf/Tgf raises the possibility of identifying candidate susceptibility genes and new targets for drug development for human type 2 diabetes. PMID- 18160997 TI - Polymorphisms of promoter and coding regions of the arylamine N-acetyltransferase 2 (NAT2) gene in the Indonesian population: proposal for a new nomenclature. AB - Polymorphisms of arylamine N-acetyltransferase 2 (NAT2) are reportedly associated with the risk of drug toxicities and development of various diseases. The present study examined NAT2 polymorphisms in both promoter and coding regions in the Indonesian population using PCR direct sequencing. The promoter and coding regions of NAT2 displayed 23 polymorphisms/variations, including eight new ones. Seven haplotypes in the promoter region and six haplotypes in the coding region were inferred. The haplotypes in promoter and coding regions showed limited combinations, and 13 combined haplotypes were inferred. The most frequent haplotypes were U1 (38.9%), U2 (33.5%) in the promoter region and NAT2*4 (37.3%), NAT2*6A (36.8%) in the coding region. When converted to predicted phenotypes, the studied population comprised 65.4% rapid acetylators and 35.6% slow acetylators according to bimodal distribution. According to trimodal distribution, frequencies of predicted phenotypes were 13.6, 50.8 and 35.6% for rapid, intermediate and slow acetylators, respectively. Frequencies of NAT2 alleles for the Indonesian population resembled those of other Southeast Asian populations. We also propose a new NAT2 nomenclature composed of haplotypes in the promoter region and conventional NAT2 haplotypes in the coding region, symbolized by NAT2*4.U1, NAT2*4.U2, NAT2*4.U3, NAT2*4.U5, NAT2*4.U6, NAT2*4.U7, NAT2*6A.U1, NAT2*7B.U2, NAT2*7B.U3, NAT2*5B.U1, NAT2*5B.U4, NAT2*12A.U4 and NAT2*13.U1. PMID- 18160998 TI - Association of an intronic haplotype of the LIPC gene with hyperalphalipoproteinemia in two independent populations. AB - Hepatic lipase (HL) plays a major role in the regulation of plasma lipids. Several groups seeking to find association between the gene encoding HL (LIPC) and plasma concentrations of high-density lipoprotein cholesterol (HDLc) using various methods and populations have reported conflicting results. We have approached the problem of demonstrating a relationship between the LIPC locus and HDLc by means of haplotype association using four single nucleotide polymorphisms (SNPs) (rs12594375G/A, rs8023503C/T, rs4775047C/T, and rs11634134T/A) located in intron 1 of the LIPC gene in two independent Japanese populations consisting of 2,970 and 1,638 individuals, respectively. Significant association between hyperalphalipoproteinemia and a specific haplotype in this intron was detected in both populations. When HDLc levels among the three haplotypic categories were analyzed [haplotype rs8023503C/rs12594375G (haplotype-1; H1) homozygotes (H1H1), haplotype rs8023503T/rs12594375A (haplotype-2; H2) homozygotes (H2H2), and heterozygotes (H1H2)], HDLc levels were lowest among H1H1 [mean +/- standard error (SE) = 58.4 +/- 0.4 mg/dl], highest among H2H2 (62.5 +/- 0.8 mg/dl), and intermediate among H1H2 (59.2 +/- 0.4 mg/dl) (P = 0.00011), indicating that H2 haplotype elevates plasma HDLc levels. This association was validated in the second population (n = 1,638) (P = 0.00070). The results provide convincing evidence that the LIPC locus influences HDL metabolism. PMID- 18160999 TI - Tandem duplications of two separate fragments of the dystrophin gene in a patient with Duchenne muscular dystrophy. AB - Mutations in the dystrophin gene result in the most common inherited muscle disease, Duchenne muscular dystrophy (DMD). Duplications spanning one or more exons have been found to be the second most common disease-causing mutation in the dystrophin gene. Although the duplicated exons are commonly thought to be arranged in tandem, rare noncontiguous exon duplications have been disclosed without clarifying their location or orientation. Here we present the first report that details the exact locations and orientations of noncontiguous duplications in the dystrophin gene. Multiplex ligation-dependent probe amplification analysis of the dystrophin gene of a Japanese boy with DMD revealed that his genomic DNA contained duplications of exons from two separate fragments of the gene: one from exon 45 to exon 48 and the other from exon 55 to exon 63. To clarify the locations and orientations of the duplicated exons, reverse transcription-nested PCR analysis of dystrophin mRNA was conducted. Interestingly, the extra copies of exons 45-48 and exons 55-63 were found to be properly oriented between exons 48 and 49 and exons 63 and 64, respectively. These results indicated that two tandem duplication events occurred in the dystrophin gene of this patient and should contribute to the understanding of the duplication mechanisms that contribute to the development of DMD. PMID- 18161000 TI - Family-based analysis of vitamin D receptor gene polymorphisms and type 1 diabetes in the population of South Croatia. AB - Type 1 diabetes mellitus (T1DM) is a disease characterised by the autoimmune destruction of insulin-producing pancreatic beta cells. Vitamin D is a known immune system modulator and its effects are exerted via the vitamin D receptor (VDR). Several VDR gene single nucleotide polymorphisms (SNPs) have been commonly studied in relation to T1DM. The aim of this study was to evaluate the role of VDR gene variation in T1DM susceptibility by genotyping four SNPs (FokI rs10735810, TaqI-rs731236, BsmI-rs1544410, and Tru9I-rs757343) in 160 case-parent trio samples from the population of South Croatia. We observed overtransmission of Tru9I allele G and undertransmission of the Tru9I-BsmI A-A haplotype from parents to affected children (P = 0.032, P = 0.002, respectively). These results indicate a possible role of the VDR gene in T1DM aetiology. In conclusion, this family-based study presents some evidence of association of specific VDR gene variants with T1DM in the population of South Croatia. PMID- 18161001 TI - Spontaneous pneumothorax in Wegener's granulomatosis: a case report. AB - Wegener's granulomatosis (WG) is a multiorgan system disease of unknown etiology characterized by granulomatous inflammation, tissue necrosis, and variable degrees of vasculitis in small- and medium-sized blood vessels. In patients with WG, the musculoskeletal system can be involved before other typical symptoms of the disease. Also in WG, pneumothorax can develop as a rare complication. In this paper, we report a case with WG who had early articular involvement in the absence of classic features of illness and had pneumothorax which resulted from the rupture of the sub-pleural nodule. The aim of this work is to increase the awareness of early articular involvement in WG and the pneumothorax possibility especially in cases with cavitated nodules closure to pleura. This case emphasizes that early diagnosis and treatment is very important in avoiding further complications. PMID- 18161002 TI - Successful use of etanercept in the treatment of acute lupus hemophagocytic syndrome. AB - Hemophagocytic syndrome has been reported to be associated with systemic lupus erythematosus. A 25-year-old woman with systemic lupus erythematosus developed hemophagocytic syndrome that was refractory to the combination therapy with high dose corticosteroid, cyclosporine, and high-dose intravenous immunoglobulin, and successfully treated with the tumor necrosis factor inhibitor, etanercept. This case report provided the first observation that etanercept may be useful for the treatment of hemophagocytic syndrome associated with systemic lupus erythematosus refractory to conventional therapy. PMID- 18161003 TI - Leflunomide-induced lung injury that developed after its withdrawal, coinciding with peripheral blood lymphocyte count decrease. AB - A 60-year-old rheumatoid arthritis (RA) female with lung fibrosis was treated with leflunomide (LEF) for only 12 days, and responded well. Twenty-five days after the withdrawal of the drug, she had fever, dyspnea, and an elevated serum C reactive protein level. Chest CT revealed ground-glass opacities (GGOs) and consolidations forming a mosaic pattern, in lung fields including the upper, anterior and central areas, and honeycomb patterns in the lung bases and backs. The level of plasma A771726, an active metabolite of LEF, was still as high as that usually noted under LEF therapy. After pulsed steroid and cholestyramine administration, A771726 was depleted and she recovered. The peripheral blood lymphocyte count that had been approximately 1,000/microL, decreased to 220/microL just at the onset of lung injury, and rapidly and steadily returned to the preinjury level preceding recovery from the injury. Serum albumin level decreased in association with lung injury, and gradually returned to the preinjury level. Special caution is necessary when prescribing leflunomide to elderly patients with preexisting interstitial lung disease, and remains necessary until at least 1 month after its withdrawal. PMID- 18161004 TI - PPAR-gamma ligand promotes the growth of APC-mutated HT-29 human colon cancer cells in vitro and in vivo. AB - PPAR-gamma has been known to induce suppression, differentiation and reversal of malignant changes in colon cancer in vitro. However, there are several reports that PPAR-gamma ligands enhance colon polyp development in APCmin mice in vivo. These contradictory results have not yet been thoroughly explained. To explain the contradictory results, we analyzed the effects of different concentrations of the PPAR-gamma agonist, 15-deoxy-D12, 14-prostaglandin (15-d Delta PGJ2) and pioglitazone, on APC gene-mutated colon cancer cell lines (HT-29). We measured cell growth and suppression by cell count and MTT assay and analyzed the expression of beta-catenin and c-Myc protein by Western blot. In addition, we inoculated HT-29 cells into APCmin mice to compare tumor size. High concentrations (10-100 microM/L 15-d Delta PGJ2 and pioglitazone) of PPAR-gamma ligand suppressed growth, while low concentrations (0.01-1 microM/L 15-d Delta PGJ2 and pioglitazone) of PPAR-gamma ligand promoted growth. In particular, the effects of 0.1 microM/L 15-d Delta PGJ2 and pioglitazone on cell growth were statistically significant (P = 0.003, P = 0.001, respectively). Tumor growth was associated with an increase in beta-catenin and c-Myc expression. The growth of xenograft tumors was greater in PPAR-gamma ligand-treated mice than in control mice (control vs day 14: P = 0.024, control vs day 28: P = 0.007). The expression of beta-catenin and c-Myc protein were also elevated in PPAR-gamma-treated mouse tissues. PPAR-gamma ligand can promote the growth of APC-mutated HT-29 colon cancer cells in vitro and in vivo. In addition, the tumor promoting effect seems to be associated with an increase in beta-catenin and c-Myc expression. We think that well-controlled clinical trials should be conducted to confirm our results and to verify clinical applications. PMID- 18161005 TI - Phase II trial of lenalidomide in patients with metastatic renal cell carcinoma. AB - Lenalidomide (CC-5013) is a structural derivative of thalidomide, with antiangiogenic and immunomodulatory effects. Fourteen patients with metastatic renal cell carcinoma (RCC) were enrolled on a phase 2 trial of lenalidomide administered orally at 25 mg daily for 21 days followed by a rest period of 7 days. The best response was stable disease in eight patients (57%) of the 14 evaluable patients. Toxicities included fatigue, hyperglycemia, dyspnea, and myelosuppression with decreased hemoglobin, lymphopenia, and neutropenia. Lenalidomide is tolerable, but no objective responses were observed in this clinical trial. PMID- 18161006 TI - Superior vena cava filters: hindsight, insight and foresight. AB - Upper Extremity Deep Venous Thromboses (UEDVT) are encountered with increasing frequency as the use of central lines and catheters increases. Previously disregarded as benign they are now established to contribute to Pulmonary Embolism (PE) and death. Superior vena cava filters (SVC filters) have been shown to be safe and efficacious for prevention of UEDVT in case anti-coagulation is contraindicated. Compared to Inferior Vena Cava Filters (IVC Filters), data are scarce however and long-term, randomized studies are lacking. SVC filter insertion presents unique indications, contraindications, complications and technical challenges different from their IVC counterparts. PMID- 18161007 TI - Differential expression of MYB gene (OgMYB1) determines color patterning in floral tissue of Oncidium Gower Ramsey. AB - The yellow coloration pattern in Oncidium floral lip associated with red sepal and petal tissues is an ideal model to study coordinate regulation of anthocyanin synthesis. In this study, chromatography analysis revealed that the red coloration in floral tissues was composed of malvidin-3-O-galactoside, peonidin-3 O-glucoside, delphinidin-3-O-glucoside and cyanidin-3-O-glucoside compounds. By contrary, these pigments were not detected in yellow lip tissue. Four key genes involved in anthocyanin biosynthetic pathway, i.e. chalcone synthase (OgCHS), chalcone isomerase (OgCHI), dihydroflavonol 4-reductase (OgDFR) and anthocyanidin synthase (OgANS) were isolated and their expression patterns were characterized. Northern blot analysis confirmed that although they are active during floral development, OgCHI and OgDFR genes are specifically down-regulated in yellow lip tissue. Bombardment with OgCHI and OgDFR genes into lip tissue driven by a flower specific promoter, Pchrc (chromoplast-specific carotenoid-associated gene), demonstrated that transient expression of these two genes resulted in anthocyanin production in yellow lip. Further analysis of a R2R3 MYB transcription factor, OgMYB1, revealed that although it is actively expressed during floral development, it is not expressed in yellow lip tissue. Transient expression of OgMYB1 in lip tissues by bombardment can also induce formation of red pigments through the activation of OgCHI and OgDFR transcription. These results demonstrate that differential expression of OgMYB1 is critical to determine the color pattern of floral organ in Oncidium Gower Ramsey. PMID- 18161008 TI - The genes and enzymes involved in the biosynthesis of thiamin and thiamin diphosphate in yeasts. AB - Thiamin (vitamin B1) is an essential molecule for all living organisms. Its major biologically active derivative is thiamin diphosphate, which serves as a cofactor for several enzymes involved in carbohydrate and amino acid metabolism. Important new functions for thiamin and its phosphate esters have recently been suggested, e.g. in gene expression regulation by influencing mRNA structure, in DNA repair after UV illumination, and in the protection of some organelles against reactive oxygen species. Unlike higher animals, which rely on nutritional thiamin intake, yeasts can synthesize thiamin de novo. The biosynthesis pathways include the separate synthesis of two precursors, 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate and 5-(2-hydroxyethyl)-4-methylthiazole phosphate, which are then condensed into thiamin monophosphate. Additionally, yeasts evolved salvage mechanisms to utilize thiamin and its dephosphorylated late precursors, 4-amino-5 hydroxymethyl-2-methylpyrimidine and 5-(2-hydroxyethyl)-4-methylthiazole, from the environment. The current state of knowledge on the discrete steps of thiamin biosynthesis in yeasts is far from satisfactory; many intermediates are postulated only by analogy to the much better understood biosynthesis process in bacteria. On the other hand, the genetic mechanisms regulating thiamin biosynthesis in yeasts are currently under extensive exploration. Only recently, the structures of some of the yeast enzymes involved in thiamin biosynthesis, such as thiamin diphosphokinase and thiazole synthase, were determined at the atomic resolution, and mechanistic proposals for the catalysis of particular biosynthetic steps started to emerge. PMID- 18161009 TI - Increased pressure stimulates aberrant dendritic cell maturation. AB - Patients with malignancy typically exhibit abnormal dendritic cell profiles. Interstitial tumor pressure is increased 20-50 mmHg over that in normal tissue. We hypothesized that elevated pressure in the tumor microenvironment may influence dendritic cell (DC) phenotype and function. Monocyte-derived immature and mature DC isolated from healthy human donors were exposed to either ambient or 40 mmHg increased pressure at 37 degrees C for 12 hours, then assessed for expression of CD80, CD86, CD83, CD40, MHC-I and MHC-II. IL-12 production and phagocytosis of CFSE-labeled tumor lysate were assessed in parallel. Elevated pressure significantly increased expression of all co-stimulatory and MHC molecules on mature DC. Immature DC significantly increased expression of CD80, CD86, CD83 and MHC-II, but not MHC-I and CD40, versus ambient pressure controls. Pressure-treated immature DC phenotypically resembled mature DC controls, but produced low IL-12. Phenotypic maturation correlated with decreased phagocytic capacity. These results suggest increased extracellular pressure may cause aberrant DC maturation and impair tumor immunosurveillance. PMID- 18161010 TI - The cytoplasmic domain of chondrolectin interacts with the beta-subunit of Rab geranylgeranyl transferase. AB - Mouse chondrolectin (chodl) was isolated out of the tail tip of four-day old 129/SvJ mice as a by-product of a PCR-based subtractive cDNA library screening. The gene is predominantly expressed in adult skeletal muscle, heart, testes and lungs and in embryonic stadia. Chodl is the mouse homologue of human chondrolectin (CHODL), a gene that encodes for a type Ia transmembrane protein and that is expressed in human testis, prostate, heart and skeletal muscle tissue. CHODL-splice variants (CHODL ( f ), CHODL ( f Delta E ), CHODL ( Delta E )) are detected in human leukocytes. The proteins of the chondrolectin family belong to the family of C-type lectins. As the members of this protein family are important for a wide array of biological processes, the function of chodl was investigated by searching for its protein interaction partners. The beta-subunit of Rab geranylgeranyl transferase (Rabggtb) was isolated 8 times after a complete Sos recruitment system (SRS) screen with the cytoplasmic domain of chodl. The interaction was confirmed with in vitro transcription/translation and co immunoprecipitation (co-IP) experiments. PMID- 18161011 TI - Can CYP1A1 siRNA be an effective treatment for lung cancer? AB - Previously, we identified a novel correlation between the upregulated expression of telomerase (hTERT) and cytochrome P450 1A1 (CYP1A1) in A549 human lung cancer cell line. The expression correlation was confirmed by silencing CYP1A1 expression using siRNA technology and observing a silencing of hTERT transcription. Furthermore, silencing CYP1A1 and subsequently downregulating hTERT resulted in the reduction of cancer cell viability by more than 40%, which appeared as early as 24 hours after the treatment. The concomitant downregulation of CYP1A1 and hTERT resulted in rapid cell death. This finding can be further exploited to develop new molecular targets for the treatment of lung cancer. PMID- 18161012 TI - Effects of THC and lofexidine in a human laboratory model of marijuana withdrawal and relapse. AB - INTRODUCTION: Individuals seeking treatment for their marijuana use rarely achieve sustained abstinence. OBJECTIVES: The objectives of the study are to determine if THC, a cannabinoid agonist, and lofexidine, an alpha(2)-adrenergic receptor agonist, given alone and in combination, decreased symptoms of marijuana withdrawal and relapse, defined as a return to marijuana use after a period of abstinence. MATERIALS AND METHODS: Nontreatment-seeking, male volunteers (n = 8), averaging 12 marijuana cigarettes/day, were maintained on each of four medication conditions for 7 days: placebo, tetrahydrocannabinol (THC) (60 mg/day), lofexidine (2.4 mg/day), and THC (60 mg/day) combined with lofexidine (2.4 mg/day); each inpatient phase was separated by an outpatient washout phase. During the first three inpatient days, placebo marijuana was available for self administration (withdrawal). For the next 4 days, active marijuana was available for self-administration (relapse). Participants paid for self-administered marijuana using study earnings. Self-administration, mood, task performance, food intake, and sleep were measured. RESULTS: THC reversed the anorexia and weight loss associated with marijuana withdrawal, and decreased a subset of withdrawal symptoms, but increased sleep onset latency, and did not decrease marijuana relapse. Lofexidine was sedating, worsened abstinence-related anorexia, and did not robustly attenuate withdrawal, but improved sleep and decreased marijuana relapse. The combination of lofexidine and THC produced the most robust improvements in sleep and decreased marijuana withdrawal, craving, and relapse in daily marijuana smokers relative to either medication alone. CONCLUSIONS: These data suggest the combination of lofexidine and THC warrant further testing as a potential treatment for marijuana dependence. PMID- 18161013 TI - Evaluation of mucosal damage and recovery in the gastrointestinal tract of rats by a penetration enhancer. AB - PURPOSE: To evaluate absorption barrier recovery in the gastrointestinal tract after treatment with a penetration enhancer by using a poorly absorbed marker and correlate results with morphological recovery. METHODS: Oral gavage of sodium dodecyl sulfate (SDS) was given to Wistar rats. Phenol red (PR) was given at different time points following administration of SDS. Blood samples were obtained from the jugular vein. Pharmacokinetic analysis was performed on the in vivo data using WinNonlin and MATLAB5 software. The pharmacokinetic parameters of PR were compared to the negative control to measure functional recovery. The intestinal tissues were observed using light and transmission electron microscopy. RESULTS: Absorption was highest when PR was co-administered with SDS. C (max), AUC and K (a) decreased and T (max) and MAT increased as the recovery period (time between administration of SDS and PR) increased. The pharmacokinetic parameters approached the negative control profile in one hour after treatment with 1% SDS. Microscopy results showed recovery of paracellular and transcellular barrier at this time. CONCLUSIONS: Absorption barrier recovery could be measured using a poorly absorbed marker. Functional recovery showed a good correlation with morphological recovery. The local effects of SDS were found to be temporary and reversible. PMID- 18161014 TI - Structural behaviour of 2-hydroxypropyl-beta-cyclodextrin in water: molecular dynamics simulation studies. AB - PURPOSE: To investigate the effect of 2-hydroxypropyl side group substitutions on the structure of beta-cyclodextrin (CD) in water. METHODS: Molecular dynamics simulations were carried out on four HPBCDs that broadly represent a range of degree of substitutions in order to investigate the effect of substitution of beta-cyclodextrin with 2-hydroxypropyl groups at various O2 and O6 positions of the glucose units. RESULTS: The 2-hydroxypropyl side groups located at the O2 positions widen the cavity entrance at the secondary OH position of the CD molecule. These groups are spatially more spread out but dynamically more restricted, due to the formation of a hydrogen bond network between the hydroxyl groups of the side chains and the glucose units. On the other hand, the 2 hydroxypropyl groups at the O6 positions are dynamically more flexible. CONCLUSIONS: The extent and the location of the substitution can affect the cavity structure of the CD molecule, and thus possibly the molecular encapsulation capabilities. PMID- 18161015 TI - Recognition of 'fortune of others' emotions in Asperger syndrome and high functioning autism. AB - 'Fortune of others' emotions, such as envy and gloating over the other's misfortune, are complex emotions experienced in situations where events are presumed to be desirable or undesirable for another person. The present paper explores the notion that individuals with AS and HFA are impaired in understanding of envy and gloating. We tested the ability of adults with AS/HFA to understand envy and gloating and compared their performance to that of age matched healthy controls. The 'fortune of others' emotion task and an additional theory-of-mind (ToM) task were based on a task designed to assess ToM on the basis of eye gaze direction. Individuals with AS and HFA showed no difficulty on basic ToM conditions, but were impaired in their ability to identify envy and gloating. Furthermore, the ability to recognize these emotions was related to scores on a self-rating scale of perspective-taking ability and the ToM task. PMID- 18161016 TI - Brief report: judging pain intensity in children with autism undergoing venepuncture: the influence of facial activity. AB - The biasing effect of pain sensitivity information and the impact of facial activity on observers' judgements of pain intensity of children with autism were examined. Observers received information that pain experience in children with autism is either the same as, more intense than, or less intense than children without autism. After viewing six video clips of children with autism undergoing venepuncture, observers estimated pain intensity using a visual analogue scale. Facial activity as coded by Chambers et al. (Child Facial Action Coding System Revised Manual, 1996) had a significant impact on observers' estimates of pain intensity; pain sensitivity information did not. These results have important implications for the assessment and management of pain in children with autism. PMID- 18161017 TI - Hand asymmetry in heterosexual and homosexual men and women: relationship to 2D:4D digit ratios and other sexually dimorphic anatomical traits. AB - Sexual differentiation leads to the development of distinctive anatomical structures (e.g., gonads and genitalia); it also produces less obvious anatomical shifts in brain, bones, muscles, etc. This study is a retrospective analysis of growth patterns in the hands in relation to sex and sexual orientation. Using data from three published studies, we analyzed four hand traits in adults: hand width, hand length, second digit length, and fourth digit length. Using these measurements, we derived estimates of trait laterality (directional asymmetry or DA) and developmental instability (fluctuating asymmetry or FA). High FA is a putative indicator of interference with the cellular and molecular mechanisms regulating development. We focused on how these derived variables were related to sex, sexual orientation, and putative markers of early sex steroid exposure (e.g., the second to fourth digit ratio or 2D:4D). Our data point to three principal conclusions. First, individual differences in DA appeared to be a major source of variation in the 2D:4D ratio. The 2D:4D ratios of heterosexual men differed depending on whether they had leftward or rightward DA in their digits. Homosexual women showed the same pattern. Individuals with leftward DA in both digits had lower 2D:4D ratios than those with rightward DA. This effect was absent in heterosexual women and homosexual men. This led to sex differences in 2D:4D and sexual orientation differences in 2D:4D in the leftward DA group, but not in the rightward DA group. The second conclusion was that DA in digit length and hand width varied with sex; women were more likely to have rightward asymmetry than men. Homosexual men and women were generally sex typical in DA. The third conclusion was that homosexuality is unlikely to be a result of increased developmental instability. Although limited in scope, the present evidence actually suggests that homosexuals have lower FA than heterosexuals, raising the question of whether the positive fitness components associated with low FA may contribute to selection that maintains homosexuality in a population. PMID- 18161018 TI - HIV voluntary counseling and testing and behavioral risk reduction in developing countries: a meta-analysis, 1990--2005. AB - The effectiveness of HIV voluntary counseling and testing (VCT) in reducing HIV risk behaviors in developing countries was assessed using meta-analytic methods. A standardized protocol was used for searching, acquiring, and extracting study data and meta-analyzing the results. Seven studies met the inclusion criteria. VCT recipients were significantly less likely to engage in unprotected sex when compared to behaviors before receiving VCT, or as compared to participants who had not received VCT [OR 1.69; 95%CI 1.25-2.31]. VCT had no significant effect on the number of sex partners [OR 1.22; 95%CI 0.89-1.67]. While these findings provide only moderate evidence in support of VCT as an effective prevention strategy, neither do they negate the need to expand access to HIV testing and counseling services. Such expansion, however, must be accompanied by rigorous evaluation in order to test, refine and maximize the preventive benefits of learning one's HIV infection status through HIV testing and counseling. PMID- 18161019 TI - New populations at high risk of HIV/STIs in low-income, urban coastal Peru. AB - The HIV epidemic in Peru is concentrated primarily among men who have sex with men. HIV interventions have focused exclusively on a narrowly defined group of MSM and FSW to the exclusion of other populations potentially at increased risk. Interventions targeting MSM and FSW are insufficient and there is evidence that focusing prevention efforts solely on these populations may ignore others that do not fall directly into these categories. This paper describes non-traditional, vulnerable populations within low-income neighborhoods. These populations were identified through the use of ethnographic and epidemiologic formative research methods and the results are reported in this publication. Although the traditional vulnerable groups are still in need of prevention efforts, this study provides evidence of previously unrecognized populations at increased risk that should also receive attention from HIV/STI prevention programs. PMID- 18161020 TI - Multiple sexual partnerships in a sample of African-American crack smokers. AB - The sample for this study consisted of 692 sexually active African-American crack cocaine users living in Houston, TX who reported more than one sexual partner in the previous 30 days. Participants were asked to describe each of their two most recent partners from a list of eight choices: spouse/like a spouse/lover; close friend/friend/acquaintance/customer you like/customer. Analyses were conducted on the 1,384 partners and 692 partnership combinations reported. Partnerships and partnership patterns were examined with respect to three risk behaviors unprotected sex, alcohol use to accompany sex, and drug use to accompany sex-and with respect to three affective measures-partner intimacy, condom use responsibility, and condom use self-efficacy. Results indicate that while many partnerships were based on trading sex for money or drugs, many participants reported partners they considered a spouse or friend. Risk behaviors and affective measures were found to differ by partner type. PMID- 18161021 TI - Predicting nonresponse bias from teacher ratings of mental health problems in primary school children. AB - The impact of nonresponse on estimates of mental health problems was examined in a prospective teacher screen in a community survey of 9,155 7-9 year olds. For 6,611 of the children, parents consented to participation in the actual study (Responders), while for 2,544 children parental consent was not obtained (Nonresponders). The teacher screen involved assessment of a broad set of symptoms of mental health problems and functional impairment. Calculations of non response coefficients, a function of effect sizes and non-response proportion, revealed only ignorable nonresponse bias for both mean scores and correlations. However, the results from binary logistic regressions revealed that children ascribed signs of mental health problems by their teachers were less likely to participate. This was most frequent among children with only moderate symptoms. However, it also involved children with high symptom scores related to inattention, hyperactivity, emotions and peer relationship problems. These findings suggest that measures based on effect size can underestimate the magnitude of non-response bias and that a logistic regression approach may be more appropriate for studies geared at estimating prevalence of mental health problems in children. PMID- 18161022 TI - Introduction of Kenneth I. Maton on his reciept of the 2006 SCRA Award for Distinguished Contributions to Theory and Research. PMID- 18161023 TI - Chemical conjugation of muramyl dipeptide and paclitaxel to explore the combination of immunotherapy and chemotherapy for cancer. AB - Paclitaxel (Taxol) conjugated to muramyl dipeptide (MDP) is described. Biological testing showed that the conjugation of MDP at 2'-O-paclitaxel (2'- O -MTC-01) not only has antitumor activity, but also have immunoenhancement capacity. Compared with paclitaxel or MDP alone or with a mixture of paclitaxel + MDP, 2'- O -MTC-01 significantly increases the production and expression of TNF-alpha and IL-12 from mouse peritoneal macrophages, which demonstrates a synergism of MDP and paclitaxel in one conjugated molecule. PMID- 18161024 TI - Is human placenta proteoglycan remodeling involved in pre-eclampsia? AB - Impaired placento-fetal communication is a coherent symptom of exaggerated pre eclampsia. The impact of the cellular expression of different glycosaminoglycans (GAGs) in this event on the placenta in pre-eclampsia is still obscure. This is the first study aimed at discovering the relationship between structural alterations of different sulfated GAGs at the molecular level and the development of pre-eclampsia in inflicted placenta. Sulfated GAGs were isolated and purified from control and pre-eclampsia placentas. The amount and the molecular weight of GAG in each tissue sample were measured. The polydispersity of the recovered GAG samples were determined by polyacrylamide gel electrophoresis. The disaccharide composition of chondroitin sulfate, dermatan sulfate and heparan sulfate were deduced by chondroitinase and heparinase depolymerization followed by liquid chromatography-mass spectrometry. The in vivo sulfo-modulation of GAGs in pre eclampsia and control placenta were examined using RT-PCR to determine the transcription levels of different sulfotransferases involved in GAG biosynthesis. Marked differences in GAG sulfation patterns and mRNA level of encoding selected GAG O-sulfotransferases were observed in pre-eclampsia. These data suggest a linkage between pre-eclampsia and the observed alterations in placental GAGs and could provide new insights about the modulating role of GAGs in the development and the severity of placental pre-eclampsia. PMID- 18161025 TI - TLR-independent induction of human monocyte IL-1 by phosphoglycolipids from thermophilic bacteria. AB - The structures of phosphoglycolipids PGL1 and PGL2 from the thermophilic bacteria Meiothermus taiwanensis, Meiothermus ruber, Thermus thermophilus, and Thermus oshimai are determined recently (Yang et al. in J Lipid Res. 47:1823-1932, 2006). These bacteria belong to Gram-negative bacteria that do not contain lipopolysaccharide, but high amounts of phosphoglycolipids and glycoglycerolipids. Here we show that PGL1/PGL2 mixture (PGL1: PGL2 = 10:1 ~ 10:2) from M. taiwanensis and T. oshimai, but not T. thermophilus and M. ruber, up-regulate interleukin-1beta (IL-1beta) production in human THP-1 monocytes and blood-isolated primary monocytes. PGL2 was purified after phospholipase A2 hydrolysis of PGL1 in the PGL1/PGL2 mixture followed by column chromatography. PGL2 did not induce proIL-1 production, even, partially (35-40%) inhibited PGL1 mediated proIL-1 production, showing that PGL1 is the main inducer of proIL-1 production in PGL1/PGL2 mixture. The production of proIL-1 stimulated by phosphoglycolipids was strongly inhibited by specific PKC-alpha, MEK1/2, and JNK inhibitors, but not by p38-specific inhibitor. The intracellular calcium influx was involved in phosphoglycolipids-mediated proIL-1 production. Using blocking antibody and Toll-like receptor (TLR)-linked NF-kappaB luciferase assays, we found that the cellular receptor(s) for phosphoglycolipids on proIL-1 production was TLR-independent. Further, phosphoglycolipids isolated from T. thermophilus and M. ruber did not induce proIL-1 production, even though T. thermophilus possess more PGL1 than PGL2 (6:4). Specially, the fatty acid composition of phosphoglycolipids from both T. thermophilus and M. ruber consists of a low percentage of C15 (<10%) and a high percentage of C17 (>75%). It suggests, the C15 percentage of PGL may play a critical role in PGL-mediated proIL-1 induction. PMID- 18161026 TI - Characteristics of urban sidewalks/streets and objectively measured physical activity. AB - Several studies have found significant relationships between environmental characteristics (e.g., number of destinations, aesthetics) and physical activity. While a few of these studies verified that the physical activities assessed were performed in the environments examined, none have done this in an urban, neighborhood setting. This information will help efforts to inform policy decisions regarding the design of more "physically active" communities. Fourteen environmental characteristics of 60, 305-m-long segments, located in an urban, residential setting, were directly measured using standardized procedures. The number of individuals walking, jogging, and biking in the segments was assessed using an observation technique. The segments were heterogeneous with regards to several of the environmental characteristics. A total of 473 individuals were seen walking, bicycling, or jogging in the segments during 3,600 min of observation (60 min/segment). Of the 473 seen, 315 were walking, 116 bicycling, and 42 jogging. A greater number of individuals were seen walking in segments with more traffic, sidewalk defects, graffiti, and litter and less desirable property aesthetics. Only one environmental characteristic was associated with bicycling and none were significantly related with jogging. This study provides further evidence that environmental characteristics and walking are related. It also adds new information regarding the importance of scale (e.g., micro, macro) and how some environmental characteristics of urban, residential sidewalks and streets relate to physical activity. PMID- 18161027 TI - Biological and chemical monitoring of some major Calabrian (Italy) Rivers. AB - To address the problem of whether hydrographic basins of southern Italian regions such as Calabria are exposed to "ecological stability pressures," we here provide the first results of four chemical and biological (macroinvertebrates) monitoring activities. The data accumulated within four hydrographic basins between May and November of 2002, November of 2005 and May of 2006 showed that about 1/3 of the macrobenthos sampled among a total of 14 rivers in the northern-central part of Calabria region provided optimal environmental quality values of Extended Biotic Index (E.B.I.), 1/3 denoted a slight impairment and the remaining samples exhibited very strong environmental alterations. It is interesting to note that E.B.I. is directly correlated with the taxonomic richness plus abundance, and inversely with the dominating taxon. This suggests that high environmental quality values are rather positively correlated with spatial heterogeneity and taxa eveness. In addition, the more compromised stations were located on plain segments where anthropization is reported to be at its highest level and there the macrobenthic communities seem to be made up of mostly potamon type of taxa with a high abundance of collector euriecious organisms while better quality stations, mostly located at high altitude, are characterized by a considerable presence of E.P.T. taxa. Chemical parameters did not result to be substantially altered underling that the hydrologic and geomorphological alterations may play a primary role in the control of macrobenthic communities and environmental quality. Overall the above results suggest that E.B.I. variations seem to be highly coherent with changes of main biodiversity parameters and their joined application is an efficient tool for detecting ecological state alterations of lotic environments. PMID- 18161028 TI - A new assessment method for urbanization environmental impact: urban environment entropy model and its application. AB - The thermodynamic law is one of the most widely used scientific principles. The comparability between the environmental impact of urbanization and the thermodynamic entropy was systematically analyzed. Consequently, the concept "Urban Environment Entropy" was brought forward and the "Urban Environment Entropy" model was established for urbanization environmental impact assessment in this study. The model was then utilized in a case study for the assessment of river water quality in the Pearl River Delta Economic Zone. The results indicated that the assessing results of the model are consistent to that of the equalized synthetic pollution index method. Therefore, it can be concluded that the Urban Environment Entropy model has high reliability and can be applied widely in urbanization environmental assessment research using many different environmental parameters. PMID- 18161029 TI - Application of multivariate statistical techniques to evaluation of water quality in the Mala Welna River (Western Poland). AB - The paper presents the results of determinations of physico-chemical parameters of the Mala Welna waters, a river situated in Wielkopolska voivodeship (Western Poland). Samples for the physico-chemical analysis were taken in eight gauging cross-sections once a month between May and November 2006. To assess the physico chemical composition of surface water, use was made of multivariate statistical methods of data analysis, viz. cluster analysis (CA), factor analysis (FA), principal components analysis (PCA), and discriminant analysis (DA). They made it possible to observe similarities and differences in the physico-chemical composition of water in the gauging cross-sections, to identify water quality indicators suitable for characterising its temporal and spatial variability, to uncover hidden factors accounting for the structure of the data, and to assess the impact of man-made sources of water pollution. PMID- 18161030 TI - Variable phenotypes associated with mutations in DOK7. AB - Many patients with the limb-girdle variant of congenital myasthenic syndrome (CMS) possess mutations in the human Dok-7 gene (DOK7). We identified six unrelated CMS patients with DOK7 mutations. Two patients, one mildly and the other moderately affected, were homozygous for the previously described 1263insC mutation. The common 1124_1127dupTGCC mutation was detected in the other four patients, whose clinical phenotypes range from mildly to severely affected. This striking phenotypic heterogeneity found both within and between mutational classes is made more compelling by data from our electrophysiological studies and electron microscopy of the neuromuscular junction (NMJ). Indeed, several aspects of the physiological and morphometric data do not correlate with genotype or severity of clinical phenotype. Overall, our study corroborates the findings of others and provides an additional demonstration of the considerable phenotypic variability associated with CMS due to DOK7 mutations. PMID- 18161031 TI - Practice and endpoint accuracy with the left and right hands of old adults: the right-hemisphere aging model. AB - The purpose of the study was to quantify the aging-related differences in endpoint accuracy during isometric contractions of the left and right hands based on the prediction that declines in motor performance with aging may be greater for muscles controlled by the right hemisphere. Twelve young (6 men, 25 +/- 5 years) and 12 old (6 men, 76 +/- 6 years) adults performed a task that involved matching the peak of a force-time trajectory to a target. The old adults were less accurate than the young men and exhibited greater endpoint error with the left hand than the right hand on day 1, but not on days 2 and 3. Although electromyographic amplitude was similar between hands, old adults exhibited greater timing variability. These findings indicate that given sufficient practice there was no difference in endpoint accuracy between the left and right hands of old adults, which is not consistent with the prediction of an asymmetrical decline in motor performance by the right-hemisphere aging model. Conversely, an inability by an old adult to achieve similar accuracy with both hands during such tasks likely indicates an underlying motor impairment. PMID- 18161032 TI - Self-determination theory, motivational interviewing, and the treatment of clients with acute suicidal ideation. AB - An overwhelming number of individuals who die by suicide suffer from mental disorders, but comparatively few are seen in psychological or psychiatric treatment. Given the efficacy of psychosocial interventions in decreasing suicide related behavior, the development of strategies to increase motivation for treatment is critical for clients with acute suicidal ideation. Self Determination Theory (SDT) provides a framework for understanding how the therapeutic relationship can affect clients' motivation for treatment. When working with clients with serious suicidal ideation, clinicians can use the principles of SDT or interventions that are congruent with SDT, such as motivational interviewing (MI), to increase clients' engagement in treatment and improve treatment outcome. SDT-based suggestions and an adaptation of MI are introduced to guide clinicians through this process. PMID- 18161033 TI - The relational context of aggression in borderline personality disorder: using adult attachment style to predict forms of hostility. AB - Attachment theory provides a framework for understanding and predicting critical aspects of aggression in the personality disorders. An association between borderline personality disorder (BPD) and insecure forms of adult attachment marked by high relationship anxiety has been repeatedly observed in the empirical literature. Aggression also has been linked to insecure attachment. The present study extends previous work by exploring the degree to which the underlying attachment dimensions of relationship anxiety and avoidance are associated in BPD with the following forms of hostility: (a) direct aggression (verbal or physical) initiated towards others, (b) expectation/perception of aggression from others (including "reactive" counteraggression when/if provoked), (c) aggression directed towards the self in the form of suicidality or parasuicidality, and (d) affective experience of irritability or anger. The issue was studied in a sample of 92 patients diagnosed with BPD. Results show significant association between more fearful forms of attachment (simultaneous presence of relationship anxiety and avoidance) and the more reactive form of aggression involving expectation of hostility from others. Self-harm was significantly associated only with relational avoidance while anger and irritability were associated only with relational anxiety. Implications for understanding relational aspects of BPD aggression in research and clinical work are discussed. PMID- 18161034 TI - Psychometric properties of the Beck Depression Inventory-II in nonclinical adolescent samples. AB - This study examined the factor structure and psychometric properties of the Beck Depression Inventory-II (BDI-II; A. T. Beck, Steer, & Brown, 1996) in samples of high-school adolescents (N=414; 210 boys and 204 girls, ages 14-18 years). Confirmatory factor analyses provided satisfactory fit estimates for the two- and three-factor oblique solutions reported frequently in the extant literature. The solution to a general factor with domain-specific somatic and cognitive-affective factors was examined as an alternative model to previously established models for the current high-school sample data. Results provided stronger support for the general factor model. Estimates of internal consistency for scores on this instrument were high (coefficient alpha=.92, average interitem correlation=.35). The mean BDI-II total score for the nonclinical samples (M=12.50, SD=10.50) was compared with the mean scores reported for various adolescent normative samples in the extant literature. The BDI-II total score correlated moderately and significantly with scores on self-report measures of hopelessness (r=.63), anxiety (r=.53), and suicide-related behaviors (r=.57), providing support of adequate correlates for the BDI-II. Estimates of known-groups validity were evaluated using data from a small sample of 167 clinical adolescent inpatients. Specific study findings, limitations, and recommendations are discussed. PMID- 18161036 TI - Psychological treatment of postpartum depression: a meta-analysis. AB - Postpartum depression is a widespread and disruptive depressive disorder seriously affecting the lives of new mothers and their families. We conducted a meta-analysis of controlled and comparative studies of psychological treatments of postpartum depression. Seventeen studies were included. The mean standardized effect size of all psychological treatments compared to control conditions was 0.61 (95% CI: 0.37 approximately 0.85). Several subgroup analyses were conducted. Studies with waiting list control groups had a larger mean effect size (0.96; 95% CI: 0.63 approximately 1.29) than studies with a care-as-usual control group (0.41; 95% CI: 0.25 approximately 0.58). No definite conclusions can be drawn about the longer term effects. Too few studies were available to draw conclusions about the relative effects of psychological treatments compared to pharmacological and other treatments. PMID- 18161037 TI - Higher levels of central adiposity in healthy premenopausal women with family histories of premenopausal breast cancer. AB - Research strongly suggests that lower overall adiposity and higher central adiposity are independent risk factors for premenopausal breast cancer in the general population. We aimed to test the possibility that these factors may contribute to familial risk of premenopausal breast cancer. A convenience sample of healthy women, ages 25-49, was recruited to yield three study groups: (1) Women with first-degree family histories of premenopausal breast cancer, operationally defined as being diagnosed prior to age 50 (Group FH < 50, n = 39); (2) Women with first-degree family histories of postmenopausal breast cancer, operationally defined as being diagnosed at age 50 or after (Group FH > or = 50, n = 33); and (3) Women without a history of breast cancer in first-degree relatives (Group FH-, n = 132). Multinomial logistic regression analyses, including possible confounders, waist circumference, and BMI, revealed a lower BMI among FH < 50 compared to either FH- (OR = 0.72; 95% CI = 0.59-0.87), or FH > or = 50 women (OR = 0.75; 95% CI = 0.60-0.95), and higher waist circumferences in FH < 50 compared to either FH- (OR = 1.15; 95% CI = 1.06-1.25), or FH > or = 50 women (OR = 1.16; 95% CI = 1.05-1.28). No group differences were seen for waist skinfold measures. These results support the possibility that differences in patterns of adiposity may contribute to familial risk of premenopausal breast cancer, and suggest the importance of conducting large scale, population-based studies of the link between body size characteristics and familial breast cancer risk. PMID- 18161038 TI - Active ghrelin levels across time and associations with leptin and anthropometrics in healthy ache Amerindian women of Paraguay. AB - Active (acylated) ghrelin is a peptide hormone secreted primarily by the stomach, positively associated with fasting, orexigenic, and promotes growth hormone secretion. It is therefore important to energy intake management. The objective of this pilot research was to (1) compare active ghrelin with previous measurements of leptin and anthropometrics; (2) assess the consistency of active ghrelin across time in this population; (3) extend our understanding of potential population variation in active ghrelin. Two serum samples separated by 10 days at the same time between meals were collected from healthy Ache women (n = 12, mean age 32.2 +/- 14.0 SD) to determine consistency over time, associations with leptin, and anthropmetric values. Mean active ghrelin was 72.9 +/- 23.0 pg/ml, highly correlated (r(2) = 0.95, P < 0.0001) between collections, and showed no paired mean differences (P < 0.18). There was no significant correlation with leptin, age, or anthropometric measures. Active ghrelin appears to be consistent over time in this population, perhaps reflecting regimented meal schedules and less interpopulation variation compared to leptin. PMID- 18161039 TI - A comparative analysis of estimation of age at menarche by various methods in women participating in the Krakow Longitudinal Growth Study, Poland. AB - The purpose of this study was to see whether menarcheal age assessment by means of the most frequently used methods that were conducted every time on the same group of girls would yield the same results. One hundred and one Polish girls, whose ages at menarche were recorded in a longitudinal study between 1976 and 1990, were asked to recall the age of menarche in 2004. The mean menarcheal ages of those women were calculated by means of the probit (PA), prospective (AA), retrospective method without age correction (RA), and retrospective method with the recall age corrected by 0.5 year (RcA). The PA, AA, and retrospective methods: RA and RcA revealed results: 13.14 +/- 1.1; 13.10 +/- 1.1; 13.12 +/- 1.36, and 14.39 +/- 1.34 years, respectively. The menarcheal AA was insignificantly different from the PA (95% CI) and RA. The RcA was significantly higher than the AA and RA (P = 0.05). The correlation coefficient (r) between AA and RA was 0.70. Only 16% of the interviewed women accurately remembered the date of their menarche, 63% of them missed their menarche time by about 1 year, whereas 22% were wrong by 2 and more years. (1) The PA and the AA method yield comparable results when estimating menarcheal age. (2) The menarcheal age determined by the retrospective methods is not very reliable and the application of age correction overestimates the results. PMID- 18161040 TI - Y-STR analysis in Brazilian and South Amerindian populations. AB - A sample of 203 Brazilian males from Rio Grande do Sul (RS), the Brazilian southernmost state, was typed for 11 Y-STR markers (DYS19, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS385, DYS437, DYS438, and DYS439). We also typed 42 individuals from two South Amerindian tribes (Kaingang and Guarani) to use the data as parental Amerindian contribution to our analyses. Gene and haplotypic diversities were estimated, with the South Amerindian samples showing smaller values for these parameters than Brazilians. To obtain a more comprehensive picture of the genetic structure of the Brazilian population as a whole, the Y STR data from the RS sample was compared with those already published. No genetic substructuring was observed in the comparisons performed. Multidimensional scaling confirmed the proposed European source of most Y-chromosome Brazilian patrilineages. PMID- 18161041 TI - Major gene effect and additive familial pattern of inheritance of asthma exist among families of probands with sickle cell anemia and asthma. AB - In the United States, sickle cell anemia (SCA) affects approximately 1 in 400 African-American newborns. Acute episodes of pain and acute chest syndrome (ACS) are the two leading causes of hospitalization. A relationship between the diagnosis of asthma and the incidence of pain and ACS has been established. We tested the hypothesis that a familial pattern of inheritance of asthma exists among first degree relatives of probands with SCA and asthma. Segregation analysis was performed in 104 families ascertained through affected probands. Of these, 19.7% (41/208) of the parents and 31.8% (28/88) of siblings of affected probands reported having been told by a doctor he or she had asthma at any age. Modes of inheritance were tested, using the Pedigree Analysis Package parameterized for the discrete trait of asthma affection status. A major effect was present and significant. Further tests were performed to determine whether transmission probabilities of the major effect followed Mendelian expectations. The additive mode of inheritance was the most parsimonious, while the residual heritability was found negligible. Our results support the hypothesis that a familial pattern of inheritance of asthma exists among first degree relatives of probands with SCA and asthma, suggesting that asthma is a co-morbid condition with SCA rather than a lung disease phenotype mimicking asthma. PMID- 18161042 TI - Sudden gains in the treatment of generalized anxiety disorder. AB - The objective of this study was to investigate the prevalence and timing of sudden gains over the course of brief, psychodynamically oriented treatment for generalized anxiety disorder (GAD). Data were used from two studies of brief (i.e., 16-session) supportive-expressive psychotherapy for GAD. Anxiety symptoms were measured at every weekly treatment session. Sudden gains in anxiety symptoms were defined to parallel previous research on sudden gains in major depressive disorder (MDD). Overall, sudden gains were found for 11 of 68 participants (16.2%), with 4 (36.4%) of these patients experiencing reversals of these gains and losing over 50% of the sudden gain during subsequent treatment sessions. Applying a baseline severity cutoff and a duration criteria similar to those used in previous studies of sudden gains resulted in 10 of 29 (34.5%) patients showing sudden gains. Of these sudden gainers, 4 (40.0%) experienced a reversal and 7 (70%) experienced an upwards spike in symptoms during their psychotherapy course. When defined in a parallel fashion, rates of sudden gains in GAD are similar to those found in MDD, although anxiety symptoms are highly variable. PMID- 18161043 TI - Cognitive mediational deficits and the role of coping styles in pedophile and ephebophile Roman Catholic clergy. AB - This study was designed to examine hypothesized differences between sex offending and nonoffending Roman Catholic clergy on cognitive mediation abilities as measured by the Rorschach Inkblot Test (H. Rorschach, 1921/1942). This study compared 78 priest pedophiles and 77 priest ephebophiles with 80 nonoffending priest controls on the Inkblot test using J. E. Exner's (2003) Comprehensive System. The three groups were compared on seven variables that constitute Exner's Cognitive Mediation cluster. Additionally, the groups' coping styles were compared to examine the interaction of coping style and cognitive mediational abilities. We found interactions between coping style and offending status across most of the cognitive variables indicating impairment in the mild to pathological ranges. Moreover, significantly higher unusual thinking styles (Xu%) and significantly lower conventional thinking styles (X+%) in offenders compared to nonoffenders. Those with an Extratensive style (n=31) showed significantly higher distorted thinking when compared to the Introversive (n=81), Ambitent (n=73), and Avoidant (n=50) coping styles. This study suggests that offenders display significantly higher distorted thinking styles than do nonoffenders. Possible reasons for these discrepancies and the role of coping styles in abusive behaviors were discussed. PMID- 18161044 TI - Christian religious functioning and trauma outcomes. AB - While some trauma survivors find their faith helpful in recovery, others find it a source of distress, and still others abandon their faith. More complex conceptualizations of religious functioning are needed to explore its relationship with trauma. This study explores such relationships using measures of religious action and behaviors in a community sample of 327 church-going, self identified trauma survivors. A principal components analysis of positive and negative religious coping, religious comforts and strains, and prayer functions identified two dimensions: Seeking Spiritual Support, which was positively related to posttraumatic growth, and Religious Strain, which was positively related to posttraumatic symptoms. PMID- 18161045 TI - Scared to lose control? General and health locus of control in females with a phobia of vomiting. AB - The term emetophobia (i.e., a fear of vomiting) exists as rather an elusive predicament, often eluding conventional treatment. The present study involved 149 participants, consisting of 51 emetophobics, 48 phobic controls (i.e. those who suffered from a different phobia), and 50 nonphobic controls. Participants were administered the Rotter (1966) Locus of Control Scale and the Health Locus of Control Scale by B.S. Wallston, Wallston, Kaplan, and Maides (1976). Significant differences were found among the three groups; specifically, that emetophobics had a significantly higher internal Locus of Control Scale score with regard to both general and health-related issues than did the two control groups. It is suggested that vomiting phobics may have a fear of losing control, and that their vomiting phobia is reflective of this alternative, underlying problem. More research is required to explore the association between emetophobia and issues surrounding control; however, the current study suggests that it may be helpful for therapists to consider this aspect when treating a patient with vomiting phobia. PMID- 18161046 TI - Client personality characteristics predict satisfaction with cognitive behavior therapy. AB - Ratings of treatment satisfaction are a means for cognitive behavior therapy clients to provide their unique personal perspective on their therapy experience. Treatment satisfaction is a variable of growing importance as a predictor of outcome for various medical and psychological treatments including treatments for chronic pain (D. C. Turk et al., 2003). Our goal was to determine whether satisfaction with cognitive behavior therapy sessions varied as a function of patient personality characteristics in a sample of 43 older adults (average age=72.3 years, SD=8.0) participating in a psychosocial pain management therapy program with a cognitive behavioral orientation. Participants completed the NEO Five Factor Inventory (P. T. Costa, Jr. & R. R. McCrae, 1992) prior to the commencement of treatment and a psychometrically valid questionnaire, assessing satisfaction with psychological therapy, after each therapy session. The core personality dimensions of neuroticism, openness, and agreeableness were predictive of aspects of satisfaction with therapy. These findings have the potential of being useful to clinicians concerned with the prediction of response to therapy. PMID- 18161047 TI - Model for end-stage liver disease (MELD) for liver allocation: a 5-year score card. PMID- 18161048 TI - Aberrant methylation of multiple tumor suppressor genes in aging liver, chronic hepatitis, and hepatocellular carcinoma. AB - Aberrant DNA methylation is an important epigenetic alteration in hepatocellular carcinoma (HCC). However, the molecular processes underlying the methylator phenotype and the contribution of hepatitis viruses are poorly understood. The current study is a comprehensive methylation analysis of human liver tissue specimens. A total of 176 liver tissues, including 77 pairs of HCCs and matching noncancerous liver and 22 normal livers, were analyzed for methylation. Methylation of 19 epigenetic markers was quantified, and the results were correlated with different disease states and the presence or absence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections. Based on methylation profiles, the 19 loci were categorized into 3 groups. Normal liver tissues showed methylation primarily in group 1 loci (HIC-1, CASP8, GSTP1, SOCS1, RASSF1A, p16, APC), which was significantly higher than group 2 (CDH1, RUNX3, RIZ1, SFRP2, MINT31) and group 3 markers (COX2, MINT1, CACNA1G, RASSF2, MINT2, Reprimo, DCC) (P < 0.0001). Noncancerous livers demonstrated increased methylation in both group 1 and group 2 loci. Methylation was significantly more abundant in HCV positive livers compared with normal liver tissues. Conversely, HCC showed frequent methylation at each locus investigated in all 3 groups. However, the group 3 loci showed more dense and frequent methylation in HCV-positive cancers compared with both HBV-positive cancers and virus-negative cancers (P < 0.0001). CONCLUSION: Methylation in HCC is frequent but occurs in a gene-specific and disease-specific manner. Methylation profiling allowed us to determine that aberrant methylation is commonly present in normal aging livers, and sequentially progresses with advancing stages of chronic viral infection. Finally, our data provide evidence that HCV infection may accelerate the methylation process and suggests a continuum of increasing methylation with persistent viral infection and carcinogenesis in the liver. PMID- 18161049 TI - Natural antisense transcript stabilizes inducible nitric oxide synthase messenger RNA in rat hepatocytes. AB - During inflammation, inducible nitric oxide synthase (iNOS) is induced to generate the important mediator nitric oxide (NO). Interleukin 1beta (IL-1beta) induces iNOS messenger RNA (mRNA), iNOS protein, and NO in rat hepatocytes. We found that the stability of iNOS mRNA changed during the induction and that the antisense (AS) strand corresponding to the 3'-untranslated region (3'UTR) of iNOS mRNA was transcribed from the iNOS gene. Expression levels of the iNOS AS transcript correlated with those of iNOS mRNA. The 1.5-kilobase region 3' flanking to iNOS gene exon 27 was involved in IL-1beta induction. Knockdown experiments suggest that sense oligonucleotides to iNOS mRNA significantly reduced iNOS mRNA levels in the hepatocytes by blocking the interaction between iNOS mRNA and the AS transcript. Overexpression of iNOS AS transcript stabilized the reporter luciferase mRNA through the fused iNOS mRNA 3'UTR. These results together with the data in a yeast RNA-hybrid assay suggested that the iNOS AS transcript interacted with iNOS mRNA and stabilized iNOS mRNA. The iNOS mRNA colocalized with the AU-rich element-binding protein HuR, a human homolog of embryonic lethal-abnormal visual protein, and heterogeneous nuclear ribonucleoprotein L (hnRNP L) in the cytoplasm of rat hepatocytes. Interaction assays further revealed that the iNOS AS transcript interacted with HuR, which interacted with hnRNP L, suggesting that iNOS mRNA, the AS transcript, and the RNA-binding proteins may mutually interact. CONCLUSION: The natural AS transcript of the iNOS gene interacts with iNOS mRNA and may play an important role in the stability of iNOS mRNA. This RNA-RNA interaction may be a new therapeutic target for NO-mediating inflammatory diseases. PMID- 18161050 TI - Etiology-dependent molecular mechanisms in human hepatocarcinogenesis. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and is characterized by aggressive tumor behavior coupled with poor prognosis. Various etiologies have been linked to HCC development, most prominently chronic hepatitis B and C virus infections as well as chronic alcohol consumption. In approximately 10% of HCCs, the etiology remains cryptic; however, recent epidemiological data suggest that most of these cryptogenic HCCs develop due to nonalcoholic steatohepatitis. To identify etiology-dependent DNA copy number aberrations and genes relevant to hepatocarcinogenesis, we performed array-based comparative genomic hybridization of 63 HCCs of well-defined etiology and 4 HCC cell lines followed by gene expression profiling and functional analyses of candidate genes. For a 10-megabase chromosome region on 8q24, we observed etiology-dependent copy number gains and MYC overexpression in viral and alcohol related HCCs, resulting in up-regulation of MYC target genes. Cryptogenic HCCs showed neither 8q24 gains, nor MYC overexpression, nor target gene activation, suggesting that tumors of this etiology develop by way of a distinct MYC independent pathomechanism. Furthermore, we detected several etiology-independent small chromosome aberrations, including amplification of MDM4 on 1q32.1 and frequent gains of EEF1A2 on 20q13.33. Both genes were overexpressed in approximately half the HCCs examined, and gene silencing reduced cell viability as well as proliferation and increased apoptosis rates in HCC cell lines. CONCLUSION: Our findings suggest that MDM4 and EEF1A2 act as etiology-independent oncogenes in a significant percentage of HCCs. PMID- 18161051 TI - Association of gankyrin protein expression with early clinical stages and insulin like growth factor-binding protein 5 expression in human hepatocellular carcinoma. AB - Gankyrin (also known as PSMD10) is a liver oncoprotein that interacts with multiple proteins including MDM2 and accelerates degradation of the tumor suppressors p53 and Rb. We produced a monoclonal anti-gankyrin antibody and immunohistochemically assessed the clinicopathological significance of gankyrin overexpression in 43 specimens of human hepatocellular carcinoma (HCC). Specific cytoplasmic staining for gankyrin was observed in 62.8% (27/43) of HCCs, which was significantly associated with low TNM stage (P = 0.004), no capsular invasion (P = 0.018), no portal venous invasion (P = 0.008), and no intrahepatic metastasis (P = 0.012). The cumulative survival rate of patients with gankyrin positive HCC was significantly higher than that with gankyrin-negative HCC (P = 0.037). p53 and MDM2 were positively stained by antibodies in 30.2% and 23.3%, respectively, of HCCs, but neither was inversely associated with gankyrin expression. In the Huh-7 human HCC cell line, overexpression of gankyrin up regulated expression of insulin-like growth factor binding protein 5 (IGFBP-5), whereas suppression of gankyrin expression by siRNA down-regulated it. Supression of IGFBP-5 expression inhibited proliferation of Huh-7 cells as well as U-2 OS osteosarcoma cells. In HCC specimens, positive staining for IGFBP-5 was observed by immunohistochemistry in 41.9% (18/43), and the level of expression was significantly correlated with that of gankyrin (rho = 0.629, P < 0.001). CONCLUSION: These results suggest that gankyrin plays an oncogenic role(s) mainly at the early stages of human hepatocarcinogenesis, and that IGFBP-5 inducible by gankyrin overexpression may be involved in it. PMID- 18161052 TI - Improved correction for population stratification in genome-wide association studies by identifying hidden population structures. AB - Hidden population substructure can cause population stratification and lead to false-positive findings in population-based genome-wide association (GWA) studies. Given a large panel of markers scanned in a GWA study, it becomes increasingly feasible to uncover the hidden population substructure within the study sample based on measured genotypes across the genome. Recognizing that population substructure can be displayed as clustered and/or continuous patterns of genetic variation, we propose a method that aims at the detection and correction of the confounding effect resulting from both patterns of population substructure. The proposed method is an extension of the EIGENSTRAT method (Price et al. [2006] Nat Genet 38:904-909). This approach is computationally feasible and easily applied to large-scale GWA studies. We show through simulation studies that, compared with the EIGENSTRAT method, the new method requires a smaller number of markers and yields a more appropriate correction for population stratification. PMID- 18161053 TI - Computational models of ethanol-induced neurodevelopmental toxicity across species: Implications for risk assessment. AB - Computational, systems-based approaches can provide a quantitative construct for evaluating risk in the context of mechanistic data. Previously, we developed computational models for the rat, mouse, rhesus monkey, and human, describing the acquisition of adult neuron number in the neocortex during the key neurodevelopmental processes of neurogenesis and synaptogenesis. Here we apply mechanistic data from the rat describing ethanol-induced toxicity in the developing neocortex to evaluate the utility of these models for analyzing neurodevelopmental toxicity across species. Our model can explain long-term neocortical neuronal loss in the rodent model after in utero exposure to ethanol based on inhibition of proliferation during neurogenesis. Our human model predicts a significant neuronal deficit after daily peak BECs reaching 10-20 mg/dl, which is the approximate BEC reached after drinking one standard drink within one hour. In contrast, peak daily BECs of 100 mg/dl are necessary to predict similar deficits in the rat. Our model prediction of increased sensitivity of primate species to ethanol-induced inhibition of proliferation is based on application of in vivo experimental data from primates showing a prolonged rapid growth period in the primate versus rodent neuronal progenitor population. To place our predictions into a broader context, we evaluate the evidence for functional low-dose effects across rats, monkeys, and humans. Results from this critical evaluation suggest subtle effects are evident at doses causing peak BECs of approximately 20 mg/dl daily, corroborating our model predictions. Our example highlights the utility of a systems-based modeling approach in risk assessment. PMID- 18161054 TI - Dr. Holzheimer's affirmation of the findings of McVay and Anson (1940) and Condon(1964). PMID- 18161055 TI - The clinical anatomy of congenital portosystemic venous shunts. AB - Congenital portosystemic venous shunts are rare. Their gross anatomy has not been well defined. Four different varieties of congenital portosystemic venous shunts are described in six children seen during a 10-year period, focusing on the anatomy of the shunt as determined by imaging studies and surgery. A detailed review of the literature indicates that congenital portosystemic venous shunts are best classified as: extrahepatic or intrahepatic. Extrahepatic shunts may be further subdivided into portocaval shunts (type 1 end-to-side and type 2 side-to side) and others. Intrahepatic shunts are due to an abnormal intrahepatic connection between the portal vein and hepatic vein/inferior vena cava or a persistent patent ductus venosus. Additional congenital anomalies, particularly cardiac malformations, may be associated with any type. Some congenital intrahepatic portosystemic venous shunts close spontaneously in infancy; all other congenital portosystemic venous shunts tend to remain patent. To a variable extent, depending largely on the volume and duration of the shunt, affected individuals are at risk of developing hepatic encephalopathy and/or an intrahepatic tumor. The key to understanding the pathogenesis of these shunts lies in the normal developmental mechanisms underlying the formation of the portal vein and inferior vena cava in the embryo. PMID- 18161056 TI - Alcohol and stroke risk: an elusive dose-response relationship. PMID- 18161057 TI - System for tamoxifen-inducible expression of cre-recombinase from the Foxa2 locus in mice. AB - To study the roles of key transcription factor networks, growth factors, and signaling molecules in the endoderm, notochord, and floorplate, we developed an inducible Cre-expressing system for altering gene function in this tissue. We generated an allele of Foxa2 that directs a tamoxifen-regulated Cre in the Foxa2 expression domain (Foxa2(mcm)). Activity of Foxa2(mcm) recapitulates endogenous Foxa2 expression in endoderm, notochord, and floorplate. Efficiency of the system in a given tissue type was dose- and timing-dependent. By comparing efficiency and location of Cre activity after administration of tamoxifen by oral gavage vs. intraperitoneal injection, we found that oral gavage achieves more rapid, robust recombination with less embryonic toxicity. This system will be useful for controlling the activity of floxed alleles at multiple stages of mouse embryogenesis and fetal development. PMID- 18161058 TI - Characterization of the cardiac phenotype in neonatal Ts65Dn mice. AB - The Ts65Dn mouse is the most-studied of murine models for Down syndrome. Homology between the triplicated murine genes and those on human chromosome 21 correlates with shared anomalies of Ts65Dn mice and Down syndrome patients, including congenital heart defects. Lethality is associated with inheritance of the T65Dn chromosome, and anomalies such as right aortic arch with Kommerell's diverticulum and interrupted aortic arch were found in trisomic neonates. The incidence of gross vascular abnormalities was 17% in the trisomic population. Histological analyses revealed interventricular septal defects and broad foramen ovale, while immunohistochemistry showed abnormal muscle composition in the cardiac valves of trisomic neonates. These findings confirm that the gene imbalance present in Ts65Dn disrupts crucial pathways during cardiac development. The candidate genes for congenital heart defects that are among the 104 triplicated genes in Ts65Dn mice are, therefore, implicated in the dysregulation of normal cardiogenic pathways in this model. PMID- 18161059 TI - Developmental expression patterns of the zebrafish small heat shock proteins. AB - Small heat shock proteins (sHSPs), or alpha-crystallins, are low-molecular weight proteins found in every kingdom and nearly every species examined to date. Many, if not all, sHSPs act as molecular chaperones. Several also have functions independent of their chaperone activity, and at least a few are expressed in specific spatiotemporal patterns during embryonic and/or juvenile stages, suggesting specific roles during development. To date, however, no one has systematically characterized the expression patterns of all of the sHSPs during development in any organism. We have characterized the normal heat shock-induced expression patterns of all 13 zebrafish sHSPs during development. Seven of the sHSPs are expressed in a tissue-specific manner during development, and five are upregulated by heat shock. The results of these studies provide a foundation for analysis of sHSP function during normal development and their roles in protecting cells from the effects environmental stressors. PMID- 18161060 TI - How to stem a global health crisis. PMID- 18161062 TI - NF-kB induces lung maturation during mouse lung morphogenesis. AB - Lung maturation is hallmarked by the appearance of surfactant-producing alveoli during transition from the saccular to alveolar stage of lung development. Inflammation can disrupt this process and accelerate lung maturity following intrauterine amniotic infection (chorioamnionitis). Nuclear factor kB (NF-kB) is a transcription factor central to multiple inflammatory and developmental pathways, including dorsal-ventral patterning in fruit flies, limb and mammary and submandibular gland development in mice, and branching morphogenesis in chick lungs. Given its shared role in inflammation and developmental signaling, we hypothesized that overexpression of NF-kB targeted to the lung epithelium would exert maturational effects on alveolar development. We generated transgenic mice with lung-specific overexpression of the RelA subunit of NF-kB using a surfactant protein C promoter construct. Our results showed that RelA overexpression in the lung yields increased alveolar type I and type II cells. These findings are consistent with a model whereby NF-kB may induce maturation of lung development through decreased apoptosis of epithelial cells. PMID- 18161063 TI - Comparison of molecular and cellular events during lower jaw regeneration of newt (Cynops pyrrhogaster) and West African clawed frog (Xenopus tropicalis). AB - When mammals, including humans, lose a major part of their lower jaw, they are unable to regenerate the lost structures. Urodele amphibians, especially newts, can regenerate their lower jaw after amputation, whereas most anuran amphibians, including the West African clawed frog, can not. In the present study, we investigated the difference between newts and frogs during lower jaw regeneration. One difference was the distribution of myosin heavy chain (MHC) mRNA after lower jaw amputation: MHC mRNA was immediately expressed at the tip of the amputated lower jaw in newts but not in frogs. Moreover, there were proliferating cells that expressed Pax7 in newts but not in frogs, although proliferating cells were present in both animals. These results suggest that the difference of the jaw-regenerating abilities between newts and frogs depends on the expression of MHC mRNA at the tip of the amputated jaw and the contribution of Pax7-positive cells. PMID- 18161064 TI - Construction of X-ray-inducible promoters through cis-acting element elongation and error-prone polymerase chain reaction. AB - BACKGROUND: A promoter that is activated by ionizing radiation may be a useful tool for cancer therapy since, with such a promoter, the therapeutic gene can be expressed only in cancer tissues by irradiation. An artificially constructed promoter is advantageous as natural promoters may have physiological limitations. However, reasonably designing a promoter is hampered by shortage of information about the relationship between the structure and properties of a promoter DNA. MATERIALS AND METHODS: Binding sites of four transcription factors that were activated by radiation were randomly ligated and linked to a TATA-box sequence to control the luciferase gene located downstream. Transiently transfected cancer cells with such a vector were exposed to X-ray irradiation and enhancement of luciferase expression was assessed. To improve promoter sensitivity, mutations were randomly introduced into a constructed promoter by error-prone polymerase chain reaction (epPCR). RESULTS: Of the 11 promoters constructed, the clone 11 promoter (clone 11 + TATA-box) showed a 5-fold enhancement 6 h after the 10 Gy X ray irradiation in HeLa cells. A mutant designated the clone 11-9-37 promoter generated through two steps of epPCR showed a sensitivity 4.8 times higher than the clone 11 promoter to the 10 Gy X-rays, showing 21.6-fold enhancement of luciferase expression. Clone 11 was composed of 16 cis-acting elements, and the clone 11-9-37 promoter carried six point mutations. CONCLUSION: A sensitively responsive promoter to radiation could be constructed using this method, possibly leading to the construction of a promoter of interest that could be applied for clinical use. PMID- 18161065 TI - Fate of glyphosate in soil and the possibility of leaching to ground and surface waters: a review. AB - The very wide use of glyphosate to control weeds in agricultural, silvicultural and urban areas throughout the world requires that special attention be paid to its possible transport from terrestrial to aquatic environments. The aim of this review is to present and discuss the state of knowledge on sorption, degradation and leachability of glyphosate in soils. Difficulties of drawing clear and unambiguous conclusions because of strong soil dependency and limited conclusive investigations are pointed out. Nevertheless, the risk of ground and surface water pollution by glyphosate seems limited because of sorption onto variable charge soil minerals, e.g. aluminium and iron oxides, and because of microbial degradation. Although sorption and degradation are affected by many factors that might be expected to affect glyphosate mobility in soils, glyphosate leaching seems mainly determined by soil structure and rainfall. Limited leaching has been observed in non-structured sandy soils, while subsurface leaching to drainage systems was observed in a structured soil with preferential flow in macropores, but only when high rainfall followed glyphosate application. Glyphosate in drainage water runs into surface waters but not necessarily to groundwater because it may be sorbed and degraded in deeper soil layers before reaching the groundwater. Although the transport of glyphosate from land to water environments seems very limited, knowledge about subsurface leaching and surface runoff of glyphosate as well as the importance of this transport as related to ground and surface water quality is scarce. PMID- 18161066 TI - Health economics: basic principles and application in mental health. PMID- 18161067 TI - Ebola outbreak in Uganda "atypical", say experts. PMID- 18161068 TI - Reports call for overhaul of US foreign assistance. PMID- 18161069 TI - Polish doctors fall out with new government over pay. PMID- 18161070 TI - Minimally invasive, robotic cardiac surgery. PMID- 18161071 TI - The preeminence of clusters. AB - Many cities and countries still view the foundation of a biotech sector as desirable for a high-tech, intellectually driven economy. But a discussion by seasoned biotech management and investors suggests that attaining an environment with the right mix of money, management and innovation remains a difficult and long-term challenge. PMID- 18161072 TI - Which thoughts can kill a boxer? naive theories about cognitive and emotional antecedents of suicide. AB - OBJECTIVES: We investigated naive theories regarding the association among beliefs, emotions and behaviours to test Rational Emotive Behaviour Therapy's (REBT) assumption that rational cognitions and adaptive emotions lead to functional behaviours whereas irrational cognitions and maladaptive emotions trigger dysfunctional reactions. DESIGN: We applied an experimental between subjects design. METHODS: Participants read newspaper articles about the defeat of a boxer. In one condition, the authentic article informed participants that he committed suicide and in the other, a fictitious article about the same defeat described the athlete as successfully continuing his career. Different question formats were employed to assess participants' assumptions about the stimulus person's defeat-related cognitions and emotions: rating scales, sentence completion and free responses. RESULTS: Participants assumed significantly more irrational beliefs (e.g. I absolutely have to win) on the side of the boxer in the suicide scenario than in the non-suicide version. This finding was obtained by directive and non-directive assessment methods. Additionally, participants expected the suicidal stimulus person to be experiencing maladaptive emotions (e.g. depression, guilt) whereas a successful resumption of his career lead to expectations of adaptive affects (e.g. sadness, concern). Ratings of the functionality revealed that sadness, fear, annoyance and concern were expected to be more functional than depression, anxiety, rage and guilt. CONCLUSIONS: The results show that naive psychological theories about the antecedents of dysfunctional behaviour are in accordance with theoretical assumptions of REBT: Irrational beliefs are viewed to be connected with maladaptive emotions and to result in dysfunctional behaviour, and adaptive emotions are thought to be of higher functional value than their maladaptive counterparts. The use of different question formats and a between-subject design excluded that results are due to methodological artifacts or contrast effects. PMID- 18161073 TI - Reduced awareness of executive dysfunction in Alzheimer's disease is associated with increased carer burden. PMID- 18161074 TI - "Alien face" in corticobasal degeneration syndrome: extending clinical features. PMID- 18161075 TI - Influence of receptor activator of nuclear factor (NF)-kappaB ligand (RANKL), macrophage-colony stimulating factor (M-CSF) and fetal calf serum on human osteoclast formation and activity. AB - Human osteoclast (OC) formation and activity was studied in cultures of peripheral blood mononuclear cells (PBMNC) from six healthy donors after stimulation with fetal calf serum (FCS), under the influence of the receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) and the macrophage-colony stimulating factor (M-CSF). The results showed that selected FCS could stimulate OC formation without any medium supplementation with osteoclastogenic factors. The OC formation, investigated by quantification of multinucleated tartrate resistant acid phosphatase-positive cells (TRAP+ cells), and the sensitivity of OC progenitors to RANKL and M-CSF, varied widely between individual donors. The OC resorption activity, measured in the "pit-assay" on dentine, was strictly dependent on the presence of RANKL and M-CSF in the medium and was also donor dependent. The considerable donor variability should be considered in culture studies investigating, e.g. the interactions of OC with biomaterials or the influence of cytokines, growth factors and drugs on osteoclastogenesis. PMID- 18161076 TI - [Paradigm shift for the definition and classification of osteoporosis]. PMID- 18161077 TI - [Current global trends in the prevention and treatment of osteoporosis]. PMID- 18161078 TI - [Japanese Guidelines for the Prevention and Treatment of Osteoporosis (2006 Edition) and its significance]. PMID- 18161079 TI - [Outline of Guidelines 2004 for bone marker in osteoporosis]. PMID- 18161080 TI - [Management of hip fracture in older people: a clinical guideline in Japan 2005]. PMID- 18161081 TI - [Guidelines on the management and treatment of glucocorticoid-induced osteoporosis (2004) in Japan]. PMID- 18161082 TI - [Diagnosis and management of Paget's disease of bone]. PMID- 18161083 TI - [Regulation of bone mass by PI3 kinase/Akt signaling]. PMID- 18161084 TI - [Osteoblast differentiation induced by BMP signaling and Runx2 through Cbfb regulation]. PMID- 18161085 TI - [Role of Gli family members in Indian hedgehog-induced osteoblast differentiation]. PMID- 18161086 TI - [Function of Osterix in osteoblasts]. PMID- 18161087 TI - [Functional analysis of newly identified c-Src-binding proteins in osteoclastogenesis]. PMID- 18161088 TI - [Regulation of bone formation by adiponectin through autocrine/paracrine and endocrine pathways]. PMID- 18161089 TI - [Regulation of bone metabolism by pathogenic mutations and polymorphism in the LRP5-Wnt signaling genes]. PMID- 18161090 TI - [Novel phosphate-regulating hormone, fibroblast growth factor (FGF) 23]. PMID- 18161091 TI - [Control of bone remodeling by leptin and sympathetic nervous system]. PMID- 18161092 TI - [Gamma-GTP]. PMID- 18161093 TI - [Difference in prevalence of osteoporosis diagnosed by bone density measurement within various bone sites]. PMID- 18161094 TI - [Prevalence of osteoporosis-cohort studies in Japan]. PMID- 18161095 TI - [Prospective cohort studies of incident fractures in patients with rheumatoid arthritis]. PMID- 18161096 TI - [Progress in the international cohort studies for the prevention of osteoporosis]. PMID- 18161097 TI - [Impact of osteoporosis-related fractures on mortality and the quality of life (QOL)]. PMID- 18161098 TI - [Lumbar: DXA(QDR, DPX, XR)]. PMID- 18161099 TI - [Radius measurement: DXA, pQCT]. PMID- 18161100 TI - [Second metacarpal bone: RA(CXD, DIP), DXA]. PMID- 18161101 TI - [Femur: DXA(QDR, DPX, XR)]. PMID- 18161102 TI - [QUS and DXA(heel scan) in calcaneus]. PMID- 18161103 TI - [New assessment of bone fragility--from viewpoint of microstructure]. PMID- 18161104 TI - [Evaluation of bone mechanical properties in terms of bone quality]. PMID- 18161105 TI - [Turnover in bone]. PMID- 18161106 TI - [Mineralization in bone]. PMID- 18161107 TI - [Accumulation of microdamage in bone]. PMID- 18161108 TI - [Role of enzymatic cross-links and non-enzymatic cross-link, advanced glycation end products, as a determinant of bone quality in osteoporosis and diabetes]. PMID- 18161109 TI - [Assessment of bone quality with metabolic bone markers]. PMID- 18161110 TI - [Metabolic markers of bone and clinical use]. PMID- 18161111 TI - [Biochemical markers of bone turnover: Serum P1NP]. PMID- 18161112 TI - [Biochemical markers for bone turnover: Serum beta-CTX]. PMID- 18161113 TI - [Biological marker for bone turnover: Serum osteocalcin]. PMID- 18161114 TI - [Biological markers for bone turnover: Genetic polymorphisms of tissue-non specific alkaline phosphatase (TNSALP) gene and the variation of enzymatic properties]. PMID- 18161115 TI - [Assessment of fracture risk based on bone mineral density]. PMID- 18161116 TI - [Clinical utility of bone markers in the assessment of fracture risk]. PMID- 18161117 TI - [WHO FRAT and fracture risk]. PMID- 18161118 TI - [A-TOP research group/JOINT-02]. PMID- 18161119 TI - [RECORD study]. PMID- 18161120 TI - [FACT study]. PMID- 18161121 TI - [CORE study]. PMID- 18161122 TI - [Risk and benefit assessment in hormone replacement therapy]. PMID- 18161123 TI - [Bisphosphonates for osteoporosis]. PMID- 18161124 TI - [Combination therapy for osteoporosis]. PMID- 18161125 TI - [Medical practice and problem of osteoporosis management by physicians]. PMID- 18161126 TI - [Adherence to medications for treatment of osteoporosis]. PMID- 18161127 TI - [Evidence of alendronate treatment for osteoporosis]. PMID- 18161128 TI - [Evidence of risedronate for treatment of osteoporosis]. PMID- 18161129 TI - [Influence on atherosclerosis of improvement of bone mineral density after medication for osteoporosis]. PMID- 18161130 TI - [Clinical study of bisphosphonates for bone metastasis in breast cancer]. PMID- 18161131 TI - [Clinical implications of bisphosphonate for bone metastases of prostate cancer]. PMID- 18161132 TI - [Effectiveness of bisphosphonate administration for prevention of osteopenia during Gn-RH agonist therapy of endometriosis]. PMID- 18161133 TI - [Efficacy and tolerability of once-weekly administration of 35 mg alendronate and 17.5 mg risedronate]. PMID- 18161134 TI - [New evidence of conjugated estrogen and 17beta-estradiol for treatment and prevention of osteoporosis]. PMID- 18161135 TI - [Evidence of raloxifene on postmenopausal osteoporosis]. PMID- 18161136 TI - [Effects of SERM on the cardiovascular system]. PMID- 18161137 TI - [Recent insights into mechanisms for analgesic effect by calcitonin]. PMID- 18161138 TI - [Antinociceptive effect of calcitonin]. PMID- 18161139 TI - [Latest evidence on the suppressive effect of active vitamin D3 on fractures]. PMID- 18161140 TI - [Evidence for vitamin D effect on skeletal muscle]. PMID- 18161141 TI - [Vitamin K2 treatment for osteoporosis: what is the evidence today?]. PMID- 18161142 TI - [New bone density conservation agents for osteoporosis under research and development: Minodronic acid]. PMID- 18161143 TI - [New bone density conservation agents for osteoporosis under research and development: Zoledronate]. PMID- 18161144 TI - [New bone density conservation agents for osteoporosis under research and development: Ibandronate]. PMID- 18161145 TI - [Status of novel bone-targeting SERMs in development]. PMID- 18161146 TI - [New bone density conservation agents for osteoporosis under research and development: PTH (1-34)]. PMID- 18161147 TI - [New bone density conservation agents for osteoporosis under research and development: Strontium ranelate]. PMID- 18161148 TI - [New bone density conservation agents for osteoporosis under research and development: ED-71]. PMID- 18161149 TI - [New bone density conservation agents for osteoporosis under research and development: Anti-RANKL antibody]. PMID- 18161150 TI - [Surgical treatment for osteoporotic spinal fractures]. PMID- 18161151 TI - [Percutaneous transpedicular vertebroplasty and kyphoplasty for osteoporotic vertebral compression fractures]. PMID- 18161152 TI - [Present status and a future problem in an orthopaedic treatment for hip fracture]. PMID- 18161153 TI - [Primary osteoporosis--its diagnosis and treatment]. PMID- 18161154 TI - [Diagnosis and treatment of postmenopausal osteoporosis]. PMID- 18161155 TI - [Diagnosis and treatment of osteoporosis in men]. PMID- 18161156 TI - [Diagnosis, clinical feature and treatment of glucocorticoid-induced osteoporosis in patients with rheumatoid arthritis]. PMID- 18161157 TI - [Diagnosis and treatment of glucocorticoid-induced osteoporosis in collagen vascular disease]. PMID- 18161158 TI - [Diagnosis and treatment of steroid-induced osteoporosis in respiratory diseases]. PMID- 18161159 TI - [Bone loss in hemodialysis patients]. PMID- 18161160 TI - [Osteoporosis in patients with diabetes mellitus]. PMID- 18161161 TI - [Transplantation and osteoporosis]. PMID- 18161162 TI - [Bisphosphonate treatment in osteonecrosis of the femoral head]. PMID- 18161163 TI - [Metabolic anomalies in bone metastasis of cancer]. PMID- 18161164 TI - [Characteristic features of bone disease in disseminated carcinomatosis of bone marrow]. PMID- 18161165 TI - [Treatment and prevention of fractures in stroke patients]. PMID- 18161166 TI - [Quality of life and medical complications in spinal deformity]. PMID- 18161167 TI - [Quality of life and activity of daily living are influenced by spinal deformities in Japanese elderly women]. PMID- 18161168 TI - [Impact of vertebral fractures on health related quality of life in the elderly with osteoporosis]. PMID- 18161169 TI - [Drug therapy for osteoporosis patients with low back pain]. PMID- 18161170 TI - [Effect of physical activity on bone mineral density]. PMID- 18161171 TI - [Role of nutrition in the prevention and treatment of osteoporosis]. PMID- 18161172 TI - [Fracture prevention for the elderly--fall prevention and hip protector]. PMID- 18161173 TI - [Simple screening test for risk of falls in the elderly]. PMID- 18161174 TI - [Economical viewpoint for treatment of osteoporosis]. PMID- 18161175 TI - [Cost-effectiveness on osteoporosis of HRT]. PMID- 18161176 TI - [Vitamin D insufficiency and bone mineral accrual]. PMID- 18161177 TI - [Therapeutic guidelines in prostate cancer and role for bisphosphonates in bone metastasis]. PMID- 18161178 TI - Arthur Cooper and his pet gibbon ape. PMID- 18161179 TI - Study seeks rural and remote locum nurses. PMID- 18161180 TI - Staff our greatest asset. PMID- 18161181 TI - [Guidelines for evaluation of the safety of nanomaterials]. AB - Due to the current intensive development of nanotechnology all over the world, there is a considerable increase in human exposure to nanoparticles and nanomaterials and their environmental entry and accumulation. A number of the guidelines of the Russian Consumer Surveillance Agency emphasize that the potential risk of nanomaterials to human health and the environment is to be characterized since they have quite other physicochemical properties and a biological effect that the substances in their usual physicochemical state. The paper discusses a ways of entering nanoparticles into the body and their potential toxicity, as well as the procedure for assessing a risk associated with the production, use, and recycling of nanoparticles and nanomaterials. PMID- 18161182 TI - [A system of measures to prevent and reduce the incidence of environment dependent diseases]. PMID- 18161183 TI - [Assessment of real risk of urban chemical exposure to the population's health]. AB - Improving the chemoanalytical quality control of the urban environment requires transition from the assessment considering only certain things determined by target analyses to the comprehensive assessment based on monitoring with the identification of the maximally complete spectra of substances contained in the environmental objects and coming from the sources of pollution, by using the appropriate algorithm; identification, quantifying a spectrum of pollutants as completely as possible; selection of the leading indices, by evaluating the detected composition of pollutions from the degree of their hygienic significance, by taking into account a set of criteria (detection rate, concentrations, group affiliation, specificity for a nearby source of pollution in the check of drinking water, a capacity for transformation, possible formation of more toxic transformation products); and monitoring through target tests by the chosen leading indices. PMID- 18161184 TI - [Motor transport emission, ambient air quality, and the Moscow population's health]. AB - As of 2006, the city's motor transport fleet amounted to as many as 3 million units that annually consume about 5 million tons of petrol. The use rate of all kinds of vehicles has increased, resulting in the growth of the proportion of ambient air pollutants discharged by motor transport, which surpasses the increase of the absolute size of the fleet. The contribution of traveling sources to ambient air pollution is growing steadily and it has been recently about 90% (1 million tons). Implementation of measures and developed managerial decisions, and ecological programs, improvement of Moscow town-planning measures, and environment-improving measures against motor vehicles have contributed to a reduction in chemical and physical burdens on the population. The characteristics of the capital's ambient air pollution have been recently observed to become stable and improve. There is stabilization in morbidity due to respiratory diseases in all population groups. The prevalence of chronic respiratory diseases in children is on the decrease, the increase rate was 1.4% versus 33.5% in the preceding period. Assessment of carcinogenic risk showed that ambient air pollution and drinking water contamination had a negative impact on the Moscow population. PMID- 18161185 TI - [Use of an assessment of risk to the population's health in substantiating the sanitary protective zone of OOO "Karbodin" in Orekhovo-Zuyevo, Moscow Region]. PMID- 18161186 TI - [Cryophysical method for estimating the content of structured-phase water]. AB - The paper provides evidence for the principle of calculating the content of water bound states from the quantity of gases evolved during a cryophysical process. A cryophysical procedure has been developed to estimate the degree of water structuredness, by calculating the index from the mean values and spatial variance characteristics. PMID- 18161187 TI - [Nonspecific effects of noise exposure]. PMID- 18161188 TI - [Electromagnetic radiation and its risk to the population's health in the use of cellular communication means]. PMID- 18161189 TI - [Current socio-medical problems in the maintenance of agricultural workers' health]. PMID- 18161190 TI - [Estimation of the sanitary and microbiological indices of stored and bacteriophaged milk]. AB - The paper gives data on the microbiological study of raw milk delivered by various suppliers to a reprocessor. More than 57.3% of the farms are shown to deliver abnormal milk of poor and very poor sanitary quality. The content of lactobacilli can be indirectly judged from the titer and presence of lactophages, the quantity of the latter may serve as an index of the sanitary state of raw milk. Milk bactocentrifugation considerably reduce the quantity of bacteria and their spores, facilitating a longer preservation of milk organoleptic properties. PMID- 18161191 TI - [Impact of cooking on the content of fish mercury]. AB - Changes in the levels of mercury in the fish inhabiting the water reservoirs of Russia were studied after cooking it in a variety of ways (boiling for different time periods, roasting, and smoking). To have a general idea of the impact of various procedures for cooking the fish on its mercury amount, the authors measured mercury concentrations in smoked perch and manufactured and homemade canned salt-and fresh-water fishes. The absolute content of mercury in the fish is shown to be unchanged during its cooking and not to depend on the procedure and duration of the latter. PMID- 18161192 TI - [Assessment of the structure and quality of nutrition as a factor that influences thyroid functional activity]. AB - The purpose of the study was to determine the provision of schoolchildren with vitamin A, beta-carotene, and zinc, by taking into account their mutual effect on thyroid functional activity. The study was retrospective and performed by the Adygei Branch of the Kuban State Medical University, Federal Health Care and Social Development Agency (Maikop). The nutrition pattern was studied in 103 pupils living in the Republic of Adygea. The amount of food used with- in 3 days (with one day-off being compulsorily included) was estimated to determine the actual dietary levels of micronutrients in the schoolchildren. Their provision with vitamin A and beta-carotene was determined by the dark adaptation test (Kravkov-Purkinje test). The data were processed by parametric statistical methods, by using the computer program "Statistica 5.0". Questionnaire analysis has indicated that the diet is unbalanced in the majority of the examined schoolchildren. The study revealed the insufficient dietary intake of iodine, zinc, vitamin A, and beta-carotene. About half the parents of the schoolchildren participating in the study indicated the use of iodinated salt for cooking. Insufficient vitamin A provision was revealed in 77.8% of the pupils. For prevention of deficiencies of vitamin A and traces, 37% of the schoolchildren were found to take multivitamin preparations. Vitamin A deficiency is one of the factors contributing to the development of iodine deficiencies. Along with this, by affecting vitamin A metabolism, inadequate dietary intake of zinc adversely affects thyroid functional activity. Monitoring of adequate dietary intake of vitamin A and zinc should be one of the stages in preventing iodine deficiencies and reducing the number of risk groups in an area with low environmental iodine levels. PMID- 18161193 TI - [Development of a system for the early diagnosis and prevention of children's health changes caused by exposure to heavy metals]. AB - At the moment, the making a diagnosis of an environment-induced disease in Russia is not routine and this is not relied on a rather evidence basis. Nevertheless, the choice of exposure methods from educational and preventive to therapeutic depends on the intensity and pattern of exposure to ecotoxicants and on the magnitude of appropriate specific changes. Epidemiological surveys of 5-7-year old children from 3 towns of Russia have revealed that 8.3-24.5% of the children have moderate excesses of the allowable levels of lead in blood; 4.6-12.8% have excess hair magnesium levels, and 5.9-33.8% have excess hair copper levels. The real damage to the children's neuropsychic health manifests as a significant reduction in the memory and academic achievement indices, the parameters of fine and gross motor coordination, speech expressiveness, and in the rate of minor successive movements. The prevalence of environment-dependent neuropsychic malformations is 3-7%, as calculated with reference to all the children examined. The children aged 5-7 years are shown not to be in need of therapeutic measures in all the towns compared. At the same time, 61.1-83.2% of the children need neuropsychic health-promoting measures. For this, it is expedient to use informational and educational technologies. About 4.3-17.8 and 11.5 - 21.1% need primary and secondary preventive measures against environment-dependent neuropsychic changes. The authors present approaches to developing measures to promote neuropsychic health and to prevent neuropsychic malformations. There is evidence that primary prevention is more effective for the pediatric population. There is no question that multivariate analysis of the problems associated with childhood neuropsychic development, including environmental, behavioral, sociomedical components, is of importance. These approaches may be realized at the municipal level and taken as the basis for preparing national guidelines for the promotion of health and for the prevention of environment-dependent conditions in children. PMID- 18161194 TI - [The performance of vital organs and systems in urban and rural adolescents]. PMID- 18161195 TI - [Assessment of the health of the pediatric population living in towns with varying ambient air pollution]. PMID- 18161196 TI - [Evaluation of toxicity and hazard of waste resulting from the treatment of municipal waste]. AB - Municipal waste, sand trap sediment, floatable substances, raw sedimentation sludge, biological solids, a mixture of compact and dehydrated sediments, ashes, combustion gases and waste by GOST 12.1.007-76 are shown to be classified as classes 3 and 4 hazard. The conditions contributing to the occurrence of acute and chronic intoxications, infectious diseases, and helminthic invasions in the personnel, to environmental pollution at the basic stages of municipal waste and combustion sediment utilization are considered. PMID- 18161197 TI - Universal coverage, guaranteed. My idea: an employer mandate and a new insurance plan that pays for itself. PMID- 18161198 TI - Get rid of the for-profits. Healthcare can no longer be a commodity for sale to those who can afford it. PMID- 18161199 TI - A federalist approach. I would use federal incentives to deregulate, reform state health insurance. PMID- 18161200 TI - Care is complicated...but paying for it is simple: one government program. PMID- 18161201 TI - Commissioning. The grand plan for the NHS to lead the world. PMID- 18161202 TI - Data briefing. Babies still not all born equal. PMID- 18161203 TI - Descriptive epidemiology of Vibrio parahaemolyticus and other Vibrio species infections in British Columbia: 2001-2006. PMID- 18161204 TI - Encephalitis in the summer: a case of snowshoe hare (California serogroup) virus infection in Nova Scotia. PMID- 18161205 TI - A community outbreak of travel-acquired hepatitis A transmitted by an infected food handler. PMID- 18161206 TI - Summary of the outcomes from the National Consensus Conference for Vaccine Preventable Diseases in Canada, Quebec, Quebec, June 12-14, 2005. PMID- 18161207 TI - Initial health screening results for Karen refugees: a retrospective review. PMID- 18161208 TI - [Liberalization of divorce: a Nordic model?]. PMID- 18161209 TI - Subject to empire: married women and the British nationality and Status of Aliens Act. PMID- 18161210 TI - Low-dose atorvastatin in severe chronic kidney disease patients: a randomized, controlled endpoint study. AB - OBJECTIVE: There have been no endpoint studies with statins for patients with severe renal failure. The purpose of this prospective, open, randomized, controlled study was to investigate whether atorvastatin (10 mg/day) would alter cardiovascular endpoints and the overall mortality rate of patients with chronic kidney disease stage 4 or 5 (creatinine clearance < 30 ml/min). MATERIAL AND METHODS: The study subjects comprised 143 patients who were randomized either to placebo (controls; n=73; mean age 69.5 years) or to treatment with atorvastatin (n=70; mean age 67.9 years). The patients included were either non-dialysis (n=33), haemodialysis (n=97) or peritoneal dialysis (n=13) patients. Analysis focused on the primary endpoints of all-cause mortality, non-lethal acute myocardial infarction, coronary artery bypass graft surgery and percutaneous transluminal coronary angioplasty. Statistical analysis for endpoint data was mainly by intention-to-treat. RESULTS: Primary endpoints occurred in 74% of the subjects. There was no difference in outcome between the control and atorvastatin groups. The 5-year endpoint-free survival rate from study entry was 20%. Atorvastatin was withdrawn in 20% of patients due to unacceptable side-effects. In the atorvastatin group, low-density lipoprotein (LDL) cholesterol was reduced by 35% at 1 month and then sustained. The controls showed a progressive reduction in LDL cholesterol until 36 months. CONCLUSIONS: Although atorvastatin reduced total and LDL cholesterol effectively it was not beneficial regarding the long term outcomes of cardiovascular endpoints or survival. In contrast to other patient groups, patients with severe chronic kidney disease, especially those on dialysis, seem to derive limited benefit from this lower dose of atorvastatin. PMID- 18161211 TI - [Agricultural machinery and Hungarian agriculture after World War II, 1945-48]. PMID- 18161212 TI - Good sports and right sorts: guns, gender, and imperialism in British India. PMID- 18161213 TI - Charity, rights, and entitlement: gender, labor, and welfare in early-twentieth century Chile. PMID- 18161214 TI - Colonial government, social conflict and state involvement in Africa's open economies: the origins of the Ghana cocoa marketing board, 1939-46. PMID- 18161216 TI - Race, empire and British wartime national identity, 1939-45. PMID- 18161217 TI - [Radical structural thinking and the organization of totalitarian rule: worldview elites and human scientists in the Nazi regime]. PMID- 18161218 TI - A glittering icon of Fascist femininity: Trebisonda "Ondina" Valla. AB - This essay focuses on the second decade of the Italian Fascist regime through its emblematic symbol, Trebisonda 'Ondina' Valla. Valla gained first place in the 80 metre hurdles at the Berlin Olympics of 1936, and became the first world-class female athlete in Italian history, in spite of the generally backward condition of Italian women. In those years, a paternalistic and conservative society deeply discriminated against female participation in not only sport but also other cultural activities. The Catholic Church, medical expertise, eugenics theories and the fascist regime were all opposed to female competitive sport. The Church demanded female morality, modesty and domesticity while, the medical profession recommend only basic physical exercise for female health and motherhood. While promoting the myth of the New Italy as a modern nation, Fascism wished it to be inhabited by a traditional womanhood. Paradoxically, however, Mussolini supported Valla because she epitomized a dynamic fascism and brought Italian Fascism international visibility. The serendipitous value of Valla was that she encouraged young women to attempt to force open the bars of their political cage, and at the same time forced the fascist ideology to reconsider and reconstruct fascist principles in the interest of international propaganda. Thus while Valla was a political instrument of fascist purpose, she was also an agent of female emancipation. She was a political icon that also became a gender icon. In both roles she became a symbol of congratulation but also of confrontation, contradiction and paradox. PMID- 18161219 TI - The emotions in war: fear and the British and American military, 1914-45. PMID- 18161220 TI - [The 50th Anniversary Educational Symposium of the Japanese Society for Medical Mycology: Mycological study on Malassezia]. PMID- 18161221 TI - [The 50th Anniversary Educational Symposium of the Japanese Society for Medical Mycology: Visceral mycoses]. PMID- 18161222 TI - [The 50th Anniversary Educational Symposium of the Japanese Society for Medical Mycology: Pathological study on mycoses]. PMID- 18161223 TI - [The 50th Anniversary Educational Symposium of the Japanese Society for Medical Mycology: Nocardiosis and actinomycosis]. PMID- 18161224 TI - [The 50th Anniversary Educational Symposium of the Japanese Society for Medical Mycology: sporotrichosis and chromomycosis]. PMID- 18161225 TI - Beyond the call of duty. A naval nurse has made a career--and a difference--out of serving the needs of others. PMID- 18161226 TI - Geneva conventions 101. Why medical students need to learn a lot more about what constitutes torture. PMID- 18161227 TI - Mitt Romney's defining moment. PMID- 18161228 TI - Staph on the march. The deadly bacteria are beginning to emerge in schools. PMID- 18161229 TI - Of primary importance. Primary-care physicians seek to legitimize the 'medical home' concept to improve quality, costs--but will insurers buy it? AB - As primary-care physicians seek to promote the cause of establishing a "medical home" for patients, many questions still remain over how the system would work, and how payments would be structured. "This is not going to be sustainable unless it's linked to reimbursement," says Greg Pawlson, left, executive vice president of the National Committee for Quality Assurance. PMID- 18161230 TI - Nevada eyes DaVita bills. U.S. attorneys also looking into alleged overbilling. PMID- 18161231 TI - False claims recoveries down. Experts: 2006 figure understates feds' determination. PMID- 18161232 TI - Archimedes' new benefactor. RWJ Foundation gives Kaiser for-profit $15.6 million. PMID- 18161233 TI - Longer life, less healthy. Community outreach needed: experts. PMID- 18161234 TI - Marshfield deal delayed. Wis. clinic aims to acquire first wholly owned hospital. PMID- 18161235 TI - Maryland has nurses in mind. $59 million initiative to help deal with shortage. PMID- 18161236 TI - The watchdogs are baying. Physician deals may need to be cleaned up to avoid that knock on the door. PMID- 18161237 TI - Let the consumer rule. I seek excellence, affordability in a reformed system. PMID- 18161238 TI - Caution: more scrutiny ahead. Regardless of federal standards on what constitutes adequate community benefits, hospitals still face close look from the states. PMID- 18161239 TI - A long-term solution. IT scarce in nursing homes, but likely key to future care. PMID- 18161240 TI - Med-mal merger dissent. Shareholder seeks to block Doctors Co., SCPIE deal. PMID- 18161241 TI - The power of birth order. PMID- 18161242 TI - Mechanics of the passive knee joint. Part 2: interaction between the ligaments and the articular surfaces in guiding the joint motion. AB - The aim of this study was to examine how the interaction between ligament tensions and contact forces guides the knee joint through its specific pattern of passive motion. A computer model was built based on cadaver data. The passive motion and the ligament lengthening and force patterns predicted by the model were verified with data from the literature. The contribution of each ligament and contact force was measured in terms of the rotational moment that it produced about the tibial medial plateau and the anterior-posterior (AP) force that it exerted on the tibia. The high tension of the anterior cruciate ligament (ACL) and the geometric constraints of the anterior horns of the menisci were found to be key features that stabilized the knee at full extension. The mutual effect of the cruciates was found as the reason for the screw-home mechanism at early flexion. Past 300, the AP component of contact force on the convex geometry of the lateral tibial plateau and tension of the lateral collateral ligament (LCL) were identified as elements that control the joint motion. From 60 degrees to 90 degrees, reduction in the tension of the ACL was determined as a reason for continuation of the tibial anterior translation. From 90 degrees to 120 degrees, increase in the tension of the posterior cruciate ligament and the AP component of the contact force on the convex geometry of the lateral tibial plateau pushed the tibia more anteriorly. This anterior translation was limited by the constraining effects of the ACL tension and the AP component of the contact force on the medial meniscus. The important guiding role observed for the LCL suggests that it should not be overlooked in knee models. PMID- 18161243 TI - Adhesion of endothelial cells using self-assembly peptides under precise deformation control of tissue-engineered vessels. AB - The objective of this study was to anchor endothelial cells using self-assembly peptides under precise deformation control of tissue-engineered vessels. An acelluarized vascular matrix was used as the control group to examine the function of self-assembly peptides. In the experiment group, the self-assembly peptides were added to the inner surface of tissue-engineered vessels to form a monolayer. Then the endothelial cells were injected into the vascular lumen. A deformation control system was developed which was based on real-time image analysis and feedback control system. After dynamic culture by different deformation (set points 1, 5, and 10 per cent), the endothelial cell densities of experimental and control groups were compared. Both the self-assembly peptides and the extent of deformation affected the endothelial cell density on the inner surface of tissue-engineered vessels. The construct with self-assembly peptides under 5 per cent deformation gained the highest endothelial cell density. It was concluded that the deformation of assembled peptides contributes to the development and adhesion of endothelial cells in the inner surface of tissue engineered vessels. PMID- 18161244 TI - Analytical model development for the prediction of the frictional resistance of a capsule endoscope inside an intestine. AB - For the purpose of optimizing the design of the locomotion mechanism as well as the body shape of a self-propelled capsule endoscope, an analytical model for the prediction of frictional resistance of the capsule moving inside the small intestine was first developed. The model was developed by considering the contact geometry and viscoelasticity of the intestine, based on the experimental investigations on the material properties of the intestine and the friction of the capsule inside the small intestine. In order to verify the model and to investigate the distributions of various stress components applied to the capsule, finite element (FE) analyses were carried out. The comparison of the frictional resistance between the predicted and the experimental values suggested that the proposed model could predict the frictional force of the capsule with reasonable accuracy. Also, the FE analysis results of various stress components revealed the stress relaxation of the intestine and explained that such stress relaxation characteristics of the intestine resulted in lower frictional force as the speed of the capsule decreased. These results suggested that the frontal shape of the capsule was critical to the design of the capsule with desired frictional performance. It was shown that the proposed model can provide quantitative estimation of the frictional resistance of the capsule under various moving conditions inside the intestine. The model is expected to be useful in the design optimization of the capsule locomotion inside the intestine. PMID- 18161245 TI - A method to quantify hand-transmitted vibration exposure based on the biodynamic stress concept. AB - This study generally hypothesized that the vibration-induced biodynamic stress and number of its cycles in a substructure of the hand-arm system play an important role in the development of vibration-induced disorders in the substructure. As the first step to test this hypothesis, the specific aims of this study were to develop a practical method to quantify the biodynamic stress cycle measure, to compare it with ISO-weighted and unweighted accelerations, and to assess its potential for applications. A mechanical-equivalent model of the system was established using reported experimental data. The model was used to estimate the average stresses in the fingers and palm. The frequency weightings of the stresses in these substructures were derived using the proposed stress cycle measure. This study found the frequency dependence of the average stress distributed in the fingers is different from that in the palm. Therefore, this study predicted that the frequency dependencies of finger disorders could also be different from those of the disorders in the palm, wrist, and arms. If vibration induced white finger (VWF) is correlated better with unweighted acceleration than with ISO-weighted acceleration, the biodynamic stress distributed in the fingers is likely to play a more important role in the development of VWF than is th e biodynamic stressdistributed in the other substructures of the hand-arm system. The results of this study also suggest that the ISO weighting underestimates the high-frequency effect on the finger disorder development but it may provide a reasonable risk assessment of the disorders in the wrist and arm. PMID- 18161246 TI - Finite element modelling of the Ilizarov external fixation system. AB - This study describes a computational method for predicting the mechanical response of any configuration of the Ilizarov external fixation system. Mechanical testing of each of the individual components (ring, threaded rod, and wire) of the Ilizarov system was used to determine the stiffness of each component. Finite element (FE) analysis was then used to model each of the individual components. Each model was tuned to match the mechanical testing. A modular FE modelling system, using a master input file, was then developed where the tuned FE models of the individual components could be generated, positioned, and interconnected to replicate a range of fixator configurations. The results showed that the stiffness predications from the FE modelling of the fixator configurations were consistently 10 per cent higher than the stiffness values obtained from the mechanical testing. The FE modelling system can be used to predict the characteristic response of the fixator configurations and clearly shows the relative changes in that response for variations in the number of components used. PMID- 18161247 TI - Fabrication of porous bioactive structures using the selective laser sintering technique. AB - Hydroxyapatite, a ceramic with which natural bone inherently bonds, has been incorporated into a polymer matrix to enhance the bioactivity of implant materials. In order to manufacture custom-made bioactive implants rapidly, selective laser sintering has been investigated to fabricate hydroxyapatite and polyamide composites and their properties investigated. One objective of this research was to identify the maximum hydroxyapatite content that could be incorporated into the matrix, which was sintered at various parameters. The study focused on investigating the control of porosity and pore size of the matrix by manipulating the selective laser sintering parameters of the laser power and laser scan speed. The interception method was used to analyse the internal porous morphology of the matrices which were cross-sectioned through the vertical plane. Most notably, all structures built demonstrated interconnection and penetration throughout the matrix. Liquid displacement was also used to analyse the porosity of the matrices. The laser power showed a negative relationship between porosity and variation in parameter values until a critical power value was reached. However, the same relationship for laser scan speed matrices was inconsistent. The effects of the laser power and laser scanning speed on the features of porous structures that could influence cell spreading, proliferation, and bone regeneration are presented. PMID- 18161248 TI - On the effect of dynamic flow conditions on blood microstructure investigated with optical shearing microscopy and rheometry. AB - Red blood cell (RBC) aggregation affects significantly the flow of blood at low shear rates. Increased RBC aggregation is associated with various pathological conditions; hence an accurate quantification and better understanding of the phenomenon is important. The present study aims to improve understanding of the effect of dynamic flow conditions on aggregate formation; whole blood samples from healthy volunteers, adjusted at 0.45 haematocrit were tested in different flow conditions with a plate-plate optical shearing system, image analysis, and a double-walled Couette rheometric cell. Results are presented in terms of aggregation index Aa, aggregate size index As and number of aggregates, which are shown to vary with shear rate gamma and with different shear rate variations with time gamma. The aggregation index Aa was observed to increase as the shear rate decreased between 10 and 3 s(-1). Above 10 s(-1), Aa was found to have a minimum value indicating minimal aggregation while, at approximately 3 s(-1), Aa reaches a maximum. The aggregation size index As, the number of aggregates, and the blood viscosity were found to vary considerably when the same sample was examined over the same shear rate range, but for different variations of shear rate with time, gamma. PMID- 18161249 TI - Precision of wear measurement using the shadowgraph technique. AB - The shadowgraph technique is a relatively easy-to-use and inexpensive method of wear measurement from explanted acetabular cups. In this technique, from a mould of the internal surface of the cup, measurements of linear wear and wear angle can be obtained, from which volumetric wear can be calculated. In this study the measurement precision of this technique was assessed and the influence of different observers and multiple moulds was also determined. It was found that linear wear (coefficient of variation (CV) = 1.49 per cent) can be more precisely determined than wear angle (CV = 8.18 per cent) and that both the observer and the mould can significantly influence the results obtained, although the influence of the mould is considerably less than that of the observer. PMID- 18161250 TI - A dynamic model of simulating stress distribution in the distal femur after total knee replacement. AB - The aim of this study has been to develop a dynamic model of the knee joint after total knee replacement (TKR) to analyse the stress distribution in the distal femur during daily activities. Using MSC/ADAMS and MSC/MARC software, a dynamic model of an implanted knee joint has been developed. This model consists of the components of the knee prosthesis as well as the bones and ligaments of the knee. The femur, tibia, fibula, and patella have been modelled as mixed cortico cancellous bone. The distal part of femur has been modelled as a flexible body with springs used to simulate the ligaments positioned at their anatomical insertion points. With this dynamic model a gait cycle was simulated. Stress shielding was identified in the distal femur after TKR, which is consistent with other investigators' results. Interestingly, higher stresses were found in the bone adjacent to the femoral component peg. This dynamic model can now be used to analyse the stress distribution in the distal femur with different load conditions. This will help to improve implant designs and will allow comparison of prostheses from different manufacturers. PMID- 18161251 TI - Quantification of the coupled motion that occurs with axial rotation and lateral bending of the head-neck complex: an experimental examination. AB - The vertebrae of the cervical spine exhibit out-of-plane or coupled motion during axial rotation and lateral bending. Quantifying the range of motion (ROM) of this occurrence can aid the understanding of cervical spine injury mechanisms and disorders, as well as the development of new treatment methods. Previous studies have formulated ratios to describe coupled motion obtained from in-vitro examinations. The aim of the present study was to use in-vivo test data to develop mathematical relationships to quantify the coupled motion that occurs with axial rotation and lateral bending of the head-neck complex. Using a three dimensional motion analyser it was possible to trace the coupling effect throughout the full range of unrestricted head-neck motion. Values for primary and coupled ROMs were obtained, showing no significant difference between male and female primary ROMs but a small disparity between male and female coupled ROMs. Regression equations were found to quantify coupled motion throughout the range of axial rotation and lateral bending. The present experimental study also examines the range of horizontally fixed axial rotation of the head to determine the minimum amount of coupled lateral bending that takes place, which has not been measured previously. PMID- 18161252 TI - Comparison of anatomical shoulder prostheses and the proximal humeri of Chinese people. AB - Reconstruction of normal anatomy is important to a successful proximal humeral arthroplasty, in which the surgeon needs to know multiple measured anatomical parameters during the surgery. Based on three-dimensional computed tomography scanning data, computer models of the proximal humeri of 180 Chinese people were built. They came from 77 males and 103 females; their ages ranged from 19 to 86 years. Anatomical parameters including the curvature diameter of the head, the head height, the inclination, and the retroversion were measured using the software in these three-dimensional proximal humerus models. The methodology was valid and the measured results showed marked variations in all those parameters. On average, the curvature diameter of the head was 44.6 mm, ranging from 36.4 to 55.5 mm. The head height was located at 16.7 +/- 1.9 mm (minimum, 12.4 mm; maximum, 22.0 mm). The inclination angle ranged from 122.35 degrees to 147.09 degrees (mean, 132.4 +/- 4.7 degrees). The proximal humeral retroversion was found to be extremely variable, ranging from -4.71 degrees to 52.46 degrees, and has a mean of 21.11 degrees. Then these measured results were compared with four commonly used anatomical prostheses. The wide range of variation in the normal values makes it difficult to restore anatomical characteristics individually. Thus, the use of an accurate three-dimensional model to guide surgery preoperatively can improve the outcome of some serious cases, such as gross degenerative change and large-bone-loss surgery. PMID- 18161253 TI - Can cobalt levels estimate in-vivo wear of metal-on-metal bearings used in hip arthroplasty? AB - High levels of cobalt and chromium ions are detected in the blood and urine of patients with metal-on-metal (MoM) hip replacement. These elements are released as a result of wear at the bearing surfaces. Wear rates depend on a multitude of factors, which include the bearing geometry, carbon content, manufacturing processes, lubrication, speed and direction of sliding of the surfaces, pattern of loading, and orientation of the components. In-vivo wear of MoM bearings cannot be reliably measured on X-rays because no distinction can be made between the bearing surfaces. Hip simulator studies have shown that wear rates are higher during the initial bedding-in phase and subsequently drop to very low levels. Accordingly, metal ion levels would be expected to decrease with the use of the bearing, measured as implantation time following surgery. However, several clinical studies have found that metal ion levels either gradually rise or fluctuate instead of decreasing to lower levels. Moreover, hip simulator studies predict that large-diameter bearings have lower wear rates than small-diameter bearings. In clinical studies, however, metal levels in patients with large diameter bearings are unexpectedly higher than those in patients with small diameter bearings. As a consequence, high cobalt ion levels in patients do not necessarily imply that their MoM bearings produce much wear debris at the time that their levels were measured; it may simply be due to accumulation of wear debris from the preceding time. Exercise-related cobalt rise may overcome this limitation and give a better assessment of the current wear status of a MoM bearing surface than a measure of cobalt levels only. PMID- 18161254 TI - In-vitro method for assessing femoral implant-bone micromotions in resurfacing hip implants under different loading conditions. AB - Although prosthesis-bone micromotion is known to influence the stability of total hip replacement, no protocol exists to investigate resurfacing hip implants. An in-vitro protocol was developed to measure prosthesis-bone micromotions of resurfaced femurs. In order to assess the effect of all loading directions, the protocol included a variety of in-vitro loading scenarios covering the range of directions spanned by the hip resultant force in the most typical motor tasks. Gap-opening and shear-slippage micromotions were measured in the locations where they reach the maximum value. The applicability of the protocol was assessed on two commercial designs and different head sizes. Intra-specimen repeatability and inter-specimen reproducibility were excellent (comparable with the best protocols for cemented hip stems). Results showed that the protocol is accurate enough to detect prosthesis-bone micromotions of the order of a few microns. Statistically significant differences were observed in relation to the direction of the applied force. Using the whole range of hip loads enabled detection of maximum micromotions for any design (the peak value could be different for different loading directions). Application of the protocol during a test to failure indicated that the system could track micromotion up to the last instant prior to failure. The protocol proposed is thus completely validated and can be applied for preliminary screening of new epiphyseal designs. PMID- 18161255 TI - Fabrication and evaluation of calcium phosphate cement scaffold with controlled internal channel architecture and complex shape. AB - The ability to have precise control over internal channel architecture, porosity, and external shape is essential for tissue engineering. The feasibility of using indirect stereo-lithography (SL) to produce scaffolds from calcium phosphate cement materials for bone tissue engineering has been investigated. The internal channel architecture of the scaffolds was created by removal of the negative resin moulds made with SL. Scanning electron microscopy (SEM) showed highly open, well-interconnected channel architecture. The X-ray diffraction examination revealed that the hydroxyapatite phase formed at room temperature in the cement was basically stable up to 850 degrees C. There was no phase decomposition of hydroxyapatite, although the crystallinity and grain size were different. The ability of resulting structure to support osteoblastic cells culture was tested in vitro. Cells were evenly distributed on exterior surfaces and grew into the internal channels of scaffolds. To exploit the ability of this technique, anatomically shaped femoral supracondylar scaffolds with 300-800 microm interconnected channels were produced and characterized. PMID- 18161256 TI - Increasing patients' body mass. Are the criteria for testing stemmed femoral components in total hip arthroplasty still valid? AB - The increasing incidence of obesity in the population is a topic of current interest. This trend is reflected in patients undergoing primary Charnley hip replacement. Over a 21 year period from January 1986 to October 2006, during which 6910 primary operations were performed, the body mass of patients increased at a mean rate of 0.48 kg/year. The findings suggest that the test criteria for the endurance of stemmed femoral components be modified. The maximum load should be increased from 2.3 kN to 2.5 kN to reflect patients' increasing weight. The duration of the test should be increased from 5 x 10(6) cycles during which the 'femoral component shall not fracture' to 10 x 10(6) cycles in order to reflect not only the patients' activity level but also the 4-11 year period at risk for stem fracture, and so 'reflect developing clinical experience'. PMID- 18161257 TI - Reproduction of fretting wear at the stem-cement interface in total hip replacement. AB - The stem-cement interface experiences fretting wear in vivo due to low-amplitude oscillatory micromotion under physiological loading, as a consequence it is considered to play an important part in the overall wear of cemented total hip replacement. Despite its potential significance, in-vitro simulation to reproduce fretting wear has seldom been attempted and even then with only limited success. In the present study, fretting wear was successfully reproduced at the stem cement interface through an in-vitro wear simulation, which was performed in part with reference to ISO 7206-4: 2002. The wear locations compared well with the results of retrieval studies. There was no evidence of bone cement transfer films on the stem surface and no fatigue cracks in the cement mantle. The cement surface was severely damaged in those areas in contact with the fretting zones on the stem surface, with retention of cement debris in the micropores. Furthermore, it was suggested that these micropores contributed to initiation and propagation of fretting wear. This study gave scope for further comparative study of the influence of stem geometry, stem surface finish, and bone cement brand on generation of fretting wear. PMID- 18161258 TI - [Invasive zygomycosis in patients treated for haematological malignancies]. AB - A 52-year-old man underwent haematopoietic stem-cell transplant for myelodysplastic syndrome; after treatment with voriconazole for invasive aspergillosis, he was diagnosed with invasive zygomycosis caused by Rhizopus microsporus. He died despite treatment with intravenous liposomal amphotericin B and posaconazole. A 5-year-old boy with acute lymphatic leukaemia was diagnosed with invasive zygomycosis at autopsy. In a third case, a 16-year-old boy with acute myeloid leukaemia received repeated courses of empiric antifungal therapy, although the presence of an invasive fungal infection was not demonstrated. The patient died, and disseminated invasive zygomycosis caused by Rhizomucor pusillus was found at autopsy. Invasive infections by Zygomycetes are difficult to diagnose and are associated with a high mortality rate. The incidence of invasive zygomycosis appears to be increasing. Therefore, awareness of this type of invasive fungal infection is warranted. Lipid formulations ofamphotericin B remain the first choice for therapy. PMID- 18161259 TI - [Invasive zygomycosis: notably in diabetes mellitus and iron overload]. AB - The incidence of invasive zygomycosis, a severe and often life-threatening infection, is increasing. The most common manifestations are pulmonary infection (following anti-cancer chemotherapy or haematopoietic stem-cell transplant) and invasive rhinocerebral infection (in patients with diabetes mellitus or iron overload). Iron metabolism plays an important role in the pathogenesis of infection in these high-risk populations. Rapid diagnosis, reversal of the underlying predisposition and timely surgical debridement are the underlying principles of therapy for this disease. PMID- 18161260 TI - [Prophylactic treatment with clotting factor to prevent joint damage in patients with severe haemophilia: costs versus benefits]. AB - A recent publication stated that the cost of prophylactic clotting factor replacement therapy to prevent joint damage in patients with haemophilia is USD 300,000 per patient per year. Therefore, despite the relatively small number of patients with severe haemophilia, the disease has an important socioeconomic impact. Joint damage is the most frequent and most debilitating comorbidity of haemophilia and can be prevented by adequate prophylactic treatment. But when and how should prophylaxis be started? The aforementioned study provides thorough evidence that prophylaxis is clinically beneficial despite its high cost. A reasonable approach is to start individualised prophylactic treatment after the first episode of joint bleeding early in life, because prophylaxis will not prevent progression of established joint disease evident after multiple bleeding episodes. PMID- 18161261 TI - [New treatment methods for autoimmune thrombocytopenia]. AB - --For many years, splenectomy has been considered the only therapy with proven efficacy in patients with relapsed/refractory idiopathic thrombocytopenic purpura (ITP) following corticosteroid therapy. --A broad spectrum of (mostly immunosuppressive) agents are available for patients who fail to respond to splenectomy. However, the risks associated with these agents sometimes outweigh their benefits. --Recently, several new or renewed strategies have been evaluated for chronic refractory ITP. --Short-term therapy with high-dose dexamethasone is an effective alternative to long-term treatment with corticosteroids. --Depletion of B lymphocytes with rituximab, an agent that has an established role in the treatment of non-Hodgkin's lymphoma, also appears to be effective in autoimmune disorders, particularly in ITP. --Although the main problem in ITP is the increased destruction of thrombocytes, stimulation of thrombocyte production with thrombopoietin(TPO)-receptor agonists can increase thrombocyte counts. --Two TPO receptor agonists, AMG531 and eltrombopag, induce responses in 70-80% of ITP patients and are expected to gain approval for use in ITP soon. PMID- 18161262 TI - [The endocannabinoid system, overweight, and the CB1-endocannabinoid-receptor antagonist rimonabant]. AB - --Obesity is an important healthcare issue. --Recent research has led to insights into the role of the endocannabinoid system in the regulation of body weight. - Rimonabant is a CB1-endocannabinoid-receptor antagonist. --Four trials were published recently on the efficacy and safety of rimonabant in the treatment of people with obesity. --When combined with a hypocaloric diet, rimonabant 20 mg/day was more effective than placebo in achieving and maintaining weight loss. In addition, treatment with rimonabant had beneficial effects on insulin resistance, HDL-cholesterol and hypertriglyceridaemia. --There is concern regarding the increased incidence of depression during treatment. --Whether the beneficial effects of rimonabant on weight reduction and cardiovascular risk factors translate into a reduction in cardiovascular morbidity and mortality remains to be established in large phase III trials. PMID- 18161263 TI - [New drugs; rimonabant]. AB - The endocannabinoid system controls the regulation of food intake and appetite in the brain and lipogenesis in adipose tissue. Rimonabant belongs to the new drug class of cannabinoid-1 receptor antagonists. It can decrease appetite and food intake and thus stimulate weight loss. Rimonabant is indicated for severe obesity and as an adjunct to lifestyle modifications for obese patients with type 2 diabetes or hyperlipidaemia. Safety concerns limit the clinical applicability of the drug. The drug has not been approved in the US due to its neurological and psychiatric adverse effects. Rimonabant is approved in Europe but is contraindicated in patients with major depression and those taking antidepressants. PMID- 18161264 TI - [Diagnostic image (351). A neonate with a blister]. AB - A blister was noted on the dorsal side of the right hand of an otherwise healthy female neonate: a sucking blister, which disappeared in a few days spontaneously. PMID- 18161265 TI - [Surgical drainage of the pancreatic duct in patients with chronic pancreatitis is more effective than endoscopic drainage: randomized trial]. AB - OBJECTIVE: To compare endoscopic and surgical drainage of the pancreatic duct for ductal decompression in patients with severe pain due to chronic pancreatitis and a dilated pancreatic duct. DESIGN: Randomized clinical trial. METHOD: All symptomatic patients with chronic pancreatitis and a distal obstruction of the pancreatic duct, but without an inflammatory mass, were eligible for this study. Patients were randomized to endoscopic transampullary pancreatic duct drainage or to operative pancreaticojejunostomy. The primary end point was the average Izbicki pain score, measured during 2 years of follow-up. The secondary endpoints were pain relief at the end of follow-up, physical and mental health, morbidity, mortality, hospital stay and number of procedures performed. RESULTS: Of 118 patients who were evaluated between January 2000-October 2004 39 patients were randomized; 19 were treated endoscopically (16 of whom underwent lithotripsy) and 20 by operative pancreaticojejunostomy. During 24 months of follow-up, compared with endoscopic drainage, surgery was associated with lower Izbicki pain scores (51 versus 25; p < 0.001) and better SF-36 physical health summary scores (p = 0.003). Furthermore, at the end of follow-up, pain relief was achieved in 32% of patients randomized to endoscopic drainage and 75% of patients randomized to surgical drainage (p = 0.007). Complication rates and hospital stay were similar, but endoscopic treatment required more procedures (median 8 versus 3; p < 0.001). PMID- 18161266 TI - [Relationship between rubella virus and Fuchs heterochromic uveitis; 2 patients]. AB - Two otherwise healthy men, aged 26 and 29 years, were diagnosed with Fuchs heterochromic uveitis (FHU) on the basis of the presence of iris heterochromia or iris atrophy, stellate corneal precipitates, and/or cataract. Microbiological investigation of aqueous humour demonstrated intraocular antibody production against rubella virus, but not against Toxoplasma gondii, herpes simplex virus or varicella zoster virus. Microbial nucleic acid detection was negative for all pathogens. Some time later, both patients underwent cataract surgery, which improved their vision considerably. FHU is a chronic, generally unilateral iridocyclitis, accompanied by the above-mentioned ophthalmologic manifestations in the absence of systemic disease. Little is known about the pathogenesis ofFHU, but recent publications have provided evidence for the possible involvement of the rubella virus. PMID- 18161267 TI - [The practice guideline 'atopic dermatitis']. PMID- 18161268 TI - [Electroconvulsive therapy in depressed adolescents]. PMID- 18161269 TI - [Lung neoplasm: central hospital care for multidisciplinary care]. PMID- 18161270 TI - Critics accuse government of 'reality gap' in extra GP plans. PMID- 18161271 TI - Primary care. A view to a cull: has the PCT shake-up delivered? PMID- 18161272 TI - Simon Stevens on voice over choice. PMID- 18161273 TI - Mental health. How we can find hope for the lost generation. PMID- 18161274 TI - Data briefing. Is Darzi wrong about GP access? PMID- 18161275 TI - Mental health. Unhappy old age. PMID- 18161276 TI - FESC not enough alone. AB - *Two in three commissioning directors and 44 per cent of chief executives don't know if they will use FESC. *More than 70% of chief excutives and commissioning directors would rather get other help in commissioning. *More than half of chief executives and commissioning directors think FESC will at best be 'a little' successful. PMID- 18161277 TI - Purchasing power. AB - *Primary care trusts need to be better commissioners to balance provider strength and develop new services. *The private sector has commissioning skills lacking in the NHS that need to be tapped into. *FESC provides a list of companies for PCTs to use. PMID- 18161278 TI - Mark Britnell on World Class Commissioning. PMID- 18161279 TI - Wise buys. AB - *Seven pilot organisations will use FESC companies to try to improve their commissioning performance. *Hillington PCT is the furthest forward, hoping to save 11m pounds in a three-year contract with BUPA Commissioning. *Multi-PCT deals are likely in the West Midlands, NHS East of England and NHS South Central. PMID- 18161280 TI - Staffing. The real workforce. PMID- 18161281 TI - [Advances in studies on absorption, distribution, metabolism of flavonoids]. AB - Plenty of data and tests suggested that flavonoids have strong physiological and pharmacological activities. In this paper, the absorption, distribution and metabolism of flavonoids in gaster, gut and liver were introduced. The research of absorption, distribution and metabolism on flavonoids will provide theoretical basis for developing new drugs of flavoniods. PMID- 18161282 TI - [Advances in the studies of Oroxylum indicum]. AB - According to documents of recent fifty years the article summarized the herbalogical study, processing, chemical constituent, quality standard, pharmacologic action and clinical study of Oroxylum indicum. The chemical constituents mainly are flavonoids, glycoside and volatile oil. It also contains pterocarpan and rhodioside with p-hydroxyphenylethanols and cyclohexanols. Quantitative index was added in the quality standard of china pharmacopeia since 1995, but the quantitative standard is blank at the present, and the baicalin was mostly used for quantitative standard in the documents which lack of mark. Baicalein -7-O-diglucoside didn't existed in the other plants. It's mark will do good to the enhancement of quality standard. Pharmacologic action and clinical study were mainly anti-inflammatory, anti-mutagenesis, antibiosis, anticancera,and coughing. PMID- 18161283 TI - [Phylogeny relationship and molecular identification of ten Huperzia species (Huperziaceae) based on matK gene sequences]. AB - OBJECTIVE: To study the phylogeny relationship and molecular identification of 10 species from Huperzia (Huperziaceae) based on matK gene sequences data. METHOD: Total DNA of nine species from Huperzia was extracted; matK gene sequence was amplified by PCR. PCR product was directly sequenced after purification. RESULT: The chloroplast matK gene nucleotide sequences from 9 species of Huperzia species were sequenced. The matK gene nucleotide sequences length was 1 589 bp. Analysis with Huperzia lucidula matK gene nucleotide sequences (download from GenBank) and taking Lycopodiella cernua as outgroup, Maximum Parsimony, Neighbor-Joining analyses and genetic distances were conducted using MEGA 3.1 software. 35 variable sites and 35 parsimony informative sites have been found. Pairwise genetic distances among 10 species of Huperzia was 1.59% - 0.25%. CONCLUSION: The results were consistent with the taxonomy in morphological of Huperzia. But H. longipetiolata and H. serrata were resolved into in different clade. There are 19 different sites of matK gene sequences between H. longipetiolata and H. serrata, the genetic distances is 0.121%. It is suggested H. longipetiolata should be as an independent species. PMID- 18161284 TI - [Study on seedling quality of Psoralea corylifolia and influence factors]. AB - OBJECTIVE: To study the effects of different fertilization, measure of breeding seedling, breeding and sowing density on seedlings of Psoralea corylifolia. METHOD: The combination of field sowing and indoor test was applied. RESULT: All agricultural measures showed positive effect on the growth and development of seedling of P. corylifolia. CONCLUSION: The suitable sewing time should be the first and second ten days in March. The suitable sowing density is 150 kg x hm( 2). The combined of application of organic manure and chemical fertilizer is better than using one of them alone. PMID- 18161285 TI - [Study on seedling techniques of Paris polyphylla var. yunnanensis]. AB - OBJECTIVE: To shorten the duration of seed germinating, improve seed emergence and eventurally to establish a practical method for seeding of Paris polyphylla var. yunnanensis seed. METHOD: To study the effect of some factors (temperature, sowing time, covering-soil depth, plastic film ) on seed emergence of P. polyphylla var. yunnanensis. RESULT: The most suitable temperature for embryonic post-maturity and seed germinating is 18-20 degrees C. Paris seed could emerge in April when it was transfer to a low temperature 0-10 degrees C for 2-4 months after being treated under 18-20 degrees C for 3-4 months. For seed emergence, the best sowing time was before April. The suitable soil depth was about 1 cm. It is a better way to cover the seedling bed with black plastic film for improving the emergence percentage. CONCLUSION: Above results provide seedling techniques of P. polyphylla var. yunnanensis. PMID- 18161286 TI - [Study on correlation of soil nutrients and content of active constituents in root of Rumex gmelini]. AB - OBJECTIVE: To provide scientific basis for the selection of agrotype and property fertilization for Rumex gmelini cultivated in compliance with good agricultural practice (GAP). METHOD: HPLC method was applied to determinate the content of seven active constituents (resveratrol, polydatin, chrysophanol 1-glucoside, nepodin, emodin, chrysophanol and physcion) of annual R. gmelini. And the correlation between soil nutrients and content of active constituents in the root of R. gmelini were analyzed by stepwise regression analysis. RESULT: Seven regression equation were established. The statistic significance was found in three of them. CONCLUSION: The soil with high total K level is not suitable for R. gmelini cultivation. But the higher available N, available P, available K level of soil is suitable. PMID- 18161287 TI - [Analysis on genetic diversity of different Salvia miltiorrhiza geographical populations in China]. AB - OBJECTIVE: To research on genetic diversity of different Salvia miltiorrhiza geographical populations in China. METHOD: The genetic diversity of 27 S. miltiorrhiza geographical populations from ten provinces in China was estimated using amplified fragment length polymorphism (AFLP) markers. The data of amplified bands were analyzed by the software POPGENE and SPSS. RESULT: The ten primers employed produced a total of 528 discernable and reproduceable amplified fragments. There were 476 polymorphic brands. The percentage of polymorphic bands with in different populations was 90.15%. Genetic diversity analysis showed that Neis gene diversity (He) was 0.261 2 and Shannon's genetic diversity index (1) was 0.403 3. The coefficient of gene similarity was 0.504 0-0.789 0 between populations. The cluster map including all samples were obtained by UPGMA. In the map, there were seven cluster groups and one individual outside the groups. CONCLUSION: The genetic diversity with in different geographical population of S. miltiorrhiza in China is plentiful. PMID- 18161288 TI - [Determination of tetrandrine and fangchinoline in Radix Stephaniae tetrandrae and its preparation by nonaqueous capillary chromatography]. AB - OBJECTIVE: To establish a new method for the determination of fangchinoline and tetrandrine in Stephania tetrandra and Fengtongan capsule by noanqueous capillary electrophoresis. METHOD: Separation was carried out in an uncoated fused capillary (50 cm x 75 microm i.d.) with a running buffer containing 50 mmol x L( 1) ammonium acetate, 1.0% acetic acid and 20% acetonitrile in methanol. A separation voltage of 20 kV and a UV detector wavelength at 214 nm were adopted. Sample was introduced from the anode. RESULT: The calibration ranges were 1.00, 500 mg x L(-1) for both analytes. Under the optimum conditions, the relative standard deviation (RSD, n = 6) for the migration time of each analyte were 0.09%, 1.9% (intra-day) and 0.63%, 1.9% (inter-day); The RSD for the peak area of each analyte were 0.45%, 5.9% (intra-day) and 2.3%, 5.6% (inter-day), respectively. The contents of the analytes were determined easily with average recoveries 102% for fangchinoline and 105% for tetrandrine in S. tetrandra and 94.6% for fangchinoline and 98.7% for tetrandrine in Fengtongan capsules, respectively. CONCLUSION: The proposed method is simple, rapid, accurate and higher repeatable, and can be used to control of the quality of S. tetrandra and Fengtongan capsules. PMID- 18161289 TI - [Studies on effect of microemulsion in enhancing solubility of baicalin and puerarin]. AB - OBJECTIVE: To study the effect of O/W Microemulsion in enhancing solubility of baicalin and puerarin. METHOD: The Pseudo-ternary phase diagram was employed to selected the formulations of O/W microemulsion. The partical size of microemulsion, and solubility of baicalin and puerarin in water, oil phase, surfactant, microemulsion and alcohol were determined, respectively. RESULT: 11 types of microemulsion formulations have the effect in enhancing the solubility of baicalin and puerarin; the solubility of Baicalin in microemulsion is higher as 5-15 times than it in water, the solubility of Puerarin in microemulsion is higher as 3 - 4 times than it in water. CONCLUSION: Microemulsion is good in enhancing solubility of effective compounds extracted from Chinese medical herbs. PMID- 18161290 TI - [Extraction of sex hormone from antler velvet with supercritical CO2]. AB - OBJECTIVE: To study the method of extraction of sex hormone from antler velvet with supercritical CO2. METHOD: Supercritical fluid extraction (SFE) was used to extract sex hormone from antler velvet and radioimmunoassay (RIA) was used to analyze the extracts. The chemical compositions in extracts were identified by GC MS, TLC and HPLC, respectively. RESULT: The experimental results indicated that the extraction yield was 1.56% when 85% ethanol was used as co-solvent at temperature of 65 degrees C and extraction pressure of 30 MPa. Estradiol and progesterone in the extracts were 3.07, 776.18 ng x g(-1) respectively. CONCLUSION: It is feasible to extract hormone from antler velvet with supercritical CO2. PMID- 18161291 TI - [In vitro evaluation of self-emulsifying drug delivery system of volatile oil from rhizome of Ligusticum chuanxiong]. AB - OBJECTIVE: To investigate the evaluation method for self-emulsifying drug delivery system of volatile oil from rhizome of Ligusticum chuanxiong (VOC SEDDS). METHOD: The self-emulsifying ability, the efficiency of self emulsification, the properties of emulsion, the dissolution of volatile oil from Rhizome of Ligusticum Chuanxiong and the stability of the emulsion were determined. RESULT: The optimized formulation can fully emulsify in 5 min and the particle sizes were around 102 nm. Zeta potential was about -30 mV. The O/W emulsions were stable through centrifugation with high reproducibility. In vitro dissolution test indicated that over 80% of drug dissolved in 30 min and VOC SEDDS was stable under light and high temperature in 10 d. CONCLUSION: VOC SEDDS has strong self-emulsifying ability, fine stability and high dissolution rate in vitro. PMID- 18161292 TI - [Preparation of sinomenine microemulsion and its transdermal absorption]. AB - OBJECTIVE: To screen the optimum formulation and prepare O/W sinomenine microemulsion and investigate its in vitro transdermal delivery ability. METHOD: The microemulsions were prepared with the formulation containing oleic acid-tween 80-dehydrated alcohol-water by the pseudo-ternary phase diagram. The permeation flux of sinomenine was determined in vitro by Franz diffusion cell fitted with rat skin. The sinomenine was determined by HPLC. The transdermal characteristics of sinomenine microemulsion were compared with that of sinomenine gels. RESULT: The steady state flux of sinomenine microemulsion was significantly higher than that of sinomenine gels. The average permeation rate of sinomenine microemulsion was 116. 44 microg x cm(-2) x h(-1) in vitro. CONCLUSION: These results indicated that the studied microemulsion system with high permeation rate may be a potential vehicle for the transdermal delivery of sinomenine. PMID- 18161293 TI - [Determination of trace elements in 13 traditional Chinese medicines in medicated diet for diabetes mellitus by ICP-MS]. AB - OBJECTIVE: To investigate six trace elements including Cr, Zn, Mn, Se, Cu and V of 13 traditional Chinese medicines (TCM) commonly used in diet therapy of diabetes mellitus. METHOD: The samples were prepared through digesting in a microwave oven and determined with Inductively Coupled Plasma Mass Spectrometry (ICP-MS). RESULT: The correlative coefficients of the calibration curves were above 0.999. Standard addition recovery was 88.7% - 110.0% and RSD < or = 7.0%. Under the given analytical conditions, the results obtained showed that the contents of the six trace elements varied in these TCM samples, for element Cr, ranging between 0-11.82 microg x g(-1), Mn in 4.36-564.3 microg x g(-1), Cu in 0.55-13.34 microg x g(-1), Zn in 3.19-86.23 microg x g(-1) Se in 0-0.39 microg x g(-1) and V in 0.05-12.04 microg x g(-1), respectively. CONCLUSION: The possible relationship between the content of the six trace elements and 13 TCM samples for dietetic therapy of diabetes mellitus was also discussed. PMID- 18161294 TI - [Quinovic acid triterpenoid saponins from bark of Mitragyna rotundifolia]. AB - OBJECTIVE: To study the chemical constituents from the bark of Mitragyna rotundifolia. METHOD: Column chromatographic techniques were applied to isolate constituents. A combination of IR, MS and NMR spectroscopy was used to identify structures of constituents. RESULT: Six compounds were isolated from the n-BuOH fraction and their structures were elucidated as quinovic acid-3-O-beta-D-6-deoxy glucopyranoside, 28-O-beta-D-glucopyranosyl ester (1), quinovic acid-27-O-alpha-L rhamnopyranosyl ester (2), quinovic acid-3-O-alpha-L-rhamnopyranoside (3), qunovic acid-27-O-beta-D-glucopyranosyl ester (4), quovic acid-3-O-beta-D-6-deoxy glucopyranoside (5), qunovic acid-27-O-beta-6-deoxy-D-glucopyranosyl ester (6). CONCLUSION: Compounds 1 - 6 were isolated for the first time from the plant. Compounds 1 - 4 and 6 were isolated for the first time form the genus. PMID- 18161295 TI - [Quantitative analysis of adenosine and cordycepin in Cordyceps sinensis and its substitutes with LC-MS-MS]. AB - OBJECTIVE: To develop a LC-MS-MS method for determination of adenosine and cordycepin in Cordyceps sinensis and it's substitutes. METHOD: The sawple was extracted with. 90% methanol. Multi-reactions monitoring (MRM) technique was adopted. RESULT: The regression equations and coefficients were Y = 89.04X + 506.85 (r = 0.999 7) for adenosine, Y = 99.66X + 1 251.34 (r = 0.998 8) for cordycepin respectively. The linear range was 5.0-1 000.0 microg x L(-1) for adenosine and cordycepin. The limits of detection (LOD) were 0. 44 microg x L(-1) for adenosine and 0.31 microg x L(-1) for cordycepin, respectively. The average recoveries of adenosine and cordycepin were 98.1% and 97.9%, respectively. CONCLUSION: The method was highly sensitive, selective and fast, which can be used for the determination of adenosine and cordycepin in C. sinensis and it's substitutes. This method can also be applied for the quality control of the medicinal materials. PMID- 18161296 TI - [Studies on bioassay-guided anti-inflammatory fraction in bark of Albizia julibrissin combined determination with LC-MS-MS]. AB - OBJECTIVE: To search the anti-inflammatory fraction of Albizia julibrissin. METHOD: Inflammatory model of Kunming mice ear edema induced by croton oil and determination combined with the LC-MS-MS-guided fractionation and isolation were used. RESULT: The n-butanol fraction (AJ-B) obtained from the ethanolic extract of the Cortex albiziae was the major active fraction. The lignan glycosides fraction (AJ-B-1), which was further isolated from AJ-B, showed significant anti inflammatory activity and exhibited dose-dependent relationship in the dose of 5 to 20 mg x kg(-1). CONCLUSION: The method of bioassay-guided fractionation and isolation combined with the LC-MS-MS determination may be of benefit to the logical studies on the bioactive fractions or constituents of traditional Chinese materia medica. PMID- 18161297 TI - [RP-HPLC characteristics of dragon's blood]. AB - OBJECTIVE: To study the fingerprint of dragon's blood resina draconis by high performance liquid chromatography. METHOD: The samples are extracted with methanol and separated on a Eclipse XDB-C18 column (4.6 mm x 150 mm, 5 microm) with the mobile phase of acetonitrile-H2O in gradient mode, and the flow rate was 1.0 mL x min(-1), the detection wavelength was 275 nm and the temperature of column was 40 degrees C. Loureirin B was used as the reference compound. RESULT: HPLC fingerprint of dragon's blood was established and the similarity of the fingerprint was compared. CONCLUSION: The method is simple, accurate, and can be used to control the quality of dragon's blood. PMID- 18161298 TI - [Difference of gene expression profile in human gastric cancer grafted onto nude mice treated with WCA]. AB - OBJECTIVE: Chinese jianpi herbal recipe Weichangan (WCA) could increase the survival rate of advanced gastric cancer. This study was designed to investigate the molecular mechanism of WCA in treatment of gastric cancer by cDNA array, real time quantitative PCR and immunohistochemical technique. METHOD: A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mouse was used as the animal model. The mice were divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-day period or 5-fluorouracil (5-FU) over 6-day period starting at 8th day after grafting. Control animals received saline on an identical schedule. Animals were killed 41 days after being grafted. To assess the effect of therapy tumor weight was determined by a electron balance immediately after the animals killed. SP immunohistochemical method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in xenografts. For detection of apoptotic cells, apoptotic indices (AI) were examined by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method. SP method was also used to detect the expression of cleaved Caspase-3. The expression profiles in paired WCA treated gastric cancer samples and the N. S. control samples were studied by using a cDNA array representing 14, 181 cDNA clusters. The alterations in gene expression levels were confirmed by real-time quantitative PCR. SP method was used to detect the expression of Phospho-Stat3 (Tyr705) and bcl-2. RESULT: When compared with controls, tumor growth was significantly inhibited by treatment with the WCA or 5 FU (P < 0.01, respectively). The average of tumor inhibitory rate in WCA group was (44.32 +/- 5.67)% and 5-FU (47.04 +/- 11.33)%. The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group respectively. AI of human gastric cancer xenografts in nude mice was significantly increased to (9.72 +/- 4.51)% using TUNEL method in WCA group compared with the controls (2.45 +/- 1.37)%. 5-FU group was also found a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expression ESTs among the control sample pool and WCA sample pool. There were 24 ESTs up regulated in WCA samples and 21 ESTs down-regulated. These 45 ESTs contains 35 cloned genes and 11 unknown ESTs. By using Real-time Quantitative PCR, the expression level of Stat3 (2(-deltadeltaCT) = 0.16) , RIPX (2(-deltadeltaCT) = 0.18), ROD1 (2(-deltadeltaCT) = 0.23) and bcl-2 (2 (-deltadeltaCT) = 0.10) was lower in WCA group than that in control group respectively. The expression of Phospho-Stat3 (Tyr705) and bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively. CONCLUSION: Chinese jianpi herbal recipe WCA could inhibit gastric cancer cell SGC-7901 growth in vivo. WCA could induce gastric cancer cell apoptosis and suppress proliferation. Its mechanisms might be involved in the down-regulation of Stat3, RIPX, ROD1 and bcl-2 gene. PMID- 18161299 TI - [Protection of polysaccharides-2b from mudan cortex of Paeonia suffruticosa andr on diabetic cataract in rats]. AB - OBJECTIVE: To observe the the protective effection of polysaccharides-2b of mudan cortex of Paeonia suffruticosa andr (PSM2b) on diabetic cataract. METHOD: The animal model of diabetic cataract in rats was induced by streptozotocin (STZ) and freund's adjuvant complete (CFA). The initial opacity occurrence time in lens was investigated with cranny lamp, and opacity degree of lens was compared too. The activities of glutathione peroxidase (GSH-pX), superoxide dismutase (SOD), catalase (CAT), the content of malonaldehyde (MDA) in serum and lens were detected. At the same time, the activities of Na(+) -K(+) -ATPase, the content of macromolecular weight protein and infusibility protein in lens were detected too. RESULT: The results examinated by cranny lamp showed that PSM2b could significantly postpone the occurrence and alleviate opacity degree of lens. Compared with model group, every treatment group of PSM2b could lower the level of MDA, high and middle dose groups could increase the levels of SOD, GSH-pX, CAT in serum and lens in evidence, and enhance the activity of Na(+) -K(+) -ATPase. These indexes present favorable positive correlation between dose and effect. CONCLUSION: All these results demonstrated that PSM2b had apparently protective effection on diabetic cataract in rats. PMID- 18161300 TI - [Effects of theanine and houpu extract in 7-day chick social separation-stress procedure]. AB - OBJECTIVE: To examine the abilities of theanine and houpu extracts (HE) to reverse behavioral indexes (separation vocalization, stress-induced analgesia and activity). METHOD: 7-day-old chicks received IP injection of theanine and HE 30 min before being tested in the presence of three social companions or in isolation for 3-min observation period. Dependent measures were: a) Chicks were placed into an infrared ray device to calculate their spontaneous activities by a computer program b) record the separation vocalizations for every chick. c) In the experiment of stress-induced analgesia, 50 uL of formalin (0.1%) was injected into the plantar of the animal foot to index stress-induced analgesia (i. e. foot lift frequency, foot-lift duration and peck frequency). RESULT: In the experiments, isolated chicks exhibited more vocalizations (P < 0.01) and fewer pain-related behaviors than non-isolated chicks (P < 0.01). Theanine (12.5, 25, 50 mg x kg(-1)) and HE (25 mg x kg(-1)) decrease separately the tendency (dB) of the principal frequency (P < 0.05, P < 0.01); The stress induced analgesia can be reversed by theanine in 25, 50 mg x kg(-1). Both of the materials do not affect the spontaneous activities in this chick model without causing sedation. CONCLUSION: These results suggest that theanine and HE in the dosages may be useful in modulating anxiety states. They are seems no synergism in the chick model. PMID- 18161301 TI - [Effect of Qingkailing injection on rat CYP1A2 and CYP2D6 activity]. AB - OBJECTIVE: The present study investigates the influence of Qingkailing injection on rat liver CYP1A2 and CYP2D6 activity in vivo and in vitro, respectively. METHOD: We employed HPLC to measure the metabolites of caffeine in the whole blood and calculated the ratio be between the metabolite and caffeine, which was used as index to evaluate the effect of Qingkailing injection on rat CYP1A2 activity in vivo; We also detected the CYP1A2 and CYP2D6 activity in microsomal reconstituted system by analysis of phenacetin metabolism and dextromethorphan metabolism with HPLC. RESULT: The metabolism of caffeine in treated groups was (15.9 +/- 3.8)%, (14.5 +/- 1.8)%, (12.3 +/- 1.2)%, with different concentration of Qingkailing injection (0.15, 0.3, 0.6 mL x kg(-1)) compared with (16.8 +/- 5.9)% in the control group, which was no significant difference among groups. In rat liver microsomal reconstituted system, Qingkailing injection has no inhibitory effect on CYP2D6 activity while the group with high dose has inhibitory effect on rat CYP1A2. CONCLUSION: Qingkailing injection has no inhibitory effect on rat CYP1A2 and CYP2D6 in vivo and in vitro. PMID- 18161302 TI - [Nephrotoxicity of Radix Aristolochice and it's substitution material Radix Inulae in rats]. AB - OBJECTIVE: To evaluate the toxicity of Radix Aristolochiae and Radix Inulae, and to supply the toxicity experimental data that Radix Inulae supersedes Radix Aristolochiae in clinic. METHOD: A long dose of Radix Aristolochice and Radix Inulae was given intragastrically to rats for six months, then drug withdrawal for a month. The hematology and biochemical indicators were measured, and the pathologic changes of kidney, liver, stomach and urinary bladder were examined. RESULT: The rats of Radix Aristolochice showed serious toxic responses of renal tubule atrophy and necrosis, meanwhile, the levels of BUN, Cr and NAG were increased obviously. Hepatonecrosis, renal tubular necrosis, gastric carcinoma and bladder carcinoma were discovered with pathologic assaying. But the rats of Radix Inulae did not. CONCLUSION: Radix Aristolochiae could damage kidney and liver, and cause gastric carcinoma and bladder carcinoma by intensive toxicity. Radix Inulae could take the place of Radix Aristolochiae to use in clinic. PMID- 18161303 TI - [Inhibition of curcumin on histone deacetylase and expression promotion of P21 (WAF1/CIP1) in HepG2 cells]. AB - OBJECTIVE: To investigate the effect of curcumin (Cur) on histone deacetylase (HDAC1) and P21(WAF1/CIP1), a cyclin dependent kinase inhibitor, in HepG2 cells for exploring the mechanism of Cur in anti-cancer. METHOD: The HDAC1, P21(WAF1/CIP1) proteins and P21(WAF1/CIP1) mRNA were extracted from human hepatoma cells treated with or without Cur of different concentrations at different time points. Western blot analysis was performed to determine the levels of HDAC1 and P21(WAF1/CIP1) proteins, respectively. RT-PCR was performed to detect the level of P21(WAF1/CIP1) mRNA. RESULT: The IC50 of concentration treated by Cur was 25 micromol x L(1) on HepG2 cell. The level of HDAC1 was obviously inhibited by Cur, and decreased at 4 hours at IC, and lasted for 48 h in a time-dependent manner. The inhibition of HDAC1 was significant at the Cur concentration of 12.5 micromol x L(-1) but there was no difference between 50 and 100 micromol x L(-1). The levels of P21(WAF1/CIP1) mRNA and protein were up regulated by Cur in dose and time-dependent manner, and the change of mRNA and protein was detected at 8 hours and lasted for 48 hours. CONCLUSION: Cur can inhibit the level of HDAC1 and enhance the expression of P21(WAF1/CIP1) protein and mRNA, and the results suggest that inhibiting HDAC1 and increasing P21(WAF1/CIP1) may be one of the possible mechanisms of anti-cancer by Cur. PMID- 18161304 TI - [Antitumour effect of total saponins of Rubus parvifolius on malignant melanoma]. AB - OBJECTIVE: To evaluate the antitumor effect of total saponins of R. parvifolius on malignant melanoma. METHOD: The human malignant melanoma A375 cells were regularlly subcultured in vitro, and were divided into five groups contained positive control group (CTX), high concentration (0.01 mg x mL(-1)) and middle concentration (0.001 mg x mL(-1)) and low concentration (0.000 1 mg x mL(-1)) total saponins of R. parvifolius groups and negative control group. By using MTT colorimetric method, the cell viability was measured. B16 melanoma cells were transplanted to mice, which were divided into positive control group, high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) and low dose (25 mg x kg(-1)) total saponins of R. parvifolius groups and negative control group. The inhibition effect of the tumor in vivo, mean survival time and rate of life elongation of the mice were observed. TUNEL method was used to detect the apoptosis of B16 malignant melanoma. RESULT: Antitumor assay in vitro showed that the absorbency increased in the concentration of 0.01, 0.001 mg x mL(-1) with statistical significance (P < 0.05 vs negative control). Antitumor assay in vivo showed that the tumor inhibitory rate of high dose (100 mg x kg(-1)) and middle dose (50 mg x kg(-1)) of total saponins of R. parvifolius were 37.02% and 30.61%, respectively. Loaded tumor mouse survival duration could be prolonged. The apoptosis indexes of B16 tumor cells in three treatment groups were 32.5%, 20.5% and 5.5%, respective and there was statistical significance (P < 0.05 vs negative control). CONCLUSION: The total saponins of R. parvifolius has remarkable inhibition of proliferation of malignant melanoma cells in vivo and in vitro and exerts antitumor activities through promoting tumor cell apoptosis. PMID- 18161305 TI - [Inhibitory effect of quercetin, rutin and puerarin on LDL oxidation induced by Cu2+]. AB - OBJECTIVE: To evaluate the inhibitory effect of quercetin, rutin and puerarin on the LDL oxidation induced by Cu2+ and to investigate their action on the prevention and treatment of atherosclerosis. METHOD: The serum LDL was isolated by the one step density gradient ultracentrifugation. The LDL oxidation was induced by Cu2+ in vitro for different time periods. Quercetin, rutin and puerarin at 5 micromol x L(-1) were added respectively, as the experimental groups, 3 hours before oxidation. The oxidation of LDL in experimental and control groups was identified by measuring A234, REM, TBARS and protein carbonyls content, and the values were compared between the two groups. RESULT: (1) The values of A234, REM, TBARS and protein carbonyls formation increased gradually during LDL oxidation induced by Cu2+ in vitro. (2) During LDL oxidation induced by Cu2+ in vitro and incubation with each of quercetin, rutin and puerarin, the kinetic changes of A234, REM, TBARS and protein carbonyls formation showed lag phases of 2-6 h, 2 h and 2 h respectively, and the corresponding values for each of the agents treated group were reduced by 27.7%-49.6%, 24.1%-38.6%, 19.8%-34.3% and 36.4%-56.8%; 12.8%-39.3%, 15.7%-32.0%, 19.0%-28.1% and 12.8%-50.3%; and 3.3% 19.2%, 7.0%-22.5%, 19.5%-22.8% and 8.6%-47.0%, respectively. CONCLUSION: These results suggest that quercetin, rutin and puerarin can substantially inhibit LDL oxidation, and quercetin has antioxidation ability stronger than rutin and puerarin. PMID- 18161306 TI - [Influence of effective part of Zingiber officinal on expression of monocyte chemotactic protein-1 and adhesion molecules]. AB - OBJECTIVE: To investigate the effect of effective parts of Zingiber officinal (EPZ) on the adhesion of ECV-304 cells with monocytes cultivated in vitro and on the expression of monocyte chemotactic protein-1 (MCP-1) and adhesion molecules. METHOD: The model of ECV-304 cell oxidative stress injury was established by hydrogen peroxide (H2O2). Then EPZ-contained blood serum was taken as experimental drug. The adherence of monocytes to endothelial cell were measured by method of rose Bengal. The total RNA of cells was extracted. The intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and MCP 1 mRNA expression in cells were detected by RT-PCR. MCP-1 protein expression were detected by ELISA. RESULT: EPZ could decrease the adhesion of monocytes with ECV 304 cells obviously. Meanwhile it could diminish the expression of ICAM-1, VCAM-1 and MCP-1 in injured ECV-304 cells. CONCLUSION: EPZ could inhibit H2O2-induced ICAM-1, VCAM-1 and MCP-1 expression in ECV-304 and could inhibit the adherence of monocytes to endothelial cell, which may result in the protect effect in endothelial cells. PMID- 18161307 TI - [Preparation of poly (trifluoropropyl) methylsiloxane coating for solid-phase microextraction and its application for the analysis of phenolic compounds in sea water]. AB - The poly (trifluoropropyl) methylsiloxane (PTFPMS) coating was prepared as solid phase microextraction (SPME) coating by sol-gel technique. The coating has a porous microstructure and the thickness was 10 microm. Since the PTFPMS phase contains the functional group of -C2H4CF3, it exhibits high affinity to compounds containing electron-attracting groups. The PTFPMS phase can withstand desorption temperature up to 280 degrees C. The lifetime of the fiber SPME is more than 150 times. This coating was used to analyse the phenolic compounds in water samples by head-space solid-phase microextraction (HS-SPME) method. The parameters affecting the extraction efficiency, such as extraction mode, extraction temperature, extraction time, ion strength and pH, were optimized. The limits of detection (LODs) of the method were 0.02 - 1.2 microg/L. The relative standard deviations (RSDs) of single fiber and fiber-to-fiber (n = 5 ) were less than 4.4%, and 15.7%, respectively. The results indicated that the sol-gel method has good repeatability for the preparation of the PTFPMS coating. PMID- 18161308 TI - [Determination of Sudan Red dyes in hot chili products by humic acid-bonded silica solid-phase extraction coupled with high performance liquid chromatography]. AB - Humic acid-bonded silica was prepared and used as solid-phase extraction material for the pretreatment of Sudan Red dyes in hot chili products, which was followed by high performance liquid chromatographic determination. This sorbent exhibits an excellent adsorption capacity for Sudan Red dyes. A sample was first extracted with n-hexane. Then the extract was cleaned up by humic acid-bonded silica cartridge and eluted with ethanol/dichloromethane (3:7, v/v). Some interferences in hot chili products such as natural pigments and oil were removed by washing step. Good linearities were achieved in the range from 0.05 to 5 mg/L for four Sudan dyes, with linear coefficient r2 values above 0.999. The limits of detection (LOD) in chili powder and chili oil were from 19.8 to 28.8 ng/g and 7.9 to 11.5 ng/g, respectively. Excellent method reproducibility was found by intra- and inter-day precisions (the relative standard deviations less than 9.1% and 8.8%, respectively). Recoveries for them in the real samples were between 82.6% and 94.0%. The proposed method is proved to be simple, rapid, and sensitive in the regular analysis of Sudan Red I, II, III and IV in foods. PMID- 18161309 TI - [Sample pre-processing for mass spectrometric analysis of glycoprotein structure]. AB - A novel, simple and fast method based on a quadrupole ion trap (QIT) and a time of-flight mass spectrograph was developed for the glycosylation site and glycan structure analysis of glycoprotein by in-solution digestion and in-gel digestion which did not need to enrich and label the glycopeptide. Horseradish peroxidase (HRP) and RNase B were utilised to optimize the conditions of sample pre processing. The effects of the conditions were investigated separately and the optimal conditions were: heat denaturation of RNase B, chemical denaturation of HRP, RNase B digestion with endoproteinase Lys-C, HRP digestion with trypsin, 12 16 h of digestion, 50% acetonitrile-5% trifluoroacetic acid of the extraction solution, sandwich spotting method of the sample. PMID- 18161310 TI - [Application of accelerated solvent extraction technique for analysis of active components in traditional Chinese medicinal herbs]. AB - The application of accelerated solvent extraction (ASE) technique for the Analysis of active components in traditional Chinese medicinal herbs was introduced by using two kinds of herbs as examples. The conditions including extraction temperature, static extraction time, the ratio of material to solvent and solvent of ASE for extraction of salvianolic acid B in Salvia miltiorrhiza were optimized by orthogonal experiments, and the optimal conditions were obtained. Different extraction methods (ASE, steam distillation, ultrasonic wave and Soxhlet extraction) were used to extract volatile oil in Aucklandia lappa Decne. Results of the comparative experiments indicated that ASE was the most effective method in this case. All the results from these studies demonstrate that ASE is indeed a powerful tool in the preparation of herbal extracts for downstream chromatographic analysis. PMID- 18161311 TI - [Sample pretreatment in ion chromatography]. AB - Some sample pretreatment techniques used in ion chromatographic analysis such as absorption, extraction, dialysis, combustion, alkaline fusion, on-line preconcentration, neutralization and precipitation are summarized. A total of 130 references are cited. PMID- 18161312 TI - [Determination of amphetamine and ketamine in human urines using high performance liquid chromatography coupled with liquid-liquid-liquid phase microextraction]. AB - Liquid-liquid-liquid phase microextraction (LLLME) coupled with high performance liquid chromatography was developed for the determination of amphetamine and ketamine in human urines. The effects of the extraction solvent, the pH value of the donor solution, the stirring rate, the extraction time and HCl concentration in the acceptor phase on the enrichment factors of analytes were investigated. The optimal experimental conditions, 300 microL of toluene as the organic phase, pH 11 of the donor phase, 1.0 microL of 0.1 mol/L HCl as the acceptor phase, 600 r/min of the stirring rate, and 50 min of the extraction time, were obtained. Under the above conditions, high enrichment factors were reached. The linear ranges were from 0.01 to 10 microg/mL for amphetamine, and 0.01 to 5 microg/mL for ketamine. The relative standard deviations were lower than 2%. The limits of detection were 5 ng/mL for the two target compounds at signal-to-noise ratio of 3. The method with little solvent consumption may provide high analyte pre concentration and excellent sample clean-up, and it is a sensitive and suitable method for the simultaneous determination of above two analytes in human urines. PMID- 18161313 TI - [Development of on-line sample preparation coupled with liquid chromatography mass spectrometry for analysis of small molecules in biofluids]. AB - On-line liquid chromatography-mass spectrometry (LC-MS) is a highly sensitive, highly selective and high throughput method for the analysis of small molecules in biofluids. The developments, characterizations and applications of pre-column, on-line LC-MS method and its configuration in the recent five years are reviewed. PMID- 18161314 TI - [A multi-step distillation method for sample preparation in gas chromatographic analysis]. AB - A sample pretreatment device based on multi-step distillation principle was constructed and evaluated. Trace components in the sample were extracted, separated and enriched at the cold receptor of the molecular diffusion distillation device. The cold receptor was designed as an exchangeable part to avoid the cross interferences from one sample to another one. A model sample containing four kinds of organophosphorus pesticides in ethyl actate was tested in the device, and the distillated and enriched solution was analyzed by a gas chromatograph. It was proven that the interferential components to gas chromatography such as protein, lipid and pigment could be removed effectively, and the enrichment factor for trace target components was more than 150 times. The recoveries of the method were from 90% to 117%, and the relative standard deviations (RSDs) from 1.3% to 10%. The limit of detection (LOD) of this method was 1 ng/mL without further concentration. The reused ratio of the organic reagents was more than 95%. Practical application was performed by using a spiked tea sample. The device can treat more than 100 g of samples with a boiling point range of 120 - 600 degrees C. PMID- 18161315 TI - [Recent advances in enantiomeric separation of chiral environmental pollutants by capillary electrophoresis]. AB - Capillary electrophoresis (CE) is an effective analytical tool in chiral separation. This review article is focused on the major developments in the recent applications of CE to the enantioseparation of the chiral environmental pollutants in the past decade. Developments are classified according to the types of chiral pollutants, i. e. phenoxyalkanoic acid herbicides, organophosphorus/organochlorine/pyrethroid pesticides, triazole fungicides, polychlorinated biphenyls (PCBs). The advances in enantiomeric separation of the above-mentioned chiral pollutants by CE are reviewed. In addition, the emerging chiral pollutants are highlighted and the prospects of CE in the analysis of chiral environmental pollutants are also drafted. PMID- 18161316 TI - [Non-equilibrium thermodynamic separation theory of nonlinear chromatograph I. Local Lagrangian approach for nonlinear equilibrium-dispersive processes]. AB - The matrix forms of local Lagrangian approach (LLA) are developed based on Lagrangian description for single-component in nonlinear, non-ideal chromatography. A local thermodynamic path (LTP) is designed based on essential physical principles, such as the Lagrangian description, the local equilibrium assumption and the thermodynamic state functions. With the LTP, the iteration equations of fully thermodynamic states on time sequence in the matrix forms are obtained with the Markov character. And the convergence, compatibility and stability of the LLA based on the LTP are discussed with some theoretical analysis and numerical experiments, and the stability condition of the LLA is given. The algorithm of the LLA in the vector form is shown as the computer program to simulate the elution profiles affected by a few of factors, space distribution, axial diffusions, injection samples, etc. According to the LLA, the corresponding relationships are established between the trajectories of discrete time state and discrete time control vectors in the ergodic space. And a compendium algorithm of multistage decision problems concerning the optimal control of nonlinear, non-idea chromatography is given with Bellman's dynamic programming to find the optimal trajectories of state vector and control vector. The matrix forms of the LLA remove the gap between preparative chromatography theories and Markov decision processes or optimal control approaches based on discrete time states. PMID- 18161317 TI - [Determination of hydroxyl radical in CuSO4-vitamin C reaction system and scavenging activities of Chrysanthemum using capillary electrophoresis with electrochemical detection]. AB - High performance capillary electrophoresis (CE) with electrochemical detection (ECD) was employed to determine the amount of hydroxyl radical in CuSO4-vitamin C reaction system (pH 7.4). The effects of some important factors, such as the acidity of the running buffer, separation voltage, injection time and the potential applied to the working electrode were investigated to choose the CE-ECD optimum conditions. The reaction system to produce hydroxyl radical was also optimized such as the effects of reactant concentration and reaction time on the concentration of 3,4-dihydroxy-benzoic acid. Operated in a wall-jet configuration, a 300 microm diameter carbon-disc electrode was used as the working electrode. Excellent linearity was obtained in the concentration ranging from 1.5 x 10(-4) mol/L to 6.0 x 10(-6) mol/L for the reaction product. The detection limit was 1.5 x 10(-6) mol/L (S/N = 3). This method was successfully applied to determine scavenging activities of Chrysanthemum for hydroxyl radical. PMID- 18161318 TI - [Separation of water-soluble active components in Salvia miltiorrhiza bge. f. alba using capillary zone electrophoresis]. AB - To establish a high performance capillary zone electrophoretic method for the separation of water-soluble active components, protocatechuic aldehyde (PAH), danshensu (DSS), and protocatechuic acid (PA), in Salvia miltiorrhiza bge. f. alba, many factors that can affect the resolution and the migration time, including the concentration of phosphate-borax buffer, pH value, separation voltage and detection wavelength were investigated. The optimized conditions were as follows: uncoated capillary column, 75 microm x 60.2 cm (effective length of 50 cm); column temperature, 25 degrees C; detection wavelength of the photo-diode array detector, 210 nm; injection, 3.45 kPa x 4 s; separation voltage, 27.5 kV; running buffer, 5 mmol/L borax (pH 7.5). Under the optimized conditions, good separation of PAH, DSS and PA can be achieved in less than 8 min. PMID- 18161319 TI - [A novel method for the direct measurement of electroosmotic flow velocity on microfluidic chips]. AB - As microfluidic technologies mature, accurate methods for the measurement of electroosmotic flow velocity on microfluidic chips are becoming increasingly important. A change of migration time of Rhodamine 123 (Rh123) at different pH values of the buffer was observed during the electrophoresis, and it was deduced that there is uncharged molecular form of Rh123 in alkaline solution because of deprotonation. Therefore, a method of direct measurement of electroosmotic flow velocity on microfluidic chip was developed based on the detection of migration time of the uncharged Rh123, because the migration speed of uncharged molecule is equal to the electroosmotic flow velocity. The electroosmotic flow velocity of glass microchip at pH 9.3 and 10.1 of the buffer was detected to be 3.9 x 10(-4) cm2/(s V) and 4.1 x 10(-4) cm2/(s V), respectively, which were in agreement with those obtained by the traditional method. PMID- 18161320 TI - [Determination of oxymatrine in urine samples by capillary electrophoresis with stacking induced by moving reaction boundary]. AB - A stacking system based on moving reaction boundary (MRB) for stacking and quantitative determination of oxymatrine (OMT) in urine samples was developed. The experimental conditions were optimized for the stacking of OMT as well as its separation. The optimized conditions were 20 mmol/L HCOONa (pH 10.70 adjusted by weak alkali of ammonia rather than strong alkali of sodium hydroxide) as sample buffer, 40 mmol/L HCOOH-HCOONa (pH 2.60) as stacking buffer, 100 mmol/L HCOOH HCOONa (pH 4.80) as separation buffer, 1.4 kPa (3 min) sample phase injection and 1.4 kPa (7 min) stacking phase injection, 210 nm of detection wavelength, 21 kV of voltage. The linear response of OMT concentration ranged from 2.2 to 65 mg/L with high correlation coefficient (r = 0. 9991), the limit of detection (LOD) for OMT was 0.74 mg/L, and sensitivity was enhanced by 70 times. This method can be well used for quantification of OMT in urine samples with high sensitivity and can be further applied in the investigation of pharmacokinetics. PMID- 18161321 TI - [Separation of cefoperazone and its S-isomer and other related substances by micellar electrokinetic capillary chromatography]. AB - The separation of cefoperazone, its S-isomer, impurity A and other unknown related substances by micellar electrokinetic capillary chromatography (MECC) using sodium dodecyl sulphate (SDS) as the micellar phase was investigated. The effects of pH, concentration of phosphate buffer solution, SDS micelle concentration, methanol volume fraction, applied voltage and temperature on the separation were studied. It was found that the migration of these compounds was affected by these factors, especially by pH of the solution. The elution, as well as the migration time and separation efficiency of cefoperazone, its S-isomer, impurity A and other related substances changed with the acidity of the solution. The optimized separation conditions consisted of a running buffer of 70 mmol/L sodium phosphate buffer, at pH 6.5, containing 100 mmol/L SDS, with an applied voltage of 15 kV and a temperature of 25 degrees C. An uncoated fused-silica capillary of 51.0 cm x 75 microm (42.5 cm of effective length) was used. The sample was injected into the column by pressure (5 kPa) for 5 s. The detection wavelength was set at 254 nm. Twenty-eight impurities in cefoperazone sodium could be detected. Cefoperazone sodium and the degradation products could be separated well. The method was applied to separate and determine cefoperazone and its related substances successfully. PMID- 18161322 TI - [Determination of Sudan Red I-IV in duck egg yolk using ultra performance liquid chromatography-tandem mass spectrometry]. AB - A method of the simultaneous analysis of Sudan Red I - IV in duck egg yolk was developed with an ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Acetonitrile was used to extract the residues from duck egg yolk, and water was added into the extract to precipitate impurities such as protein and fat. The supernatant was detected by the UPLC-MS/MS after refrigeration and centrifugation. The sample was separated on a Waters Acquity BHE C18 column, and detected by MS/MS with the multiple reaction monitoring (MRM) mode. The limits of detection (LOD) were 0.05 microg/L in the standard solution, and 10 microg/kg of the four substances in the sample spiked with Sudan Red I - IV. The average recoveries were between 50.2% - 101.3% at three spiking levels of 100.0, 200.0, 300.0 microg/kg. The experiment results show that this method is of high sensitivity, low LOD, better determination capacity and shorter analysis time. The method could meet the high-throughout detection of Sudan Red in food samples. PMID- 18161323 TI - [Determination of the molecular mass of modified citrus pectin using high performance size exclusion chromatography]. AB - Modified citrus pectin (MCP) is a polysaccharide with anti-tumor growth, anti invasiveness and anti-metastasis activities. It is of significance to determine the molecular mass and polydispersity of polysaccharide, which could influence pharmacological activities of polysaccharide. To the best of our knowledge, no study has been reported on calculating the molecular mass of MCP by high performance size exclusion chromatography-refractive index detection (HPSEC-RID). A TSK G3000PW(XL) column was used at 40 degrees C, and the mobile phase was composed of 50 mmol/L NH4Ac solution. Pullulan with known molecular mass was chosen as calibration standard to obtain regression equation. Relative molecular mass of MCP can be derived from the regression equation. Relative molecular masses of MCP were obtained: M(n), 21000; M(w), 43000; M(p), 46000; M(z), 66000. The polydispersity index of MCP was 2. The established method is simple, rapid and repeatable for quality control in preparing MCP and it would be also applicable to the study of composition-activity relationship of MCP in the future. PMID- 18161324 TI - Determination of pitavastatin from human plasma using high performance liquid chromatography with fluorescence detection. AB - Pitavastatin belongs to the class of coenzyme A reductase inhibitors. Very few methods of assaying pitavastatin from human plasma are available in literature. An analytical method is presented for the determination of the drug from human plasma making use of the fluorescent property of the drug. The drug is extracted from plasma using ethyl acetate under neutral condition and then analyzed by reversed-phase high performance liquid chromatography (HPLC) with fluorescence detection (lambda(Ex) 245 nm; lambda(Em) 420 nm). Analysis of pitavastatin was carried out on a C18 HPLC column using a gradient flow of mobile phase (0.01 mol/L monobasic potassium phosphate (pH 3.20) -acetonitrile, 63:37, v/v). Fluorescein isothiocyanate was used as internal standard. The dynamic range of assay was 3 to 50 ng/mL. The intraday precision was less than 10% and accuracy ranged from 95.2% to 112.6%. The same for interday check was less than 12% and 92.8% to 105.1%, respectively. The drug was found to be stable under the assay conditions. The developed method is simple, precise, accurate, and stable. This indicates that it can be applied to routine analysis of this drug in human subjects where there are large numbers of samples without the need of specialized instruments like column switching. PMID- 18161325 TI - [Separation and quantification of theanine enantiomers using high performance liquid chromatography coupled with chiral derivatization]. AB - L-Theanine is a unique non-protein amino acid in tea, and shows many physiological functions. To directly extract L-theanine from tea is a lengthy and very costly process. Although the chemical synthesis of theanines is simple, the product is unfortunately a mixture of DL-enantiomers. A chiral derivatization method was developed for the separation and quantification of theanine enantiomers by reversed-phase high performance liquid chromatography (RP-HPLC). 1 Fluoro-2,4-dinitrophenyl-5-L-alanine amide (FDAA)was used as the chiral reagent. This method showed good linearity for both L-theanine (ranging from 1.732 x 10( 3) to 2.077 microg) and D-theanine (ranging from 1.696 x 10(-3) to 2.044 microg). The recoveries were in the range of 97.3% - 102.0% for L-theanine and 97.2% - 103.2% for D-theanine. This method also showed excellent limit of detection (approximately 5 x 10(-4) microg) and limit of quantification (approximately 1 x 10(-3) microg) for both L-theanine and D-theanine. The results demonstrated that this method is precise, accurate and can be used for the determination of theanine enantiomers. PMID- 18161326 TI - [Ligand exchange chromatographic separation of DL-amino acids based on monodisperse resin-bonded chiral stationary phase]. AB - L-Proline chiral stationary phase for ligand exchange chromatography was prepared in the following steps. First, the particles were completely hydrolyzed. Second, the hydrolyzed particles reacted with epichlorhydrin to obtain chlorinated beads. Third, the chlorinated beads reacted with L-proline. The chiral stationary phase was characterized by elemental analysis. Chromatographic resolutions of DL-amino acids were achieved on the chiral stationary phases by using 0.1 mol/L NaAc with 0.1 mmol/L Cu(Ac)2 solution as mobile phase and detection at 254 nm. The elution order of D-isomer before L-isomer was observed for all the DL-amino acids resolved except DL-proline. For DL-proline the elution order was different from the others because of its five membered ring structure. The influences of the mobile phase pH, concentration of Cu(II), flow rate of eluent and temperature of column on the resolution of DL-amino acids by ligand exchange chromatography were investigated for the optimization of chromatographic conditions. The results showed that enantioseparation of some DL-amino acids was performed by ligand exchange chromatography on this chiral stationary phase with satisfactory results. PMID- 18161327 TI - [Simultaneous determination of nine sulfonamide residues in milk using solid phase extraction and high performance liquid chromatography]. AB - A simple, rapid, sensitive and reliable high performance liquid chromatographic method for the simultaneous determination of nine sulfonamides (SAs) (sulfadiazine, sulfathiazole, sulfamethazine, sulfamethoxypyridazine, sufamonomethoxine, sulfachloropyridazine, sulfamethoxazole, sulfadimethoxine and sulfaquinoxaline) in milk was developed. The method includes the shaking of the milk with phosphate buffer, centrifugation, and cleaning up on a C18 solid-phase extraction cartridge. The C18 cartridge was washed with water, and the SAs were eluted with methanol. After the evaporation, the residue was dissolved in ethyl acetate, and cleaned up on an amino solid-phase extraction column. Analytical separation was performed on an Inertsil ODS-3 column with photodiode-array detection at 270 nm under the gradient condition. The mean recoveries of SAs at the spiked levels of 10, 20, 40 microg/L were between 72.1% and 88.3%. The relative standard deviations were between 2.3% and 5.0%. The detection limits were 1.7 - 2.8 microg/L, and the quantification limits wer 5.7 - 9.2 microg/L. PMID- 18161328 TI - [Chiral separation of tenatoprazole enantiomers using high performance liquid chromatography on vacomycin-bonded chiral stationary phase]. AB - Vacomycin-bonded chiral stationary phase was used for the direct chiral separation of tenatoprazole enantiomers using reversed-phase high performance liquid chromatography (HPLC). The influences of the kinds and concentration of buffer and organic modifier, the pH value of buffer, column length and column temperature on the separation were examined. The chiral HPLC method for the separation of tenatoprazole enantiomers on a Chirobiotic V column (150 mm x 4.6 mm, 5 microm) was established with simplicity and good reproducibility using 0.02 mol/L ammonium acetate buffer (pH 6.0)-tetrahydrofuran (93:7, v/v) as the mobile phase at a flow rate of 0.5 mL/min and 20 degrees C. Under the above conditions, the enantiomers were separated on baseline with the resolution of 1.68. The relative standard deviations (RSDs) for the retention times of tenatoprazole enantiomers were 0.48% and 0.49% (n = 6). The RSDs for the peak areas of tenatoprazole enantiomers were 0.45% and 0.55% (n = 6). PMID- 18161329 TI - [Isolation and preparation of flavones from the leaves of Lindera aggregata using high speed counter-current chromatography]. AB - Lindera aggregata (Sims) Kosterm. belongs to Lauraceae family and is distributed in the southern parts of China, Japan and southeastern Asia. The leaves of this plant are frequently used in folk medicine, as antioxidant, antiinflammatory and antibacterial. In previous reports, eleven flavones were isolated from this plant. The study on the flavone constituents of the leaves of this plant led to the isolation of five flavone compounds, as quercetin-3-O-beta-D-glucoside (1), quercetin-5-O-beta-D-glucoside (2), quercetin-3-O-beta-D-arabinfuranoside (3), quercetin-3-O-rhamnopyranoside (4) and kaempferol-7-O-alpha-L-rhamnopyranoside (5), by means of high speed counter-current chromatographic (HSCCC) procedures. A two-phase solvent system composed of hexane-ethyl acetate-n-butanol-acetic acid water (2: 4: 2: 1.5: 6, v/v) was used at a flow rate of 2.0 mL/min, while the apparatus rotated at 800 r/min and the effluent was detected at 280 nm. Compounds 1, 2, 3 and 5 were for the first time obtained from Lindera aggregata (Sims) Kosterm. PMID- 18161330 TI - [Isolation and determination of ecliptal in Herba Ecliptae by reversed-phase high performance liquid chromatography]. AB - The ecliptal was isolated from Herba Ecliptae by high performance liquid chromatography and identified by mass spectrometry and nuclear magnetic resonance (1H NMR). A method for determination of ecliptal in Herba Ecliptae was developed by using a Kromasil C18 column (200 mm x 4.6 mm, 5 microm) with a mixture of acetonitrile and 0.1% phosphoric acid solution (65:35, v/v) as mobile phase at a flow rate of 1.0 mL/min. The detection wavelength was set at 365 nm and temperature was set at 30 degrees C. The volume of injection was 10 microL. There was good linear relationship (r = 0. 9993) between the mass concentration and the peak area of ecliptal in the range of 3.0 - 60.0 microg/mL. The recovery was found to be in the range of 96.7% - 100.0%. The results of the experiments have demonstrated that the established method is rapid and simple with good accuracy and reproducibility. The method is suitable for the quality control of Herba Ecliptae from different sources. PMID- 18161331 TI - [Simultaneous determination of seven sulfonamides and metronidazole and chloramphenicol in cosmetics by high performance liquid chromatography]. AB - A method for the simultaneous determination of seven sulfonamides (sulfacetamide, sulfapyridine, sulfamerazine, sulfamethazine, sulfamater, sulfamonomethoxine, sulfamethoxazole) and metronidazole, chloramphenicol in cosmetics has been developed by high performance liquid chromatography with a photodiode array detector (HPLC-PDA). The chromatographic column Atlantis dC18 (150 mm x 4.6 mm, 5 microm) was used with acetonitrile-0.1% formic acid (2:8, v/v) aqueous solution as mobile phase at a flow rate of 1.0 mL/min. Samples were extracted by mobile phase and detected by HPLC-PDA at 268 nm. The limits of quantification were 3 - 80 microg/g. The correlation coefficients of linear calibration curves were over 0. 999 3 within the sulfonamide concentration range of 20 - 200 microg/mL and metronidazole and chloramphenicol concentration range of 40 - 400 microg/mL. Average recoveries were 83.8% - 105.3% at the spiked levels of 50 microg/mL and 150 microg/mL for sulfonamides, and 100 microg/mL and 300 microg/mL for metronidazole and chloramphenicol. The relative standard deviations were below 5%. This method can be routinely used for the determination of seven sulfonamides and metronidazole, chloramphenicol in cosmetics. PMID- 18161332 TI - [Determination of polycyclic aromatic hydrocarbons in Dianchi Lake using stir bar sorptive extraction-thermal desorption-gas chromatography/mass spectrometry]. AB - A fast method for determination of polycyclic aromatic hydrocarbons (PAHs) in water samples was established. The samples from the Dianchi Lake and Panlong River were analyzed directly using stir bar sorptive extraction-thermal desorption system-gas chromatography/ mass spectrometry (SBSE-TDS-GC/MS). Polydimethylsiloxane (PDMS) was used as the extraction phase. The samples were extracted with stir bars at 1100 r/min for 60 min at ambient temperature (25 degrees C). Desorption was carried out at 300 degrees C for 5 min under a helium flow of 50 mL/min in the splitless mode while maintaining a cryofocusing temperature of 0 degrees C in the programmed-temperature vaporization (PTV) injector of the GC/MS system. The limits of detection of PAHs ranged from 1.0 pg to 468.8 pg. The theoretical recoveries of PAHs were greater than 90% (except that naphthalene was 64.90%), and the recoveries of 16 PAHs from water samples ranged from 83.1% to 109.4%, and the relative standard deviations (RSDs) ranged from 1.2% to 9.3% (n = 6). PAHs in Dianchi Lake and Panlong River samples were quantified rapidly by this method. The results show that, the total content of these 16 PAHs is 89.16 ng/L in Dianchi Lake, 65.41 ng/L in the up-stream of Panlong River, 339.22 ng/L in the middle-stream of Panlong River and 62.25 ng/L in the down-stream of Panlong River. It is demonstrated that the Dianchi Lake and Panlong River were polluted by PAHs to some degree. PMID- 18161333 TI - [Determination of acetanilide herbicide residues in tea by gas chromatography mass spectrometry with two different ionization techniques]. AB - An analytical method of solid phase extraction-gas chromatography-mass spectrometry with two different ionization techniques was established for simultaneous determination of 12 acetanilide herbicide residues in tea-leaves. Herbicides were extracted from tea-leaf samples with ethyl acetate. The extract was cleaned-up on an active carbon SPE column connected to a Florisil SPE column. Analytical screening was determined by the technique of gas chromatography (GC) mass spectrometry (MS) in the selected ion monitoring (SIM) mode with either electron impact ionization (EI) or negative chemical ionization (NCI). It is reliable and stable that the recoveries of all herbicides were in the range from 50% to 110% at three spiked levels, 10 microg/kg, 20 microg/kg and 40 microg/kg, and the relative standard deviations (RSDs) were no more than 10.9%. The two different ionization techniques are complementary as more ion fragmentation information can be obtained from the EI mode while more molecular ion information from the NCI mode. By comparison of the two techniques, the selectivity of NCI SIM was much better than that of EI-SIM method. The sensitivities of the both techniques were high, the limit of quantitative (LOQ) for each herbicide was no more than 2.0 microg/kg, and the limit of detection (LOD) with NCI-SIM technique was much lower than that of EI-SIM when analyzing herbicides with several halogen atoms in the molecule. PMID- 18161334 TI - [Determination of atrazine and its metabolites in human urines using gas chromatography]. AB - Atrazine (ATZ) was widely used to control broadleaf weeds. Numerous animal experiments have proved that atrazine is a suspicious endocrine disruptor. Thus, further development of the ability to estimate low-dose human exposure to atrazine is requested in epidemiologic studies to correlate the toxicological effects associated with the concentrations of ATZ and its metabolites in human body. A method for detecting ATZ and its metabolites (deethylatrazine (DEA), deisopropylatrazine (DIA), deethyldeisopropylatrazine (DEDIA)) in human urines using gas chromatography was established. A urine sample was extracted by ethyl acetate, and purified using a Florisil column. Final concentrated extract was analyzed by a gas chromatograph-electron capture detector. The conditions of this method were optimized. The limits of detection were 0. 002 5 mg/L for DEDIA, 0. 005 mg/L for DEA, DIA and ATZ. The linear ranges were from 0.2 to 8 ng for all analytes. The atrazine concentrations in urine samples of the workers collected from an atrazine plant were determined by this method. The concentration ranges were 0.003 -0.301 mg/L for DEDIA, 0.005 -0.011 mg/L for DEA, 0.006 -0.276 mg/L for DIA, and 0.005 -0.012 mg/L for ATZ. PMID- 18161335 TI - [Classification of blue gel inks and investigation of change rule of the ink with writing age using pyrolysis gas chromatography]. AB - Gel ink pen (neutral pen) is a new popular pen. Its coloring matters are usually pigments which could not be extracted by water or organic solvents. Therefore, the methods of gas chromatography and high performance liquid chromatography for analyzing the inks of ball-point pen and water-based pen cannot be used here. There are no effective methods to identify gel inks in forensic science up to now. Pyrolysis gas chromatography (Py-GC) was used to analyze 65 different blue gel inks in this report. The pyrolysis was performed with a CZ-100 pyroprobe system using a coil filament with a quartz sample tube at 770 degrees C for 10 s. Agilent-6890N Gas Chromatograph was used with a nitrogen-phosphorus detector. A DM-5 Dikma Technology column (30 m x 0.32 mm x 0.25 microm) was used. Based upon the number and retention time of peaks in the pyrograms, the gel inks were separated into 3 classes. The most of the gel inks fell into the second group which have two characteristic peaks of copper phthalocyanine pigment. The results showed that the retention times and ratio of the peak areas were stable and reproducible. The writing age of blue gel ink was primarily investigated by the relative change rule of the main components in the ink, and the curves of writing age was obtained. PMID- 18161336 TI - [Determination of phthalates in drinks by HPLC-ESI/MS]. PMID- 18161337 TI - [Simultaneous determination of 8-methoxypsoralen and 5-methoxypsoralen in cosmetics using LC-MS/MS]. PMID- 18161338 TI - [Separation and determination of furanocoumarins in shatian pomelo juice by HPLC MS]. PMID- 18161339 TI - [Determination of rivanol in amniotic fluid using HPLC]. PMID- 18161340 TI - [In situ separation and determination of raw material, products and by-products during A series of reactions of alkylated 3-bromthiophene with HPLC]. PMID- 18161341 TI - [Simultaneous determination of chrysoidine and auramine O in bean products by HPLC]. PMID- 18161342 TI - [Study on chemical constituents of the essential oil from Artemisia selengensis in Dongting Lake Area]. PMID- 18161343 TI - [Determination of swainsonine in A. variabilis bunge by gas chromatography with methyl-alpha-D-mannopyranoside internal standard method]. PMID- 18161344 TI - [Reduction mammaplasty with the superomedial glandular pedicle]. AB - OBJECTIVE: To explore a new technique that keep function and good aesthetic shape after reduction mammaplasty. METHODS: The superomedial glandular pedicle technique was applied with differing patterns of skin incision according to the degree of the mammary hypertrophy. The extra glandular tissue was removed from the lateral and inferior segment of the breast. The remaining superomedial glandular tissue was remodeled a conical shape. The skin incision was sutured after excision of excess skin. RESULTS: 36 cases (72 breasts) were treated with this technique. The results were satisfactory and there were no complications. The breasts maintained a lasting and aesthetically pleasing shape and the nipple areola preserved good sensation. CONCLUSIONS: The superomedial glandular pedicle is a safe and effective technique that can provide long-lasting outcome. The design of the procedure can be adapted to a variety of skin incision patterns and breast morphologies. PMID- 18161345 TI - [Modification of Lejour reduction mammaplasty--mammaplasty of L-shaped scar]. AB - OBJECTIVE: To introduce a modification of Lejour reduction mammaplasty. METHODS: With the upper pedical flap as the base of mammaplasty, the lower part of breast was resected while excess skin was pushed to lateral and formed "L"-shaped scar after it was resected. RESULTS: From October 2005 to April 2006, the modified Lejour reduction mammaplasty was applied to 10 mammahypertrophic patients with 20 breasts in sum. The result of operation was good and only "L"-shape scar was left in the lower lateral part of the breast. CONCLUSIONS: This method is easy to perform and could avoid inverted T scar caused by routine mammaplasty technique. This method is worth introducing widely. PMID- 18161346 TI - [Analysis of the complications of two kinds reduction mammaplasty]. AB - OBJECTIVE: To compare the complications of two kinds reduction mammaplasty (group A: central pedicle plus lateral incision; group B: double circle incision plus mesh) and analyse the reason of the complications to avoid or reduce their happening. METHODS: The complications in 58 patients undergone reduction mammaplasty was investigated from August 2003 to May 2006. And the kind and rate of the complications with two methods was compared. The complications include: infection, hematoma, fat liquefaction, nipple necrosis, skin necrosis, hypertrophic scars, mesh pucker, asymmetry, inadequate reduction and so on. RESULTS: Follow-up study for 4 to 33 months, 7 in group A (36 cases) got complications; 6 in group B (19 cases) got complications. CONCLUSIONS: The rate of complications in group B was higher than in group A. The complications in group A were related to destroyed blood supply, which was due to excess excision of tissue. The complications in group B were relate to the mesh, such as too hard, not smooth, too tight, thin skin covering and so on. PMID- 18161347 TI - [Local flap for reconstruction of nasal tip defect: functional and aesthetic evaluations]. AB - OBJECTIVE: To introduce the experience of reconstruction of nasal tip defect, which could obtain good functional and aesthetic results. METHODS: Based on the aesthetic subunit principle, according to the size, shape, depth of nasal tip defect, and the conditions of surrounding skin, homologous local flap (include: frontonasal flap, bilobed flap, nasolabial flap, reforming rhomboid flap and forehead flap, etc.) was designed to cover the defect wound. The secondary defect of donor area was directly sutured. RESULTS: A total of 83 patients with nasal tip defect were treated by local flap transfers. The area of the defect ranged between 0.6 x 0.6 cm and 2.5 cm x 2.5 cm. The nasal tip defect was successfully closed in all patients, and all flaps survived well without obvious complications. 76 out of these 83 patients were available for follow-up from 1 to 36 months. The repaired tissue was in good match with surrounding tissue, with nice nasal contour, and cosmetic results were satisfactory. No tumor recurrence occurred. CONCLUSIONS: Based on the nasal aesthetic subunit principle, local flap could be used to reconstruct the nasal tip defect below 2.5 cm x 2.5 cm, and obtain good color, contour and texture in match with the surrounding skin. Cosmetic results were satisfactory. PMID- 18161348 TI - [The preoperative design for remodeling inferior one-third facial contour]. AB - OBJECTIVE: Facial contour remodeling is an important part of plastic surgery. Some skeletal abnormalities, such as mandible protrude, maxillary retrusion, induce the facial asymmetry (including arrowe direction or/and coronary direction), malocclusion, etc. These abnormalities are seen commonly in the lower one third of the face. Evaluation and preoperative design is very important for the treatment of these abnormalities. The purpose of this item is to summarize our experience in the preoperative design during the treatment. METHODS AND RESULTS: Observation of the facial feature of the patient, cephalometric tracing, face-bow transfer and model reconstruction were necessary for the preoperative design. Three-dimensional direction was included in the bone movement for operation and soft tissue should be taken into account simultaneously. All the factors mentioned above were summarized, and outcomes were gathered. CONCLUSIONS: Determination of facial midline was very important, especially for asymmetry patient. The bone was moved in three-dimensional direction. The same site of osteotectomy may produce different effects by different movements. PMID- 18161349 TI - [Significance of preserving the posterior branch of the great auricular nerve in parotid surgery]. AB - OBJECTIVE: To investigate the feasibility, technique and significance of preserving the posterior branch of the great auricular nerve in parotid surgery. METHODS: 48 cases with parotid tumor who underwent regular parotid surgery with cosmetic incision were included. The sensory testing was carried out in upper auricle, lobule, infra-auricular, pre-auricular and post-auricular region preoperatively and at 10 days, 1 month, 6 months and 12 months after operation. RESULTS: The posterior branch of the great auricular nerve was preserved in 35 out of 48 patients. There was no sensory loss in the upper auricle and post auricular region. Early after operation, sensory disturbance was found in lobule, infra-auricular region, but significantly in pre-auricular region. The sensory disturbance was recovered slowly to nearly normal level at six months after operation. 13 cases with sacrifice of the nerve had a significantly higher sensory disturbance and a longer recovery time. CONCLUSIONS: Preservation of the posterior branch of the great auricular nerve during parotid surgery is technically feasible and can reduce the postoperative sensory disturbance and possible permanent sensory loss. It further helps to improve the quality of life early after operation. PMID- 18161350 TI - [Clinical significance of vascular distribution type of the expanded skin in the mastoid region in congenital microtia]. AB - OBJECTIVE: To classify the blood vessel distribution of the expanded skin in the mastoid region and its relevance to reasonable and reliable design of the expanded flap for auricular reconstruction in congenital microtia. METHODS: The blood vessel distribution of the expanded skin in the mastoid region was observed by light permeation test. The expanded flaps with different ratio of length to width were designed according to their blood vessel distribution types. RESULTS: The vascular distribution of the expanded skin in 403 cases was divided into five types. All the flaps survived completely. CONCLUSIONS: The blood vessel distribution type of the expanded skin in mastoid region has great significance for the design of post-auricular expanded flap in auricular reconstruction. PMID- 18161351 TI - [Clinical application of xenogenic (porcine) acellular dermal atrix (ADM) in scar treatment]. AB - OBJECTIVE: To investigate the role of xenogenic (porcine) ADM as dermal substitute in scar treatment. METHODS: After scar excision, the wounds were covered with composite grafts of DR procine ADM and autologous thin split thickness grafts in one stage or in two stages. RESULTS: 22 out of 47 cases were treated in two-staged procedure. After the ADMs were applied to the wound, the autologous thin split-thickness grafts were implanted 7 days later. 25 cases were treated in one-staged procedure. The survival rates of composite grafts were (88.3 +/- 3.7)% for subcutaneous recipient bed and (89.7 +/- 3.4)% for deep fascia recipient bed in group with two-staged procedure, compared with (92.5 +/- 4.1)% and (93.2 +/- 5.2)%, respectively, in group with one-staged procedure. Early after grafts taken, the grafts had a pink colour and smooth surface. The patients were followed up for 90 days at most. The survived composite grafts were durable, elastic, smooth and soft with good function and appearance like normal skin. They could even be pinched up. The scar along the edge of the grafts was slightly hypertrophic. CONCLUSIONS: The survival rate of composite graft is higher in patients with one-staged procedure. The elasticity and textural of the taken grafts are better on subcutaneous recipient bed than on deep fascia recipient bed, though the function has no difference. Xenogenic (porcine) ADM can be an optimal dermal substitute for wound coverage after scar excision. PMID- 18161352 TI - [Classification and surgical management of upperlid dermatochalasis in middle aged and elderly patients]. AB - OBJECTIVE: To investigate the classification and surgical management of upper lid dematochalasis in middle-aged and elderly patients. METHODS: 105 cases with upper lid dermatochalasis who underwent surgical treatment between May, 2000 to May, 2005 were retrospectively analyzed. The dermatochalasis was classified according to eye brow position, bulge contour and the skin amount of upper lid. The post operative cosmetic result was also assessed. RESULTS: Of the 105 treated cases, the post-operative cosmetic result was assessed as very satisfactory in 77 (87%) cases, as satisfactory in 11(10%) cases, and as dissatisfactory in 3 (2%) cases. CONCLUSIONS: Dermatochalasis in middle-aged and elderly patients has different types and should be treated with different surgical method. PMID- 18161353 TI - [Frontalis muscle fascial flap passing through the pulley of orbital septum for correction of severe blepharoptosis]. AB - OBJECTIVE: To evaluate the clinical result of the frontalis muscle fascial flap passing through the pulley of orbital septum for correction of severe blepharoptosis. METHODS: 57 eyes in 52 cases with congenital severe blepharoptosis were treated in recent two years. After the frontalis muscle fascial flap was prepared beneath the orbicularis oculi muscle, the pulley was created by two parallel horizontal incision on the orbital septum at the upper orbital rim and 1 cm under the upper rim. The frontalis muscle fascial flap was then pulled down behind the pulley and out to be attached to the upper margin of tarsal plate. RESULTS: The following-up period was 3-6 months. Satisfactory cosmetic result was achieved in 52 eyes. Three eyes had ptosis relapse and 2 eyes had unnatural contour of the palpebral margin which required another corrective operation. No other complication was observed. CONCLUSIONS: The pulley created by the orbital septum makes the traction lines of the frontalis muscle fascial flap in a similar direction as the natural movement of levator muscle. So both the postoperative static and dynamic appearance of the upper lid is more natural. The technique is very practicable in correction of blepharoptosis. PMID- 18161354 TI - [Surgical management of blepharoptosis: a report of 500 cases]. AB - OBJECTIVE: To investigate the various surgical techniques and their results for different types of blepharoptosis. METHODS: 620 eyes of 500 cases with blepharoptosis who underwent surgical treatment were retrospectively analyzed. The patients were diagnosed as congenital, neurogenic, myogenic, traumatic, senile and mechanical ptosis. The used techniques included resection of levator muscle, anastomosis of frontalis muscle and levator aponeurosis, Whitnall's ligament sling, Friedenwald- Guyton's frontalis muscle fixation, levator aponeurosis reconstruction, modified Hotz's tarsectomy. RESULTS: The overall success rate was 90.3% (560/620). 60 eyes with unsatisfactory result showed overcorrection in 5 eyes and undercorrection in 55 eyes in which the patients also suffered from combined eyelid deformities in 4 eyes, entropion in 6 eyes and ectropion in 2 eyes. The success rate of resection of levator muscle for the mild and moderate congenital ptosis was 93.8%. The success rates of resection of levator muscle and anastomosis of levator aponeurosis and frontalis muscle for the severe congenital ptosis were 72.4% and 100% respectively. The success rate of Whitnall's ligament sling for the recurrent congenital ptosis was 90%. The success rates of Friedenwald-Guyton's frontalis muscle fixation, Whitnall's ligament sling and anastomosis of levator aponeurosis and frontalis muscle for the neurogenic and myogenic ptosis was 41.6%, 80% and 90% respectively. The success rates of levator aponeurosis reconstruction for the traumatic and senile ptosis were 94.7% and 100%, respectively. The success rate of modified Hotz' tarsectomy for the mechanical ptosis was 93.3%. CONCLUSIONS: The key point for successful correction of ptosis is the selection of the right indication for each technique depended on the type and severity of the ptosis. The technique skill is also very important. PMID- 18161355 TI - [Comparison study of three methods for the treatment of microgenia]. AB - OBJECTIVE: To analyze the clinical results and complications of three methods for microgenia, including chin augmentation with silicone implant, bone autograft, and genioplasty. METHODS: The advantages and disadvantages of the three methods for microgenia were assessed through 3-D CT reconstruction, X-ray and intraoperative observation during the second operation. The indications for each technique were also studied. RESULTS: The frequently observed problems for chin augmentation with silicone implants were implant malposition, underlying bone absorption, periosteal reaction and undercorrection. Chin augmentation with bone autograft had a great long-term bone graft absorption which led to undercorrection. Genioplasty showed a satisfactory cosmetic result with no serious complication. CONCLUSIONS: Chin augmentation with silicone implant is suitable for mild microgenia with a shallow mentolabial groove, but without facial vertical insufficient and facial asymmetry. Genioplasty can be used in all kinds of microgenia in any severity, especially those with deviated chin and facial asymmetry. Chin augmentation with bone autograft can't achieve good long term result and should be applied prudently. PMID- 18161356 TI - [An experimental study on influence of concentration of fetal bovine serum on chondrogenesis of BMSCs]. AB - OBJECTIVE: To explore the chondrogenetic effect of induce media containing different concentrations of fetal bovine serum (FBS) on BMSCs differentiation in vitro and provide technical parameters for cartilage engineering in vitro. METHODS: Passage 2 BMSCs of swine were seeded at the density of 5 x 10(7) cells/cm3 to disc-shaped PGA scaffolds with a diameter of 5mm and a thickness of 2mm. After 7days, the scaffolds were induced in media with TGF-beta1, IGF-I, dexamethasone, and different concentrations of FBS: 0% in A group, 5% in B group, and 10% in C group. Specimens were collected after 8 weeks for gross observation, size evaluation, wet weight, glycosaminoglycan (GAG) content, histology assessment, and immunohistology of type II collagen. RESULTS: The compound of C group showed china-white color, hard and fine texture, no obvious change in size and shape, typical lacuna structures, cartilage specific ECM, and significantly higher wet weight and GAG content. The compound of B group showed reduced size, fewer lacuna structures and some cartilage specific ECM. And the compound of A group showed greatly reduced size, soft and loose texture, and no typical lacuna structure or cartilage specific ECM. CONCLUSIONS: FBS was indispensable to chondrogenetic media for in-vitro tissue engineering of cartilage with BMSCs. PMID- 18161357 TI - [Experimental study of construction of tissue engineered bone ectopically by human bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To study the possibility and mechanism of construction of tissue engineered bone with human bone marrow mesenchymal stem cells (hBMSCs) as seeding cells and partially demineralized bone matrix (pDBM) as scaffold. METHODS: hBMSCs are cultured and mutiplified. The 4th grade hBMSCs are seeded on the pDBM, the growth and adhesion of hBMSCs on pDBM are observed under scanning electro microscope. The adhesion efficiency is assessed. The complexes are implanted in the nude mice subcutaneously, the pDBM without cells as control. The grafts are taken out on the 8th and 12th week. RESULTS: There is new bone formation on the 8th and 12th week in complex group. There is a layer of osteoblast like cells adhered on the surface of most of the new bone, which suggest the possibility of intramembranous ossification. There is no bone formation in control group. CONCLUSIONS: Tissue engineered bone can be constructed with hBMScs and pDBM in vivo, and the mechanism of which could be intramembranous ossification. PMID- 18161358 TI - [Experimental study of adipose tissue differentiation using adipose-derived stem cells harvested from GFP transgenic mice]. AB - OBJECTIVE: To identify the expression of protein which is characteristic of stem cell, induce the adipogenic differentiation in vitro by ASCs harvested from GFP transgenic mice, and to assess the possibility of constructing adipose tissue via the attachment of ASCs to type I collagen scaffold. METHODS: Inguinal fat pads from GFP transgenic mice were used for the isolation of ASCs. Expression of CD29, CD34, CD45, CD133 and HLA-DR were detected by flow cytometry. After expansion to three passages, the ASCs were incubated in an adipogenic medium for two weeks. Then they were attached to collagen I scaffold and co-cultured for 12 hours, followed by transplantation under the dorsal skin of athymic mice for 2 months. Adipogenic differentiation of ASCs in vitro was assessed by morphological observation, Oil red O staining and newly formed tissue was detected by HE staining. RESULTS: The cultured stem cells were fibroblast-like cells and showed highly homogeneous appearance with active proliferation capacity. Stem cells' characteristic CD expression was proved. After being incubated in an adipogenic medium, they could differentiate into mature adipocytes. Accumulation of lipid droplets in the cytoplasm was testified by Oil red O staining, morphological and biological observation. 0.5 ml new tissue was formed and was confirmed by fluorescent observation and HE staining to be mature adipose tissue. CONCLUSIONS: Adipose derived stem cells can successfully differentiate into mature adipocytes exhibiting an adipose-like morphology and expression of intracytoplasmic lipid droplet. It was an efficient model for adipose tissue engineering with ASCs and type I collagen scaffold. PMID- 18161359 TI - [Morphologic observation of the regenerated nerve in reconstructed penis with sensory nerve implantation in rabbit]. AB - OBJECTIVE: To investigate the mechanism of sensory nerve regeneration of the reconstructed penis with sensory nerve implantation and to explore a new surgical technique to improve the postoperative sensory function in phallic reconstruction. METHODS: Adult male New Zealand rabbits were randomly divided into experimental group (n = 20, with sensory nerve implantation) and control group (n = 20, without sensory nerve implantation), which were both performed phalloplasty with a superficial epigastric faciovascular pedicle flap. Postoperatively, the nerve regeneration process of the reconstructed penis was observed histologically. RESULTS: In experimental group, the amount of CGRP positive nerve fibers increased markedly with the time prolonged, whereas merely a few CGRP positive fibers scattered in deep dermis 6 months later in the other group. The cutaneous sensory nerve regeneration of the reconstructed penis in experimental group shows the procedure that the myelinated axon began to exist within 3 months, thereafter the myelinated axon and unmyelinated axon were both observed under the electron microscope. CONCLUSION: These findings show that the rabbit model of phalloplasty with sensory nerve implantation can acquire well sensory reinnervation, and bring a light to clinical application for restoration of sensory function in reconstructed penis. PMID- 18161360 TI - [An experimental study of the effect of burying testis in thigh pocket on spermatogenesis]. AB - OBJECTIVE: To explore the effect of burying testis in thigh pocket on spermatogenesis. METHODS: Guizhou miniature male pigs at child-bearing period were randomly divided to receive operation of scrotum incision and dissection with the testis burying in thigh pocket (experimental group) or without (control group). 3 months later, testis biopsy was performed on 2 pigs from each group for pathological examination. Then every male pig from both experimental (n = 6) and control group (n = 6) got a mating partner and lived together for 3 months. The fertility of the male pigs was observed. 6 months after operation, testis biopsy was performed again on all the animals from both the groups. RESULTS: Both at 3 months and 6 months after operation, the pathological examination showed the spermatogenic cells of all stage in contorted seminiferous tubules markedly decreased with no mature sperm in experimental group, while normal spermatogenic cells with mature sperm in control group. After the male pigs lived with mating partners for 3 months, no female pigs staying with the experimental group became pregnant, but the male pigs in control group had a normal fertility. CONCLUSIONS: Burying testis in thigh pocket impedes spermatogenesis in the miniature male pig. So burying testis in thigh pocket is not recommended for patients with scrotum skin defect who wish to remain fertile. PMID- 18161361 TI - [A long-term study of regeneration of mechanical sensory fibers after free nerve transplantation to the rabbit reconstructed penis]. AB - OBJECTIVE: To explore the regeneration of mechanical sensory fibers after free nerve transplantation. METHOD: Neuroelectrophysiological technique (single nerve fiber recording) was used to test the regeneration rate of mechanical sensory fibers, the proportion of rapidly and slowly adapting receptors, the stimulating thresholds of regenerated mechanoreceptors and conduction velocity of regenerated fibers. The regeneration pattern of the mechanoreceptors after free nerve transplantation to the rabbit reconstructed penis was also analyzed. RESULTS: 9 months after operation, the number of regenerated mechanical sensory fiber was almost normal. The regenerated rapidly adapting receptors had a higher proportion with higher mature degree than the regenerated slowly adapting receptors. 9 months after nerve transplantation the stimulating thresholds of regenerated mechanoreceptors and conduction velocity of regenerated fibers remained below normal. CONCLUSION: After free nerve transplantation to the rabbit reconstructed penis, the function of both rapidly and slowly adapting sensory nerve fiber partially recovered, but in different extent. PMID- 18161362 TI - [Effect of HMME-PDT on hyperplastic scar in rabbit ear model]. AB - OBJECTIVE: To investigate the effect of HMME-PDT (Hematoporphyrin Monomethyl Ether-Photodynamic therapy) on Hyperplastic scar in the rabbit ear. METHODS: The acute model of dermal Hyperplastic scar in the rabbit ear was established. 24 scars were randomly divided into 2 groups: the experimental group (n = 12) received HMME-PDT treatment, and the controlled group (n = 12) received no special treatment. Specimens were harvested from scars on postoperative 28 day. Scar hyper plasty and collagen fibers were observed by haematoxylin-eosin staining and Van-Gieson staining respectively. The microvessel density was calculated under microscope. RESULTS: Compared with the controlled group, HMME PDT treatment in the experimental group reduced scar formation, decreased the microvessel density and prevented excess collagen deposition at the wound site. CONCLUSIONS: HMME-PDT may play a role in inhibiting hyperplastic scar in rabbit ear. PMID- 18161363 TI - [Association between p53 gene codon 72 polymorphism and keloid in Chinese population]. AB - OBJECTIVE: To investigate the relationship between p53 gene codon 72 polymorphism and genetic predisposition to keloid in Chinese population. METHODS: PCR-based restriction fragment length polymorphism (PCR-RFLP) analysis was used to detect p53 gene codon 72 genotypes of 60 keloid samples and 102 whole blood samples from healthy controls in China. RESULTS: There was no significant difference in the distribution of p53 gene codon 72 polymorphism between the keloid patients and the healthy controls (X2 = 2.910, P = 0.233), nor did the frequencies for Pro and Arg alleles (X2 = 0.882, P = 0.348), and there was no significant difference in the distribution of p53 gene codon 72 polymorphism in keloid patients and normal controls from China and Japan respectively (X2 = 3.942, P = 0.139; X2 = 3.260, P = 0.196). But the Arg/Arg genotype was significantly higher than the Pro/Pro genotype among the patients with keloid in shoulder and back (P < 0.01). CONCLUSIONS: There was no significant association between the distribution of p53 gene codon 72 polymorphism and keloid in Chinese population, but Arg/Arg genotype may affect the formation of keloids in shoulder and back compared to others. Further research should be done to investigate the relationship between p53 gene codon 72 polymorphism and keloids in different sites. PMID- 18161364 TI - [Experimental study on the mechanism of nonablative skin photorejuvenation]. AB - OBJECTIVE: To explore the Mechanism of nonablative skin rejuvenation. METHOD: The Kunming mice be used as subjects and divided into three groups (A, B, C). A, B, C groups were irradiated with 1 320 nm cooltouch laser (20 J/cm2) in the skin of left back; B and C groups were irradiated two and three times respectively; the skin of right back of A, B, C groups was adopted as control. The expression of bFGF and TGF-beta1 in the mouse skin was examined by the immunohistochemistry . The fibroblasts were isolated from the foreskin and cultured. One group is a control and other three ones are low, intermediate and high energetic groups respectively. The fibroblasts were irradiated by laser with 15 J/cm2 ,20 J/cm2 and 24 J/cm2 energy for three times. We examined the levels of bFGF and TGF-beta1 by ELISA in 0, 24, 48 and 72 hours. RESULTS: According to this research on immunohistochemistry result, there are significant differences in the expression of bFGF and TGF-beta1 between the group irradiated by three times and others (P < 0.01). The number of fibroblasts get increased after being irradiated by laser. The ELISA result indicates that the secretion of bFGF increased in the group of intermediate and high energetic level after laser irradiating and may reach the peak at 24 hours (P < 0.01). The amount of TGF-beta1 secretion, however, seems to get decreased in each group at all energetic levels, and at 24 hours it can reach the top level as well. CONCLUSION: The direct influence of laser on the fibroblasts is to promote secretion of bFGF and to inhibit secretion of TGF beta1, while its influence on the tissue is to promote the secretions of the both. Nonablative skin rejuvenation not only can induce fibroblasts to secrete more bFGF but also induce the blood vessels to release cytokines which stimulate endothelial cell to express more of bFGF and TGF-beta1. Furthermore, fibroblastic proliferation can accelerate by laser's irradiating. PMID- 18161365 TI - [Anatomy of buccal and marginal mandibular branches of facial nerve and its clinical significance]. AB - OBJECTIVE: To study the course and distribution of buccal and marginal mandibular branches of facial nerve, and its relevance to the treatment of facial paralysis and the protection of facial nerve during surgery. METHODS: 12 cadaver heads were dissected (24 specimens). The course of the buccal and marginal mandibular branch and the interconnections between them were observed. The relationship of buccal branch to parotid duct, marginal mandibular branch to the inferior border of mandible were studied. With modified Sihler's staining technique, the distribution of facial nerve branches in innervated mimetic muscles was displayed. These anatomic relationships mentioned above were further confirmed during the operation of 40 patients with facial paralysis. RESULTS: Parotid duct had a constant surface landmark. Buccal branch mainly consisted of 2-3 ramifications in 87.5% of the specimens, while marginal mandibular branch was double or single in 95.9% of the specimens. The buccal branch coursed within the distance between 10.7 mm above and 9.3 mm below the parotid duct, and innervated mimetic muscles of midface. The marginal mandibular branch coursed within the distance between 13.4 mm above and 4.8 mm below the lower border of mandible, crossed superiorly the facial artery and innervated mimetic muscles of lower lip. CONCLUSIONS: There is a close relationship of buccal branch to parotid duct and marginal mandibular branch to facial artery and lower border of mandible. With modified Sihler's staining technique, the original 3-dimensional picture of the intramuscular nerve distribution in human mimetic muscles. PMID- 18161366 TI - [Digital medicine and computer-aided surgery]. AB - Digital medicine is an important development of the modern medicine and the computer-aided surgery is an important part of the digital medicine. Applications of digital techniques in the computer aided preoperative planning are elucidated in the paper. Techniques of computer-aided design and computer aided manufacture have been widely used in the fields of customized prosthesis replacement, design and manufacture of customized surgical tools and medical auxiliary instruments and here some clinical cases are introduced. The fundamental technologies used in the surgical navigation and related clinical applications are also explained. The development trend of the computer aided surgery is described and it is concluded that the computer-aided surgery has a bright future. PMID- 18161367 TI - [A positioning error measurement method in radiotherapy based on 3D visualization]. AB - The positioning error in radiotherapy is one of the most important factors that influence the location precision of the tumor. Based on the CT-on-rails technology, this paper describes the research on measuring the positioning error in radiotherapy by comparing the planning CT images with the treatment CT images using 3-dimension (3D) methods. It can help doctors to measure positioning errors more accurately than 2D methods. It also supports the powerful 3D interaction such as drag-dropping, rotating and picking-up the object, so that doctors can visualize and measure the positioning errors intuitively. PMID- 18161368 TI - [Digital interpolation beamformer in medical ultrasound imaging]. AB - A digital beamformer in medical ultrasound imaging systems conventionally requires that the returning signals received by transducers are sampled at a rate significantly higher than the Nyquist rate. This paper presents a digital beamformer that employs zero padding followed by digital interpolation filtering to achieve vernier beamformer delays. The experimental results show that the digital interpolation beamformer can greatly decrease the complexity of A/D converter circuitry. PMID- 18161369 TI - [Methods of extracting AEP for monitoring the anesthetic depth]. AB - Many researches have proved that auditory evoked potentials (AEP) can indicate the depth of anesthesia in the clinical practice. This paper introduces two methods of extracting AEP: the moving time average (MTA) model and the autoregressive model with exogenous input (ARX). In our study we used both the MTA model and the ARX model to extract AEP and the results show that using the ARX model can get the results significantly faster than using the MTA model, and the ARX model is more suitable for the real-time monitoring of the anesthetic depth. PMID- 18161370 TI - [Research on data management of medical equipments in HIFU]. AB - Based on Model-JC Focused Ultrasound Tumor Therapeutic System developed by Chongqing HAIFU (HIFU) Technology Co., Ltd., this paper presents a equip-data discrete management and multi-process equip-data input system along with its design profile and experimental data. And a time-efficiency model is finally set up for multi-process processing. PMID- 18161371 TI - [Construction and research for a web-based economic PACS]. AB - A Java program known as the Applet Viewer is embedded in the browser of the client PC to access the web server which connects with DICOM server. The images can be viewed, queried and diagnosed remotely with the browser. PMID- 18161372 TI - [Study of medical instrument sterilization based on plasma]. AB - In this paper, the generation of the plasma, its principle and its application in the sterilization of medical instruments are described. The relation between discharge voltage and air pressure, frequency, electrode distance is analysed and discussed in glow discharge so as to get a new formula of discharge voltage in the whole frequency range and a design has been made to produce a plasma RF power. The result of the study has a great significance to the application of plasma sterilization of medical instruments. PMID- 18161373 TI - [Design of the wireless medical information system based on PDA]. AB - This paper introduces the design of the wireless medical information management system based on PDA and its model whose experiment results show that it can realize the functional requirements of the design. PMID- 18161374 TI - [Design and practice of the Clinical Information System for ICU]. AB - This paper presents a typical Clinical Information System for ICU and its design and implementation. This system is able to capture and archive vital data from the monitor network, providing a whole digital solution in ICU. These vital data can be used in quantitative analysis in the computer-assisted decision support. PMID- 18161375 TI - [Design of a novel fuzzy FHR monitor and its applications]. AB - The paper presents a novel fetal heart rate (FHR) detecting system which is a fuzzy counter using the auto-correlation and fuzzy mathematic algorithm. This system is able to deal with the increase and decrease phenomena of FHR, caused by the various interference and noises. It has a higher detection level than ever and a better robust system based on the DSP. PMID- 18161376 TI - [Design of a high-voltage insulation testing system of X-ray high frequency generators]. AB - In this paper, we analyze the transformer of X-ray high-voltage high-frequency generators and, have designed and implemented a high-voltage insulation testing system for its oil tank using full-bridge series resonant soft switching PFM DC DC converter. PMID- 18161377 TI - [Comments on anesthesia machines' and monitors' designs]. AB - From a view point of an anesthesiologist, the article points out some design issues of modern monitors and anesthesia machines. User interface problems such as auto-adjusted amplitude display, excessively complicated menus, unreasonable switch arrangement, unsuitable alarm settings or identical cylinder connectors, may affect patient safety and increase anesthesiologists' workload or mental burden. PMID- 18161378 TI - [An overview of robot-assisted percutaneous surgery systems]. AB - This paper introduces a new robot-assisted percutaneous surgery system, its structure and the functions of each subsystems. In addition, a discussion about some foreign systems is made and the technical developing trend of robot-assisted percutaneous surgery systems is given. PMID- 18161379 TI - [Transformer winding's temperature rising and an analysis of its uncertainty]. AB - This paper introduces the temperature rising experimental process and some matters needing attention when the transformer is normally loading. And an analysis of the uncertainty for transformer's temperature rising is also made based on the practical examples' data. PMID- 18161380 TI - Exploring an unfamiliar world. Advice for novices delving into the research arena. PMID- 18161381 TI - What challenges do new researchers face when applying for funding? Doctoral candidate offers advice for overcoming barriers. PMID- 18161382 TI - ONS advocates visit with their policy makers at home. PMID- 18161383 TI - Patient care goes paperless as telemedicine moves mainstream. PMID- 18161384 TI - Put evidence into practice to treat chemotherapy-induced peripheral neuropathy. PMID- 18161385 TI - Healthful eating and exercise may lower mortality after breast cancer. PMID- 18161386 TI - Are your drugs safe? PMID- 18161387 TI - Oncology and specialty drug management in managed care. PMID- 18161388 TI - Letrozole as adjuvant therapy for postmenopausal women with early breast cancer. PMID- 18161389 TI - A conversation with Cary Presant. PMID- 18161390 TI - Oncology and managed care. PMID- 18161391 TI - Economic effect of augmentation strategies in patients with type 2 diabetes initiated on sulfonylureas. AB - This study aimed to assess the economic effect of adding either a thiazolidinedione (TZD) or metformin to the initial sulfonylurea therapy in patients with type 2 diabetes. Researchers identified patients newly diagnosed with type 2 diabetes who initiated therapy with a sulfonylurea during a four-year period from a Medicaid claims database. They then followed these patients after physicians added either a TZD or metformin to the regimen and assessed type 2 diabetes-related costs in the 12-month follow-up period. Multivariate results indicated that patients augmented with metformin incurred 33.3% lower total diabetes-related costs, compared with those augmented with a TZD (P < .001). The study results can aid in making formulary decisions and developing treatment algorithms for step-wise management of type 2 diabetes. PMID- 18161392 TI - Disease management. A condition no longer heard. PMID- 18161393 TI - The effect of the doughnut hole. PMID- 18161394 TI - The Medicare Part D doughnut hole: effect on pharmacy utilization. AB - The Medicare Prescription Drug Improvement and Modernization Act of 2003 offers prescription drug coverage through the Medicare part D program. However, the standard benefit does include a gap in coverage, commonly known as the "doughnut hole". This study, which included 90,615 subjects, aimed to evaluate the effect of the prescription drug coverage gap on drug utilization and expenditures. Beneficiaries in the study group were older (76.34 vs. 73.04 yr, P < .0001) and sicker (5.39 vs. 3.66 disease conditions, P < .0001) than those in the control group. They also incurred substantially higher out-of-pocket expenses ($2534 vs. $598, P < .0001) than the individuals in the control group. From the preperiod to the postperiod, the study group's average days of therapy decreased by 15.85% (from 1104 to 929, P < .0001), and total costs fell 28.02% (from $2441 to $1757, P < .0001). The average out-of-pocket costs increased by 88.94% (from $877 to $1657, P < .0001); in the control group, however, the average days of therapy increased by 1.77% (from 680 to 692), and total costs rose by 2.19% (from $1322 to $1351). Out-of-pocket costs decreased by 5.54% ($307 to $290). Using difference-indifference models, the Medicare part D prescription drug coverage gap was estimated to have reduced medication utilization by 187.49 days of therapy (P < .0001) while raising out-of-pocket costs by $796.49 (P < .0001) and increasing the generic utilization rate by 7.33% (P < .0001). Regular Medicare part D beneficiaries reduced medication utilization after they reached the coverage gap, which raises concerns those beneficiaries may face an increased risk of adverse health events. PMID- 18161395 TI - Reducing errors and risk in a prenatal network with an electronic medical record. AB - In 2002 the Institute of Medicine called for implementation of information technologies in health care settings to improve quality of care and reduce the incidence of medical errors. Nowhere is this need more critical than in obstetrics. In recent years numerous electronic prenatal medical records have become available. To date there has been little literature to identify what constitutes the important features in these systems, nor research into whether these systems actually improve the quality of care or the outcome of pregnancies. In this article we will elucidate some of the features that we feel are critical if we are to achieve these goals. Some of these features are: (1) availability across a computer network so that providers can access the record in a variety of settings, (2) simplicity of the user interface to ensure provider compliance with the system, (3) an intelligent system to encourage completeness of documentation in the medical record, (4) a problem-oriented obstetric chart so that no issue is overlooked and each is adequately addressed, and (5) administrative features to allow evaluation to ensure improved quality of care. These features together, we believe, will help to minimize medical errors, improve patient outcomes and reduce liability exposure. PMID- 18161396 TI - An interactive computer program for teaching residents pap smear classification, screening and management guidelines: a pilot study. AB - OBJECTIVE: To assess the feasibility, utility and resident attitudes towards an interactive, Web-based computer learning program. STUDY DESIGN: Pretest/ posttest evaluation of an interactive curriculum to teach current recommendations for cervical cancer screening and classification and management of abnormal Pap smears. Thirty-five Obstetrics and Gynecology (OB/ GYN) residents were encouraged to complete a Web-based computer program that consisted of a pretest, 9 educational sections (3 didactic and 6 case based), a posttest and an exit survey. Clinical cases involved virtual case management of patients' cytologic, histologic and colposcopic images. Mastery was set at 82% (or 23/28 correct answers). RESULTS: Thirty-one of the 35 residents (89%) completed the curriculum. No resident demonstrated mastery (82%) on the pretest; 15 of 31 residents demonstrated mastery on the posttest (p = 0.04). Mean improvement for all residents was 9 additional questions answered correctly. All the residents thought that the Web site was useful and covered all the learning objectives and would recommend it to other residents. CONCLUSION: A program to teach current cervical cancer screening, classification and abnormal Pap smear management guidelines was useful, feasible and well accepted by residents in this population. PMID- 18161397 TI - Diagnostic criteria for uterine smooth muscle tumors: leiomyoma variants associated with malignant behavior. AB - OBJECTIVE: To determine the prognostic accuracy of current diagnostic criteria for uterine smooth muscle tumors. STUDY DESIGN: Cases of uterine leiomyosarcoma (LMS) treated from 1976 to 1999 were analyzed retrospectively. Uterine LMS specimens were reevaluated using current criteria by a pathologist specializing in gynecologic diseases. Kaplan-Meier survival curves were evaluated. RESULTS: Specimens were available from 67 patients diagnosed with uterine LMS. On rereview, only 47 specimens were thought to represent uterine LMS. The 20 other patients were deemed to have leiomyomas or leiomyoma variants, including 13 cellular leiomyomas, 5 atypical leiomyomas and 2 leiomyomas. Median survival for patients with uterine LMS was 2.1 years. (Ninety-seven percent of disease specific deaths occurred within 6 years after the diagnosis.) With leiomyoma variants, median survival was > 25 years. Among these 18 women were 3 disease specific deaths (all > 6 years after diagnosis). CONCLUSION: Diagnostic criteria for uterine smooth muscle tumors require continued refinement. A small but significant number of patients diagnosed with leiomyoma variants will die of the disease. In contrast to the aggressive behavior of uterine LMS, disease-specific deaths attributed to leiomyoma variants occurred later. With this potential for delayed recurrence, these patients warrant close clinical surveillance. PMID- 18161398 TI - Metformin and insulin in the management of gestational diabetes mellitus: preliminary results of a comparison. AB - OBJECTIVE: To compare glycemic control and neonatal outcomes in women with gestational diabetes mellitus (GDM) treated with metformin vs. insulin. STUDY DESIGN: Women with GDM not controlled with diet and exercise were randomized to metformin (n = 32) or insulin (n = 31). The levels of glycemic control as well as maternal/neonatal complications were evaluated. RESULTS: The mean (+/- SD) fasting and 2-hour postprandial blood glucose did not differ statistically between the 2 treatment groups. No patient failed metformin and required insulin. The majority (27/32) were easily controlled on the initial dosage (500 mg twice a day). Gestational age at entry and delivery (p = 0.077, 0.412) were similar. The difference in the rate of cesarean delivery was not statistically significant between the 2 groups (p = 0.102). Neonatal statistics were also not different between the metformin and insulin groups: birth weight, Apgar score at 5 minutes, respiratory distress syndrome, hyperbilirubinemia, neonatal hypoglycemia and neonatal intensive care unit admission (p = 0.144-0.373). CONCLUSION: Based on these preliminary data, metformin appears to be an effective alternative to insulin in the treatment of GDM. PMID- 18161399 TI - Value of 3D ultrasound in the management of suspected Asherman's syndrome. AB - OBJECTIVE: To assess the value of 3-dimensional (3D) ultrasound in the management of patients with suspected Asherman's syndrome. STUDY DESIGN: A case series of 54 infertile patients who presented to a tertiary care center between 1998 and 2004 with suspected Asherman's syndrome underwent both hysterosalpingography and 3D ultrasound prior to hysteroscopy. Sensitivity and the ability to attain fertility postoperatively were calculated. RESULTS: Intrauterine adhesions (IUAs) were demonstrated on 3D ultrasound and HSG in all cases and confirmed by hysteroscopy. However, 3D ultrasound had a sensitivity of 100% and HSG a sensitivity of 66.7% for correctly grading the extent of IUAs. In 61.1% of cases in which HSG results were inconsistent with hysteroscopy, lower uterine segment outflow obstruction was present, and HSG misclassified findings as severe Asherman's with complete cavity obstruction. Postoperatively, 90% of patients conceived. CONCLUSION: 3D ultrasound provides a more accurate depiction of adhesions and extent of cavity damage than HSG in patients with suspected Asherman's syndrome, particularly when differentiating severe IUAs from lower uterine segment outflow obstruction. Therefore, grading systems utilizing HSG to classify severity of disease should be revised to include 3D ultrasound findings. PMID- 18161400 TI - Analgesic effect of etoricoxib in secondary dysmenorrhea: a randomized, double blind, crossover, controlled trial. AB - OBJECTIVE: To determine the efficacy of etoricoxib in the treatment of secondary dysmenorrhea. STUDY DESIGN: A single daily dose of etoricoxib, 120 mg, was compared with mefenamic acid (500 mg) every 8 hours and a placebo in a randomized, double-blind, crossover, controlled trial for the relief of moderate and severe pain from secondary dysmenorrhea in 22 women treated over 3 menstrual periods. Each treatment lasted up to 3 days. The first-dose analysis was based on an 8-hour observation period. RESULTS: Etoricoxib, 120 mg, provided analgesic efficacy superior to that of the placebo for the summed pain intensity difference over 8 hours (p = 0.005), and total pain relief over 8 hours (p < 0.05), and all primary and secondary end points were similar to those of mefenamic acid. No serious or unexpected adverse events were reported, and no subjects withdrew from the study because of an adverse event. However, the incidence of nausea and epigastic pain in the mefenamic acid group was significantly higher than in the etoricoxib group. CONCLUSION: A single daily dose of etoricoxib, 120 mg, was as efficacious as mefenamic acid in the management of secondary dysmenorrhea, with a lower incidence of epigastric pain, and was well tolerated for the treatment of secondary dysmenorrhea. PMID- 18161401 TI - Effects of low-dose oral contraceptives on body weight: results of a randomized study of up to 13 cycles of use. AB - OBJECTIVE: To compare the effect of 2 oral contraceptives (OCs) on body weight. STUDY DESIGN: A randomized, parallel-group, multicenter study of 1,723 women taking an OC with norgestimate (NGM) 180/215/250 microg/ethinyl estradiol (EE) 25 microg vs. 1,171 women taking on OC with norethindrone acetate 1 mg/EE 20 microg for 6-13 cycles was performed. Body weight changes between baseline and cycle 6 and baseline and cycle 13 were analyzed. Analysis included not only changes in mean body weight but also the distribution of changes that were within 5% of baseline weight, 5-10% of baseline weight and > 10% of baseline weight. Only the 10% change was felt to be clinically significant. RESULTS: The distribution of body weight changes did not statistically differ between the 2 OC groups for any parameter measured. The mean weight change after 6 months for the NGM/EE and norethindrone acetate/EE groups was +0.71 kg and +0.57 kg, respectively. At 13 cycles for the NGM/EE and norethindrone acetate/ EE groups, the mean body weight change was +0.93 kg and +0.62 kg, respectively. Only 0.3% of subjects in both OC groups experienced a 10% change in weight. CONCLUSION: Use of OCs does not substantially affect body weight for most women. PMID- 18161402 TI - Behavior of vascular endothelial growth factor and erythropoietin throughout the menstrual cycle in healthy women. AB - OBJECTIVE: To evaluate any correlations between erythropoietin (EPO) and vascular endothelial growth factor (VEGF) levels in the serum and the menstrual fluid of healthy women during the different phases of the menstrual cycle. STUDY DESIGN: Blood samples from 25 healthy female volunteers were obtained for serum VEGF and EPO detection on the 1st, 7th, 14th, 21st and 25th days of the menstrual cycle. Menstrual fluid samples for VEGF and EPO detection were obtained on the 1st and 4th days of menstruation. RESULTS: Circulating VEGF levels were found to increase in a stage-dependent cyclic manner. The mean VEGF concentration in menstrual blood on the 1st day of the cycle was significantly higher than the mean plasma value and was reduced to a significant extent on the 4th day of the cycle. We found no significant changes in serum EPO levels. Mean EPO concentration detected in menstrual blood was comparable to those in serum blood either on the 1st or 4th day of the menstrual cycle. CONCLUSION: During menstruation, a local production of VEGF occurs independent of systemic production, thus sustaining angiogenic activity in autonomous, independent ways. Our findings demonstrate the presence of an "open compartment" that reflects the systemic pattern of EPO at the uterine level that allows us to speculate on different effects beyond the angiogenic action of EPO. PMID- 18161403 TI - Patterns of care for cervical cancer patients treated in the U.S. military health care system. AB - OBJECTIVE: To determine whether women diagnosed with cervical cancer within the U.S. Department of Defense (DOD) Military Health Care System received treatment within standard guidelines and whether survival was influenced by implementation of these guidelines. STUDY DESIGN: We identified 621 women treated from 1994 to 2002. Guideline therapy was defined as hysterectomy or radiation for FIGO stage < or = IB, and hysterectomy or chemoradiation for advanced localized FIGO stage > IB. Survival analysis was performed using Cox Proportional Hazards models. RESULTS: Of those 621 women, 25% received no surgery, while 41% received radiation therapy. 6% of all patients received chemoradiation therapy prior to the Clinical Announcement received versus 26% of patients after the announcement. Variables associated with significant increased risk of death were advanced age, advanced stage and poorly differentiated lesions, p < 0.01. The lack of guideline therapy was associated with an increased risk of death, p < 0.005. Those patients who did not receive guideline therapy were twice as likely to die (HR 2.11, p = 0.005). CONCLUSION: Our study suggests that chemoradiation was rapidly introduced in the DOD care system after the 1999 National Cancer Institute's Clinical Announcement. However, some patients did not receive guideline therapy. PMID- 18161404 TI - Pregnancy outcomes in women with chronic hypertension: a population-based study. AB - OBJECTIVE: To determine the pregnancy outcomes associated with maternal chronic hypertension. STUDY DESIGN: Retrospective, population-based cohort study of maternal and infant discharge records linked to birth records in California from 1991 to 2001 were examined for demographics and pregnancy outcomes, and comparisons were made between those with and without chronic hypertension. One randomly selected pregnancy per subject was included. RESULTS: The number of women who delivered with chronic hypertension (0.69% incidence) was 29,842. As compared to non-chronic hypertensive patients, fetal and neonatal mortality and in-hospital maternal mortality were increased (ORs and 95% CIs 2.3, (2.1, 2.6); 2.3, (2.0, 2.7); and 4.8, (3.1, 7.6) respectively). Major maternal morbidity was increased: stroke, OR 5.3, (3.7, 7.5); renal failure, OR 6.0, (4.4, 8.1); pulmonary edema, OR 5.2, (3.9, 6.7); severe preeclampsia, OR 2.7, (2.5, 2.9); and placental abruption OR 2.1, (2.0, 2.3). Neonatal morbidity was increased as well: fetal growth restriction, OR 4.9, (4.7, 5.2); prematurity, OR 3.2, (3.1, 3.3); low birth weight, OR 5.4, (5.2, 5.5); very low birth weight, OR 6.5, (6.2, 6.8); and respiratory distress syndrome, OR 4.0, (3.8, 4.2). CONCLUSION: Pregnant women with chronic hypertension have significantly increased risks of maternal and perinatal morbidity and mortality. Women with this condition should be treated as high risk with appropriate maternal and fetal surveillance. PMID- 18161405 TI - Common iliac artery injury during the abdominal entry phase of gynecologic laparoscopy: a case report. AB - BACKGROUND: Although not frequent, major vascular injury (MVI) during laparoscopy in gynecologic surgery can cause a sharp increase in morbidity and mortality. CASE: During the abdominal entry stage of operative laparoscopy, perforation of the left common iliac artery occurred. Prolonged hiccups might have played a role. CONCLUSION: Entry into the abdominal cavity during laparoscopy should be performed with the safer, controlled technique. In spite of the fact that no surgical procedure or de- * vice is absolutely foolproof, the laparoscopic abdominal approach should be performed with the technique that maximally reduces the likelihood of MVI. Once a major vascular injury is recognized or suspected, immediate conversion to laparotomy should be considered. PMID- 18161406 TI - Symptomatic large bowel endometriosis in a woman with a hormonal intrauterine device: a case report. AB - BACKGROUND: Extragenital endometriosis can occur in the rectum and sigmoid causing cyclic rectal bleeding. A hormonal intrauterine device (IUD) (20 microg/24 h levonorgestrel releasing), originally developed as an easily reversible contraceptive method, is a therapeutic option for bleeding disorders. CASE: A 34-year-old woman using depot progesterone injection (crys-talline suspension of 150 mg medroxyprogesterone acetate) for contraception was amenorrheic and asymptomatic. After switching to a levonorgestrel-releasing IUD the patient experienced irregular bleeding with concomitant dysmenorrhea and rectal bleeding. Colonoscopy revealed a sigmoid mass. Laparotomy with resection of the sigmoidal mass and ovarian cyst was performed. Histopathologic analysis confirmed the suspected diagnosis of large bowel endometriosis. CONCLUSION: In our patient, large bowel endometriosis became symptomatic 2 years after insertion of hormonal IUD. The suppressive effect of the hormonal IUD seemed to be insufficient for the control of extragenital endometriosis. PMID- 18161407 TI - Prolonged vaginal and oral metronidazole for refractory Trichomonas vaginalis: a case report. AB - BACKGROUND: Resistant Trichomonas vaginalis infection is typically treated with increased doses of oral metronidazole. Because of side effects that limit compliance, such treatment may be ineffective. CASE: A young woman had an 8-year history of T vaginalis that was refractory to multiple courses of metronidazole, tinidazole and intravaginal paromomycin. After microscopic examination confirmed trichomonads, she began an extended, 14-day course of combined oral and vaginal metronidazole. Six weeks later her symptoms were unchanged, but microscopy revealed fewer trichomonads. After a 5% acetic acid vaginal wash and dilute silver nitrate bladder instillation, she was given the same treatment for 21 days. Symptoms and trichomonads resolved, and 3 years later she remained asymptomatic. CONCLUSION: Prolonged vaginal and oral metronidazole, in combination with bladder and vaginal irrigation, should be considered in women with refractory T vaginalis. PMID- 18161408 TI - Myocardial infarction in pregnancy associated with clomiphene citrate for ovulation induction: a case report. AB - BACKGROUND: Clomiphene citrate (CC) is commonly prescribed for ovulation induction. It is considered safe, with minimal side effects. Thromboembolism is a rare but life-threatening complication that has been reported after ovulation induction with CC. Spontaneous coronary thrombosis or thromboembolism with subsequent clot lysis has been suggested as one of the most common causes of myocardial infarction (MI) during pregnancy, with a subsequently normal coronary angiogram. CASE: A 33-year-old woman with a 5-week gestation had recently received CC for ovulation induction and presented with chest pain. An electrocardiogram showed a lateral and anterior wall myocardial infarction. Cardiac enzymes showed a peak rise in troponin I to 9.10 ng/mL. An initial exercise stress test was normal. At the time of admission, the patient was at high risk of radiation injury to the fetus, so a coronary angiogram was postponed until the second trimester. It showed normal coronary vessels. CONCLUSION: This appears to be the first reported case documenting a possible association between CC and myocardial infarction. Thrombosis might be a rare but hazardous complication of CC. Given this life-threatening complication, appropriate prophylactic measures should be used in high-risk woman undergoing ovarian stimulation. PMID- 18161409 TI - Chuvash-type polycythemia in pregnancy managed by repeated venisections and heparin therapy: a case report. AB - BACKGROUND: Chuvash-type polycythemia is a rare blood disease associated with an increased tendency toward thrombosis and its resulting problems, which at times can be life threatening. The main goal of treatment is to prevent thrombosis and minimize the complications of therapy. CASE: A 20-year-old, Asian woman with her first pregnancy was admitted to the hospital at 16 weeks' gestation. Her initial blood work showed a hemoglobin level of 19 g/dL, hematocrit of 55% and erythropoietin level of 21.9 IU/L (laboratory reference range, 5-25). The results of a molecular analysis of the blood specimen confirmed the diagnosis of Chuvash type polycythemia due to von Hippel-Lindau gene mutation. Weekly venisection until delivery was planned. Subsequently, the hemoglobin dropped to 15 g/dL and hematocrit to 44.6%. A 30-week scan demonstrated static growth below the third centile and oligohydramnios but normal Doppler findings. Prophylactic low molecular-weight heparin was started. The next growth scan, at 32 weeks, showed an improvement in abdominal circumference. Three days later, fetal heart monitoring showed an abnormal pattern requiring an emergency cesarean section. CONCLUSION: Early institution of heparin is beneficial in the management of Chuvash-type polycythemia in pregnancy as the condition is thrombogenic. PMID- 18161410 TI - Vulvar metastasis as the initial presentation of non-Hodgkin's lymphoma: a case report. AB - BACKGROUND: Non-Hodgkin's lymphoma constitutes 40% of malignant lymphomas. Involvement of the vulva, either primarily or secondarily, appears to be very rare. CASE: An 87-year-old woman with a 2-cm, left vulvar mass was initially treated with antibiotics due to a suspicion of Bartholin's gland abscess. The mass grew rapidly, from 2 to 10 cm, within 3 months. Biopsy showed a diffuse, large, B-cell lymphoma. After 1 cycle (5 days) of treatment with 200 mg endoxan and 60 mg prednisolone, the mass significantly decreased in size. The decrease in size occurred within 2 days after the beginning of treatment and resolved completely 7 days later. CONCLUSION: If a vulvar mass is found in an elderly woman, the physician should consider the possibility of malignancy, especially metastasis. PMID- 18161411 TI - Massive vulvar edema in a woman with preeclampsia: a case report. AB - BACKGROUND: Massive vulvar edema in a woman with preeclampsia preceded the development of massive ascites and impending eclampsia. CASE: A 17-year-old preeclamptic, primiparous woman was admitted with preeclampsia and massive vulvar edema. Other causes were excluded. The vulvar edema increased as the blood pressure and ascites increased, and a severe headache developed. Cesarean section for increasing preclampsia was performed. In the puerperium, the blood pressure improved and vulvar edema resolved. The clinical picture of the vulvar edema correlated with the severity of the preeclampsia. CONCLUSION: The presence of vulvar edema in women with preeclampsia should indicate immediate admission to the hospital. These patients must be considered as at high risk, and close monitoring must be instituted. In our case, vulvar edema preceded massive ascites development. We assume a common development mechanism for these signs in preeclampsia, due mainly to increased capillary permeability and hypoalbuminemia. The attending physician must be prepared for immediate delivery and possible preeclampsia complications in these patients. PMID- 18161412 TI - Chemical activation using an open-end coaxial applicator. AB - This article gives an overview of a novel, experimentally simple and versatile method of activation of chemical processes with microwaves without resorting to an oven. It is based on the use of a microwave antenna, namely an open-end coaxial dipole applicator immersed in an ordinary reaction vessel. Accounts of the apparatus at 2450 MHz and of the procedures adopted in reactions of organic synthesis, extraction of essential oils from plants and photo catalytic mineralization of liquid pollutants are given, discussing the necessary safety measures. The method is of practical interest for scientific and industrial applications. Thus this article is meant for a broad audience of scientists, engineers and technicians interested in new technologies for chemistry. PMID- 18161413 TI - The effect of different formulations on physical properties of cakes baked with microwave and near infrared-microwave combinations. AB - The aim of this study was to determine the effect of different formulations on color and textural characteristics of different cakes during baking in microwave and near infrared-microwave combination ovens. For comparison, cakes were also baked in conventional ovens. Color and hardness for both types of baking schemes were found to be dependent on formulation. Cakes containing Simplesse, a fat replacer consisting mostly of whey protein, baked in microwave and near infrared microwave combination ovens were found to be the firmest cakes. PMID- 18161414 TI - Carbon loaded Teflon (CLT): a power density meter for biological experiments using millimeter waves. AB - The standard technique for measurement of millimeter wave fields utilizes an open ended waveguide attached to a HP power meter. The alignment of the waveguide with the propagation (K) vector is critical to making accurate measurements. Using this technique, it is difficult and time consuming to make a detailed map of average incident power density over areas of biological interest and the spatial resolution of this instrument does not allow accurate measurements in non-uniform fields. For biological experiments, it is important to know the center field average incident power density and the distribution over the exposed area. Two 4 ft x 4 ft x 1/32 inch sheets of carbon loaded Teflon (CLT) (one 15% carbon and one 25% carbon) were procured and a series of tests to determine the usefulness of CLT in defining fields in the millimeter wavelength range was initiated. Since the CLT was to be used both in the laboratory, where the environment was well controlled, and in the field, where the environment could not be controlled, tests were made to determine effects of change in environmental conditions on ability to use CLT as a millimeter wave dosimeter. The empirical results of this study indicate CLT to be an effective dosimeter for biological experiments both in the laboratory and in the field. PMID- 18161415 TI - Comparison of conventional extraction under reflux conditions and microwave assisted extraction of oil from popcorn. AB - Popcorn offers an environmentally friendly alternative to the commercial synthetic loose-fill packing materials. Popcorn could be used for cushioning purposes if the oil is extracted after the popping process. Conventional and microwave-assisted extraction methods were used for oil extraction from whole and ground, popped and unpopped kernels. The conventional extraction method achieved 68.5% oil recovery from whole popped kernels. However, whole unpopped kernels were not efficiently de-oiled with either of the methods. Extraction of oil from popped kernels is recommended; corn varieties with higher starch content and lower oil content should be used. PMID- 18161416 TI - Review on non-thermal effects of microwave irradiation in organic synthesis. AB - The aim of this review is to show the occurrence of non-thermal effects in organic synthesis. The effect of microwave irradiation is a consequence of the interaction of radiation with matter and a combination of thermal and non-thermal effects. Thermal effects are well-described and arise from the heating rate, sometimes non accessible by conventional heating, superheating, "hot spots" and the selective absorption of radiation by polar substances. The existence of non thermal effects of the highly polarized radiation is still a controversial topic, and one usually masked by thermal effects. Separation and identification of thermal and non-thermal effects is a complex matter, but essential to the study of non-thermal effects. Some predictive models have also been described. PMID- 18161417 TI - Microwave enhanced polarization in a carbon dioxide molecule. AB - This paper presents the results of the dielectric response of carbon dioxide measured using a loaded microwave cavity operating in the TE011, mode of a cylindrical cavity near the frequencies 8.8, 9.7 and 10.2 GHz. The temperature dependence of the dielectric response of gas phase CO2 over the range of 160 to 213 degrees K (-113 to -60 degrees C) was measured. Slater perturbation equations for loaded resonant cavities were used to relate the macroscopic parameters deltaf and delta(1/Q) to the real and imaginary parts, epsilon' and epsilon", respectively, hence to calculate the dielectric parameters at each temperature and frequency. Selected peaks in the dielectric response were identified to indicate the frequencies at which strong coupling between the microwave field and the CO2 molecules can be achieved. PMID- 18161418 TI - Analysis of a modified variable-length tuning post in a rectangular waveguide. AB - A modified variable-length post for use in high power waveguide tuning networks is analyzed to determine its reactance properties. Theoretical predictions are shown to be in good agreement with experimental results. An investigation of the reactance characteristics and the aperture voltage characteristics of the modified post, as a function of its dimensions, is also reported. PMID- 18161419 TI - Permittivity of naphthenic acid-water mixture. AB - Naphthenic acid (NA) is predominantly a mono-carboxylic acid obtained as a by product of petroleum refining with variable composition and ingredients. It is reported that water affected by processes in the petroleum industries generally contains 40-120 mg IL of naphthenic acid which is considered to be in the range of toxicity to human consumption [Clemente et. al, 2005; McMartin, 2003]. This contaminated water needs treatment before its use as drinking water by remote communities. Recent literature suggests that NAs could be separated from diesel fuel using microwave radiation [Lingzhao et. al, 2004]. Removal of naphthenic acid from vacuum cut #1 distillate oil of Daqing using microwaves has also been reported by Huang et. al [2006]. The microwave treatment can be applied to drinking water containing small concentrations of naphthenic acid. In this case permittivity information is useful in designing a microwave applicator and modeling studies. Permittivity measurements were done using a HP 8510 Vector Network Analyzer and coaxial probe reflection method to study the dielectric properties of naphthenic acid in water. The effects of process variables such as frequency, concentration and temperature on dielectric properties were determined. PMID- 18161420 TI - Dielectric properties of human colostrum at microwave frequencies. AB - This article communicates the study of both the dielectric properties of human colostrums and breast milk at microwave frequencies. The colostrum samples were taken immediately after child birth and breast milk samples were collected at weekly intervals following the delivery. Rectangular cavity perturbation technique is used for the measurements of dielectric properties at the S-band of microwave frequency. The dielectric constants of the colostrums samples and breast milk samples are found to increase as weeks elapse, which is attributed to the reduced fat content and increased lactose concentration. The conductivity of these samples is similarly found to increase due to the increased dilution. PMID- 18161421 TI - Microwave vacuum dryer setup and preliminary drying studies on strawberries and carrots. AB - A laboratory scale microwave vacuum dryer with the ability to record temporal variation of mass and temperature of a drying product was designed and built. The initial study was set up to investigate the effect of the position of a vacuum pressure control valve at two vacuum pressure levels, 6.5 and 13.3 kPa, with a fixed microwave power input of 1.5 W/g. Then, strawberry halves and carrot cubes (10 x 10 x 10 mm) were used for a preliminary study to investigate the effect on drying product temperature and the effect of input microwave powers (1, 1.5 and 2 W/g) at a fixed level of vacuum pressure (6.5 kPa). The position of the valve which allows air to pass through the vacuum container was found to provide shorter drying time and reduced the occurrence of water vapor condensation. The product temperature at the end stage of drying under continuous microwave mode was too high to ensure quality for both dried strawberry halves and carrot cubes. PMID- 18161422 TI - Dielectric properties of honeydew melons and correlation with quality. AB - Dielectric properties of three honeydew melon cultivars, grown and harvested to provide a range of maturities, were measured with an open-ended coaxial-line probe and impedance analyzer over the frequency range from 10 MHz to 1.8 GHz. Probe measurements were made on the external surface of the melons and also on tissue samples from the edible internal tissue. Moisture content and soluble solids content (SSC) were measured for internal tissue samples, and SSC (sweetness) was used as the quality factor for correlation with the dielectric properties. Individual dielectric constant and loss factor correlations with SSC were low, but a high correlation was obtained between the SSC and permittivity from a complex-plane plot of dielectric constant and loss factor, each divided by SSC, for both the external surface and internal tissue measurements. However, SSC prediction from the dielectric properties by these relationships was not as high as expected. Permittivity data (dielectric constant and loss factor) for the melons are presented graphically to show their relationships with frequency for external surface and internal tissue measurements. A dielectric relaxation for the external surface measurements, which is attributable to bound water and Maxwell-Wagner relaxations, is also illustrated. Coefficients of determination for complex-plane plots, moisture content and SSC relationship, and penetration depth are also shown graphically. Further studies are needed for determining the practicality of sensing melon quality from the dielectric properties. PMID- 18161423 TI - Drying and color characteristics of coriander foliage using convective thin-layer and microwave drying. AB - Heat sensitive properties (aromatic, medicinal, color) provide herbs and spices with their high market value. In order to prevent extreme loss of heat sensitive properties when drying herbs, they are normally dried at low temperatures for longer periods of time to preserve these sensory properties. High energy consumption often results from drying herbs over a long period. Coriander (Coriandrum sativum L., Umbelliferae) was dehydrated in two different drying units (thin layer convection and microwave dryers) in order to compare the drying and final product quality (color) characteristics. Microwave drying of the coriander foliage was faster than convective drying. The entire drying process took place in the falling rate period for both microwave and convective dried samples. The drying rate for the microwave dried samples ranged from 42.3 to 48.2% db/min and that of the convective dried samples ranged from 7.1 to 12.5% db/min. The fresh sample color had the lowest L value at 26.83 with higher L values for all dried samples. The results show that convective thin layer dried coriander samples exhibited a significantly greater color change than microwave dried coriander samples. The color change index values for the microwave dried samples ranged from 2.67 to 3.27 and that of the convective dried samples varied from 4.59 to 6.58. PMID- 18161424 TI - Medicine off the battlefield. VA hospitals, community-based clinics meet special challenges in trying to provide care to veterans living in rural areas. AB - The VA faces special challenges in providing care for veterans in remote areas, who often live long distances from its medical centers. A network of community based clinics and contracts with local hospitals help fill the gap. But the VA should also be contracting with other providers to broaden the scope of its services, says Hilda Heady, left, of the West Virginia Rural Education Partnerships. PMID- 18161425 TI - Investors staying alert. UHS and CHS provide updates on federal probes. PMID- 18161426 TI - Hospitals' code blues. MS-DRG switch may initially delay pay. PMID- 18161427 TI - When we're 65 ... Few are looking ahead at how to care for a coming horde of new seniors. PMID- 18161428 TI - By the numbers. Most profitable critical-access hospitals ranked by operating margin, based on 2005 Medicare cost report data. PMID- 18161429 TI - Not a happy ending. Stand-alone N.J. hospital likely to close doors after affiliation talks fail. PMID- 18161430 TI - Improve reimbursement: MGMA. Group urges members to voice concerns to Congress. PMID- 18161431 TI - [Therapy without success. Every 3d powder inhaler is used improperly (interview by Maria Weiss)]. PMID- 18161432 TI - [Depressed patients in family practice. Take the positive approach!]. PMID- 18161433 TI - [Medical care in retirement and nursing homes. How bad is the situation in reality?]. PMID- 18161434 TI - [Patient care in nursing homes by the family physician]. AB - When treating the acute and chronic diseases of nursing home residents, the family physician must consider the patient's decreased ability to make decisions and shortened life expectancy in each case. Nurses tend the elderly people, help them wherever it is necessary, have time to listen to the hardships and worries of their charges and are an irreplaceable help to the family physician for observing patients and assessing the measures taken based on these observations. PMID- 18161435 TI - [Control of symptoms during the final stage of the patient's life]. AB - Nursing homes have taken on more and more the character of a hospice where palliative competence has significantly become more important. The aim of palliative treatment is not to heal but to control the symptoms during the final stage. The treatment of pain is definitely the main priority. Diagnosing and estimating the severity of the pain are basic requirements of an effective analgesic treatment. In addition to the pain, there are other symptoms such as nausea and emesis, dyspnoea, death rattle,fear, delirium and obstipation in the final stages of a disease which should be controlled very carefully. Not only patients suffering from cancer, but also very sick multimorbide patients and those suffering from dementia in geriatric environments will profit from palliative care. PMID- 18161436 TI - [Hypertension, CHD and heart failure are the sequelae. Sleep apnoea. Premature death is to be expected]. PMID- 18161437 TI - [Effect of weight loss on blood pressure]. AB - The treatment of arterial hypertension is based on changes in lifestyle and above all weight loss. The expected reduction of systolic and diastolic blood pressure through weight loss is greater the higher the starting blood pressure. For patients who are not successful through calorie reduction and sport, weight loss can be enhanced with pharmacotherapy. Various substance-dependent blood pressure reactions have been observed that must be monitored. The blood pressure of patients who have undergone surgical intervention for weight loss decreases significantly to the upper normal range. PMID- 18161438 TI - [Emergency checklist: biceps tendon rupture]. PMID- 18161439 TI - [New study demonstrates dual effect of an oxycodone/naloxone combination. Strong pain palliation and preserving intestinal function]. PMID- 18161440 TI - [Chickenpox misjudged for a long time. No real pustules, but still very dangerous ]. PMID- 18161441 TI - Attention to detail helps Clarian measure ROI. AB - Clarian Health takes a risk and embarks on a branding strategy. Taking a multi integrated approach, with The Heavy Weights in Indianapolis, Clarian started managing its marketing by tracking return on investment (ROI) and making the most of a CRM database. In a groundbreaking approach, it found that it was actually able to turn its marketing effort into a money-maker, leading higher-ups to see its outreach efforts as necessity. PMID- 18161442 TI - Halloween bash brings more treats than tricks. AB - Advocate Hope Children's Hospital in Oak Lawn, IL, chose to host an event in the spirit of fall and Halloween. More than 300 guests attended the annual Hearts for Hope Halloween bash, a costume gala that is held each year to raise funds. This year, the fund-raiser benefited the construction of a Ronald McDonald House near the facility. PMID- 18161443 TI - Fund-raising aficionados: measuring out a recipe for success. AB - A five-year capital campaign comes to a close with positive results and lessons learned. Having held dozens of annual fund-raising events throughout the year, Children's Healthcare of Atlanta has learned how to put on an effective event with full community support. PMID- 18161444 TI - Going green: Metro Health starts health commune. AB - Taking on a total rebranding and environmentally friendly overhaul, Metro Health looks forward as it moves into a new facility. Incorporating green building details, Metro is managing environmental impact to improve its future in its Grand Rapids, MI, community. PMID- 18161445 TI - Darling surpasses expectations with a 4 per cent funding boost. PMID- 18161446 TI - Infection likely cause of 90 deaths. PMID- 18161447 TI - Diagnostics firm found inadequate. PMID- 18161448 TI - Workforce. Managers stuck in the middle feel pressure from all sides. PMID- 18161449 TI - Simon Stevens on hidden shallows. PMID- 18161450 TI - Mental health. Down should not mean out of a job in the NHS. PMID- 18161451 TI - Data briefing. New era of mental healthcare insights. PMID- 18161452 TI - Patient involvement. How does it feel to be our patient? PMID- 18161453 TI - More than half of trusts languish in bottom performance grades. PMID- 18161454 TI - Trusts miss hygiene goals. PMID- 18161455 TI - PCTs score worst on 'things that matter'. PMID- 18161456 TI - It's good but is it good enough? Darling's NHS spend laid bare. PMID- 18161457 TI - Sophia Christie on collective commissioning. PMID- 18161458 TI - Recruitment. No room for doctors who raised capacity. PMID- 18161459 TI - Data briefing. Four-hour target fuels admissions. PMID- 18161460 TI - Migration. Foreign affairs. AB - Many in the NHS are unwilling to discuss migrant matters, possibly for fear of being thought racist--or because they do not have accurate data. The allocation system to PCTs may be inadequate in areas where there is rapid population growth. Differences in language, health needs and lifestyle between ethnic groups are challenging the NHS. PMID- 18161461 TI - Commissioning. Four pillars for the future. PMID- 18161462 TI - Embarrassment for the DoH as NHS underspends by 1.8bn pounds sterling. PMID- 18161463 TI - DoH team to be broken up and procurement localised. PMID- 18161464 TI - Older people. Is this the answer to the age-old question of social care funding? PMID- 18161465 TI - Sophia Christie on deja vu. PMID- 18161466 TI - Geriatric medicine. 'How do you stand working with all these old people?'. PMID- 18161467 TI - Barometer. Acute trusts November 2007. PMID- 18161468 TI - Equal access. New generation of perinatal care. PMID- 18161469 TI - Banks threaten to hit FTs with rate hike over debt guarantees. PMID- 18161470 TI - Progress at outbreak trust but doctor hygiene rapped. PMID- 18161471 TI - More staff may get right to parent-friendly hours. PMID- 18161472 TI - Private finance initiative. PFI debt fears as district generals feel the squeeze. PMID- 18161473 TI - Malcolm Lowe-Lauri on new leaders. PMID- 18161474 TI - Reform. Can a constitution seal the service's freedom? PMID- 18161475 TI - Barometer. Mental health November 2007. PMID- 18161476 TI - Long-term conditions. Help patients to help themselves. PMID- 18161477 TI - The evolution of the relaxin peptide family and their receptors. AB - The relaxin peptide family in humans consists of relaxin-1, 2 and 3 and the insulin-like peptides (INSL)-3, 4, 5 and 6. The evolution of this family has been controversial; points of contention include the existence of an invertebrate relaxin and the absence of a ruminant relaxin. Over the past four years we have performed a comprehensive analysis of the relaxin peptide family using all available vertebrate and invertebrate genomes. Contrary to previous reports an invertebrate relaxin was not found; sequence similarity searches indicate the family emerged during early vertebrate evolution. Phylogenetic analyses revealed the presence ofpotential relaxin-3, relaxin and INSL5 homologs in fish; dating their emergence far earlier than previously believed. There are four known relaxin peptide family receptors; the relaxin and INSL3 receptors, the leucine rich repeat containing G protein-coupled receptors (LGR), LGR7 and LGR8 respectively; and the two relaxin-3 receptors, GPCR135 and GPCR142. Database searching identified several invertebrate ancestors of LGR7 and LGR8; the absence of an invertebrate relaxin suggests the presence of an unidentified invertebrate ligand for these receptors. No invertebrate ancestors of GPCR135 or GPCR142 were found. Based on the theory that interacting proteins co-evolve together, phylogenetic analyses of the relaxin peptide family receptors were performed to provide insight into interactions within the relaxin system. Co-evolution between INSL5 and GPCR142, as evidenced by the loss of both genes in the rat and dog and their similar expression profiles, predicted GPCR142 to be the endogeneous INSL5 receptor. This interaction has since been confirmed experimentally. The emergence and presence of multiple GPCR135 homologs in fish reflected similar findings for relaxin-3. It seems likely the ancestral relaxin system was relaxin-3 acting through GPCR135, before LGR7 was "acquired" as a relaxin receptor early in vertebrate development. PMID- 18161478 TI - Relaxin, the relaxin-like factor and their receptors. AB - In 1926 Frederick Hisaw discovered a blood-borne factor in pregnant guinea pigs that would cause relaxation of the pubic symphysis in virgin females of the species. The relaxin-like factor gene (RLF), also known as insulin-like 3 (INSL3), was recovered from a library of testicular cDNA. The function of RLF as the mediator of testicular positioning in mice was discovered by gene deletion experiments. The report that deletion of a G-protein-coupled receptor in a mouse mutant caused cryptorchidism and that relaxin and RLF and their receptors were structurally and functionally similar may well have inspired Drs. Hsueh and Sherwood to put LGR7 and relaxin together and thus, after many agonizing years of uncertainty, the relaxin receptor had yielded its identity. LGR8 was recognized as the human version of the RLF receptor and together LGR7 and LGR8, with their respective ligands, opened to detailed investigation the large and important field of G-protein activated leucine-rich repeat receptors. In the process RLF and LGR8 have yielded some general information that might contribute to our knowledge of receptor/ligand interaction, in particular the enigmatic signal initiation process. PMID- 18161479 TI - Diverse signalling mechanisms used by relaxin in natural cells and tissues: the evolution of a "neohormone". AB - The small peptide hormone relaxin is a member of a rapidly evolving family of hormones and growth factors, whose mode of action appears to be particularly adapted to purely mammalian physiology. It is representative of a new category of hormones, referred to as neohormones, which appear to have evolved specifically to accommodate the needs of viviparity, lactation and wound repair. The mechanism of receptor signalling has also evolved in this family, with older members using receptor tyrosine kinases and new members such as relaxin adopting 7 transmembrane G-protein coupled receptors. Although relaxin primarily generates cAMP as second messenger, studies of relaxin signalling show that this does not conform to a classic G-protein dependent activation of adenylate cyclase: it requires additional cytoplasmic components, it can involve further coupling to PI3-kinase and PKCzeta and it is absolutely dependent on a tyrosine kinase activity linked closely to the relaxin receptor. Relaxin may also independently activate glucocorticoid receptors. This diversity of signalling leads to a broad range of possible downstream transcriptional effects. Finally, in tissues where relaxin is known to be effective, there is often also local relaxin induction, amplifying the effects of the endocrine hormone. PMID- 18161480 TI - Relaxin physiology in the female reproductive tract during pregnancy. AB - The characteristic functions of relaxin are associated with female reproductive tract physiology. These include the regulation of biochemical processes involved in remodeling the extracellular matrix of the cervix and vagina during pregnancy and rupture of the fetal membranes at term. Such modifications enable the young to move unimpeded through the birth canal and prevent dystocia. However, relaxin's physiological actions are not limited to late gestation. New functions for this peptide hormone in implantation and placentation are also emerging. Relaxin promotes uterine and placental growth and influences vascular development and proliferation in the endometrium. This chapter provides an overview of the current literature on relaxin physiology in the uterus, cervix and vagina of pregnant females and the impact on fetal health. It also outlines the potential mechanisms of relaxin action, particularly in the cervical extracellular matrix and uterine endometrium. PMID- 18161481 TI - Relaxin and related peptides in male reproduction. AB - The relaxin hormone is renowned for its function in pregnancy, parturition and other aspects of female reproduction. At the same time, the role of relaxin in male reproduction is still debated. Relaxin is prominently expressed in prostate and its receptors are found in several male reproductive organs; however, the data indicative of its contribution to differentiation and functioning of prostate or testis are contradictory. Prostate relaxin is a main source of this peptide in the seminal plasma. The relaxin effects on sperm motility and fertilization have been reported. The expression of other relaxin related peptides, such as INSL5 and INSL6 was described in testis; yet, currently there are no experimental data to pinpoint their biological functions. The other member of relaxin peptide family, insulin-like 3 peptide (INSL3), is a major player in male development. The INSL3 peptide is expressed in testicular fetal and adult Leydig cells and is directly responsible for the process of abdominal testicular descent (migration of the testes towards the scrotum during male development). Genetic targeting of the Insl3 gene or INSL3 GPCR receptor Lgr8/Rxfp2 causes high intra-abdominal cryptorchidism due to a differentiation failure of testicular ligaments, the gubernacula. Several mutations of these two genes rendering nonfunctional proteins have been described in human patients with testicular maldescent. Thus, in this chapter we review the data related to the expression and function of relaxin and related peptides in male reproduction. PMID- 18161482 TI - The vascular actions of relaxin. AB - Relaxin is emerging as a hormone with important vascular actions. Much of our recently gained knowledge of relaxin in this context has stemmed from investigations of maternal vascular adaptations to pregnancy in which the hormone is turning out to be an important mediator. This chapter is separated into three parts. In Part 1, we discuss relaxin in the setting of normal vascular function and focus on systemic hemodynamics and arterial mechanical properties, renal and other peripheral circulations, angiogenesis, as well as the cellular mechanisms of the vasodilatory actions of relaxin. In this section, we also summarize the evidence for an arterial-derived relaxin ligand-receptor system. In Part 2, we present relaxin in the context of vascular dysfunction and the implications for relaxin as a therapeutic agent in renal and cardiac diseases, ischemia and reperfusion injury, pulmonary hypertension, vascular inflammation and preeclampsia. Finally, in Part 3, we highlight some of the controversies and unresolved issues, as well as suggest a general direction for future relaxin research that is urgently needed. PMID- 18161483 TI - The effects of relaxin on extracellular matrix remodeling in health and fibrotic disease. AB - Since its discovery as a reproductive hormone 80 years ago, relaxin has been implicated in a number of pregnancy-related functions involving extracellular matrix (ECM) turnover and collagen degradation. It is now becoming evident that relaxin's ability to reduce matrix synthesis and increase ECM degradation has important implications in several nonreproductive organs, including the heart, lung, kidney, liver and skin. The identification of relaxin and RXFP1 (Relaxin family peptide receptor-1) mRNA and/or binding sites in cells or vessels of these nonreproductive tissues, has confirmed them as targets for relaxin binding and activity. Recent studies on Rln1 and Rxfp1 gene-knockout mice have established relaxin as an important naturally occurring and protective moderator of collagen turnover, leading to improved organ structure and function. Furthermore, through its ability to regulate the ECM and in particular, collagen at multiple levels, relaxin has emerged as a potent anti-fibrotic therapy, with rapid-occurring efficacy. It not only prevents fibrogenesis, but also reduces established scarring (fibrosis), which is a leading cause of organ failure and affects several tissues regardless of etiology. This chapter will summarize these coherent findings as a means of highlighting the significance and therapeutic potential of relaxin. PMID- 18161484 TI - Relaxin-like ligand-receptor systems are autocrine/paracrine effectors in tumor cells and modulate cancer progression and tissue invasiveness. AB - Relaxin and INSL3 are novel autocrine/paracrine insulin-like hormones in tumor biology. Both effectors can bind to and activate the leucine-rich G-protein coupled receptors LGR7 relaxin receptor) or LGR8 (relaxin/INSL3 receptor). These relaxin-like ligand-receptor systems modulate cellular functions and activate signaling cascades in a tumor-specific context leading to changes in tumor cell proliferation, altered motility/migration and enhanced production/secretion ofpotent proteolytic enzymes. Matrix-metalloproteinases (MMP), tissue inhibitors of metalloproteinases (TIMP) and acid hydrolases such as cathepsins can facilitate tissue degradation and represent important proteolytic mediators of relaxin-like actions on tumor cell invasion and metastasis. This review presents recent new findings and emphasises the important functions of the relaxin/INSL3 ligand-receptor system as novel autocrine/paracrine effectors influencing tumor progression and tissue invasiveness. PMID- 18161485 TI - Relaxin-family peptide and receptor systems in brain: insights from recent anatomical and functional studies. AB - Relaxin was for many years considered primarily a hormone active within the reproductive tract with overwhelming evidence for its important roles in mammalian parturition. More recent research, however, has clearly indicated additional physiological and/or therapeutic roles for relaxin in the cardiovascular, renal and respiratory systems (see other Chapters); while a few studies have also described possible physiological effects of relaxin in the central nervous system, perhaps unsurprisingly associated with the regulation of osmotic homeostasis, blood pressure and neurohormone secretion during pregnancy and parturition. Research on relaxin and subsequently discovered, related peptides has also been particularly productive in the last five years, with some milestone discoveries (see elsewhere in this volume), including the long-awaited identification of the native receptors for relaxin and a related peptide, INSL3- the leucine-rich repeat-containing G-protein-coupled receptors-7 and -8 (LGR7/8); and the identification of a new relaxin family peptide, known as relaxin 3 and its type I G-protein-coupled receptor--GPCR135. Relaxin 3 was subsequently found to be highly conserved throughout evolution and to be the likely ancestral gene/peptide that gave rise to the current relaxin family of genes and peptides in mammals including higher primates. Interestingly, relaxin 3 and its receptor are found in highest abundance in brain, suggesting important central functions for relaxin 3/GPCR135 signaling. In this Chapter we will primarily review what is currently known about the central distribution of relaxin family peptides and their receptors and what has been described so far regarding their effects in the brain. Lastly, we will discuss likely future directions in this interesting, expanding field of research. PMID- 18161486 TI - Nuclear transformation of eukaryotic microalgae: historical overview, achievements and problems. AB - Transformation of microalgae is a first step in their use for biotechnological applications involving foreign protein production or molecular modifications of specific cell metabolic pathways. Since the first reliable achievements of nuclear transformation in Chlamydomonas, other eukaryotic microalgae have become transformed with molecular markers that allow a direct selection. Different methods--glass beads, electroporation, particle bombardment, or Agrobacterium- and constructions have been set up in several organisms and successfully used. However, some problems associated with efficiency, integration, or stability of the transgenes still persist and are analysed herein. Though the number of microalgae species successfully transformed is not very high, prospects for transformation of many more are good enough on the basis of what has been achieved so far. PMID- 18161487 TI - Transformation of cyanobacteria. AB - Cyanobacteria are a diverse and successful group of bacteria defined by their ability to carry out oxygenic photosynthesis. They occupy diverse ecological niches and are important primary producers in the oceans. Cyanobacteria are amenable to genetic manipulation. Some strains are naturally transformable. Many others have been transformed in the lab by conjugation or electroporation. The ability to transform cyanobacteria has been determinant in the development of the molecular biology of these organisms and has been the basis of many of their biotechnological applications. Cyanobacteria are the source of natural products and toxins of potential use and can be engineered to synthesize substances of biotechnological interest. Their high protein and vitamin content makes them useful as a dietary supplement. Because of their ability to occupy diverse ecological niches, they can be used to deliver to the medium substances of interest or as biosensors. PMID- 18161488 TI - Molecular biology and the biotechnological potential of diatoms. AB - Diatoms are unicellular photoautotrophic eukaryotes that play an important role in ecology by fixing large amounts of CO2 in the oceans. Because they evolved by secondary endocytobiosis--a process of uptake of a eukaryotic alga into another eukaryotic cell--they have a rather unusual cell biology and genetic constitution. Diatoms are also of biotechnological interest since they produce highly unsaturated fatty acids. In addition they are able to form delicately ornate cell walls made of amorphous silica. Understanding and modifying the processes of biomineralization in diatoms might result in new nanotechnological processes. Therefore recent advances in molecular genomics and the development of genetic tools for diatoms might pave the way for biotechnological modification and utilization of diatoms. In this chapter I will briefly characterize these extraordinary organisms, give some insights into the actual advances in molecular biology of diatoms and present some examples for the potential future use of diatoms in algal biotechnology. PMID- 18161489 TI - Tools and techniques for chloroplast transformation of Chlamydomonas. AB - The chloroplast organelle of plant and algal cells contains its own genetic system with a genome of a hundred or so genes. Stable transformation of the chloroplast was first achieved in 1988, using the newly developed biolistic method of DNA delivery to introduce cloned DNA into the genome of the green unicellular alga Chlamydomonas reinhardtii. Since that time there have been significant developments in chloroplast genetic engineering using this versatile organism, and it is probable that the next few years will see increasing interest in commercial applications whereby high-value therapeutic proteins and other recombinant products are synthesized in the Chlamydomonas chloroplast. In this chapter I review the basic methodology of chloroplast transformation, the current techniques and applications, and the future possibilities for using the Chlamydomonas chloroplast as a green organelle factory. PMID- 18161490 TI - Influence of codon bias on the expression of foreign genes in microalgae. AB - The expression of functional proteins in heterologous hosts is a core technique of modern biotechnology. The transfer to a suitable expression system is not always achieved easily because of several reasons: genes from different origins might contain codons that are rarely used in the desired host or even bear noncanonical codons, or the genes might hide expression-limiting regulatory elements within their coding sequence. These problems can also be observed when introducing foreign genes into genomes of microalgae as described in a growing number of detailed studies on transgene expression in these organisms. Particularly important for the use of algae as photosynthetic cell factories is a fundamental understanding of the influence of a foreign gene's codon composition on its expression efficiency. Therefore, the effect of codon usage of a chimeric protein on expression frequency and product accumulation in the green alga Chlamydomonas reinhardtii was analyzed. This fusion protein combines a constant region encoding the zeocin binding protein Ble with two different gene variants for the green fluorescent protein (GFP). It is shown that codon bias significantly affects the expression, but barely influences the final protein accumulation in this case. PMID- 18161491 TI - In the grip of algal genomics. AB - Algae are dominant primary producers on the Earth and have a major impact on global productivity and biogeochemical cycling. There are still few algal genomes that have been completely characterized, and resources directed toward algal genomic sequencing are limited. However, it is also becoming evident that algae and prokaryotic picoplankton have a critical role in the fixation and sequestration of carbon, and so the interest in algal genomics is expanding. There are some algae for which full or near-full genome sequences have been secured; these genomes include those of the red alga Cyanidioschyzon merolae, the green algae or chlorophytes Chlamydomonas reinhardtii and Volvox carteri, the marine picoeukaryote Ostreococcus tauri (two different strains of O. tauri have been sequenced), the diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum, and the haptophyte Emiliania huxleyi. There is also a full sequence for the vestigal 'red' algal genome of the nucleomorph of the Cyptomonad Guillardia theta. In addition, numerous genomes of photosynthetic microbes, including marine Synechococcus and Prochlorococcus species have been sequenced. There have also been projects developed to define algal transcriptomes as determined by cDNA analysis, full genome sequences of numerous plastids, and the genomes of a variety of viruses that infect marine and freshwater algae. The recent efforts focused on acquiring and analyzing algal genome sequences have generated an influx of exciting data to a field that is in its infancy. In this review I discuss potential criteria for determining which organisms should be targeted for genome projects, successful forays into algal genomic sequencing, and some of the inferences generated from the analysis of the sequence information. PMID- 18161492 TI - Insertional mutagenesis as a tool to study genes/functions in Chlamydomonas. AB - The unicellular alga Chlamydomonas reinhardtii has emerged during the last decades as a model system to understand gene functions, many of them shared by bacteria, fungi, plants, animals and humans. A powerful resource for the research community is the availability of complete collections of stable mutants for studying whole genome function. In the meantime other strategies might be developed; insertional mutagenesis has become currently the best strategy to disrupt and tag nuclear genes in Chlamydomonas allowing forward and reverse genetic approaches. Here, we outline the mutagenesis technique stressing the idea of generating databases for ordered mutant libraries, and also of improving efficient methods for reverse genetics to identify mutants defective in a particular gene. PMID- 18161493 TI - Optimization of recombinant protein expression in the chloroplasts of green algae. AB - Through advances in molecular and genetic techniques, protein expression in the chloroplasts of green algae has been optimized for high-level expression. Recombinant proteins expressed in algae have the potential to provide novel and safe treatment of disease and infection where current, high-cost drugs are the only option, or worse, where therapeutic drugs are not available due to their prohibitively high-cost to manufacture. Optimization of recombinant protein expression in Chlamydomonas reinhardtii chloroplasts has been accomplished by employing chloroplast codon bias and combinatorial examination of promoter and UTR combinations. In addition, as displayed by the expression of an anti-herpes antibody, the C. reinhardtii chloroplast is capable of correctly folding and assembling complex mammalian proteins. These data establish algal chloroplasts as a system for the production of complex human therapeutic proteins in soluble and active form, and at significantly reduced time and cost compared to existing production systems. Production of recombinant proteins in algal chloroplasts may enable further development of safe, efficacious and cost-effective protein therapeutics. PMID- 18161494 TI - Phycoremediation of heavy metals using transgenic microalgae. AB - Microalgae account for most of the biologically sequestered trace metals in aquatic environments. Their ability to adsorb and metabolize trace metals is associated with their large surface:volume ratios, the presence of high-affinity, metal-binding groups on their cell surfaces, and efficient metal uptake and storage systems. Microalgae may bind up to 10% of their biomass as metals. In addition to essential trace metals required for metabolism, microalgae can efficiently sequester toxic heavy metals. Toxic heavy metals often compete with essential trace metals for binding to and uptake into cells. Recently, transgenic approaches have been developed to further enhance the heavy metal specificity and binding capacity of microalgae with the objective of using these microalgae for the treatment of heavy metal contaminated wastewaters and sediments. These transgenic strategies have included the over expression of enzymes whose metabolic products ameliorate the effects of heavy metal-induced stress, and the expression of high-affinity, heavy metal binding proteins on the surface and in the cytoplasm of transgenic cells. The most effective strategies have substantially reduced the toxicity of heavy metals allowing transgenic cells to grow at wild-type rates in the presence of lethal concentrations of heavy metals. In addition, the metal binding capacity of transgenic algae has been increased five-fold relative to wild-type cells. Recently, fluorescent heavy metal biosensors have been developed for expression in transgenic Chlamydomonas. These fluorescent biosensor strains can be used for the detection and quantification of bioavailable heavy metals in aquatic environments. The use of transgenic microalgae to monitor and remediate heavy metals in aquatic environments is not without risk, however. Strategies to prevent the release of live microalgae having enhanced metal binding properties are described. PMID- 18161495 TI - Hydrogen fuel production by transgenic microalgae. AB - This chapter summarizes the state-of-art in the field of green algal H2 production and examines physiological and genetic engineering approaches by which to improve the hydrogen metabolism characteristics of these microalgae. Included in this chapter are emerging topics pertaining to the application of sulfur nutrient deprivation to attenuate O2-evolution and to promote H2-production, as well as the genetic engineering of sulfate uptake through manipulation of a newly reported sulfate permease in the chloroplast of the model green alga Chlamydomonas reinhardtii. Application of the green algal hydrogenase assembly genes is examined in efforts to confer H2-production capacity to other commercially significant unicellular green algae. Engineering a solution to the O2 sensitivity of the green algal hydrogenase is discussed as an alternative approach to sulfur nutrient deprivation, along with starch accumulation in microalgae for enhanced H2-production. Lastly, current efforts aiming to optimize light utilization in transgenic microalgae for enhanced H2-production under mass culture conditions are presented. It is evident that application of genetic engineering technologies and the use of transgenic green algae will improve prospects for commercial exploitation of these photosynthetic micro-organisms in the generation of H2, a clean and renewable fuel. PMID- 18161496 TI - Microalgal vaccines. AB - A variety of recombinant vaccines and vaccine delivery systems are currently under development as alternatives to vaccines produced in animals that are primarily administered by injections. These nonanimal alternatives do not transmit animal pathogens, are often rapid to develop, and can be produced on a large scale at low costs. Many of these new vaccine technologies are based on oral delivery systems and avoid the risks of disease transmission associated with the use of syringes for injectable vaccines. In addition, many of these novel systems have extended shelf life, often not requiring refrigeration and thus are applicable in developing countries or remote locations. Here we describe the development of microalgal-based immunization systems. Antigens expressed in the chloroplast or anchored to the surface of plasma membrane are shown to effectively immunize fish and rabbits. The effective oral delivery of antigens by microalgae provides a safe and inexpensive mechanism to immunize animals. The applications of microalgal vaccines are currently being investigated. PMID- 18161497 TI - What is the role of motor simulation in action and object recognition? Evidence from apraxia. AB - An important issue in contemporary cognitive neuroscience concerns the role of motor production processes in perceptual and conceptual analysis. To address this issue, we studied the performance of a large group of unilateral stroke patients across a range of tasks using the same set of common manipulable objects. All patients (n = 37) were tested for their ability to demonstrate the use of the objects, recognize the objects, recognize the corresponding object-associated pantomimes, and imitate those same pantomimes. At the group level we observed reliable correlations between object use and pantomime recognition, object use and object recognition, and pantomime imitation and pantomime recognition. At the single-case level, we document that the ability to recognize actions and objects dissociates from the ability to use those same objects. These data are problematic for the hypothesis that motor processes are constitutively involved in the recognition of actions and objects and frame new questions about the inferences that are merited by recent findings in cognitive neuroscience. PMID- 18161498 TI - Foveal crowding in posterior cortical atrophy: a specific early-visual-processing deficit affecting word reading. AB - Visual crowding is a form of masking in which single-letter identification is compromised by the presence of additional letters or other simple visual forms in close proximity. This behavioural phenomenon has been studied most frequently in the context of amblyopic and normal peripheral vision. In the current study, we investigate this phenomenon in the context of two patients with peripheral dyslexia and a third with visual disorientation consequent to bilateral posterior cortical atrophy. In one case, reading showed the effects of word length typical of letter-by-letter reading, whereas the second case was unable to read any whole words. In a series of letter identification tasks, recognition accuracy was shown to decrease significantly in the presence of a range of flanking stimuli (e.g., letters, digits, letter fragments). Compatible with previous reports of the crowding phenomenon, the flanking effect was strengthened by increasing flanker proximity but was unaffected by target or flank size, flank contrast, target flank lexicality, or flank category. One patient also showed amelioration of the flanking effect when the target and flankers were of opposite contrast polarity. To the best of our knowledge, this is the first demonstration of visual crowding in individuals with posterior cortical atrophy. We consider the relevance of these empirical findings to accounts of the letter-by-letter reading form of peripheral dyslexia. In particular, we suggest that crowding constitutes one specific form of early-visual-processing deficit, which impairs the reading process. PMID- 18161499 TI - General intelligence and memory span: evidence for a common neuroanatomic framework. AB - General intelligence (g) is highly correlated with working-memory capacity (WMC). It has been argued that these central psychological constructs should share common neural systems. The present study examines this hypothesis using structural magnetic resonance imaging to determine any overlap in brain areas where regional grey matter volumes are correlated to measures of general intelligence and to memory span. In normal volunteers (N = 48) the results (p < .05, corrected for multiple comparisons) indicate that a common anatomic framework for these constructs implicates mainly frontal grey matter regions belonging to Brodmann area (BA) 10 (right superior frontal gyrus and left middle frontal gyrus) and, to a lesser degree, the right inferior parietal lobule (BA 40). These findings support the nuclear role of a discrete parieto-frontal network. PMID- 18161500 TI - The general/specific breakdown of semantic memory and the nature of superordinate knowledge: insights from superordinate and basic-level feature norms. AB - The deterioration of semantic memory usually proceeds from more specific to more general superordinate categories, although rarer cases of superordinate knowledge impairment have also been reported. The nature of superordinate knowledge and the explanation of these two semantic impairments were evaluated from the analysis of superordinate and basic-level feature norms. The results show that, in comparison to basic-level concepts, superordinate concepts are not generally less informative and have similar feature distinctiveness and proportion of individual sensory features, but their features are less shared by their members. Results are in accord with explanations based on feature connection weights and/or concept confusability for the superordinate advantage cases. Results especially support an explanation for superordinate impairments in terms of higher semantic control requirements as related to features being less shared between concept members. Implications for patients with semantic impairments are also discussed. PMID- 18161501 TI - Lipid vesicles as membrane models for toxicological assessment of xenobiotics. AB - Traditionally animals and cell cultures have been used to assess the toxic potential of xenobiotics on cell membranes. In search for more reproducible, quantitative, cost- and time-effective assays, toxicologists have recently become interested in biomimetic lipid vesicle-based test systems. Lipid vesicles (liposomes) have long been appreciated as simple cell membrane models in biochemical and biophysical studies providing a good understanding of the physicochemical properties of liposome systems. More recently a number of reports have been published on the interactions of toxic substances with vesicles. Literature reports on liposome assays have appeared for widely different classes of xenobiotics, such as dental materials, antibiotics, detergents, and peptides. In this review we focus on those reports that contain a quantitative and significant correlation with more established toxicological tests like cell culture assays. We provide an introduction to the structure and main characteristics of vesicles and related lipid aggregates. The two main assays presented are leakage of fluorescence dyes and differential scanning calorimetry (DSC) measurements of the solid-ordered/liquid-disordered main phase transition temperature (Tm). PMID- 18161502 TI - Paraquat poisonings: mechanisms of lung toxicity, clinical features, and treatment. AB - Paraquat dichloride (methyl viologen; PQ) is an effective and widely used herbicide that has a proven safety record when appropriately applied to eliminate weeds. However, over the last decades, there have been numerous fatalities, mainly caused by accidental or voluntary ingestion. PQ poisoning is an extremely frustrating condition to manage clinically, due to the elevated morbidity and mortality observed so far and due to the lack of effective treatments to be used in humans. PQ mainly accumulates in the lung (pulmonary concentrations can be 6 to 10 times higher than those in the plasma), where it is retained even when blood levels start to decrease. The pulmonary effects can be explained by the participation of the polyamine transport system abundantly expressed in the membrane of alveolar cells type I, II, and Clara cells. Further downstream at the toxicodynamic level, the main molecular mechanism of PQ toxicity is based on redox cycling and intracellular oxidative stress generation. With this review we aimed to collect and describe the most pertinent and significant findings published in established scientific publications since the discovery of PQ, focusing on the most recent developments related to PQ lung toxicity and their relevance to the treatment of human poisonings. Considerable space is also dedicated to techniques for prognosis prediction, since these could allow development of rigorous clinical protocols that may produce comparable data for the evaluation of proposed therapies. PMID- 18161503 TI - Neurotoxins from Australo-Papuan elapids: a biochemical and pharmacological perspective. AB - Most of the medically important snakes in Papua New Guinea and Australia belong to the family Elapidae and are referred to as "Australo-Papuan" elapids. Neurotoxicity is often a life-threatening symptom of envenoming by these snakes; therefore, much attention has been paid to the isolation and detailed pharmacological and biochemical characterization of the presynaptic (beta) and postsynaptic (alpha) neurotoxins from these elapid venoms. These studies have highlighted the potential for these toxins to be used as highly potent and selective probes for biomedical research and, perhaps, the potential for their use as lead compounds for the development of pharmaceutical agents. Historically, the potency of neurotoxins/crude venoms has been determined using murine LD50 (lethal dose) assays. However, a different rank order of potency often results when crude venoms/toxins are ranked based on their in vitro pharmacological parameters (e.g., t90 values). The lack of neurotoxicity following envenoming by brown snakes, despite the presence of a potent neurotoxin in their venom, has puzzled clinical toxinologists for years. This paradox also appears to include envenoming by the Stephen's banded snake. Lastly, the in vitro studies examining the effectiveness of antivenoms as well as the potential for alternative compounds to reverse/prevent neurotoxicity are discussed. This review presents for the first time a detailed comparative analysis of the pharmacology and biochemistry of neurotoxins isolated from the Australo-Papuan elapids, placing emphasis on the time taken for onset of action, receptor binding parameters, reversibility, and the methods for determining potency. PMID- 18161504 TI - Potency of novel oximes to reactivate sarin inhibited human cholinesterases. AB - Class of monoquaternary pyridinium oximes was in vitro tested as potential reactivators of acetylcholinesterase (AChE; EC 3.1.1.7) inhibited by nerve agent sarin. Human brain homogenate was used as an appropriate source of cholinesterases. Reactivation potency of novel oximes was compared with currently available reactivators - pralidoxime, obidoxime, and HI-6. According to the obtained results, only five reactivators were able to satisfactorily renew cholinesterase potency (pralidoxime, obidoxime, HI-6, 4-PAM, and K119). Unfortunately, none of the novel tested reactivators surpassed the reactivation potency of the currently most promising reactivator, HI-6. This study shows that monoquaternary reactivators are unable to reactivate nerve agent-inhibited AChE. Due to this, in future, only bisquaternary compounds derived from HI-6 or obidoxime should be designed as new potential cholinesterase reactivators. PMID- 18161505 TI - Gentamicin attenuates gentamicin-induced ototoxicity - self-protection. AB - Aminoglycoside antibiotics cause considerable toxicity to the inner ear. A progressive hearing loss at high frequencies resulted from the loss of hair cells in the base of the cochlea and a constant preoccupation with finding a treatment that protects against their toxic effects. A self-protection phenomenon to high ototoxic doses of gentamicin is proposed in this paper. Thirty-eight adult guinea pigs with normal hearing were tested using Preyer's reflex and the distortion product otoacoustic emission (DPOAE) test, and their cochleae were analyzed by scanning electron microscopy. To the four groups investigated, group I (control) and group II (low dose, 10 mg/kg/day for 30 days) showed a normal DPOEA and normal outer hair cells; group III (high dose, 160 mg/kg/day for 10 days) showed the absence of DPOEA and damage to the outer hair cells; and group IV (low dose, 10 mg/kg/day for 30 days followed by a high dose of 160 mg/kg/day for 10 days) showed a normal DPOEA and normal outer hair cells. These results demonstrate that there was a considerable self-protection phenomenon by gentamicin. PMID- 18161506 TI - In vitro antiplatelet activity of flavonoids from Leuzea carthamoides. AB - Plants and their secondary metabolites, including flavonoids, exhibit a wide range of biological effects. Consequently, natural substances are receiving an increased attention in medicinal research. Owing to these facts, in vitro antiplatelet activity of ethanol summary extract and four flavonoids from Leuzea carthamoides was determined in human platelet-rich plasma. Arachidonic acid (AA), adenosine diphosphate (ADP), collagen (COL), and thrombin were used as agonists of platelet aggregation. The summary extract showed a significant inhibition of the aggregation induced by COL and ADP. Of the tested flavonoids, eriodictyol (1) and patuletin (2) influenced COL- and AA-induced aggregation. Their IC(50) values are presented. Flavonoid glycosides eriodictyol-7-beta-glucopyranoside (3) and 6 hydroxykaempferol-7-O-(6''-O-acetyl-beta-D[small cap]-glucopyranoside) (4) were found to be weak antiplatelet agents. These results confirmed the fact that glucosylation decreases the antiplatelet activity. Quantitative composition of tested flavonoids in L. carthamoides extract was also determined. Though two of the tested flavonoids inhibited platelet aggregation, further evaluation of L. carthamoides, in order to discover other antiplatelet active compounds and possible adverse health effects, is needed. PMID- 18161507 TI - Mutagenic, antimutagenic, cytotoxic, and apoptotic activities of extracts from Pituranthos tortuosus. AB - Mutagenic and antimutagenic activities against direct acting mutagens, nifuroxazide (NF) and sodium azide (SA), and indirect acting mutagen aflatoxin B1 (AFB1) of extracts prepared from aerial parts of Pituranthos tortuosus were investigated in bacterial assay systems (i.e., the Ames test with Salmonella typhimurium TA100, TA98, TA1538, TA1535, and the SOS chromotest with Escherichia coli PQ 37). It was found that all extracts obtained from P. tortuosus decreased the mutagenicity induced by AFB1 (10 microg/assay), SA (1.5 microg/assay), and NF (20 microg/assay). Ethyl acetate, acetone, methanol, and total oligomer flavenoid extracts exhibited the highest inhibition level of mutagenicity induced by the indirect mutagen AFB1. In addition, antiproliferative and apoptotic properties of these extracts have also been reported using two leukemia cell lines, L1210 and K562. The results revealed that all extracts showed a significant cytotoxic effect on these cell lines, and the effect was greater in the presence of human K562 chronic myelogenous leukemia cells, whereas they do not induce apoptosis. PMID- 18161508 TI - Antiproliferative, antioxidant, and antimutagenic activities of flavonoid enriched extracts from (Tunisian) Rhamnus alaternus L.: combination with the phytochemical composition. AB - A pronounced antiproliferative effect on human leukemia K562 cells was shown with flavonoid-enriched extracts from Rhamnus alaternus roots and leaves, with, respectively, IC(50) values of 165 and 210.73 microg/mL. High DPPH radical scavenging activity (7.21 and 18.84 microg/mL, respectively) and antioxidative effects using the xanthine oxidase assay (IC(50) values of 83.33 and 103.96 microg/mL, respectively) were detected in the presence of the two tested extracts. Although no mutagenic effect was observed when using the Salmonella typhimurium assay system with TA1535 and TA100 strains, the two tested extracts exhibited a high-level protection toward the direct mutagen, sodium azide-induced response. PMID- 18161509 TI - A 52-week repeated dose toxicity study of ultraviolet absorber 2-(2'-hydroxy 3',5'-di-tert-butylphenyl)benzotriazole in rats. AB - A 52-week repeated dose toxicity study of an ultraviolet absorber, 2-(2'-hydroxy 3',5' -di-tert-butylphenyl)benzotriazole (HDBB), was conducted according to OECD TG 452 under GLP. CD(SD)IGS rats were given HDBB by gavage at 0, 0.1, 0.5, or 2.5 mg/kg/day in males and 0, 0.5, 2.5, or 12.5 mg/kg/day in females. No substance related deaths or clinical signs of toxicity were observed in any group; however, a lowered body weight was found from day 36 to the end of the 52-week administration period at 2.5 mg/kg in males. At the completion of the dosing period, a decrease in red blood cells at 0.5 mg/kg and higher, and in hematocrit at 2.5 mg/kg, was detected in males. Blood biochemical changes, including increases in the levels of alkaline phosphatase and glucose and the A/G ratio, were also found at 0.5 mg/kg and higher in males and at 12.5 mg/kg in females. At necropsy, absolute and relative liver weight was increased at 0.5 mg/kg and higher in males and at 12.5 mg/kg in females. Histopathological changes were observed in the liver; centrilobular hypertrophy of hepatocytes at 0.5 mg/kg and higher in males, and at 12.5 mg/kg in females, and altered hepatocellular foci at 0.5 mg/kg and higher, and cystic degeneration and lipofuscin deposition in hepatocytes at 2.5 mg/kg in males. Based on these findings, the no observed adverse effect level was concluded to be 0.1 mg/kg/day in male rats and 2.5 mg/kg/day in female rats. PMID- 18161510 TI - Oxime silanes: structure/toxicity relationships. AB - Acute and repeated oral and dermal rat toxicology studies of standard designs were conducted on four methyl ethyl ketoxime (MEKO) silanes and four methyl isobutyl ketoxime (MIBKO) silanes. Each compound contained either MEKO or MIBKO groups (but not both) and either a single methyl, vinyl, or phenyl group (trifunctional oxime silane), two methyl groups or a methyl and vinyl group (difunctional oxime silane), or no nonoxime group (tetrafunctional oxime silane) attached to the central silicon atom. All compounds caused transient narcosis and anemia, with oral exposure associated with the hydrolyzed oxime groups. Difunctional oxime silanes, containing both a methyl and a vinyl group, caused degeneration of the seminiferous tubules of the testes following oral administration. Serial testicular histopathology indicated the effect originated at the level of the spermatocyte, resulting in a wave of cellular depletion of later maturation stages of spermatogenesis. Spermatogenesis gradually recovered but function was not evaluated. Tetrafunctional oxime silanes, trifunctional oxime silanes, including those containing a single methyl or vinyl group, or difunctional oxime silane containing two methyl groups did not affect the testes, indicating that both a methyl and vinyl group needs to be present on the oxime silane molecule for testicular toxicity. The testicular toxicity appears to be associated with the methyl/vinyl silane portion and not the oxime portion of the oxime silane molecule. With the exception of the methyl/vinyl difunctional oxime silanes, the silane portion of oxime silanes does not appear to contribute any significant toxicity to these compounds. PMID- 18161511 TI - Gonadal influence on the toxicity of 2-(2'-hydroxy-3',5'-di-tert-butylphenyl) benzotriazole in rats. AB - Previously, we showed that susceptibility of male rats to the toxicity of an ultraviolet absorber, 2-(2'-hydroxy-3',5'-di-tert-butylphenyl)benzotriazole (HDBB), was nearly 25 times higher than that of females. In the current study, we investigated the role of sex steroids in the mediation of the gender-related difference using castrated rats. Male and female castrated CD(SD) rats were given HDBB by gavage at 0, 0.5, 2.5, or 12.5 mg/kg/day for 28 days. No deaths, clinical signs of toxicity, or changes in body weight or food consumption were found at any doses. Blood biochemical changes suggestive of hepatic damage, such as increased levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase, were detected at 12.5 mg/kg/day in males. Absolute and relative liver weight increased at 0.5 mg/kg/day and above in males and at 12.5 mg/kg/day in females. In the liver, histopathological changes, such as nucleolar enlargement, increased mitosis, hypertrophy in hepatocytes, and/or focal necrosis were observed at 0.5 mg/kg/day and above in males, and at 2.5 mg/kg/day and above in females. These findings indicate that castration markedly reduced the gender-related differences in toxicity of HDBB in rats. PMID- 18161512 TI - A comparison of the potency of newly developed oximes (K074, K075) and currently available oximes (obidoxime, trimedoxime, HI-6) to counteract acute toxic effects of tabun and cyclosarin in mice. AB - The potency of newly developed oximes (K074, K075) and commonly used oximes (obidoxime, trimedoxime, and HI-6) to counteract tabun or cyclosarin-induced acute toxic effects was studied in mice. The therapeutic efficacy of trimedoxime and both newly developed oximes (K074, K075) was significantly higher than the potency of obidoxime and the oxime HI-6 in the case of acute tabun poisonings. On the other hand, the oxime HI-6 was significantly more efficacious than other studied oximes when mice were intoxicated with cyclosarin. The findings support the hypothesis that the therapeutic efficacy of oximes depends on the type of nerve agent. Due to their therapeutic efficacy, both newly developed K oximes can be considered to be promising oximes for the antidotal treatment of acute tabun poisonings, while the oxime HI-6 is still the most promising oxime for the treatment of acute cyclosarin poisonings due to its high potency to counteract cyclosarin-induced acute toxic effects. PMID- 18161513 TI - Sulfur mustard induces apoptosis in cultured normal human airway epithelial cells: evidence of a dominant caspase-8-mediated pathway and differential cellular responses. AB - We have shown that sulfur mustard (SM; bis-(2-chloroethyl) sulfide), an alkylating, vesicating chemical warfare agent, causes dermal toxicity, including skin microblisters, via the induction of both death receptor (DR) and mitochondrial pathways of apoptosis in human epidermal keratinocytes. While SM is known for its skin-vesicating properties, respiratory tract lesions are the main source of morbidity and mortality after inhalation exposure. We, therefore, investigated whether SM induces apoptotic cell death in normal human bronchial epithelial (NHBE) cells and small airway epithelial cells (SAEC) in vitro. Cells were exposed to various concentrations of SM (0, 50, 100, and 300 muM for 16 h) in the culture medium and then tested for the activation of apoptotic executioner caspase-3 and initiator caspases-8 and -9. Caspases-8 and -3 were activated by SM in both airway cell types, indicating the induction of a DR pathway of apoptosis in these cells; however, the levels of enzyme activation were different, depending on the cell type and the SM concentrations used. Consistent with enzyme activity results, immunoblot analyses revealed the proteolytic processing of the proenzymes to the active forms of caspases-8 and -3 in these cells after SM exposure. Interestingly, NHBE cells were found to be exquisitely sensitive to SM, compared to SAEC, with caspase-3 activities in SM-exposed NHBE cells approximately 2-fold higher and caspase-8 activities approximately 10-fold higher than in SAEC. Furthermore, SM activated caspase-9 in NHBE cells, but not in SAEC, indicating a possible role of the mitochondrial pathway only in the NHBE cells. The present study shows that both upper airway (NHBE cells) and deep lung (SAEC) epithelial cells undergo SM-induced apoptotic death in vitro, but distinct cell type specific responses can be elicited, which may be attributed to intrinsic properties that characterize the response of these cells to SM. These findings need to be taken into consideration in the search for modulators of these pathways for the therapeutic intervention to reduce SM injury due to respiratory tract lesions. PMID- 18161514 TI - In vitro and in vivo evaluation of various carbonyl compounds against cyanide toxicity with particular reference to alpha-ketoglutaric acid. AB - Cyanide is a rapidly acting neurotoxin that necessitates immediate, vigorous therapy. The commonly used treatment regimen for cyanide includes the intravenous administration of sodium nitrite (SN) and sodium thiosulphate (STS). Due to many limitations of these antidotes, a search for more effective, safer molecules continues. Cyanide is known to react with carbonyl compounds to form the cyanohydrin complex. The present study addresses the efficacy of several carbonyl compounds and their metabolites or nutrients with alpha-ketoglutaric acid (A-KG), citric acid, succinic acid, maleic acid, malic acid, fumaric and oxaloacetic acid, glucose, sucrose, fructose, mannitol, sorbitol, dihydroxyacetone, and glyoxal (5 or 10 mM; -10 min) against toxicity of potassium cyanide (KCN; 10 mM) in rat thymocytes in vitro. Six hours after KCN, cell viability measured by MTT assay and crystal violet dye exclusion revealed maximum cytoprotection by A-KG, followed by oxaloacetic acid. A-KG also resolved the leakage of intracellular lactate dehydrogenase, loss in nuclear integrity (propidium iodide staining), and altered mitochondrial membrane potential (rhodamine 123 assay) as a result of cyanide toxicity. Protection Index (ratio of LD(50) of KCN in protected and unprotected animals; PI) of all the compounds (oral; 1.0 g/kg; -10 min) determined in male mice, revealed that maximum protection was afforded by A-KG (7.6 PI), followed by oxaloacetic acid (6.4 PI). Comparative evaluation of various salts of A-KG alone or with STS (intraperitoneal; 1.0 g/kg; -15 min) showed that maximum protection was conferred by disodium anhydrous salt of A-KG, which also significantly prevented the inhibition of brain cytochrome oxidase caused by 0.75 LD(50) KCN. This study indicates the potential of A-KG as alternative cyanide antidote. PMID- 18161515 TI - Effect of zinc on hepatic drug metabolism under ethanol toxicity. AB - The effects of zinc on drug-metabolizing enzymes in the liver were examined in male Wistar rats following ethanol intoxication. Rats were orally fed 3 mL of 30% ethanol daily for either two, four, or eight weeks and were orally administered zinc sulfate (ZnSO4.7H2O) at a dose level of 227 mg/L. Levels of reduced glutathione (GSH) and the activities of cytochrome P-450, cytochrome b(5), NADPH cytochrome-C-reductase and glutathione-S-transferase (GST) were determined in liver after two, four, and eight weeks. Significant elevation was observed in the activities of the enzymes of the mixed function oxidase system in response to toxicity induced by ethanol at all the intervals. These effects of were ascribed to the enhanced activity of the microsomal ethanol oxidizing system and the associated increase in reactive oxygen species production. Zinc supplementation to these ethanol-intoxicated animals resulted in normalization of these elevated values significantly, but still they do not attain normal levels. Significant increase was observed in reduced glutathione content in animals after four and eight weeks of ethanol feeding, which appeared to be further elevated in combined zinc and ethanol treatment. Significant elevation in the activity of GST was illustrated on ethanol-fed animals at all the three treatment intervals. Furthermore, the activity of this enzyme was only moderately normalized following zinc treatment. This was accredited to the antioxidant potential of zinc, as well as its ability to induce metallothionein content, which provide protection against the toxic effects of ethanol. To conclude, zinc was able to normalize the effects of ethanol in the liver. PMID- 18161516 TI - Methanol extract of Biophytum sensitivum alters the cytokine profile and inhibits iNOS and COX-2 expression in LPS/Con A stimulated macrophages. AB - Biophytum sensitivum has been used in traditional folk medicine to treat numerous diseases. The molecular mechanism of B. sensitivum pharmacological and biochemical actions of macrophages in inflammation has not been clearly elucidated. We examined how the methanol extract of B. sensitivum regulates the production of interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha, IL-6, and nitric oxide (NO) in vitro and in vivo. The extract inhibits the production of NO and proinflammatory cytokines in lipopolysaccharide (LPS) or Concanavalin (Con) A-stimulated primary macrophages. In vitro L929 bioassay revealed the inhibition of TNF-alpha production by B. sensitivum treatment. Moreover, the extract could suppress the inducible nitric oxide synthase and cyclo-oxygenase-2 mRNA expression in LPS or Con A-stimulated macrophages. These findings provide evidence that B. sensitivum possesses potential anti-inflammatory activity and may be beneficial for the treatment of endotoxin shock or sepsis. PMID- 18161518 TI - Cell tracking 2007: a proliferation of probes and applications. AB - The articles in this thematic issue, entitled "Tracking Cell Proliferation and Function," illustrate some of the choices made by authors pushing the envelope for cell tracking applications in their areas of interest. Over the past decade there has been a proliferation in the range of commercially available probes for these studies, the capabilities of the instrumentation used to detect them, and in the biological systems being studied. This introductory to the thematic issue presents the advantages and limitations of the more commonly used probes such as CFSE and PKH26, as well as emerging probes that expand the range of fluorescence available, including quantum dots and the new CellVue dyes. Appropriate method and instrument setup controls and possible data analysis strategies are discussed with the goal of urging experienced investigators to include all critical information and controls when publishing their data and of aiding researchers new to cell tracking to make informed decisions on which cell tracking reagent(s) are best suited for their particular application. All cell tracking assays have the common goal of determining the fate of a particular cell population within a heterogeneous environment, whether in vivo or in vitro. Some of the common themes among the contributions found in this issue include how various probes are used to track (i) cell proliferation, (ii) regulatory and effector immune cell function and (iii) membrane transfer and antigen presentation. Although these represent only a small fraction of the large and growing list of applications for cell tracking, clearly illustrate the growing trend toward the use of multiple tracking reagents and multiple detection modalities to address complex biological questions. PMID- 18161519 TI - A flow cytometric assay for quantitation of rare antigen-specific T cells: using cell-tracking dyes to calculate precursor frequencies for proliferation. AB - Cell-tracking dyes stain cells with bright fluorescence which is partitioned between daughter cells after each cell division, so that the daughter cells have closely half the intensity of the parent cells from which they were derived. Therefore, the intensity of a cell, relative to its intensity at the time of staining, provides information about how many divisions have occurred. Knowing the number of division cycles that have occurred, one can calculate the number of cells in the original population (before culture) that were going to go on to proliferate. In this way, cell-tracking dyes provide a flow cytometric method for determining the proportion of cells in a population that will go on to proliferate in response to stimulation. This dye-dilution methodology can, therefore, detect proliferation of rare antigen-specific cells that increase in number during division and, importantly, can be used to back-calculate the precursor frequency of these rare cells. Simultaneously, the phenotype of these cells can be determined, as well as their ability to synthesize cytokines, and to express or not relevant antigen-binding receptors and activation markers. PMID- 18161520 TI - CellVue Claret, a new far-red dye, facilitates polychromatic assessment of immune cell proliferation. AB - Flow cytometric analyses of immune cell proliferation, differentiation, and function are limited by the number of different fluorochromes that can be resolved simultaneously. Additional colors to expand functional analytic capability will facilitate higher dimensional analyses of heterogeneous cell populations by basic and clinical scientists. Our aim in these studies was to evaluate CellVue Claret, a fluorescent, far-red emitting, membrane intercalating dye (excitation maximum: 655 nm, emission maximum 677 nm), as an alternative and/or complementary probe to PKH26 and CFSE(1) for polychromatic studies of immune cell proliferation and function. Using a BD FACSCalibur and human peripheral blood mononuclear cells (PBMCs) from 8 different donors (2 donors studied twice), we compared CellVue Claret with the two most commonly used visible-emitting proliferation dyes, PKH26 and CFSE, in terms of: (1) compatibility with 7-Amino-actinomycin D (7-AAD) as a viability marker; (2) effect of dye labeling on lymphocyte viability; and (3) the proliferative response of CD3+ T lymphocytes from 0-96 hours as assessed by dilution of each of the 3 cell tracking dyes in cultures stimulated with anti-CD3 plus IL-2. Post labeling recoveries and viabilities were similar for all 3 dyes, with modestly higher initial staining intensities and coefficients of variation for CellVue Claret than for CFSE or PKH26. Lymphocyte viabilities in stimulated or unstimulated cultures were also unaffected by choice of dye. Proliferative responses of viable CD3+ lymphocytes were comparable for all three dyes, whether results were reported as Proliferative Fraction (percent of cells that had divided one or more times) or as Precursor Frequency (percent of parent population that had gone on to proliferate in response to anti-CD3 plus IL-2). In summary, T cell proliferation analysis using CellVue Claret gives results equivalent to those obtained with PKH26 or CFSE, expanding the choice of proliferation dyes suitable for use in high dimensional polychromatic studies on flow cytometers with far red (633 nm-658 nm) excitation capabilities. PMID- 18161521 TI - Assessing the in vitro suppressive capacity of regulatory T cells. AB - Regulatory T cells (Treg) play a vital role in controlling peripheral immune responses in order to prevent autoimmunity and control inflammation. Altered Treg activities have been associated with the pathogenesis of multiple disorders including autoimmunity, allergy, cancer, and infection with persistent pathogens. As such, a great deal of interest has recently been directed towards developing additional tools and methods to better understand the mechanisms of suppression employed by Treg. The in vitro suppression assay has emerged as a valuable means by which to assess the functional capacity and activity of Treg. In this review, we summarize the merits and limitations of the various in vitro assays that have been utilized to assess Treg activity and present a novel two color proliferation assay that allows simultaneous monitoring of both regulatory and effector T cell activity. As further immunomodulatory therapies are explored, the need for additional methodologies to understand the cellular and molecular mechanisms of immune regulation conferred by Treg will play an increasingly important role. PMID- 18161522 TI - Use of CFSE to monitor ex vivo regulatory T-cell suppression of CD4+ and CD8+ T cell proliferation within unseparated mononuclear cells from malignant and non malignant human lymph node biopsies. AB - Regulatory T-cells (Tregs) play a critical role in the inhibition of self reactive immune responses and as such have been implicated in the suppression of anti-tumor immunity. A clearer understanding of the mechanisms by which Tregs suppress effector T-cell responses within the context of anti-tumor immunity may lead to more effective treatments. The study of Tregs, particularly in the context of ongoing active immune responses, has been challenging due to the lack of surface molecules truly unique to these cells. Several surface markers have been shown to be constitutively expressed by Tregs, such as high levels of CD25, GITR and CTLA-4, and thus have been useful for their study. However, the heterogeneity of surface marker expression still makes identifying Tregs ex vivo challenging. As such, the only means available, currently, to accurately identify Tregs ex vivo is through functional suppression assays. Tregs have been shown to inhibit a variety of cellular functions including T-cell proliferation and as such, in vitro inhibition of proliferation is routinely used as a measure of Treg mediated suppression. Several assays currently exist to assay cellular proliferation, including [(3)H]thymidine incorporation and CFSE dilution. However, a limitation of using [(3)H]thymidine is the difficulty differentiating between proliferation of the target cells and that of the Tregs themselves. Due to the ability to differentiate by flow cytometric analysis between labeled and unlabelled cells using CFSE, in contrast to [(3)H]thymidine, it is possible to analyze the proliferation of labeled target cells separate from unlabeled Tregs in co-culture experiments. In addition, the use of multi-color flow cytometry allows for the analysis of different T-cell subsets simultaneously without the necessity to separate these cells. Thus, CFSE has several advantages to [(3)H]thymidine for analysis of cellular proliferation. Herein we describe our work utilizing CFSE labeling to assess, (1) proliferative responses of CD4(+) and CD8(+) T-cells in unseparated single cell suspensions from human lymph nodes and, (2) the ability of tumor infiltrating suppressive populations, including Tregs, isolated from neoplastic lymph nodes to suppress in vitro proliferation of allogeneic CD4(+) and CD8(+) T-cells isolated from peripheral blood of healthy donors. PMID- 18161523 TI - Dye dilution proliferation assay: application of the DDPA to identify tumor specific T cell precursor frequencies in clinical trials. AB - A better understanding of immune effector and regulatory pathways has led to innovative, and complex, immunotherapy strategies. CD8(+) cytolytic T lymphocytes (CTL) provide one common pathway of tumor cell destruction. The peripheral blood CTL compartment typically comprises a minority of anti-tumor CD8(+) lymphocytes and the determination of their number during clinical trials is the focus of various laboratory methods. We have monitored tumor specific CD8(+) as well as CD4(+) lymphocyte precursor frequencies in the peripheral blood using a Dye Dilution Proliferation Assay (DDPA). We summarize our experience applying DDPA in a multi-parameter, antigen-specific assay, detailing some of its complexities and advantages. We provide examples of our clinical trial results showing tumor specific CD8(+) and CD4(+) precursor frequency (PF) data in patients being treated on novel immunotherapy trials. PMID- 18161524 TI - Lipophilic fluorochrome trackers of membrane transfers between immune cells. AB - Cell-to-cell transfers of membrane molecules between lymphoid cells (sometimes referred to as trogocytosis) is frequent and has multiple physiological consequences. Although difficult to visualize through the tracking of defined cell surface proteins, this process can be readily monitored by inserting PKH or CellVue Maroon fluorochromes into the plasma membranes of donor cells. We discuss here parameters that determine its detection by a flow-cytometry-based in vitro assay and present examples of application, including time-lapse video-microscopy analysis of transfers at the immunological synapse. By combining detection of cell-to-cell transfer and of cell surface CD107, it is possible to discriminate lymphoid cells binding target cells with and without perforin release. This could prove useful for identifying cells that destruct known target cells in autoimmune pathologies. PMID- 18161525 TI - Chemical labels metabolically installed into the glycoconjugates of the target cell surface can be used to track lymphocyte/target cell interplay via trogocytosis: comparisons with lipophilic dyes and biotin. AB - Trogocytosis, the process whereby lymphocytes capture membrane components from the cells they interact with, is classically evidenced by the transfer of fluorescent lipophilic compounds or biotinylated proteins from target cells to T or B cells. A particular class of molecules, not studied explicitly so far in the context of trogocytosis is glycoconjugates. Here, we used a method to metabolically install chemical labels in target cell glycoconjugates. Working with those target cells, we describe the conditions allowing CTL to be detected based on glycoconjugate trogocytosis triggered by antigen or stimulatory antibodies. Accordingly, we used this method to monitor the CTL response triggered in mice after vaccination. In addition, we documented the applicability of this approach to the detection of CD4(+) T and B cells. Overall, glycoconjugates were transferred between target cells and lymphocytes during trogocytosis with efficiencies comparable or higher than measured for biotinylated proteins or lipophilic dyes incorporated into general membrane lipids. From a technological point of view, our approach can be employed to detect reactive lymphocytes via glycoconjugate trogocytosis. More generally, we believe that the ever-growing ability to employ chemistry in living systems to label particular compounds will be powerful in unraveling the contributions of glycosylation to various aspects of T and B cells biology. PMID- 18161526 TI - Imaging approaches to hematopoietic stem and progenitor cell function and engraftment. AB - Cell tracking in vivo continues to provide significant insights into hematopoietic cell function and donor cell engraftment after transplantation. The combination of proliferation tracking dyes and induced expression of reporters with advanced imaging modalities has led to better understanding of qualitative and quantitative aspects of hematopoietic cells' homing, seeding and engraftment. Currently, there is no single technique that allows in vivo tracking of cells with molecular resolution, thus several techniques need to be combined. Recent developments promise better implementation of non-invasive imaging modalities to study functional and molecular characteristics of stem cells. PMID- 18161527 TI - Image-based study of interferongenic interactions between plasmacytoid dendritic cells and HSV-infected monocyte-derived dendritic cells. AB - Plasmacytoid dendritic cells (pDC) are well-known for their ability to produce large quantities of interferon-alpha (IFN-alpha) in response to viruses. In addition, pDC produce IFN-alpha in response to HSV-infected cells. We demonstrate that both tonsil and PBMC contain pDC that respond to stimulation with HSV either in suspension or in tonsil tissue-fragment culture. We hypothesized that other DC subsets acquire virus in the periphery and deliver the interferongenic signals to the pDC in the draining lymphoid tissue. As a model for pDC/myeloid DC interaction, we studied the interaction of pDC derived from blood with HSV infected and uninfected monocyte derived dendritic cells (MDDC). Infected, but not uninfected, MDDC induced IFN-alpha in pDC. To further study pDC/infected MDDC interactions, we labeled MDDC with fluorescent cell trackers PKH67 or CFSE prior to infection with HSV and co-cultured with pDC. Cells were then analyzed using conventional and imaging flow cytometry. In addition, we infected MDDC with a GFP expressing HSV prior to co-culture with pDC. Using traditional flow cytometry, we observed that pDC became fluorescent after co-incubation with uninfected or infected, fluorescently labeled MDDC, indicating that MDDC transferred fluorescent protein and membrane to pDC. By imaging flow cytometry, we observed formation of conjugates between pDC and MDDC as well as transfer and internalization of cellular components from the labeled MDDC by pDC, with preferential uptake from, and association with, infected vs. uninfected MDDC. These studies demonstrate that MDDC infected with HSV are able to stimulate IFN alpha and chemokine production by pDC through the transfer of cellular materials from the HSV-infected MDDC to the pDC. Together, these observations indicate that heterogeneous populations of DC interact to generate an effective IFN-alpha response. PMID- 18161528 TI - Long-distance three-color neuronal tracing in fixed tissue using NeuroVue dyes. AB - Dissecting development of neuronal connections is critical for understanding neuronal function in both normal and diseased states. Charting the development of the multitude of connections is a monumental task, since a given neuron typically receives hundreds of convergent inputs from other neurons and provides divergent outputs for hundreds of other neurons. Although progress is being made utilizing various mutants and/or genetic constructs expressing fluorescent proteins like GFP, substantial work remains before a database documenting the development and final location of the neuronal pathways in an adult animal is completed. The vast majority of developing neurons cannot be specifically labeled with antibodies and making specific GFP-expressing constructs to tag each of them is an overwhelming task. Fortunately, fluorescent lipophilic dyes have emerged as very useful tools to systematically compare changes in neuronal networks between wild-type and mutant mice. These dyes diffuse laterally along nerve cell membranes in fixed preparations, allowing tracing of the position of a given neuron within the neuronal network in murine mutants fixed at various stages of development. Until recently, however, most evaluations have been limited to one, or at most, two color analyses. We have previously reported three color neuronal profiling using the novel lipophilic dyes NeuroVue (NV) Green, Red and Maroon (Fritzsch et al., Brain. Res. Bull. 66: 249-258, 2005). Unfortunately such three color experiments have been limited by the fact that NV Green and its brighter successor, NV Emerald, both exhibit substantially decreased signal intensities when times greater than 48 hours at 37 degrees C are required to achieve neuronal profile filling (unpublished observations). Here we describe a standardized test system developed to allow comparison of candidate dyes and its use to evaluate a series of 488 nm-excited green-emitting lipophilic dyes. The best of these, NV Jade, has spectral properties well matched to NV Red and NV Maroon, better solubility in DMF than DiO or DiA, improved thermostability compared with NV Emerald, and the ability to fill neuronal profiles at rates of 1 mm per day for periods of at least 5 days. Use of NV Jade in combination with NV Red and NV Maroon substantially improves the efficiency of connectional analysis in complex mutants and transgenic models where limited numbers of specimens are available. PMID- 18161529 TI - Characterizing host immune responses by monitoring fluorescently marked cells. PMID- 18161530 TI - Tracking the elusive lymphocyte: methods of detection during adoptive immunotherapy. AB - Adoptive immunotherapy is an attractive cancer treatment modality due to its capacity to target primary and metastatic lesions with large numbers of tumor reactive, cytotoxic lymphocytes. The inability of fully armed lymphocytes to traffic into sites of tumor has been proposed as a causal factor for the minimal success observed clinically with this type of immunotherapy. The study of lymphocyte trafficking during adoptive immunotherapy has been limited, despite the existence of a variety of tracking methods. In murine models that simulate adoptive immunotherapy, the use of congenic mice and cell tracking dyes can be used to elucidate lymphocyte trafficking behavior. The continued development of novel technologies will further contribute to this expanding area of research. PMID- 18161531 TI - Simultaneous analysis of in vivo CD8+ T cell cytotoxicity against multiple epitopes using multicolor flow cytometry. AB - CD8+ T cells play a critical role in host defense against infections and tumors. Analysis of cytotoxic function of antigen-specific CD8+ T cells in animal models would be important in optimizing vaccine design against infections and tumors. In vivo cytotoxicity assays using fluorescent cellular dyes have been used as a popular alternative to traditionally used in vitro (51)Cr-release assays. With the identification of multiple epitopes in various pathogen models, methods to simultaneously analyze cytotoxicity of CD8+ T cells to multiple epitopes in vivo would assist studies which aim to generate protective CD8+ T cell immunity to multiple epitopes. In this study, we evaluate the use of multiple fluorescent cellular dyes for the in vivo cytotoxicity assay. The use of 3 dyes allowed us to analyze the cytotoxicity of antigen-specific CD8+ T cell populations to multiple epitopes generated by virus infections, as well as their functional avidity, in vivo. Our studies extend the use of in vivo cytotoxicity assays to allow direct comparisons of cytotoxicity to various epitopes in the same animal and may also be applicable to assessment of in vitro cytotoxicity of human CD8+ T cells specific for multiple viral or tumor antigens in clinical settings. PMID- 18161532 TI - Antiproliferative activity of cisplatin detected by CFSE in p53-proficient and p53-deficient cells. AB - We have previously developed experimental and data analysis procedures to measure the antiproliferative activity of drugs in continuously proliferating cancer cell lines using carboxyfluorescein diacetate succinimidyl ester (CFSE). The method was applied here to analyze the role of p53 in the effect of the anticancer drug cisplatin, distinguishing events occurring in the first generation of cells from those in the second and subsequent generations. A CFSE-loaded colon carcinoma cell line expressing functional wild-type p53 was treated for 1 with cisplatin in parallel with its p53-deficient counterpart, collecting frequency distributions of DNA and CFSE content up to 72 h after treatment. At a sublethal cisplatin concentration proliferation was temporarily inhibited but then the block was overcome and most cells were able to divide several times. The initial block was stronger in HCTp53-/- cells, resulting in a larger proportion of undivided cells at 24 h. This was confirmed and amplified at a higher, lethal concentration, where undivided G(2)M-blocked p53-deficient cells eventually died by non apoptotic mechanisms, while p53-proficient cells avoided this with a less stringent block. This gave p53-proficient cells more time to repair and eventually decide on survival or apoptotic death before traversing the cycle into their second generation. PMID- 18161533 TI - Novel lipophilic tracking dyes for monitoring cell proliferation. AB - The advent of contemporary digital instrumentation has enhanced both the potential and the complexity of flow cytometric experiments, allowing for the detailed dissection of immune cell subsets and their functions. The use of cell tracking labels such as PKH26 and CFSE has been important in observing such cellular functions, but their visible emission characteristics have limited the design of such analyses. As the demand for multiparametric flow cytometry intensifies, it will become increasingly important to utilize a broader range of cell tracking reagents to optimize the measurement of fluorescence signals and to provide flexibility in the use of commercially available fluorochrome - antibody combinations. We report on the evaluation of three lipophilic membrane dyes, CellVue Lavender, CellVue Plum and CellVue NIR780; with fluorescence emissions in the violet, far-red and near infrared wavelength regions, respectively. These reagents are similar to established tracking dyes such as PKH26 and CFSE in terms of staining procedure, membrane stability, optimal concentration, and lack of effect on cellular proliferation. The CellVue dyes however, exhibit different spectral characteristics than existing tracking compounds, and capitalize upon the increased number of lasers incorporated into commercially available instrumentation; thus permitting measurement of labeled populations in underexploited regions of the spectrum. PMID- 18161534 TI - Veterinary applications for monitoring mononuclear cell proliferation using cell tracking dyes. AB - Veterinary species offer unique opportunities for the study of immune responses during natural host/pathogen interactions. Experimental studies can be used to characterize the response to infection, vaccination, and influence of vaccination on the response to infection. The intent of this review is to demonstrate the use of cell tracking dyes to monitor and characterize in vitro proliferative responses by mononuclear cell subsets from veterinary species as a correlate to the in vivo response. Selected examples are provided to illustrate the usefulness of this approach to characterize various tissue dendritic cell populations, CD8 alpha alpha(+) T cells, gammadelta T cells, and CD172a(+) cells. Comparative approaches provide unique and comprehensive insights into mononuclear cell biology that may be applicable to similarly described cell populations in humans. PMID- 18161538 TI - Statistics in the world of medical devices: the contrast with pharmaceuticals. AB - Medical devices play a vital role in people's lives as these products are revolutionizing medicine with breathtaking advances in both the treatment and the detection of many diseases. While a similar, primarily therapeutic, revolution is ongoing in the pharmaceutical world; the focus here is the effect this device revolution is having on the statistical world. The similarities and differences between medical devices and pharmaceutical drugs are explored in terms of their natures, industries, and how they are regulated in the U.S. and globally. Statistical issues concerning the evaluation of devices versus those of drugs are compared and contrasted. These trends are creating new challenges for the statistical world in the development and evaluation of these new medical products. PMID- 18161539 TI - Statistical and regulatory issues in nonrandomized medical device clinical studies. AB - While randomized, well-controlled, clinical trials have been viewed as the gold standard in the evaluation of medical products, it is not uncommon for medical device clinical studies to depart from the paradigm of randomized trials, due to ethical or practical reasons. In nonrandomized studies, the advantages of well designed and conducted randomized clinical trials are no longer available, and consequently the statistical inference obtained from such studies may carry a lower level of scientific assurance, compared to randomized trials. This paper provides a brief overview of nonrandomized medical device clinical studies in terms of design and statistical analysis as well as regulatory issues, including some challenges that frequently arise in those endeavors. PMID- 18161540 TI - Design effects for highly clustered count data with varying length of follow-up- applying to the analysis of ICD shocks. AB - Highly clustered count data are commonly seen in medical device clinical studies such as cardiac rhythm management. For instance, anti-arrhythmia shocks delivered from an implantable cardioverter-defibrillator (ICD) often occur as "storms", i.e., multiple shocks within a short period of time. There are unique challenges in designing and analyzing clinical studies with such response data. In general, there is less amount of data information available when events occur as "storms" as compared to independent events, and the efficiency loss may be substantial in practice. In addition, event incidences of subjects may be highly unbalanced and length of follow-up may vary from subject to subject. In this paper, the design effect for such highly clustered and unbalanced count data is studied. Varying length of subject follow-up is taken into account by the use of a random variable that may depend on external covariates. Sample size adjustment for event correlation and imbalance is discussed. The practical application of the strategy is illustrated through a clinical study of ICD shock therapy. PMID- 18161541 TI - Longitudinal data analysis in medical device clinical studies. AB - Several statistical approaches are discussed for analysis of longitudinal clinical data from a two independent groups, randomized, prospective, multicenter study of patients with benign prostate hyperplasia (BPH) who are treated with either a new transurethral microwave thermotherapy device (TUMT, n = 147) or sham control (n = 73) at baseline and at 1, 3, and 6 months follow up. The primary clinical endpoint, based on the American Urological Association (AUA) symptom score from 7 questions (each scored 0-5) with 35 as the worst total score, is illustrated in this paper. The primary effectiveness null hypothesis is no treatment effect at any follow-up time against the alternative hypothesis of a favorable treatment effect at one or more follow-up times. Based on the prespecified clinical input of TUMT improvement in AUA mean score as a possible early effect, the expected change of slope between baseline to month 1 (baseline slope) and from month 1 to the end of month 6 (longitudinal slope), with knot at one month, were analyzed. The primary effectiveness endpoint was analyzed by a piecewise linear spline regression model with knot at month 1. Statistically significant lower mean AUA score was found in the TUMT group compared to the sham group at month 3 (P = 0.01) and at month 6 (P < 0.0001) follow-up; no significant difference in mean AUA score was found at baseline or at month 1 follow-up, after an overall statistically significant difference in mean AUA score (P = 0.004) was found between the TUMT and sham groups. PMID- 18161542 TI - Practical Bayesian design and analysis for drug and device clinical trials. AB - Perhaps the most valuable contribution of Bayesian methods to health care evaluation involves study design. Drug and medical device clinical trialists are increasingly confronted with data that feature complex correlation structures, and are costly and difficult to obtain. In such settings, Bayesian trial designs are attractive since they can incorporate historical data or information from published literature, thus saving time and expense and minimizing the number of subjects exposed to an inferior treatment. Bayesian designs can also adapt to unexpected changes in the protocol, and allow the investigator to explore the plausibility of various outcome scenarios before any patients are enrolled in the trial. Recently, the FDA Center for Devices has encouraged hierarchical Bayesian statistical approaches which allow for the incorporation of such valuable historical data into the design and analysis of new device trials. The practical application of these methods has only become feasible in the last decade due to advances in computing via Markov chain Monte Carlo (MCMC) methods, especially as implemented in the popular BUGS software package. In this paper we illustrate Bayesian analysis and sample size calculations using BRugs, a function for calling BUGS from R. We provide illustrations in two applied settings where incorporation of available historical information is crucial, one concerning an AIDS drug trial and the other a comparison of left ventricular assist devices (LVADs). PMID- 18161543 TI - Experience with reviewing Bayesian medical device trials. AB - The purpose of this paper is to present a statistical reviewer's perspective on some technical aspects of reviewing Bayesian medical device trials submitted to the Food and Drug Administration. The discussion reflects the experiences of the authors and should not be misconstrued as official guidance by the FDA. A variety of applications are described, reflecting our experience with therapeutic and diagnostic devices. In addition to Bayesian analysis of trials, Bayesian trial design and Bayesian monitoring are discussed. Analyses were implemented in WinBUGS (http://www.mrc-bsu.cam.ac.uk/bugs/winbugs/contents.shtml), with the code provided. PMID- 18161544 TI - An adaptive approach to designing comparative diagnostic accuracy studies. AB - Comparative diagnostic studies usually involve comparison of the area under receiver operating characteristic curves when biomarkers are measured on a continuous or ordinal scales. In designing such studies, specification of a number of nuisance parameters is often required to compute sample sizes. When these parameters are incorrectly specified, statistical power to detect a meaningful difference in area can be substantially adversely affected. We propose an adaptive method to calculate the sample size and show these procedures to be effective in controlling error rates. PMID- 18161545 TI - Overview of agreement statistics for medical devices. AB - This paper is an overview that summarizes recently developed tools in assessing agreement for methods comparison and instrument/assay validation in medical devices. This paper emphasizes concept, sample sizes, and examples more than analytical formulas. We have considered a unified approach of evaluating agreement among multiple instruments (k), each with multiple replicates (m) for both continuous and categorical data. We start with the basic scenario of two instruments (k = 2), each with only one measurement (m = 1). In this basic scenario for continuous data, we also consider if the target values are considered random (values of a gold standard instrument) or fixed (known values). In the more general case, we will not consider when the target values are fixed. We discuss the simplified sample size calculations. When there is a disagreement between methods, one needs to know if the source of the disagreement was due to a systematic shift (bias) or random error. The coefficients of accuracy and precision will be discussed to characterize these sources. This is important because a systematic shift usually can be easily fixed through calibration, while a random error usually is a more cumbersome variation reduction exercise. For categorical variables, we consider scaled agreement statistics. For continuous variables, we use scaled or unscaled agreement statistics. For variables with proportional error, we can simply apply a log transformation to the data. Finally, three examples are given: one for assay validation, one for a lab proficiency assessment, and one for a lab comparison on categorical assay. PMID- 18161546 TI - Comparing two medical tests when results of reference standard are unavailable for those negative via both tests. AB - In studies for comparing the diagnostic accuracy of two qualitative tests, very often the reference standard to confirm the disease status is not applied to all study subjects. We considered a situation when all subjects with a positive result by at least one of the tests had a verified disease status and none of the subjects with both tests negative results had a verification of disease status. In this paper, we discuss whether the information about the ratio of true positive rates and the ratio of false positive rates of two qualitative tests, T(New) and T(Old), is sufficient to draw a conclusion about effectiveness of the T(New). We show that if there is a statistically significant increase in true positive rates and the increase in true positive rates is statistically larger than increase in false positive rates, then a conclusion about effectiveness of test T(New) can be made and this does not require application of the reference standard to the subjects with negative results by both tests. An application of ratio of true positive rates and ratio of false positive rates to post-market studies is also presented. PMID- 18161547 TI - Statistical issues on the diagnostic multivariate index assay for targeted clinical trials. AB - In the past decade, pharmacogenomics and microarrays are considered two of the most important scientific breakthroughs for detection and treatment of diseases with many other applications. After completion of the Human Genome Project (HGP), the importance of diagnostic tests for identification of molecular targets increases as more targeted clinical trials are conducted for the individualized treatment of patients in the post-genomic era. As a result, the co-development of drug-device has become the foundation for achieving the ultimate goal of personalized medicine. One of the diagnostic devices for detection of molecular targets is the in vitro diagnostic multivariate index assays (IVDMIA) based on the genomic composite biomarker (GCB) classifiers. Thus, the quality of the IVDMIA, innovative designs, and the evaluation of efficacy for targeted clinical trials are vital for achieving this goal. However, before personalized medicine becomes a reality, many challenges for assurance of the accuracy and precision of the IVDMIA and the estimates of the treatment effect in the population with the molecular targets are to be resolved. In this paper, we identify the following issues on the IVDMIA and targeted clinical trials: (i) the selection of the differentially expressed genes, (ii) the optimal representation and algorithm for the genomic composite biomarker (GCB) classifier for the best diagnostic accuracy of the molecular target, (iii) the validation of the IVDMIA, and (iv) the evaluation of effectiveness and sample size estimation for targeted clinical trials. For each issue, the problem and possible resolutions are discussed. An overall assessment and some concluding remarks are also provided. PMID- 18161548 TI - Challenges in DNA microarray studies from the regulatory perspective. AB - Genomic classifiers using DNA microarrays are becoming powerful tools in the medical community with the potential to revolutionize the diagnosis and treatment of disease. However, despite the tremendous interest in using these classifiers in diagnosis and the management of disease, few genomic classifiers have made it into clinical practice. Some of the major challenges for the development and validation of genomic classifiers will be discussed in this article together with some of their difficulties. PMID- 18161552 TI - Effect of soil depth on phytoremediation efficiency for petroleum contaminants. AB - Biodegradation of organic contaminants in soil may be enhanced by the presence of vegetation. Evaluating the effect of soil depth on phytoremediation efficiency may provide researchers and regulators with a clearer understanding of contaminant clean-up. A column study with polycyclic aromatic hydrocarbons (PAHs) and diesel-contaminated soil was conducted over a 147-day period of switchgrass (Panicum virgatum) growth. Analysis of the contaminants and plant biomass was conducted along with microbial enumeration at three soil depths in 49-day intervals. Remediation proceeded rapidly near the surface of the soil (0-20 cm) for both vegetated and unvegetated columns, but the effect of vegetation relative to an unvegetated control only was significant in the lower soil depths. Contaminant dissipation in the 20-40 and 40-60 cm layers was not significantly different between vegetated and unvegetated soil. PMID- 18161553 TI - Application of hydroxypropyl[beta]cyclodextrin to evaluation of polycyclic aromatic hydrocarbon losses during sewage sludges composting. AB - The aims of the present study were to test the application of the non-exhaustive extraction technique (NEET) with hydroxypropyl[beta]cyclodextrin (HPCD) for the evaluation of the content of potentially bioavailable polycyclic aromatic hydrocarbon (PAH) fraction as well as the evaluation of changes in the content of this fraction during composting of municipal sewage sludges. A possibility to apply this method for the forecasting of PAH losses during composting was also studied. Four sludges (from different sewage treatment plants) with varied properties have been selected for the present experiment. The sludges were composted for 76 days. PAH content was determined as extracted with dichloromethane (exhaustive extraction technique) and potentially bioavailable PAH content by means of HPCD (non-exhaustive extraction technique) in both sewage sludges and composts obtained from them. The content of fraction extracted with HPCD ranged from 63.5 to 83.1% depending on the sludge. After composting, a significant lowering of this fraction contribution was noted for 3 sludges. However, in the composts obtained, the fraction extracted with HPCD still constituted 54.7-62.1% of the total PAH content. The composting process clearly influenced a decrease in the bioavailable fraction in the case of 3- and 5-ring PAHs. However, the 4-ring PAHs content of bioavailable fraction did not undergo any significant changes. No satisfactory results were obtained when HPCD was used as a tool to forecast PAH losses. In a number of cases only, PAH remaining after HPCD extraction achieved a value similar to that of PAH after the composting process. In the case of a number of PAH, the method with HPCD application changed the values of the bioavailable fraction. PMID- 18161554 TI - Feasibility of using microbial fuel cell technology for bioremediation of hydrocarbons in groundwater. AB - A single-cell microbial fuel cell (MFC) design was used to study anaerobic microbes that utilize petroleum contaminants as a sole substrate to produce power during remediation. Additionally, we tested various proton bridge designs to physically separate the anode and cathode chambers of a two-cell MFC by approximately 9 m (approximately 30 ft.). This separation enables the potential use of MFC technology for in situ bioremediation of petroleum hydrocarbons in the groundwater, in which oxygen is usually depleted and oxygen availability only exists at or near the surface. Sustained power generation (as high as 120 mW/m(2) cathode) was recorded for approximately 6 d in a single-cell MFC utilizing a mixture of refinery waste (containing various concentrations of hydrocarbon contaminants) and cell growth media. MFC cell potential (1KOmega external resistance) decreased by approximately 55% over the length of the 9 m proton bridge with a 6.9% decrease in potential per m of bridge. This preliminary data indicates that using MFC technology (with our modifications) may enhance bioremediation of petroleum contaminants in groundwater under anaerobic conditions. Because oxygen is eventually used as the terminal electron acceptor for anaerobic biodegradation inside an MFC, this technology may be a cost effective innovation to enhanced biodegradation in groundwater, by substituting or eliminating conventional in situ aeration. To our knowledge, this is the first report on power generation from MFCs utilizing mixed hydrocarbon substrates. In addition, this study is the first to show the applicability of using extended proton bridges for the physical separation of anode and cathode chambers over extended distances that may be encountered in the field. PMID- 18161555 TI - Remediation of TCE-contaminated groundwater by a permeable reactive barrier filled with plant mulch (Biowall). AB - A pilot-scale permeable reactive barrier filled with plant mulch was installed at Altus Air Force Base in Oklahoma, USA to treat trichloroethylene (TCE) contamination in groundwater emanating from a landfill. The barrier was constructed in June 2002. It was 139 meters long, 7 meters deep, and 0.5 meters wide. The barrier is also called a Biowall because one of the mechanisms for removal of TCE is anaerobic biodegradation. This study aimed at evaluating the performance of the pilot-scale Biowall after its installation. Data from over four years' monitoring indicated that the Biowall greatly changed geochemistry in the study area and stimulated TCE removal. The concentration of TCE in the Biowall and downgradient of the Biowall was greatly reduced as compared to that in ground water upgradient of the Biowall, while the concentration of cis-DCE in the Biowall and downgradient of the Biowall was much higher than that observed upgradient of the Biowall. Over time, the concentration of vinyl chloride in the Biowall and downgradient of the Biowall increased. Dehalococcoides DNA was detected within and downgradient of the Biowall, corresponding to the observation that vinyl chloride was produced at these locations. Results from a tracer study indicated that the regional groundwater flow pattern ultimately determined the flow direction in the area around the Biowall. The natural groundwater velocity was estimated at an average of 0.060 +/- 0.015 m/d. PMID- 18161556 TI - Phytotoxicity of oil sands naphthenic acids and dissipation from systems planted with emergent aquatic macrophytes. AB - Differences in dissipation and phytotoxicity were measured for two naphthenic acid mixtures in hydroponically grown emergent macrophytes (Typha latifolia, Phragmites australis, and Scirpus acutus). One of the naphthenic acid (NA) mixtures was extracted from tailings pond water of an oil sands operation in Fort McMurray, Alberta, Canada. The other mixture was a commercially available NA mixture. While the oil sands NA mixture was less phytotoxic to wetland plants compared to the commercially available NA mixture, they were not sequestered by wetland plants like their commercial NA counterparts. The small loss of commercial NAs from the spiked hydroponic system appeared to be selective and dependant on the specific NA compound. The results of this study indicate that plants alone may not mitigate NAs from oil sands tailings pond water. In addition, caution should be taken when making predictions on the environmental fate of oil sands naphthenic acids when using commercial NAs as surrogates. PMID- 18161557 TI - Time-dependence in mixture toxicity with soft-electrophiles: 2. Effects of relative reactivity level on time-dependent toxicity and combined effects for selected Michael acceptors. AB - Toxicity assessments for organic chemical mixtures are often described as being approximately additive. Recent mixture studies with soft electrophiles have suggested that agents with less-than fully time-dependent toxicity (TDT) may actually induce toxicity by more than one mode of toxic action within the same series of concentrations. To evaluate this concept further, four Michael acceptor electrophiles, each with a different rate of in chemico reactivity and different level of TDT, were tested with each other and in sham combinations (a single chemical tested as if it were a binary mixture) using the Microtox system. For each binary combination, each agent was tested alone and in a mixture, with toxicity assessed as inhibition of bioluminescence at 15-, 30- and 45-min of exposure. Each single agent and mixture test included seven duplicated concentrations and a duplicated control treatment. To evaluate relative reactivity, each agent was also tested with the model nucleophile glutathione (GSH). Agents with greater in chemico reactivity (mean RC(50) mM) showed greater toxicity (mean 45-min EC(50) - mM) but these were inversely related to the TDT levels of the agents. Combined effects for the sham combinations, as quantified by additivity quotient values for the EC(50) of the mixture, tended to be close to 1.00 (i.e., the dose-addition EC(50)-AQ). For true binary combinations (i.e., two chemicals tested together), the EC(50)-AQ tended to be increasingly above 1.00 when TDT levels of the agents in the mixture were more disparate. The results of this study with Michael acceptors suggested that: (i) when reactivity was fast, there was most likely a single prominent mode of toxic action, i.e., electro(nucleo)philic reactivity, leading to time-dependent toxicity at the full or high levels, (ii) when the reaction rate for a chemical was slower, two modes of action, electro(nucleo)philic reactivity and narcosis, were apparent such that the time-dependent toxicity level was lower as well, (iii) mixtures of the former agents show a combined effect that was strictly dose-additive, whereas (iv) mixtures which included one (or more) agent with a lower reaction rate had a combined effect that was approximately additive rather than strictly dose additive. PMID- 18161558 TI - Removal characteristics of endocrine-disrupting chemicals by laccase from white rot fungi. AB - Laccase from 5 white-rot fungal strains (4 Trametes and 1 Pycnoporus strains) were evaluated in the removal spectra with/without mediators against 11 EDCs. Purified laccase from Trametes sp. was also used to reveal the precise degradation spectra and degradation profiles in time course against 20 EDCs with/without mediators. In addition, effectivity of laccase for the purification of complex EDCs contamination was evaluated combining several EDCs. The removal characteristics among tested strains were almost the same and crude/purified laccase could remove various EDCs. Bis(4-hydroxyphenyl)sulfone, diethylhexylphthalate (DEHP), pyrene (PY), anthracene, 3,5-dichlorophenol and pentachlorophenol could not be removed by laccase. DEHP and PY could not be removed even with mediators. Vanillin and vanillic acid revealed to be possible naturally occurring mediators. Laccase-mediator system could expand the degradation spectrum and enhance the EDCs removal ratio and rate. When complex mixtures of EDCs were treated with laccase, the removal ratio was enhanced in comparison to that of single application. Some coexisting EDCs could act as mediators. Thus, the availability of laccase and the effectivity of a mediator on EDCs treatment were indicated. PMID- 18161559 TI - Photocatalytic ozonation of phenolic wastewaters: Syringic acid, tyrosol and gallic acid. AB - The photocatalytic ozonation of a mixture of 3 phenols (gallic acid, tyrosol and syringic acid) has been conducted under different operating conditions. The individual adsorption of the phenol type compounds onto titanium dioxide (photocatalyst) has been first evaluated. Equilibrium conditions are attained in less than an hour while the isotherm curves reveal that adsorption intensity increases in order: syringic acid < tyrosol < gallic acid. When the photocatalytic ozonation is applied, an optimum in titanium dioxide concentration is experienced (1.5 g L(-1)). Direct comparison of the photocatalytic ozonation to other less sophisticated oxidation systems (i.e., single ozonation, catalytic ozonation, photo-ozonation, etc.) indicates a higher efficiency of the former in terms of ozone uptake. PMID- 18161560 TI - Laser-enhanced photocatalytic degradation of organic pollutants from water using ZnO semiconductor catalyst. AB - Wastewater contaminated with organic pollutants creates serious environmental problems. Laser induced photocatalysis has been investigated for removal of organic pollutant like phenol from water. The important mechanism for degradation of phenol by UV laser irradiation is redox reactions triggered by semiconductor catalyst in presence of water. In this study the potential of a ZnO semiconductor catalyst has been explored as an effective catalyst for the photodegradation of phenol using a Nd:YAG laser-emitting at 355 nm in UV-radiation in a batch reactor. The influences of different parameters such as catalyst concentration, UV-radiation intensity, and pH on the extent of photo degradation have been investigated. Overall degradation of phenol achieved was 70% after 60 minutes of laser irradiation of 100 ppm phenol solution in water. The reaction rate constant fit a first-order reaction model for phenol degradation and was estimated as 0.0088 min(-1). This study proved that laser induced photocatalytic removal of phenol is highly efficient as compared with the broadband spectral sources such as lamps being applied in majority of setups worldwide. PMID- 18161561 TI - Application of cytogenetic endpoints and comet assay on human lymphocytes treated with atorvastatin in vitro. AB - This study investigated the genotoxic potential of atorvastatin on human lymphocytes using comet assay, structural chromosome aberrations (CA) and sister chromatid exchange (SCE) analysis. Lymphocyte cultures were treated with a single drug at a concentration of 30.21 ng/mL. For comet assay, cells exposed to atorvastatin for 24 h, 48 h and 72 h were embedded in agarose slides, lysed with alkaline lysis solution and exposed to an electric field. DNA migrated within the agarose and formed comets whose length depends on the amount of DNA damage. For analysis of structural CA, cells were grown on medium for 48 h and for SCE analysis for 72 h. Structural CA did not induce significant damage to the genome, although a higher CA frequency was observed in cells treated with atorvastatin for 3 h, 20 h and 48 h than in control samples. Results of the SCE analysis did show statistically significant differences in the mean SCE number between atorvastatin-exposed and control human lymphocytes and between different exposure times. Comet assay also showed increased DNA damage caused in atorvastatin exposed human lymphocytes than in corresponding control cells for exposure times of 24 h, 48 h and 72 h for the tail length and for 72 h for the tail moment. Results obtained in this study point to the significance of biological indicators providing information on the primary genome damage after long-term exposure, which can help to establish drug therapeutic concentrations that do not put patients with high blood cholesterol to a greater treatment-related risk. PMID- 18161562 TI - The potential and realistic hazards after a solar-driven chemical treatment of benzene using a health risk assessment at a gas station site in Korea. AB - In order to evaluate the potential use for ex situ remediation, a solar-driven, photocatalyzed reactor system was constructed and applied for the treatment of groundwater contaminated with benzene using selected advanced oxidation processes (AOP) processes, such as H(2)O(2)/solar light, TiO(2) slurry/solar light and immobilized TiO(2)/solar light. However; to date, there have been few attempts to characterize the potential impact of residual levels of benzene on human health after treatment. Some papers have focused on the application of treatment methods of benzene, but most have not considered the effects of realistic hazards and human health. Therefore, potential and realistic hazards of benzene to human health were investigated at a gas station site using a risk-based assessment approach. Among the different remediation actions, the solar light/TiO(2) slurry/H(2)O(2) system (Action 5) showed higher removal efficiency than the solar light/TiO(2) slurry (Action 3) and the solar light/immobilized TiO(2) (Action 2) systems for the treatment of benzene. The Action 5 remediation method achieved 98% degradation, and lead to a substantial increase in the removal of benzene due to the synergetic effect of TiO(2) with the oxidant, H(2)O(2). Also, using the realistic and potential hazard assessment instead of the point estimation of concentration after benzene treatment, the total health risk exceeded the target risk value (1 x 10(-6)). However, the 95th percentile target cancer risk, found using a probabilistic analysis (Monte Carlo method), was around 1 x 10(-6), indicating a low potential carcinogenic risk. Therefore, it was concluded that no adverse health risk was unlikely to be posed if the Action 5 system, which included the addition of TiO(2) and H(2)O(2), or if an increased reaction time was applied. In addition, continuous efforts and proper actions must be taken on the "Soil and Groundwater Remediation Action" based on the risk assessment in Korea. PMID- 18161563 TI - Dissolved VOC concentrations and salt contents affecting air-sweat equilibrium partition of hydrophilic and hydrophobic VOCs. AB - Workers have frequently disregarded long-term dermal exposure to low concentration of gaseous volatile organic compounds (VOCs). To assess dermal exposure risk to gaseous VOCs, equilibrium partitioning coefficients (p(c)) at the air-sweat interface on human skin surface must be examined. This study analyzed the p(c) values of hydrophilic iso-propanol (IPA), methyl ethyl ketone (MEK), and hydrophobic benzene, toluene, ethylbenzene, o-xylene, m-xylene, and p xylene (BTEXs) at the air-water and air-sweat interfaces at 27-47 degrees C. The hydrophilic VOCs were dissolved in pure water and artificial human sweat liquors at approximately 10-125 mg/L, and hydrophobic VOCs were at approximately 0.55 mg/L. According to experimental results, the dissolved VOC concentration and salt contents simultaneously have a co-effect on p(c) during human dermal exposure to gaseous VOCs. The salt effect resulted in increase of p(c) for hydrophilic and hydrophobic VOCs, and the dissolved VOC concentration effect resulted in a reduction in p(c), which is dominant for hydrophilic compounds of high concentrations of aqueous VOCs. The p(c) data were utilized for further assessment of risk due to dermal exposure to VOCs. PMID- 18161564 TI - Use of dewatered alum sludge as a substrate in reed bed treatment systems for wastewater treatment. AB - In this paper, two laboratory-scale simulated reed beds were investigated for the purpose of assessing the feasibility and effectiveness of using dewatered alum sludge as a possible substrate for wastewater treatment reed bed systems. One horizontal subsurface flow setup and one vertical flow setup were used. The horizontal flow system was planted with Phragmites australis while the vertical flow system was left unplanted. Thus, the latter was more akin to a sand filter system, but was examined with the potential use as a planted vertical reed bed system. The influent source used was farmyard wastewater. It is expected that the present study will provide the basis for long-term and large-scale trials in realizing the concept of integrating "waste" into treatment processes. Results obtained so far have shown that the dewatered alum sludge holds great promise as a low-cost resource media for use in reed bed treatment systems. Appreciable and stable performance was obtained during the continuous operation at high hydraulic, organic and phosphorus loadings. In particular, both wetlands achieved over 90% phosphorus removal, reflecting the significant advantage of this novel approach over conventional reed bed treatment systems. However, extensive research into possible surface clogging and possible release of some substances from the sludge to the treated effluent is necessary to ensure reliability of the system. This will help to make the alum sludge-based reed bed environmentally and economically justifiable. PMID- 18161565 TI - Comments on "Removal of aluminium by constructed wetlands with water hyacinth (Eichhornia crassipes (Mart.) Solms) grown under different nutritional conditions". PMID- 18161567 TI - Transformation kinetics of 6-methoxybenzoxazolin-2-one in soil. AB - Wheat (Triticum aestivum L.) and other cereals produce allelochemicals as natural defense compounds against weeds, fungi, insects and soil-borne diseases. The main benzoxazinoid allelochemical of wheat is 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3 one (DIMBOA), bound as beta-glucoside and released upon plant injury. When leached from wheat to soil, DIMBOA is microbially transformed to 6-methoxy benzoxazolin-2-one (MBOA). Exploiting benzoxazinoids and their degradation products as substitutes for synthetic pesticides depends on knowledge of transformation pathways and kinetics. In an MBOA degradation experiment at a concentration of 2400 nmol g(-1) soil, the previously identified transformation products 2-amino-7-methoxy-phenoxazin-3-one (AMPO) and 2-acetylamino-7-methoxy phenoxazin-3-one (AAMPO) were quantified. Three different kinetic models were applied to MBOA transformation kinetics; single first-order (SFO), first-order multi-compartment, and double first-order in parallel. SFO proved to be adequate and was subsequently applied to the transformations of MBOA, AMPO and AAMPO. Degradation endpoints, expressed as degradation time (DT), were calculated for MBOA, AMPO and AAMPO to test whether the maximum values for synthetic pesticides set by the European Commission and the Danish Environmental Protection Agency were exceeded. DT(50) values for MBOA and AMPO were 5.4 d and 321.5 d, respectively, and DT(90) values were 18.1 d and 1068 d, respectively. The DT(50) value for AMPO exceeded the maximum value. The persistence, concentrations and toxicity of metabolites such as AMPO should be considered when breeding cereal crops with increased levels of benzoxazinoids. PMID- 18161568 TI - Fate in soil of 14C-sulfadiazine residues contained in the manure of young pigs treated with a veterinary antibiotic. AB - The fate of (14)C-labeled sulfadiazine ((14)C-SDZ) residues was studied in time course experiments for 218 days of incubation using two soils (A(p) horizon of loamy sand, orthic luvisol; A(p) horizon of silt loam, cambisol) amended with fresh and aged (6 months) (14)C-manure [40 g kg(-1) of soil; 6.36 mg of sulfadiazine (SDZ) equivalents per kg of soil], which was derived from two shoats treated with (14)C-SDZ. Mineralization of (14)C-SDZ residues was below 2% after 218 days depending little on soil type. Portions of extractable (14)C (ethanol water, 9:1, v/v) decreased with time to 4-13% after 218 days of incubation with fresh and aged (14)C-manure and both soils. Non-extractable residues were the main route of the fate of the (14)C-SDZ residues (above 90% of total recovered (14)C after 218 days). These residues were high immediately after amendment depending on soil type and aging of the (14)C-manure, and were stable and not remobilized throughout 218 days of incubation. Bioavailable portions (extraction using CaCl(2) solution) also decreased with increasing incubation period (5-7% after 218 days). Due to thin-layer chromatography (TLC), 500 microg of (14)C-SDZ per kg soil were found in the ethanol-water extracts immediately after amendment with fresh (14)C-manure, and about 50 microg kg(-1) after 218 days. Bioavailable (14)C-SDZ portions present in the CaCl(2) extracts were about 350 microg kg(-1) with amendment. Higher concentrations were initially detected with aged (14)C manure (ethanol-water extracts: 1,920 microg kg(-1); CaCl(2) extracts: 1,020 microg kg(-1)), probably due to release of (14)C-SDZ from bound forms during storage. Consistent results were obtained by extraction of the (14)C-manure-soil samples with ethyl acetate; portions of N-acetylated SDZ were additionally determined. All soluble (14)C-SDZ residues contained in (14)C-manure contributed to the formation of non-extractable residues; a tendency for persistence or accumulation was not observed. SDZ's non-extractable soil residues were associated with the soluble HCl, fulvic acids and humic acids fractions, and the insoluble humin fraction. The majority of the non-extractable residues appeared to be due to stable covalent binding to soil organic matter. PMID- 18161569 TI - Atrazine sorption by hydroxy-interlayered clays and their organic complexes. AB - This study examined the sorption of atrazine by hydroxy-Fe interlayered montmorillonite (FeMt) and its hydroquinone (FeMtHQ), citrate (FeMtCt) and catechol (FeMtCC) complexes as well as by hydroxy-Al interlayered montmorillonite (AlMt) and its hydroquinone (AlMtHQ) and citrate (AlMtCt) complexes. Found among the clays were sorption distribution coefficients (K(d)) ranging from 24 to 123 mL g(-1) and maximum sorption (M) ranging from 2.2 to 16.8 microg g(-1). Both K(d) and M decreased in the order of FeMtCC > FeMtHQ > AlMtHQ > (AlMt = FeMt) > (AlMtCt = FeMtCt). The pH was negatively correlated with both K(d) (r = -0.90, p < 0.001) and M (r = -0.81, p < 0.001). When interlayered clays were associated with humified material (FeMtCC, FeMtHQ, AlMtHQ), both K(d) (r > 0.96, p < 0.01) and M (r > 0.94, p < 0.01) were highly positively correlated with total organic C and alkali-soluble C. However, clays with non-humified organic compounds (FeMtCt and AlMtCt) sorbed less atrazine than clays without any organic C (FeMt and AlMt). This suggests that functional groups of Fe-OH and Al-OH in FeMt and AlMt reduced the available sorption sites for atrazine by making complexes with citrate ions while forming FeMtCt and AlMtCt. The atrazine was sorbed through the hydrophobic interactions with organic compound surfaces as well as through H bonding and ionic bonding with clay-mineral surfaces. PMID- 18161570 TI - Degradation of atrazine by an acclimatized soil fungal isolate. AB - A fungal strain able to use atrazine (2-chloro-4-ethylamino-5-isopropylamino 1,3,5-triazine) as a source of nitrogen was isolated from a corn field soil that has been previously treated with the herbicide. This strain was purified and acclimatized to atrazine at a higher level in the laboratory. A supplemented N was required to trigger the reaction. Atrazine was degraded at a faster rate in inoculated mineral salt medium (MSM) than non-inoculated MSM. Within 20 days, nearly 34% of the atrazine was degraded in inoculated medium while only 2% of the herbicide was degraded in non-inoculated medium. Degradation of atrazine by the isolated fungal strain was also studied in sterile and non-sterile soil to determine the compatibility of the isolated strain with native microorganisms in soil. The degradation of atrazine was found to be more in inoculated sterile soil than in inoculated non-sterile soil. Cell free extract (CFE) of fungal mycelium degraded about 50% of the atrazine in buffer in 96 hours compared to the control. Four atrazine metabolites were isolated and characterized by LCMS. On the basis of morphological parameters the isolate was identified as Penicillium species. Results indicated that the microorganism may be useful for remediation of atrazine-contaminated soil. PMID- 18161571 TI - 2,4-D mineralization in unsaturated and near-saturated surface soils of an undulating, cultivated Canadian prairie landscape. AB - The herbicide 2,4-D [2,4-(dichlorophenoxy) acetic acid] is one of the most widely used pesticides in the Canadian prairies and is frequently detected as a ground and surface water contaminant. The objective of this paper was to determine the magnitude and extent of variation of 2,4-D mineralization in a cultivated undulating prairie landscape. Microcosm incubation experiments, using a 4 x 3 x 2 factorial experimental design (soil moisture, 4 levels: 60, 85, 110, 135% of field capacity; slope position, 3 levels: upper-, mid- and lower-slopes; soil depth, 2 levels: 0-5 and 5-15 cm), were used to assess 2,4-D mineralization. The first-order mineralization rate constant (k(1)) varied from 0.03 to 0.22 day(- 1), while total 2,4-D mineralization varied from 31 to 52%. At near-saturated conditions (110 and 135% of field capacity), the onset of 2,4-D degradation was delayed in soil obtained from the upper- and mid-slopes but not in soils obtained from the lower-slope position. The k(1) and total 2,4-D mineralization was significantly influenced by all three factors and their interactions. The Freundlich sorption coefficient of 2,4-D ranged from 0.83 to 2.46 microg (1 1/n)g(- 1) mL(1/n) and was significantly influenced by variations in soil organic carbon content across slope positions. The infield variability of 2,4-D sorption and mineralization observed across slope positions in this undulating field was comparable in magnitude and extent to the regional variability of 2,4-D sorption and mineralization observed in surface soils across Manitoba. The large variability of 2,4-D mineralization and sorption at different slope positions in this cultivated undulating field suggests that landform segmentation models, which are used to delineate slope positions, are important considerations in pesticide fate studies. PMID- 18161572 TI - Development of enzyme immunoassay for detection of DDT. AB - Dichlorodiphenyltrichloroethane (DDT) is one of the persistent organic pollutants (POPs) widely found in the environment and in the general population. In this study, a direct competitive enzyme immunoassay (EIA) has been developed for the quantitative analysis of DDT. To generate a specific polyclonal antibody for EIA, p, p'-DDT was conjugated to porcine thyroglobulin for rabbit immunization. At optimized EIA conditions, the standard curves ranged from 0.137 to 100 ng/mL with the quantification limit of 0.41 ng/mL. The coefficients of variation (CV%) were 5.42-10.53% for intra-assay and 6.04-7.26% for inter-assay. Cross-reactivities with DDT metabolites (DDTs, including o, p'-DDT, p, p'-DDD, o, p'-DDD, p, p'-DDE, o, p'-DDE, p, p'-dichlorobenzophenone (DCBP), o, p'-DCBP) were investigated. The polyclonal antibody showed relatively low and/or no cross-reactivity with these compounds, and the assay was seen to be highly selective for p, p'-DDT. Moreover, the DDTs could be ranked by their reactivity: DDT > DDD > DDE > DCBP. In addition, the characterization of the polyclonal antibody indicated that the antiserum possesses a high specificity for p, p'-isomers. The results indicated that the developed EIA using this antibody could be a convenient and supplemental analytical tool for monitoring DDT. PMID- 18161573 TI - Effects of the anti-fouling herbicide Irgarol 1051 on two life stages of the grass shrimp, Palaemonetes pugio. AB - This study investigated lethal and sublethal effects (glutathione, lipid peroxidation, cholesterol, and acetylcholinesterase) of the anti-fouling herbicide Irgarol 1051 on larval and adult grass shrimp (Palaemonetes pugio). The 96-hour LC50 test for larvae resulted in an estimated LC50 of 1.52 mg/L (95% confidence interval [CI] 1.26-1.85 mg/L). The adult 96-h LC50 was 2.46 mg/L (95% CI = 2.07-2.93 mg/L). Glutathione, lipid peroxidation, cholesterol and acetylcholinesterase levels were not significantly affected in adult grass shrimp by exposure of up to 3.00 mg/L irgarol. Lipid peroxidation and acetylcholinesterase levels in the larvae were significantly higher than controls in the highest irgarol exposures of 1.0 and 2.0 mg/L, respectively. Cholesterol levels were significantly reduced in larvae in all four irgarol concentrations tested while glutathione levels were not significantly affected in larvae. Both lethal and sublethal effects associated with irgarol exposure were only observed at concentrations well above those reported in the environment. PMID- 18161574 TI - Aquatic toxicity of cartap and cypermethrin to different life stages of Daphnia magna and Oryzias latipes. AB - Cartap and cypermethrin, which are among the most widely used pesticides in many countries, are considered safe because of their low mammalian toxicity and their low persistence in the environment. However, recent findings of endocrine disrupting effects and developmental neurotoxicity have raised concerns about the potential ecological impacts of these pesticides. We evaluated the aquatic toxicity of cartap [S,S'-(2-dimethylaminotrimethylene) bis(thiocarbamate), unspecified hydrochloride] and cypermethrin [(RS)-alpha-cyano-3-phenoxybenzyl (1RS,3RS,1RS,3SR)-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylate], both individually and combined, on different life stages of the freshwater cladoceran Daphnia magna and a freshwater teleost, Japanese medaka (Oryzias latipes). The 96-hr Daphnia median effective concentrations (EC50s) for cartap and cypermethrin were 91.0 microg/L and 0.00061 microg/L, respectively. Rapid recovery of Daphnia was observed after short-term pulsed exposure to cartap and cypermethrin; there were no adverse effects on reproduction or survival 20 d after a 24 hr exposure to cartap up to 1240 microg/L and cypermethrin up to 1.9 microg/L. Chronic continuous exposure (for 21 d) of 7-d-old Daphnia to cypermethrin significantly reduced the intrinsic population growth rate in a concentration-dependent manner. However, because the intrinsic population growth rates were all above zero, populations did not decrease even at the highest experimental concentration of 200 ng/L. Exposure of Daphnia neonates (< 24 hr old) to cypermethrin for 21 d caused significant, sub-lethal reproduction-related problems, such as increased time to first brood, reduced brood size, and reduced total brood number, at 0.0002, 0.002, and 0.2 ng/L cypermethrin, but the intrinsic population growth rate was not significantly affected. Oryzias latipes was relatively more resistant to both pesticides. In particular, embryos appeared to be more resistant than juveniles or adults, which may be partly due to the protective role of the chorion. The incidence of larval fish deformity was significantly higher after a 96 hr exposure to as low as 250 microg/L of cartap or 40 microg/L of cypermethrin. The mixture of both compounds showed no synergistic toxicity. The extremely high acute-to-chronic ratio suggests that the standard acute lethal toxicity assessment might not reflect the true environmental hazards of these frequently used pesticides. Ecological hazard assessments of long-term low dose or pulsed exposures to cartap and cypermethrin may reveal more realistic consequences of these compounds in surface water. PMID- 18161575 TI - Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.). AB - The aim of this study was to characterize biomarker responses in three-spined sticklebacks exposed to prochloraz (Pcz). For this purpose, adult sticklebacks were exposed for 2 weeks to prochloraz at 0, 10, 50, 100 and 500 microg/L prior to one week of depuration in clean water. At days 7, 14 and 21, several hepatic biomarkers were measured including 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), total glutathione (GSH) content and thiobarbituric acid reactive substances (TBARS). Pcz induced a transient increase of antioxidant enzymes and a depletion of glutathione content during the first 7 days of exposure. This study showed that EROD activity and antioxidants were disrupted in a transient manner. GST was rapidly induced in a dose-dependent manner and this induction was persistent and observed also after depuration. GST appeared as a valuable biomarker to assess the exposure to Pcz. PMID- 18161576 TI - Determination of cytotoxicity of nephrotoxins on murine and human kidney cell lines. AB - The present study investigates renal inner medullary collecting duct (mIMCD3) cells and human embryonic kidney cells (HEK293) for evaluation of cytotoxicity of nephrotoxic compounds. The 24 h LC(50) values for cisplatin, paraquat and ibuprofen in mIMCD3 cells were 135, 155 and 3600 microM, respectively. The 24 h LC(50) values for paraquat and ibuprofen in HEK293 cells were 180 and 1000 microM, respectively. Effects of hyperosmolality on cytotoxicity of paraquat were additive in mIMCD3 cells. These data demonstrate that renal hyperosmolality has an additive effect on cytoxicity of paraquat. PMID- 18161577 TI - Alteration of acetylcholinesterase activity in Semele solida (MOLLUSCA: SEMELIDAE) as a biochemical response to coastal anthropogenic impact. AB - Acetylcholinesterase (AChE) activity was analyzed as a molecular marker indicative of exposure to organophosphorus pesticide residues in individuals of the endemic clam species Semele solida in selected coastal locations of Chile's VIII Region. AChE activity was assayed in clams from (i) Penco Beach near the mouth of the Andalien River (Concepcion Bay), (ii) Lenga Beach, near the mouth of the Lenga Estuary (San Vicente Bay), and (iii) Coliumo Beach, near the mouth of the Coliumo Estuary (Coliumo Bay). We also analyzed variations in protein content of clam hemolymph, and variability in the activity of AChE in relation to the sizes of the individual clams sampled. Collection of the clams was done using routine methods, during the spring of 2005, the period during which the use of pesticides is typically intensified in the surrounding forestry and agriculture. The results showed no significant correlation of AChE activity with either the size of the clam, or with the concentration of proteins in the hemolymph. The lowest AChE activity was observed in clams from mouth of the Andalien River (187.5 +/- 34.9 Umin(-1)) which was significantly less than that measured in specimens collected near the mouth of the Coliumo Estuary and the mouth of the Lenga Estuary. A very close relation was observed between the degree of anthropogenic inputs, ocean dynamics, and alterations in AChE activity in S. solida. This clam appears to be a useful indicator species, and AChE activity a sensitive marker for the presence of xenobiotics. PMID- 18161578 TI - The conversion of chicken manure to bio-oil by fast pyrolysis. III. Analyses of chicken manure, bio-oils and char by Py-FIMS and Py-FDMS. AB - Fast pyrolysis of chicken manure produced the following three fractions: bio-oil Fraction I, bio-oil Fraction II, and a char. In a previous investigation we analyzed each of the four materials by curie-point pyrolysis-gas chromatography/mass spectrometry (CpPy-FDMS). The objective of this article is to report on the analyses of the same chicken manure and the three fractions derived from it by fast pyrolysis. We now used pyrolysis-field ionization mass spectrometry (Py-FIMS) to characterize the three fractions. In addition, the two bio-oil materials were analyzed by pyrolysis-field desorption mass spectrometry (Py-FDMS). The use of both Py-FIMS and Py-FDMS produced signals over significantly wider mass ranges than did CpPy-GC/MS, and so allowed us to identify considerably larger numbers of constituents in each material. Individual compounds identified in the mass spectra were classified into the following twelve compound classes: (a) low molecular weight compounds (< m/z 62); (b) carbohydrates; (c) phenols + lignin monomers; (d) lignin dimers; (e) n alkylbenzenes; (f) N-heterocyclics; (g) n-fatty acids; (h) n-alkanes; (i) alkenes; (j) sterols; (k) n-diols and (l) high molecular weight compounds (> m/z 562). Of special interest were the high abundances of low-molecular weight compounds in the two bio-oils which constituted close to one half of the two bio oils. Prominent among these compounds were water, ammonia, acetic acid, acetamide, propyl radical, formamide and hydrogen cyanide. The main quantitative differences between the two bio-oils was that bio-oil Fraction I, as analyzed by the two mass spectrometric methods, contained lower concentrations of low molecular weight compounds, carbohydrates, and N-heterocyclics than bio-oil Fraction II but was richer in lignin dimers, n-alkylbenzenes and aliphatics (n fatty acids, n-alkanes, alkenes, and n-diols). Of special interest were the N heterocyclics in the two bio-oils such as pyrazole, pyrazoline, substituted pyrroles, pyridine and substituted pyridines, substituted methoxazole, substituted pyrazines, indole and substituted indoles. Fatty acids in all four materials ranged from n-C(9) to n-C(33), alkanes from n-C(9) to n-C(40), alkenes from C(10:1) to C(40:1) and diols from n-C(7) to n-C(29). The chicken manure, bio oil Fraction I, and char each contained about 4% sterols with cholesterol, ethylcholestriene, ergosterol, ethylcholestene, ethylcholesterol and beta sitosterol as major components. Semi-quantitative estimates of the total materials identified by Py-FIMS were: chicken manure: 61.1%; bio-oil Fraction I: 81.3%; bio-oil Fraction II: 78.6%; char: 61.3%; and by Py-FDMS were: bio-oil Fraction I: 65.4%; bio-oil Fraction II: 70.0%. PMID- 18161579 TI - Vegetative buffers for fan emissions from poultry farms: 2. ammonia, dust and foliar nitrogen. AB - This study evaluated the potential of trees planted around commercial poultry farms to trap ammonia (NH(3)) and dust or particulate matter (PM). Norway spruce, Spike hybrid poplar, hybrid willow, and Streamco purpleosier willow were planted on five commercial farms from 2003 to 2004. Plant foliage was sampled in front of the exhaust fans and at a control distance away from the fans on one turkey, two laying hen, and two broiler chicken farms between June and July 2006. Samples were analyzed for dry matter (DM), nitrogen (N), and PM content. In addition, NH(3) concentrations were measured downwind of the exhaust fans among the trees and at a control distance using NH(3) passive dosi-tubes. Foliage samples were taken and analyzed separately based on plant species. The two layer farms had both spruce and poplar plantings whereas the two broiler farms had hybrid willow and Streamco willow plantings which allowed sampling and species comparisons with the effect of plant location (control vs. fan). The results showed that NH(3) concentration h(- 1) was reduced by distance from housing fans (P < or = 0.0001), especially between 0 m (12.01 ppm), 11.4 m (2.59 ppm), 15 m (2.03 ppm), and 30 m (0.31 ppm). Foliar N of plants near the fans was greater than those sampled away from the fans for poplar (3.87 vs. 2.56%; P < or = 0.0005) and hybrid willow (3.41 vs. 3.02%; P < or = 0.05). The trends for foliar N in spruce (1.91 vs. 1.77%; P = 0.26) and Streamco willow (3.85 vs. 3.33; P = 0.07) were not significant. Pooling results of the four plant species indicated greater N concentration from foliage sampled near the fans than of that away from the fans (3.27 vs. 2.67%; P < or = 0.0001). Foliar DM concentration was not affected by plant location, and when pooled the foliar DM of the four plant species near the fans was 51.3% in comparison with 48.5% at a control distance. There was a significant effect of plant location on foliar N and DM on the two layer farms with greater N and DM adjacent to fans than at a control distance (2.95 vs. 2.15% N and 45.4 vs. 38.2% DM, respectively). There were also significant plant species effects on foliar N and DM with poplar retaining greater N (3.22 vs. 1.88%) and DM (43.7 vs. 39.9%) than spruce. The interaction of location by species (P < or = 0.005) indicated that poplar was more responsive in terms of foliar N, but less responsive for DM than spruce. The effect of location and species on foliar N and DM were not clear among the two willow species on the broiler farms. Plant location had no effect on plant foliar PM weight, but plant species significantly influenced the ability of the plant foliage to trap PM with spruce and hybrid willow showing greater potential than poplar and Streamco willow for PM(2.5)(0.0054, 0.0054, 0.0005, and 0.0016 mg cm(- 2); P < or = 0.05) and total PM (0.0309, 0.0102, 0.0038, and 0.0046 mg cm(- 2), respectively; P < or = 0.001). Spruce trapped more dust compared to the other three species (hybrid willow, poplar, and Streamco willow) for PM(10) (0.0248 vs. 0.0036 mg cm(- 2); P < or = 0.0001) and PM(> 10) (0.0033 vs. 0.0003 mg cm(- 2); P = 0.052). This study indicates that poplar, hybrid willow, and Streamco willow are appropriate species to absorb poultry house aerial NH(3)-N, whereas spruce and hybrid willow are effective traps for dust and its associated odors. PMID- 18161580 TI - The origins of neurogenetics. PMID- 18161581 TI - Tributes to Jeff Hall. PMID- 18161582 TI - Fiber number in the mushroom bodies of adult Drosophila melanogaster depends on age, sex and experience. AB - The mushroom bodies are two characteristically shaped structures of the insect central brain. In Drosophila melanogaster they contain more fibers in females than in males. Within the first week of adult life the total number of fibers increases by about 15% and decreases again in flies older than 3-4 weeks. The number of mushroom body fibers is significantly reduced in flies kept under social isolation or deprived of their antennal input, but not in flies subjected to visual deprivation. PMID- 18161583 TI - A quantitative-genetic analysis of hippocampal variation in the mouse. AB - This report analyses the genetic underpinnings of the proportions of the hippocampal terminal fields in the mouse at the midseptotemporal level. We used 5 inbred strains and all possible F(1) crosses between them (diallel cross). Broad heritabilities ranged from 11 to 53%. Additive genetic variation was present for all phenotypes analyzed. Directional dominance was found for the relative size of the suprapyramidal mossy fiber terminal field only. For the stratum lacunosum moleculare, ambidirectional dominance emerged. These findings suggest that, in evolutionary history, directional selection has operated for a proportionally large suprapyramidal terminal field. For all other hippocampal variables (viz. the relative sizes for the strata oriens, pyramidale, radiatum, lacunosum moleculare, CA4, intra- and infrapyramidal mossy fiber terminal field and the absolute size of the regio inferior) past stabilizing selection was inferred. PMID- 18161584 TI - Are the structural changes in adult Drosophila mushroom bodies memory traces? Studies on biochemical learning mutants. AB - The pre-imaginal development of Drosophila mushroom bodies is under the influence of an unknown variable which causes populations of wild-type flies at eclosion to differ in the average number of Kenyon cell fibers. During the first week of adult life the number adjusts to an intermediate level which depends upon the experience of the flies. Under olfactory deprivation or social isolation it reaches a lower level than under favorable rearing conditions (J. Neurogenet., 1 (1984) 113-126). The biochemical learning mutants dance and rutabaga show no experience-dependent modulation of fiber number (Fig. 2). In both strains the mushroom bodies of young adults seem to develop abnormally; in dance a loss of about 600 fibers is observed, in rutabaga fiber number is low at eclosion and does not increase (Fig. 1a). The following model for long-term memory is proposed: in mushroom bodies outgrowth and decay of Kenyon cell fibers occur simultaneously. The fibers randomly form transient synapses onto extrinsic output neurons of the mushroom bodies and receive synapses from modulating neurons. Experience consolidates certain synapses, thus prolonging survival of the respective Kenyon cell fibers and increasing the steady state level of fiber number (Fig. 3). PMID- 18161585 TI - Mosaic analysis in the nematode Caenorhabditis elegans. PMID- 18161586 TI - Reciprocal behaviour associated with altered homeostasis and photosensitivity of Drosophila clock mutants. AB - The circadian oscillators of genetically short-period and long-period Drosophila exhibit reciprocal behaviour in four distinct ways: (1) with respect to the dependence of period on temperature, (2) in the change of period during constant darkness after ten days of constant light, (3) in the change of period during the second ten days of darkness as compared with the period during the first ten days, and (4) in the period change resulting from exposure to low-intensity constant light. The homeostatic control of the dependence of period length on temperature is impaired in the mutants as compared with wild-type files. The normal Drosophila pacemaker may comprise two mutually coupled oscillators, whereas the mutants may represent a reduction in activity of one or the other constituent oscillator. PMID- 18161587 TI - Persistent hypersynchronization of neocortical neurons in the mocha mutant of mouse. AB - A recessive mutation in the mouse at the mocha locus (mh, chromosome 10) modulates the synchronous synaptic activation of neocortical neurons, resulting in a constant 6-7 Hz (theta) wave pattern in the electrocorticogram. The gene linked brain rhythm is unaffected by motor behavior and cannot be desynchronized by sensory stimuli. This exemplary neurological mutation affecting cortical excitability is the first to reveal clearly that the predominance of a specific pattern of spontaneous brain wave activity can be inherited as a recessive trait. PMID- 18161588 TI - Cell loss in the inferior olive of the staggerer mutant mouse is an indirect effect of the gene. AB - Staggerer (sg) is an autosomal recessive mutation in mouse that causes severe cerebellar atrophy. In this mutant, the Purkinje cell (PC) number is reduced by about 75% and the remaining Purkinje cells have a reduced dendritic arbor and an ectopic location. Previous analysis of staggerer chimeras has demonstrated that the Purkinje cell phenotypes are all direct consequences of the cell-autonomous action of the staggerer gene. The two major afferents to the Purkinje cell are also affected. Virtually all of the granule cells die by the end of the first postnatal month. This death, however has been shown to be an indirect consequence of mutant gene action. The second major afferent system is from the cells of the inferior olive that projects to the main trunks of the Purkinje cell dendrite via the climbing fiber system. Quantitative studies of cell number in the inferior olive have shown that the number of cells is reduced by about 62% in adult sg/sg mutants. We report here the results of our quantitative analysis of three staggerer chimeras. beta-glucuronidase activity was used as an independent cell marker. Our findings demonstrate that inferior olive cell death in staggerer mutant mice is an indirect effect of staggerer gene action. Thus as for the granule cells, the loss of olivary neurons most likely results from a target related cell death. PMID- 18161589 TI - Examination of neurons in wild type and mutants of Caenorhabditis elegans using antibodies to horseradish peroxidase. AB - Antibodies to horseradish peroxidase (HRP) recognize 27 of 302 neurons and several non-neuronal cells in adult hermaphrodites of the soil nematode Caenorhabditis elegans and can be used to label these cells for cytological analysis in whole animals. The antibodies bind to the anterior members, but not to the posterior members of a set of mechanosensory neurons in wild type animals. Binding to one of the posterior mechanosensory neurons (PVM) occurs when this neuron migrates to an abnormal anterior position in mab-5 mutant animals, suggesting that expression of the epitope recognized by these antibodies is position dependent or that mab-5 mutations transform PVM into AVM intrinsically. The antibodies were used to characterize morphologies of two pairs of lumbar neurons (PHC and PVN) in uncoordinated mutants representing 95 unc genes. PHC and PVN morphologies were normal in most of the unc mutants examined, however, in mutants of 9 unc genes (unc-6, unc-13, unc-33, unc-44, unc-51, unc-61, unc-71, unc-73, and unc-98), misdirected PHC and/or PVN processes were observed at a high frequency. The morphologies of 2 other lumbar neurons, PHA and PHB, were determined previously in these mutants (Hedgecock et al., 1985). Mutations in most, but not all of these 9 unc genes affect the growth of the embryonic lumbar neurons PHA and PHB differently than they affect the growth of the postembryonic lumbar neurons PHC and PVN, indicating that these neurons require different, but overlapping sets of genes for different stages of normal growth and guidance. PMID- 18161590 TI - The rdgB gene of Drosophila: a link between vision and olfaction. AB - otal (ota=olfactory trap abnormal), an X-linked mutation of Drosophila isolated by virtue of abnormal olfactory behavior, is shown to be an allele of rdgB (retinal degeneration B), a gene required for normal visual system physiology. rdgB function is shown to be necessary for olfactory response of both adult files and larvae, which have distinct olfactory systems. Electrophysiological recordings from the adult antenna indicate that rdgB is required for normal response in the peripheral olfactory system: some rdgB mutants show a delayed return to the resting potential following stimulation with ethyl acetate vapor. These results indicate that rdgB is required for both visual and olfactory physiology, and they suggest that visual and olfactory transduction may share at least one common molecular step in Drosophila. PMID- 18161591 TI - Identification of linotte, a new gene affecting learning and memory in Drosophila melanogaster. AB - We describe the identification of linotte, a new autosomal gene in Drosophila involved with learning and memory. The linotte(1) mutant was derived from a PlacW transposan mutagenesis and was screened for three-hour memory deficits after classical conditioning of an olfactory avoidance response. Sensory and motor systems (olfactory acuity and shock reactivity) required for the classical conditioning experiments were normal in mutant linotte(1) files--indicating that the mutation disrupts learning/memory specifically. A chromosomal deficiency of the 37D region, where the linotte(1)P insert was localized in situ, failed to complement linotte(1)'s memory defect, and files from two lines homozygous for independent PlacW excisions show normal memory--indicating that the P insertion is responsible for the mutant phenotype. Additional behavior-genetic data suggest that linotte gene is non-vital. PMID- 18161592 TI - The central complex of Drosophila melanogaster is involved in flight control: studies on mutants and mosaics of the gene ellipsoid body open. AB - Visual flight control is studied in three mutant alleles of the gene ellipsoid body open (ebo) of Drosophila melanogaster. In mutant ebo flies the central complex is disturbed to varying degrees. Defects range from a small opening in the ellipsoid body to the dissociation of the ring into two parts, a cleft in the fan-shaped body and hypoplasia in the protocerebral bridge. Other parts of the brain are not visibly affected. Flight behavior is normal with respect to the amplitude of the optomotor response and to the object response (single rotating stripe). A reduced amplitude in the small random oscillations of the torque trace (yaw torque activity), however, is found in all three alleles. In two of them the frequency of torque spikes is reduced. In the allele ebo(678) the dynamics of the optomotor response is altered. Upon reversal of the direction of rotation mutant flies take longer than wild type to shift their yaw torque to the new response level (optomotor reversal time). Finally, these flies also behave abnormally in the flight simulator in which their yaw torque controls the angular velocity of the panorama. Many ebo(678) flies fixate a single stripe less persistently than normal flies, some even trying to fly away from it (antifixation). In ebo(678) gynandromorphs the four behavioral phenotypes ("yaw torque activity", "torque spike frequency", "on-target-fixation" and "optomotor reversal time") are all highly correlated with the phenotype of the ellipsoid body. Yaw torque activity and torque spike frequency in addition are correlated with the phenotype of the thorax suggesting that these behavioral defects are in part caused by mutant influences on the ventral ganglion. The results support the hypothesis that the central complex is involved in the control of flight behavior. PMID- 18161593 TI - Kant and the magnitude of sensation: a neglected prologue to modern psychophysics. AB - Quantitative relations between the sensations and the stimuli that produce them are the domain of psychophysics, a branch of natural science not yet known at the time of Immanuel Kant (1724-1804). But Kant's philosophical doctrines of perception imply that sensations can be quantified. Accordingly, he proposed not only to consider the magnitude of both sensations and stimuli but also to work out an appropriate mathematics that would relate these magnitudes to each other. This part of Kant's work received almost no attention up to the present time although it contains some essential elements of modern psychophysics. PMID- 18161594 TI - Animal electricity at the end of the eighteenth century: the many facets of a great scientific controversy. AB - In the 1790s, Luigi Galvani and Alessandro Volta were the main protagonists of a lively debate on the role of electricity in animal organisms. Significant developments originated from this debate, leading to the foundation of two new disciplines, electrodynamics and electrophysiology, that were to play a crucial role in the scientific and technological progress of the last two centuries. The Galvani-Volta controversy has been repeatedly reconstructed, sometimes in an attempt to identify the merits and the errors of one or the other of the two protagonists, sometimes with the aim of demonstrating that the theories elaborated by the two Italian scholars were irreconcilable, reflecting completely different ways of looking at phenomena and conceiving of scientific research. In this article a different interpretation is offered, based on a discussion of the scientific issues that were central to Galvani's and Volta's research, and with reference to the context of science and society of the eighteenth century. PMID- 18161595 TI - "A twist in the tale" of a surgeon and his patient: an Australian first in seizure localization. AB - In 1894 at St. Vincent's Hospital Melbourne, George Adlington Syme removed a meningioma from a patient with symptomatic focal epilepsy. The operation stands as the first surgery based on seizure localization in Australia. It is also the country's first documented successful resection of an intracranial meningioma. It followed William Macewen's landmark cerebral localization case on the boy John McKinley by 18 years and Victor Horsley's first epilepsy case on Hughlings Jackson's patient James B. by a mere 8 years. Syme's patient, Constable John G., survived the operation by some 23 years, dying from a gunshot wound to the head in 1917. Newly discovered inquest papers reveal that the coroner's judgment that the death was accidental completely fails to address the more credible scenario of suicide. The story makes for a fascinating epilogue to an important landmark in Australia's neurosurgical history. PMID- 18161596 TI - Vinken and Bruyn's Handbook of Clinical Neurology. A witness of late-twentieth century neurological progress. AB - Vinken and Bruyn's Handbook of Clinical Neurology (HCN) is best characterized as an encyclopedia. In this paper we describe the origin, production, and reception of HCN. Data were gathered from a literature search, by screening of HCN-volumes, interviewing key-role persons and a study of an HCN-archive. The initiative for HCN was taken by two Excerpta Medica staff members, the one a strategist with expertise in information systems, the other a gifted neurologist with an expert knowledge of who is who in the world of neurological literature. Within a period of 38 years, 2799 authors, 28 volume editors, the two initiators, and a third chief editor for the American continent described the whole of neurology in 1909 chapters on all together 46,025 pages (excluding index volumes). HCN was sold mainly to medical institutes in affluent countries. A digital version of the revised edition was proposed by the editors but refused by the publisher for commercial reasons. HCN was in general well received by book reviewers. The main criticisms concerned the price of the volumes, lack of editorial control, inadequacy of indexes, and lack of cross references. HCN offers unrivalled information on the state of the art of the clinical neurosciences in the second half of the twentieth century. In addition, it contains extensive reviews of the history of neurological diseases in the volumes of the original edition. PMID- 18161597 TI - Enrico Sereni: research on the nervous system of cephalopods. AB - This essay focuses on a paradigmatic moment in neurobiological studies of invertebrates: the research on the nervous system of cephalopods carried out by Enrico Sereni at the Naples Zoological Station between 1925 and 1931. Although he remained unknown on the historiographic scenario, probably due to his early death, he contributed to Italian science of the first half of the twentieth century. In my paper particular attention will be given to Sereni's study on the pigmentary-effector, neurohumoral, and peripheral nervous systems, since they also accounted for the historical foundation of the experimental vein that, through the years, would lead John Zachary Young, Sereni's follower, to the most well-known discovery of the giant nerve fibers. PMID- 18161598 TI - Instrument transfer as knowledge transfer in neurophysiology: Francois Magendie's (1783-1855) early attempts to measure cerebrospinal fluid pressure. AB - Francois Magendie's (1783-1855) experimental model for measuring blood pressure in animals, which he developed in 1838, had a major impact on French physiology in the nineteenth century, especially upon Etienne-Jules Marey (1830-1904) in Paris. In due course it was also adopted by other European investigators, such as the Leipzig physiologist Carl Ludwig (1816-1895), and by clinicians who developed it into a major measuring tool. Historians of science, however, have paid hardly any attention to Magendie's further laboratory investigations conducted with the assistance of Jean-Louis Marie Poiseuille's (1799-1869) sphygmometre (blood pressure meter). After having used the apparatus to conduct his experiments on a variety of blood vessels, Magendie also applied the sphygmometre in 1840 to the ventricular system of the brain in order to measure cerebrospinal fluid (CSF) pressure. But the scope of this new procedure had yet to be defined: the new measuring device invited many speculative interpretations about the meaning of CSF flow for the physiology of the ventricular system in healthy and diseased brain function. As such, Magendie's experiments produced phenomena in very heterogeneous knowledge areas, and CSF measurement was situated at the interface of quite disparate investigative spaces regarding the structure and function of the brain. In his textbook Lecons sur les Fonctions et les Maladies du Systeme Nerveux (Lectures on the Functions and Diseases of the Nervous System), Magendie described extending application of the measuring "apparatus of Poiseuille" from blood vessels to parts of the brain. The instrument thus became something of a liquidodynamometre (liquor dynamometer), that paved the way for later applications, including (after 1896) diagnostic intracranial pressure (ICP) measurement by Theodor Kocher (1841-1917) and Harvey Cushing (1869-1939). The current paper focuses on the experimental contingencies that prompted the instrument transfer in Magendie's laboratory and opened up new epistemological perspectives for research in neurophysiology. PMID- 18161599 TI - Walter Moxon, MD, FCRP (1836-1886): the cerebro-vascular system and the syndrome of "congestion of the brain": an analysis of his 1881 Croonian Lectures. AB - Walter Moxon, MD, FRCP lived, practiced medicine, taught and wrote in the mid- to late- nineteenth-century Victorian England, mostly at Guy's Hospital, London. He was widely informed in the "Art of Physic," writing on a range of issues from cerebral lateralization of articulate speech to angina pectoris. The present paper will trace briefly his contributions to the newly discovered asymmetry of articulate speech in the left frontal lobe (1866) and will in more detail trace and analyze his 1881 Croonian Lectures at the Royal College of Physicians on a medical shibboleth referred to as "congestion of the brain." In a series of ingenious and rhetorically creative arguments with imaginative tropes, demonstrations, evolutionary accounts of cognition and blood metabolism for human/biped cognition, and cogent citations from the medical literature of the day, Moxon skillfully instructs his medical audience against the misleading notion of cerebral "congestion" as an underlying pathology for cognitive, motor, and sensory deficits seen in the clinic. In so doing, he provides the medical community with an in-depth glimpse at the circulatory system, its flow dynamics, and how they serve to meet the cognitive, motor, and sensory demands of upright bipedal man. PMID- 18161600 TI - Area postrema: one of the terms described by Magnus Gustaf Retzius. PMID- 18161601 TI - Neurognostics question: American Civil War surgeon, pioneer of brain localization, and neurosurgery. PMID- 18161602 TI - Neuroanniversary 2008. PMID- 18161603 TI - NeurHistAlert 7. PMID- 18161604 TI - Neurognostics answer. PMID- 18161605 TI - Cataract surgery in Andhra Pradesh state, India: an investigation into uptake following outreach screening camps. AB - PURPOSE: Prevalence of cataract related blindness and visual impairment is extremely high in India. Service utilization, on the contrary, is sub-optimal. Services offered through outreach screening are often not taken up. Against this background, the decision making processes leading to the uptake of cataract surgery services offered during outreach screening camps were investigated in people with cataract in India. METHODS: The study used an exploratory, qualitative approach. Sixty one cataract patients from two eye hospitals in Hyderabad, India, were interviewed using convenience sampling. Two focus group discussions with cataract patients and interviews with seven key informants helped triangulate the information. RESULTS: Lack of access to personal funds limited and delayed the utilization of offered services by patients who had been advised of surgery at outreach screening camps or eye hospitals. Stigma, fatalism and ageism were other limiting factors. The majority of patients did not make the decision regarding uptake of services themselves. Hearsay reports of the outcome of surgery and quality of services had a strong influence on service uptake. Women, particularly widows and those from rural areas, were least likely to use cataract surgery services. CONCLUSION: Provision of affordable and accessible eye care does not guarantee that the care or service will be utilized. Within the decision making process about cataract surgery, dependency on and importance of the wider family was found to be an often cited context of patients' decisions. Future research and programming activities need to put an emphasis on underserved groups, especially those who may be marginalized or disempowered. PMID- 18161606 TI - The association between visual acuity and functional limitations: findings from a nationally representative population survey. AB - PURPOSE: To determine the independent effect of visual acuity on individual activities of daily living (ADL), instrumental activities of daily living (IADL) and mobility. METHODS: Cross-sectional survey on a sample representing the Finnish population aged 55 years and above. Of the 3392 eligible people, 3185 (93.9%) were interviewed, 2870 (84.6%) attended a comprehensive health examination, and 2781 (82.0%) had distance visual acuity (VA) assessed. A home interview included assessment of ADL, IADL and mobility, demographic variables and chronic conditions. Mobility measurements and binocular VA were assessed during the examination. RESULTS: Prevalence of ADL, IADL, and mobility limitations increased with decreasing VA (p<0.001). Visually impaired persons (VA< or =0.25) had ADL disabilities four times more likely than those with good VA (VA> or =0.8) after adjustment for socio-demographic and behavioral factors, and chronic conditions (OR 4.36, 95%CI 2.44-7.78). Limitations in IADL and measured mobility were five times as likely (OR 4.82, 95%CI 2.38-9.76 and OR 5.37, 95%CI 2.44-7.78, respectively), and self-reported mobility limitations were three times as likely (OR 3.07, 95%CI 1.67-9.63) as in persons with good VA. CONCLUSIONS: Decreased VA is strongly associated with functional limitations, and even a slight decrease in VA was found to be associated with limitations in functioning. PMID- 18161607 TI - Surveillance of contact lens related microbial keratitis in Australia and New Zealand: multi-source case-capture and cost-effectiveness. AB - PURPOSE: To evaluate a multi-source surveillance system used in a 12-month study of contact lens related microbial keratitis in Australia and New Zealand. METHODS: All practicing ophthalmologists and optometrists were surveyed on 6 occasions over 12 months via post or the Internet. Participation was defined as reporting at least once during the study period and the response rates represented those who responded on all six occasions. Cases were also detected through hospital audit. All ophthalmologists and a sub-group of optometrists were contacted by phone to elicit a response (active surveillance). The utilization and cost-effectiveness of active surveillance were compared to reports received via the post or the Internet. Case ascertainment and cost-effectiveness were compared for different sources of case capture. RESULTS: The rate of participation for ophthalmologists was 95.8% (711/742) and 88.5% (657/742) responded for all reporting periods. Active surveillance was required for 63% (416/661) of responses in Australia (AU) and 73% (59/81) in New Zealand (NZ) at AUD23.14 per practitioner. Internet reporting was more widely used in New Zealand (NZ: 31% vs. AU:17%, p=0.006) and was the most cost effective mode of reporting (AUD1.43 per practitioner). Postal reporting (AUD; AU:3.54,NZ:9.84 per practitioner) was under-utilized (3% of responses). Average start-up costs comprised 50% of study costs followed by active follow-up (42%), postal (6%) and Internet reporting (2%). Ophthalmologists (50.4%, 144/286 of cases) were the most cost-effective source of cases, followed by hospital audit (24.5%, 70/286) and optometry (25.1%, 72/286). Duplicate reporting occurred in 13% (37/286) of cases. CONCLUSIONS: High response rates were obtained by substantial resource commitment to active follow-up. Internet reporting was widely used and was cost-effective. Hospital audit and supplementary reporting by optometry were used for the first time in a study of contact lens related microbial keratitis, and contributed significantly to case capture. PMID- 18161608 TI - The association between maternal smoking in pregnancy, other early life characteristics and childhood vision: the Twins Eye Study in Tasmania. AB - PURPOSE: To investigate the association between maternal smoking in pregnancy, early-life environment and childhood vision. METHODS: Twin and triplet children enrolled in the Twins Eye Study in Tasmania underwent a comprehensive ophthalmic examination and their parents/guardians retrospectively answered a questionnaire regarding crawling, walking and other measures. A subset of these twins was also in the Tasmanian Infant Health Survey, which prospectively collected data on antenatal smoking, gestation, birth weight and other factors. RESULTS: The mean age of the 346 individuals (172 multiple birth sets) at the time of examination was 9.25+/-2.4 years. Mean unaided visual acuity was 0.0 (6/6). The mean spherical equivalent was +0.87D, and decreased with increasing child age (p<0.01). A prospective analysis, accounting for birth set clustering and relevant confounders, showed increasing levels of maternal smoking in the third trimester was associated with poor stereoacuity on the Titmus test (worse (>) than 100'', p=0.05) and Lang test (p=0.001) and also with the presence of esotropia (p=0.02). These associations persisted after adjustment for infant postnatal smoke exposure at one month of age. Poor stereoacuity on Titmus stereo test circles was associated with late age of first crawling (RR=1.23 (1.06, 1.42) p=0.005 per month) and late age of first walking (RR 1.18 (1.05, 1.22) p=0.001 per month). CONCLUSIONS: Antenatal smoking was independently associated with poor stereovision and the presence of esotropia. Poor stereoacuity may be associated with delayed age at first crawling or walking. PMID- 18161609 TI - Blindness and eye disease in Cambodia. AB - PURPOSE: To assess the prevalence and etiology of blindness and low vision and to assess the prevalence of common eye diseases in central Cambodia. METHODS: In this cross-sectional, population-based study, 6,558 residents of Kandal Province, Cambodia were registered, and 5,803 (88.5%) were interviewed and examined. This house-to-house survey was conducted by a team consisting of a senior ophthalmologist, a Cambodian eye doctor, and eight Cambodian eyecare workers. RESULTS: The prevalence of bilateral blindness (visual acuity <3/60) is 1.1% (95% confidence interval [CI], 0.9-1.4), and an additional 4.4% (95% CI, 3.9-5.0) have low vision (visual acuity < 6/18, > or =3/60 in the better eye). The major causes of bilateral blindness are cataract (67.4%), phthisis (6.1%), uncorrected refractive error (6.1%), corneal scar (5.3%), uncorrected aphakia (3.0%), trachoma corneal scar (3.0%), optic atrophy (3.0%), and others (6.1%). The major causes of low vision are uncorrected refractive error (49.8%) and cataract (42.7%). The prevalence of unilateral blindness is 1.2% (95% CI, 0.9-1.4), often caused by cataract, corneal scar, or phthisis. Trauma due to landmine explosions and war-related injuries was frequently the underlying etiology in subjects with phthisis, corneal scarring, or other pathology. CONCLUSIONS: The prevalence of blindness and low vision in Cambodia is relatively high compared to other developing countries. Most of the causes of blindness and low vision are treatable or preventable. Landmines and other war-related injuries are an important cause of ocular injury. These results will assist in developing a national plan for the prevention of blindness in Cambodia. PMID- 18161610 TI - Use of lipid-lowering agents for the prevention of age-related macular degeneration: a meta-analysis of observational studies. AB - PURPOSE: To examine the effect of lipid-lowering agents in the development of age related macular degeneration (AMD) through the techniques of meta-analysis. METHODS: Case-control and cohort studies presenting relative risks and 95% confidence intervals were identified through a literature review. Inclusion was limited to studies where both the exposure of interest (lipid-lowering agents) and outcome (AMD) were explicitly defined. Pooled estimates were computed using the random effects model. To quantify heterogeneity we calculated the proportion of total variance of between study variance using the Ri statistic. The Q statistic for heterogeneity was also calculated. RESULTS: Eight studies were identified. The pooled relative risk (RR) for all studies was 0.74 (95% CI, 0.55 1.00). When only those studies examining the use of statins were pooled (n=7), the RR was 0.70 (95% CI, 0.48-1.03). Using the Ri statistic, the heterogeneity between studies was found to be 0.85 for all studies and 0.89 for studies examining statins. CONCLUSION: Lipid-lowering agents, including statins, do not appear to lower the risk of developing AMD, although clinically significant effects cannot be excluded. The use of these agents in the prevention of AMD cannot be recommended until well designed prospective studies with long follow up have demonstrated a benefit. PMID- 18161611 TI - A qualitative study in the United Kingdom of factors influencing attendance by patients with diabetes at ophthalmic outpatient clinics. AB - INTRODUCTION: Many diabetics do not have regular eye examinations, although it is known that early diagnosis and treatment of sight-threatening retinopathy reduces the risk of blindness. The barriers that prevent diabetics from attending eye clinics are poorly understood. PURPOSE: To determine what factors may influence diabetic patients' attendance at eye clinics. METHODS: A qualitative survey of both patients and eye care providers was undertaken in a rural district general hospital and in an urban tertiary teaching hospital. Methods employed included semi-structured interviews, focus groups and non-participatory observations. All interviews and discussions were recorded and transcribed. The transcripts were then analyzed to detect emerging themes. Data collection continued until no new themes emerged. RESULTS: Lack of awareness was seen as the greatest barrier by both patients and providers. Patients were aware that diabetes could affect the eye, but not that it could lead to blindness, nor that severe retinopathy could be asymptomatic. Patients reported that fundus images used for screening were a valuable educational resource. Fear, particularly of laser treatment, and guilt, about poor control causing retinopathy, deterred patients from attending. Both patients and providers recognized that regular attendance was inconvenient; however, providers underestimated the difficulties patients faced in obtaining time off work to attend clinics. CONCLUSION: Providing more complete information about diabetic retinopathy, and making eye clinic attendance more convenient for patients, may increase the number of diabetics who have regular eye examinations. PMID- 18161612 TI - Lens opacities in adults in Pakistan: prevalence and risk factors. AB - PURPOSE: To investigate the prevalence and risk factors for lens opacity (LO) amongst a nationally representative sample of the adult population of Pakistan. METHODS: This national study of blindness and visual impairment (adults > or =30 years) used multistage, stratified, cluster random sampling. Grading of LO was conducted using the Mehra/Minassian classification system. LO, partly or wholly obscuring the red reflex, or previous cataract surgery were indicators of opacity. RESULTS: 16,402 (94.7%) adults were included in this analysis (study conducted 2002-2003). A total of 4,096 (standardized prevalence 20.9%, 95%CI: 20.3, 21.5%) adults were found to have LO. The highest prevalence of LO was found in Punjab province (22.2%), the lowest in Balochistan Province (18.0%). Significant positive associations were increasing age (multivariable odds ratio (OR) 3.2: 95%CI: 3.1, 3.4), hypertension (OR 1.2, 95%CI: 1.1, 1.3), history of diabetes (OR 2.6: 95%CI 2.0, 3.2) and smoking (OR 1.3: 95%CI: 1.1, 1.5). Higher body mass index (BMI) (OR 0.8: 95%CI 0.7, 0.9, heavy vs. normal BMI) and attendance to school (OR 0.6: 95%CI: 0.5, 0.8) were associated with lower risk of LO. Individuals in districts classified as hot were at significantly increased risk (OR 1.3: 95%CI: 1.1, 1.5), and those in wet districts (> 600 mm annual rainfall) had lower odds than individuals living in dry districts (OR 0.7: 95%CI: 0.6, 0.8). CONCLUSIONS: Almost a fifth of the adult population had LO. Significant positive associations were age, smoking status, hypertension, diabetes, and increased deprivation level. Protective factors included high BMI and educational achievement. The climatic associations offer novel hypothesis for further research into cataractogenesis. PMID- 18161613 TI - Quality of reporting of key methodological items of randomized controlled trials in clinical ophthalmic journals. AB - PURPOSE: To evaluate the reporting quality of key methodological items in randomized controlled trials (RCTs) in four general clinical ophthalmology journals. METHODS: The reporting of 11 key methodological items in RCTs published in American Journal of Ophthalmology, Archives of Ophthalmology, British Journal of Ophthalmology and Ophthalmology in the year 2005 was assessed. RESULTS: Sixty seven eligible RCTs were assessed and the mean number of items reported was 6.3 per RCT. No significant difference in the mean number of items reported was found between the four journals (P=0.20). The most frequently reported item was ethics approval and informed consent (97.0%), followed by masking status (85.1%), description of withdrawals (76.1%), adverse events (73.1%), and intention-to treat analysis (71.6%). Details on sequence generation, randomization restriction, allocation concealment, allocation implementation, patient flow diagrams, and sample size calculation were reported in <50% of the RCTs assessed. Both sample size and page length of the RCTs correlated with the number of methodological items reported (P=0.024 and P=0.008, respectively). CONCLUSIONS: Similar to other specialties, rooms for improvement exist in the reporting of key methodological items of RCTs in clinical ophthalmic journals. Stricter adoption of the CONSORT statement might enhance the reporting quality of RCTs in ophthalmic journals. PMID- 18161614 TI - The epidemiology of age-related eye diseases in Mainland China. AB - While many papers have been published regarding age-related eye diseases in Mainland China in the past two decades, the variable quality of those reports limit the conclusions that can be drawn. Many of these studies assessed blindness and low vision rates, and these estimates are likely accurate. However, due to lack of standardization of techniques for assessing cataract, glaucoma, age related macular degeneration, and diabetic retinopathy, estimates of the burden of these diseases on the population are less reliable. Owing to the rapid economic development of China in the last decade, resources to address eyecare problems are more likely to be available. Therefore, an accurate assessment of the burden of various eye diseases is needed in order to improve blindness prevention planning and program development. PMID- 18161615 TI - Australian prospective study of cataract surgery and age-related macular degeneration: rationale and methodology. AB - BACKGROUND: Cataract surgery is the most frequently performed ophthalmic procedure worldwide. While benefits gained from cataract surgery outweigh surgical risks, there have been concerns that older persons may have an increased risk of developing age-related macular degeneration (AMD) after cataract surgery. OBJECTIVE: The Australian Prospective Study of Cataract Surgery and Age-Related Macular Degeneration Study aims to assess the risk of AMD in a large cohort of older patients following cataract surgery. The current report describes the study rationale, design and methodology. DESIGN: Longitudinal study. PARTICIPANTS: Approximately 2000 cataract surgical patients aged 65 years or older are being recruited from both public and private sources in western Sydney, Australia. METHODS: At study visits, participants are interviewed using standardized questionnaires to obtain information on demographic, medical, and ocular conditions and AMD risk factors, together with data on general health and vision related quality of life. Eye examinations include visual acuity, intraocular pressure, keratometry and A-scan measurements, plus lens and retinal photography, following pupil dilatation. Retinal photographs taken before cataract surgery, and at 1, 6, 12, and 24 months after surgery are graded for early and late AMD lesions, using the Wisconsin age-related maculopathy grading system. The 1-month post-operative retinal photographs supplement the baseline macular assessment for cases in which cataract occludes a clear view of the macula pre-operatively. It is intended that study participants will be followed for up to five years post operatively to clarify the question of whether aphakic or pseudophakic, compared to phakic eyes, have a greater risk of developing AMD. PMID- 18161616 TI - A review of the molecular genetics of congenital Idiopathic Nystagmus (CIN). AB - Congenital Idiopathic Nystagmus (CIN) is genetically heterogeneous. Autosomal dominant, autosomal recessive and X-linked patterns of inheritance have been reported. Linkage analysis has suggested the existence of at least three distinct loci for both autosomal dominant and X-linked forms, although only one disease gene has been identified (FRMD7, Xq26.2). The pathophysiological mechanisms underlying nystagmus are poorly understood but it is anticipated that characterization of the FRMD7 gene and identification of novel nystagmus genes will provide insights into this condition and the functioning and development of the visual pathways in general. PMID- 18161617 TI - Pseudo-vitelliform macular detachment and cuticular drusen: exclusion of 6 candidate genes. AB - PURPOSE: The etiology and genetic cause of pseudo-vitelliform macular detachment with cuticular drusen (PVMD/CD) are unknown; nor is it clear if this phenotype represents a separate disease entity, or is a sub-phenotype of disorders with overlapping clinical presentation. To answer this question, we screened a cohort of patients affected with PVMD/CD for variation in six plausible candidate genes (ABCA4, VMD2, TIMP-3, peripherin/RDS, fibulin 5 (FIBL5) and complement factor H (CFH)) associated with diseases of overlapping phenotypes. METHODS: Twenty-eight patients, diagnosed with pseudo-vitelliform macular detachment and cuticular drusen, were evaluated by clinical examination, fundus photography, fluorescein angiography and autofluorescence imaging. DNA from all study subjects were screened for variants in the ABCA4, VMD2, TIMP-3, peripherin/RDS, FIBL5 and CFH genes by a combination of DHPLC, array screening and direct sequencing. RESULTS: All patients presented with cuticular drusen; pseudo-vitelliform detachment was seen in 21 cases, while atrophic changes following regression of the detachment were seen in the remaining 7 subjects. Visual acuity ranged from 20/20 to CF. The screening revealed an I32V mutation in peripherin/RDS in one patient and 2ABCA4 variants, T897I and G1961E, in 2 more patients. No amino acid-altering variants were detected in VMD2, TIMP-3, and FIBL5 genes. The frequency of the CFH Y402H variant in this cohort corresponded to that detected in the general population. CONCLUSIONS: Screening of 6 candidate genes detected possibly disease-associated mutations in only 3/28 (10.7%) of patients presenting with PVMD/CD, eliminating these genes as causal for this phenotype. PMID- 18161618 TI - A new GJA1 (connexin 43) mutation causing oculodentodigital dysplasia associated to uncommon features. AB - Oculodentodigital dysplasia (ODDD) is an autosomal dominant disorder that includes a clinical spectrum of craniofacial, neurologic, limb, and ocular malformations. The disease is caused by heterozygous mutations in the 6q22-q23 located GJA1 gene, that encodes connexin 43 (Cx43). In this paper we describe a novel Cx43 mutation (G2V) found in a Mexican eight-year-old boy. This de novo mutation predicts a missense substitution at the second amino acid of Cx43, in the first intracellular domain, and is the most amino-terminal located mutation reported so far. Umbilical hernia and congenital optociliary veins, two uncommon ODDD-associated features, were recognized in our patient. The phenotype of three previously described patients with Cx43 first intracellular domain mutation is discussed and compared with that observed in our patient. This case expands the phenotypic and genotypic spectrum of ODDD. PMID- 18161619 TI - LOC387715/HTRA1 and complement factor H variants in patients with age-related macular degeneration seen at the mayo clinic. AB - PURPOSE: To confirm association of the complement factor H allelic variant (CFH Y402H) and the LOC387715/HTRA1 (LOC387715 A69S) risk alleles with age-related macular degeneration (AMD). STUDY POPULATION: Study of 89 Caucasian patients with neovascular (exudative) AMD and 232 Caucasian controls. METHODS: The Y402H variant of CFH gene and A69S variant of LOC387715/HTRA1 gene locus were examined. RESULTS: For CFH, the odds ratio for the homozygous variant was 4.97 (CI 2.52 to 9.79). For LOC387715/HTRA1 the odds ratio for the homozygous risk variant was 7.75 (CI 3.46 to 17.35). The odds ratio for heterozygous carriers was 3.35 (CI 1.91 to 5.90). PMID- 18161620 TI - Lens coloboma associated with a ciliary body cyst. AB - Isolated congenital lens coloboma is a sectoral indentation of the crystalline lens due to zonular weakness or absence. The purpose of this report is to describe the association of lens coloboma with an adjacent cyst in the ciliary body and to suggest that ciliary body cysts may be an under-recognized cause of congenital lens coloboma. PMID- 18161621 TI - Chorioretinopathy and microcephaly with normal development. AB - PURPOSE: To report a pediatric patient with bilateral chorioretinopathy and microcephaly who from birth to 2 years of age is reaching appropriate developmental milestones. DESIGN: Retrospective case report with clinical findings and literature review. MAIN OUTCOME MEASURES: Clinical findings and visual acuity estimated by sweep visual evoked potentials (VEP), electroretinogram (ERG) and fundoscopic exam. RESULTS: A microcephalic child with normal motor and cognitive development had improving sweep VEP despite atypical fundoscopic findings of bilateral chorioretinopathy, attenuated retinal vessels, and anomalous optic nerves. The etiology for these collective findings despite extensive workup, including prenatal TORCH titers and neuro-imaging, has remained unidentified. CONCLUSIONS: Most published cases of microcephaly with chorioretinopathy have described patients with mild to severe mental retardation. Patients with chorioretinopathy and microcephaly may, however, reach all developmental milestones with improvement in visual development as was seen in this case. The long-term cognitive and visual prognosis may be better than previously reported. PMID- 18161622 TI - Two novel deletions (array CGH findings) in pigment dispersion syndrome. AB - PURPOSE: We report the first male with pigment dispersion syndrome and a balanced translocation t(10;15)(p11.1;q11.1). METHODS: Cytogenetic analyses using Giemsa banding and FISH methods, and array CGH were performed. RESULTS: Array CGH analyses did not show altered DNA sequences in the breakpoints of the translocation, but revealed two novel deletions in 2q22.1 and 18q22.1. CONCLUSION: We suppose that the coexistence of t(10;15) and pigment dispersion syndrome in our patient is a coincidence. The deletion in 2q22.1, where the gene LRP1B has been located, may play a major role in the dysembryogenesis of the eye and cause the disorder. PMID- 18161623 TI - Severe form of familial exudative vitreoretinopathy caused by homozygous R417Q mutation in frizzled-4 gene. AB - PURPOSE: To report the clinical features of a patient with familial exudative vitreoretinopathy (FEVR) associated with homozygous R417Q mutation in the frizzled-4 gene (FZD4). METHODS: Clinical examination and mutation analysis by direct sequencing. RESULTS: A five-month-old girl was found to have leukocoria associated with retrolental fibroplasia in the right eye and a severe falciform retinal fold in the left eye. Mutational analysis revealed a homozygous R417Q mutation in the FZD4 gene. Her parents who carried the same mutation heterozygously exhibited milder ocular phenotype. CONCLUSIONS: Homozygous state for the FZD4 gene is possibly involved in the severity of the FEVR phenotype. PMID- 18161624 TI - Genetic screening of leber congenital amaurosis in a large consanguineous Iranian family. AB - The molecular defect of one large consanguineous Iranian kindred with Leber Congenital Amaurosis (LCA) is presented. The phenotype mapped to 17p13.1 (LCA1) and excluded from five other LCA loci. Sequence analysis of the GUCY2D gene identified a novel homozygous missense mutation (I816S) that segregated with the inherited disease-haplotype in six affected, eight parents, and two normal gene carriers. This mutation was absent in three other normal family members and 92 normal control subjects. In silico analysis predicted that alteration of the highly conserved isoleucine residue at position 816 to serine is deleterious by affecting secondary structure of the GUCY2D protein. PMID- 18161625 TI - Reassessment of the pathologic significance of the 9438 mitochondrial DNA mutation associated with LHON. PMID- 18161627 TI - Effect of product temperature during primary drying on the long-term stability of lyophilized proteins. AB - Our objective was to investigate the effect of performing primary drying at product temperatures below and above Tg' (glass transition temperature of the freeze-concentrated phase) on the long-term stability of lyophilized proteins. Two protective media differing in the nature of the bulking agent used (amorphous or crystalline) were selected. Several lyophilization cycles were performed by using various combinations of shelf temperature and chamber pressure to obtain different values of product temperature during primary drying. The antigenic activity of the proteins was measured after lyophilization and after 6 months of storage at 4 degrees C and 25 degrees C. After 6 months of storage and regardless of the protective medium, the losses of antigenic activity of both toxins increased from 0% when primary drying was performed at a product temperature lower than Tg' and to 25% when the product temperature was higher than Tg'. The use of partially crystalline systems makes it possible to withstand high primary drying temperatures (above Tg'). However, the shelf life of lyophilized proteins may be decreased when the amorphous phase including the protein and the stabilizing molecule changes to the viscous state. PMID- 18161628 TI - Water sorption behavior and swelling characteristics of starches subjected to dielectric heating. AB - The aim of this study was to investigate the effects of microwave irradiation and storage on the moisture content, adsorption behavior and swelling properties of potato (B-type) and maize starches (A-type). Volumetric heating resulted in reversible moisture loss from both types of samples. The crystallinity of potato starch was decreased, whereas its water retention capacity and swelling power were increased irreversibly, and its swelling capacity was increased reversibly by the thermal process applied. The corresponding parameters of maize starch were not influenced significantly by dielectric heating; this may be related to its special structure resulting in the thermal resistance of this polymer. Our results allow the conclusion that microwave irradiation offers an appropriate and selective alternative for the physicochemical modification of potato starch. In consequence of its low susceptibility to thermal processes, maize starch may be used for the microwave drying of pharmaceutical formulations containing starch. PMID- 18161629 TI - Effect of divalent cations on pore formation and degradation of poly(D,L-lactide co-glycolide). AB - Poly(D,L-lactide-co-glycolide) (PLG) is probably the biodegradable polymer most often used for polymeric controlled-release formulations. Different salts have been shown to affect the swelling and degradation of PLG, which, in turn, affect the release of encapsulated drugs. In this investigation the effect of divalent cations was especially investigated. Films of PLG were incubated in phosphate buffer saline (PBS), a buffer containing salts similar to plasma, Hepes buffer, and Hepes buffer with ZnCl2, CaCl2, MgCl2, or Na2CO3 added. Pore formation at the surface and inside the film was analyzed by scanning electron microscopy. The samples were also analyzed gravimetrically at predetermined intervals to determine the mass loss, and for some samples the pH within the PLG films was determined by confocal microscopy. Pores were formed faster in the presence of all divalent cations, and the results indicated a greater degradation rate in the presence of Zn2+. The catalyzing effect of the divalent cations on degradation was attributed to their ability to act as Lewis acids. Pores were formed more slowly in PBS than in a buffer containing salts similar to plasma, which should be considered when choosing the in vitro release medium. PMID- 18161630 TI - Simultaneous determination of levodopa and carbidopa in levodopa-carbidopa tablets by ATR-FTIR spectrometry. AB - A novel analytical procedure has been developed for quantitative determination of levodopa and carbidopa in aqueous binary solutions acidified by HCl and without any other sample pretreatment. The method is based partially on least squares treatment of data obtained by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectrometry in 1211-1315 cm(-1) and 1488-1550 cm(-1) spectral regions. The simple, rapid, and accurate proposed method was applied to determine levodopa and carbidopa in Levodopa-C tablets. The statistical parameters, such as R2, RSD, SEE, SECV, LOD, and recovery were evaluated. Number of factors, scans, and resolution were optimized. In this method R2 and RSD for levodopa and carbidopa were (0.9965, 1.209) and (0.9537, 0.813), respectively. PMID- 18161631 TI - In vitro release--in vivo microbiological and toxicological studies on ketoconazole lipid granules. AB - In some multidrug therapy programs, ketoconazole (KTZ) may be administered with some antacids that could modify its dissolution rate and reduce its absorption, thus leading to therapeutic failures. The primary aim of this study was to evaluate the influence of Compritol HD5 ATO and Compritol 888 ATO on this interaction in comparison with commercial KTZ tablets. The second aim was to prepare lipid granules of KTZ that could be an alternative to the commercial formulation. Therefore, six KTZ sustained-release granules were prepared with different lipid concentrations, because they were found to be more suitable than tablets that are dissolved only in gastric medium. The results confirmed that the dissolution rate of KTZ granules was significantly reduced in the presence of antacids. The ideal formulation was selected as granules including 5% of Compritol lipids in relation to the suitability of the target profile. Therapeutic effects of orally administered, ideal KTZ granule formulations, and commercial tablets were evaluated in vivo by the experimental model of murine vulvo-vaginal candidiasis (VVC) with and without antacids. It was found that formulations were very effective on VVC, and the therapeutic effect decreased significantly in the presence of antacids. Histopathological studies were carried out for vagina, stomach, and liver tissues and hepatoxicity was also examined. The levels of reduced glutathione (GSH) were measured to assess the oxidative stress induced by KTZ and function of the liver. It was observed that orally administered formulations of KTZ were successful in treating candidiasis in mice without irritancy in stomach. However, liver tissues were damaged. The decreased GSH levels indicated toxicity in our study. This study suggested that in vitro release and in vivo microbiological-toxicological properties of KTZ were affected by antacids and drug-excipient interactions. Lipid granules of KTZ prepared with Compritol 888 ATO could be proposed as a new KTZ solid dosage form with optimum dissolution and therapeutic characteristics. PMID- 18161632 TI - Development of liposomal systems of finasteride for topical applications: design, characterization, and in vitro evaluation. AB - Finasteride (FNS) is a "drug of choice" for benign prostate hypertrophy and prostate cancer. The drug has also been reported to be useful orally in the treatment of some difficult-to-treat androgen-dependent skin disorders, such as seborrhea, acne, hirsutism, and androgenetic alopecia. However, the ideal route for its administration (i.e., topical) remains unexplored. This has logically suggested the search for strategic formulation approaches to make the drug effective on topical applications, hitherto unexplored. The present study targets the encasement of drug molecules in the interiors of vesicular compartments (liposomes) made up of hydrogenated phospholipids, as an attempt toward the development of a trans-epidermal therapeutic system of FNS. Multilamellar drug loaded liposomes were prepared by thin-film hydration with sonication method and optimized with respect to drug payload, entrapment efficiency, and size by formulating different vesicular compositions under different process conditions. The vesicular systems consisting of saturated phospholipid (100 mg), cholesterol (50 mg), and FNS (5 mg) showed highest drug payload (2.9 mg/100 mg of total lipids), and drug entrapment efficiency (88.6%). Mean (+/-SD) vesicle size of the prepared liposomes was found to be 3.66+/-1.6 microm. Significantly higher skin permeation of FNS through excised abdominal mice skin of FNS was achieved from the liposomal formulations vis-a-vis corresponding solution and conventional gels. Liposomal FNS formulations also showed more than fivefold higher deposition of drug in skin than the corresponding plain drug solution and conventional gel. Stability studies indicated that the liposomal formulations were quite stable in the refrigerated conditions for 2 months with negligible drug leakage or vesicle size alteration during the storage period. Results of the current studies with FNS-loaded vesicular systems project the high plausibility of a topical liposomal formulation for effective localized delivery of Finasteride. PMID- 18161633 TI - Investigation of serrated roll surface differences on ribbon thickness during roller compaction. AB - Many parameters are considered when developing a roller compaction process. However, roll serration volume is generally not considered when using serrated rolls. It was hypothesized that serration volume of concavo-convex rolls can have a significant effect on ribbon thickness due to differences in nip angle. Experiments were conducted to investigate ribbon thickness using different roll sets having a range of serration volumes. Roll sets with approximately twice the serration volume produce ribbons 20-25% greater in thickness. These results correlated with physical measurements taken of the serrations from each set. Thus, differences in roll serration volume can significantly affect ribbon thickness. PMID- 18161634 TI - Stability of benzoyl peroxide in aromatic ester-containing topical formulations. AB - The chemical stability of benzoyl peroxide (BPO) was studied in solutions and gels. The solutions (1% w/v) were prepared in single solvents (alcohol USP, isopropyl alcohol USP, ethyl benzoate, C12-15 alkyl benzoate, dimethyl isosorbide, propylene carbonate, and acetone) and in binary and tertiary combinations of these solvents, with and without the addition of antioxidant(s) (BHT, BHA, eugenol, tert-butyl hydroquinone, Tenox-2, vitamin E, and vitamin C). The solutions were stored at 37 degrees C for 5 weeks, and each week were analyzed for remaining BPO. Using first-order kinetics, the stability of BPO in solution was found to decrease in the order: ternary>binary>single solvent systems. Regardless of the number of solvents present, the highest stability of BPO (t1/2>7.5 weeks) was attained in the presence of ethyl benzoate and C12-15 alkyl benzoate. The stability of BPO in solution did not change significantly with the addition of most antioxidants. The solutions in which BPO remained most stable were one in alcohol USP-ethyl benzoate-C12-15 alkyl benzoate (60:20:20; t1/2=18.15 weeks) and another in alcohol USP-C12-15 alkyl benzoate-isopropanol plus 0.1% BHT (65:20:15; t1/2=12.44 weeks). In turn, these two solutions were converted to homogeneous gels by the addition of Cab-O-Sil. The chemical stability of BPO in these gels was evaluated at 37 degrees, 45 degrees, 50 degrees, and 55 degrees C for 5 weeks. Parallel experiments were conducted with two commercial BPO products, a 2.5% tinted gel and 5% vanishing lotion. BPO was less stable in commercial products (t1/217 repeats) (chi2=20.21, p=0.0005, OR=2.30). This contrasts with Asian populations where allele 16 was identified as the risk allele, showing allele heterogeneity depending on ethnicity. PMID- 18161657 TI - Matrix metalloproteinase-19 is a predictive marker for tumor invasiveness in patients with oropharyngeal squamous cell carcinoma. AB - AIMS: To evaluate the expression of matrix metalloproteinase-19 (MMP-19) in oropharyngeal squamous cell carcinoma along with its association with structural features of invasiveness. To investigate whether MMP-19 expression correlates with lymphatic or systemic metastasis and prognosis in patients who have received definitive radiotherapy. METHODS AND RESULTS: The histological evaluation of the invasive front was based on Bryne's malignancy grading system. We correlated the immunohistochemical expression pattern with morphological parameters which characterize tumor invasiveness such as keratinization, nuclear polymorphism, invasion pattern, and the host inflammatory response. Local immunoreactivity for MMP-19 was positively correlated with tumor invasiveness as reflected in its structural characteristics and the degree of nuclear polymorphism, and negatively correlated with the inflammatory response of the host. No correlation existed between MMP-19 expression and clinicopathological features (TNM stage, grade of differentiation) or a patient''s outcome and prognosis. CONCLUSIONS: This latter finding probably reflects the unique change for MMPs from high immunoreactivity within healthy tissue areas and non-invasive tumor parts, through absence in the least invasive neoplastic regions, to strong re-expression at a highly invasive front of the same tumor. Our findings indicate that MMP-19 can be used as a marker for tumor invasiveness in patients with oropharyngeal squamous cell carcinoma. PMID- 18161658 TI - Altered salivary profile in heavy smokers and its possible connection to oral cancer. AB - Saliva is the first biological fluid to encounter inhaled cigarette smoke, whose numerous carcinogens and oxidants are responsible for the oral cancer so prevalent among smokers. Whole saliva, collected from 25 consenting heavy smokers and from a control group of 25 age- and gender-matched non-smokers, was subjected to sialochemical, biochemical, immunological and oxidative analyses. The mean flow rate was significantly higher in smokers than in non-smokers, as were the median activity value of superoxide dismutase (SOD) and the total salivary antioxidant capacity (ImAnOx) (by 32% and 12%, respectively, p=0.05). The salivary carbonyl concentration (an oxidative stress indicator) was significantly higher by 126% (p=0.0006) among smokers, while lactate dehydrogenase, albumin, total immunoglobulin G, and the metalloproteinases MMP-2 and MMP-9 concentrations were significantly lower in the smokers, by 86% (p=0.003), 65% (p=0.003), 61% (p=0.048), 35% (p=0.005) and 55% (p=0.035), respectively. Apparently, the oral cavity''s salivary antioxidant system fails to cope with the severe attack of reactive oxygen species originating in cigarette smoke. Moreover, various other salivary functional and protective parameters also decreased among the smokers. Hence, further research aimed at examining the possibility of administration of agents as antioxidants or saliva substitutes to the oral cavity of smokers should be considered. PMID- 18161659 TI - Diagnostic application of serum proteomic patterns in gastric cancer patients by ProteinChip surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. AB - Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS) is one of thecurrently used techniques to identify biomarkers for cancers. This study was planned to establish a system to accurately distinguish gastric cancer patients by using SELDI-TOF-MS. A total of 100 serum samples obtained from 60 individuals with gastric cancer and 40 healthy individuals were screened. Protein expression profiles were expressed on CM10 ProteinChip arrays and analyzed. Peak intensities were analyzed with the Biomarker Wizard software to identify peaks showing significantly different intensities between normal and cancer groups. Classification analysis and construction of decision trees were done with the Biomarker Pattern software 5.0. Seventeen protein peaks showed significant differences between the two groups. The decision tree which gave the highest discrimination included four peaks at mass 5,919, 8,583, 10,286, and 13,758 as splitters. The sensitivity and specificity for classification of the decision tree were 96.7% (58/60) and 97.5% (39/40), respectively. When the protein biomarker pattern was tested on a blinded test set, it yielded a sensitivity of 93.3% (28/30) and a specificity of 90% (18/20). These results suggest that serum protein profiling by the SELDI system may distinguish gastric cancer patients from healthy controls with relatively high sensitivity and specificity. PMID- 18161660 TI - Urinary cell-free DNA as a potential tumor marker for bladder cancer. AB - The objective was to assess the possibility of measuring urine creatinine (UCr) adjusted urinary cell-free (ucf) DNA concentration as a noninvasive screening tool for bladder cancer. Using PicoGreen-based detection, the ucf-DNA/UCr concentration was quantified in urine supernatant specimens from 46 bladder cancer patients and 98 controls and compared to 400-bp real-time PCR-based detection, which detected the amplification of 400-bp beta-actin (named 400-bp ucf-DNA/UCr). The mean concentrations for both PicoGreen and 400-bp ucf-DNA (ng/mL)/UCr (mg/dL) were significantly higher in bladder cancer patients than in controls: 15.28 vs 6.68 (p<0.001, t-test) and 14.98 vs 1.07 (p<0.001), respectively. Among different stages and grades, no significant difference was found between these two methods. The areas under the ROC curves of PicoGreen and 400-bp ucf-DNA/UCr were 0.571 (95% confidence interval, 0.451-0.692) and 0.805 (95% confidence interval, 0.713-0.896), respectively. In 400-bp ucf-DNA/UCr, the best sensitivity and specificity were 86.1% and 72.0% at the cutoff value of 0.0645. These data indicated that 400-bp ucf-DNA/UCr is more reliable for bladder cancer detection than PicoGreen. In conclusion, our results suggest that ucf DNA/UCr can be used as a potential tumor marker for bladder cancer, especially for detecting longer DNA fragments. PMID- 18161661 TI - Standardization of PSA measures: a reappraisal and an experience with WHO calibration of Beckman Coulter Access Hybritech total and free PSA. AB - Prostate-specific antigen (PSA) is the serum biomarker most widely used in prostate diseases. Since there is significant variation in PSA results among non equimolar assays, the 90:10 ratio of complexed PSA to free PSA (the Stanford standard) was proposed as standardization; this became the basis for the PSA mass standards WHO 96/670 for tPSA and 96/668 for fPSA. Nevertheless, recent publications underlined the lack of interchangeability between different commercial assays, all claimed to be equimolar and calibrated to the WHO standard. Importantly, the WHO calibration yields about 16-20% lower PSA results. Manufacturers that have chosen to calibrate existing assays to the mass value of WHO 96/760 have introduced a significant negative bias compared to the Hybritech assay calibration; this bias is transferred to clinical evaluation if the cutoff of 4 ng/mL, clinically validated for the Hybritech assay, is maintained with the WHO calibration. Beckman Coulter recently provided the option of calibrating the Access Hybritech PSA and Free PSA assays to the WHO standard introducing a different clinical cutoff. Using two different reagent lots, we tested about 200 routine patients for tPSA and fPSA with both calibrations; we also calculated the f/tPSA ratio with both calibrated methods. Moreover, we verified the analytical sensitivity and inter- and intra-assay variability. In accordance with the claim of the manufacturer, the results obtained with the WHO calibration showed a negative bias of about 25% and, as expected, no significant difference was found for % f/tPSA. The same bias was found when retesting samples of the External Quality Assessment Scheme of the Institute of Clinical Physiology of the National Research Council in Pisa. Based on this experience we decided for the moment to keep the Hybritech calibration, in order to avoid cutoff changes during patient follow-up. Moreover, we have started to provide information to clinicians aimed at the alignment of our results with the WHO standardization. PMID- 18161662 TI - Serum anti-p53 antibodies as a useful marker for prognosis of gastric carcinoma. AB - Mutations in the TP53 gene are the most common genetic alterations in cancer. Accumulation of mutated protein may induce circulating anti-p53 antibodies (anti p53Ab) in sera of cancer patients. The aim of our work was to evaluate the presence and prognostic value of anti-p53Ab in gastric cancer patients and to investigate whether their presence is related to p53 overexpression in tumor tissue. Anti-p53Ab were analyzed in sera from 111 patients with gastric carcinoma and from 64 healthy donors by ELISA. p53 expression was also quantified by ELISA in biopsies of 54 gastric cancers and 22 healthy gastric mucosas. Significant anti-p53Ab levels were found in 15.3% of patients, whereas none of the 64 donor sera were positive. High levels of p53 expression were detected only in tumor tissue, in 72.2% of cases. A significant correlation was observed between anti p53Ab and high levels of mutated p53 in tissue (p<0.05). The survival time of serum-positive patients was significantly longer than that of patients with low/negative serum levels, with a survival rate of 41.2% and 14.9%, respectively, over 48 months (p<0.05). Thus, detection of serum anti-p53Ab in gastric cancer patients can be useful to identify a subset of patients with better prognosis. PMID- 18161663 TI - MUC-1 (CA 15-3 antigen) as a highly reliable predictor of response to EGFR inhibitors in patients with bronchioloalveolar carcinoma: an experience on 26 patients. AB - BACKGROUND: Bronchioloalveolar carcinoma (BAC) is a histological subtype of non small cell lung cancer (NSCLC), particularly of adenocarcinoma. Given its multifocality and the poor activity of chemotherapy, there is no established treatment for BAC, although promising results have been achieved with inhibitors of the epidermal growth factor receptor (EGFR). No tumor marker has been validated in the diagnosis and follow-up of lung cancer, in particular to predict the outcome of treatment with EGFR inhibitors. PURPOSE: As CA 15-3 antigen serum levels are reported to be pathologically abnormal in adenocarcinoma of the lung, we chose this tumor marker to monitor treatment with EGFR inhibitors of patients affected by adenocarcinoma with BAC features or pure BAC. PATIENTS AND METHODS: We collected data from 26 consecutive Caucasian patients with BAC, mostly women and never smokers, who received EGFR inhibitors. RESULTS: We noticed that all patients with normal CA 15-3 serum levels at baseline (15/26, 57.7%) showed a response to EGFR inhibitors, whereas all patients with abnormal CA 15-3 serum levels (11/26, 42.3%) did not. CONCLUSION: Our data suggest that CA 15-3 levels might be a predictive factor for the response to EGFR inhibitors in patients with BAC. PMID- 18161665 TI - A two-staged technique for basilic vein transposition. AB - The dilemma of creating a fistula in patients without a use-able cephalic vein can be addressed by basilic vein transposition, yet results of the classic single stage procedure are inconsistent and surgeon utilization of this procedure is variable. This article describes a two-staged technique for basilic vein transposition. The two-staged technique is likely to facilitate higher fistula rates in patients unable to have a direct fistula, and warrants consideration by surgeons striving to achieve higher fistula rates. PMID- 18161666 TI - Percutaneous placement of a constrained stent for the treatment of dialysis associated arteriovenous graft steal syndrome. AB - Though rare, dialysis associated steal syndrome (DASS) can cause debilitating symptoms. Surgical revision of the dialysis access is typically required. We describe a percutaneous technique to alleviate steal syndrome utilizing a constrained stent within an arteriovenous graft. A brief review of the incidence, pathophysiology, and standard treatment of DASS is also provided. PMID- 18161667 TI - Survival of arteriovenous fistula for dialysis at different centers in the North of England. AB - BACKGROUND: Renal failure patients rely on their vascular access for hemodialysis. Surgery for construction of arteriovenous fistulae is provided by a range of specialists. The aim of this review was to assess the survival of arteriovenous fistulae for hemodialysis patients in different centers of Northern England. METHODS: Data was collected on 473 hemodialysis patients in the North of England. Risk factors for failure were determined for each patient (age, sex, diabetes), together with their current mode of dialysis and history of surgical access procedures. This was expressed against their duration of dialysis. The dialysis units were then compared for fistula survival using the Kaplan Meier method. RESULTS: 68.3% (323) patients were dialysed through via arteriovenous fistulae and 31.7% (150) via neck line. Overall fistula survival rates were 85.1% at 1 year, 82.5% at 2 years and 72.7% at 3 years. The best 1 year survival was 91.6% and worst 76.1%. These were 74.4% and 53.1% at 5 years and 74.4% and 29.5% at 10 years; these differences were highly statistically significant (p = 0.0033). CONCLUSION: Though graft survival is affected by many things, surgical training in access surgery is not mandatory and a review of surgical practice is urgently needed. PMID- 18161668 TI - Primary thromboprophylaxis with heparins for arteriovenous fistula failure in pediatric patients. AB - BACKGROUND: To reduce the incidence of early arteriovenous fistula failure (AVF) due to thrombosis in pediatric hemodialysis (HD) patients, a primary thromboprophylaxis (PTP) protocol was initiated at author's institution in June 2005. The goal of this study is to report author's experience with this protocol one year later. METHODS AND RESULTS: 19 AVFs (14 patients, Historical group) and 8 AVFs (7 patients, PTP group) were created prior to and after initiation of PTP respectively. PTP consisted of unfractionated heparin (5-10 units/kg/hr) infusion postoperatively, followed by subcutaneous low molecular weigh heparin (LMWH) until AVF was matured. LMWH dosing was 'Prophylactic' (0.5 mg/kg/d, anti-factor Xa levels: peak 0.25-0.5 and trough < 0.3 units/mL) and 'Therapeutic' (1 mg/kg/d, anti-factor Xa level: peak 0.5-1 and trough < 0.5 units/mL) based on thrombosis predisposition. In Historical group, 12 AVFs did not receive thromboprophylaxis (No-treatment group), 5 received 81 mg aspirin/day (Aspirin group), and 2 received LMWH. In No-treatment group 10/12 AVFs failed: 9 thromboses and 1 stenosis. In Aspirin group 1/5 AVFs failed due to thrombosis. In PTP group 1/8 AVFs failed due to stenosis; the first 2 AVFs developed hematoma prompting a reduction in LMWH dose and monitoring trough anti-factor Xa levels, one AVF required thrombectomy after LMWH was transiently held. The incidence of thrombosis was less in PTP group (12.5%) when comparing to No-treatment group (83%) (p < 0.05). CONCLUSION: PTP is a feasible option to prevent early thrombosis at AVF. Close clinical and laboratory monitoring including trough anti factor Xa levels is required to adjust optimum anticoagulation. Larger studies are needed to clarify safety and efficacy of our PTP protocol. PMID- 18161669 TI - Fluoroscopically guided vs modified traditional placement of tunneled hemodialysis catheters: clinical outcomes and cost analysis. AB - Tunneled cuffed internal jugular vein catheters are widely used to provide short to medium-term vascular access for hemodialysis. The NKF-K/DOQI guidelines state that fluoroscopy is mandatory for insertion of all cuffed dialysis catheters. The KDOQI recommendation makes it difficult for Nephrologists to perform this procedure without access to fluoroscopy. This results in unnecessary waiting times and the inappropriate use of acute, non-tunneled catheters. The purpose of this study is: 1) to compare the outcomes of fluoroscopically guided vs modified traditional catheter placement technique, and 2) to perform a cost analysis of the two techniques. We performed a retrospective investigation of 202 tunneled hemodialysis catheters performed at our tertiary care hospital. Procedural data were obtained from the University of Wisconsin Department of Medicine, Nephrology Section Interventional Nephrology procedural database. Patient demographics, laboratory tests were obtained from the University of Wisconsin Hospital electronic medical record (EMR). Logistic regression was used to evaluate the effect of blind vs fluoro-guided placement on clinical outcomes, corrected for side of procedure, age, gender, previous history of catheter placement, diabetes mellitus (DM), and pre-procedural coagulation parameters. Baseline characteristics of 'blind' vs fluoro-guided groups differed with respect to side of procedure and DM (91.0% vs 79.6%, p = 0.02 and 43.30% vs 58.40%, p = 0.02, respectively). Non-fluoroscopic placement of catheters was associated with a decreased odds ratio of immediate success (OR = 0.1298, CI = 0.02 - 0.71). No difference in major or minor bleeding complications was discovered between the blind vs fluoro-guided group. Cost analysis revealed that performing the non fluoroscopic technique as the preferred initial procedure would represent a substantial reduction in total bills submitted to third-party payers, including Medicare. PMID- 18161670 TI - High blood flow rates with adjustment of needle diameter do not increase hemolysis during hemodialysis treatment. AB - BACKGROUND: Higher blood flow in dialysis therapy is often avoided due to concerns about shear-induced blood damage despite the lack of reliable data. OBJECTIVE: This study investigated the influence of higher blood flow rates on plasma free hemoglobin (Hb) concentration after hemodialysis (HD) treatment. METHODS: Thirty-two chronic HD patients were treated once with a blood flow rate of 250 mL/min using a 17G needle, and once with a blood flow rate of 500 mL/min using a 14G needle. Arterial and venous pressure and blood pressure (BP) were recorded before and after treatment. Blood samples were taken before and after treatment for analysis of plasma free Hb, pH, HCO3, base excess, hematocrit value, urea, sodium, potassium and calcium. RESULTS: HD treatment at blood flow rates of 500 mL/min did not increase plasma free Hb compared to treatments at blood flow rates of 250 mL/min. Frequency of intradialytic BP drops was not different either. By adaptation of the needle size, negative arterial pressure could be kept at a similar level. Urea reduction rates were significantly higher during treatments with higher blood flow rates. CONCLUSION: Higher blood flow rates can be applied without an increased hemolysis risk provided that needle sizes are adapted accordingly. PMID- 18161671 TI - Venae comitantes fistulae: an option in patients with difficult hemodialysis access. AB - OBJECTIVES: To evaluate the outcome of use of venae comitantes vessels in the formation of arterio-venous fistulae as vascular access for hemodialysis, in patients with limited venous anatomy. METHODS: Twenty patients who underwent arterio-venous anastomosis between brachial artery and venae comitantes were identified (2002 - 2005) and the notes reviewed. RESULTS: There was early failure in two (10%) patients (immediate postoperative period) and a further four (20%) failed late (mean 26 weeks, range 7-60). One patient developed a steal syndrome with radial nerve dysfunction requiring ligation of the fistula. Six (30%) patients utilised their fistulae for dialysis successfully without additional surgery and a further 6(30%) required surgical intervention to exteriorise the fistulae by the use of interposition grafts to allow successful use. CONCLUSION: Venae Comitantes arterial fistulae offer an option in patients with limited venous anatomy for standard reconstruction. If access surgery utilizes such veins second stage procedures are often required with overall 70% use. PMID- 18161672 TI - Effect of ethanol/trisodium citrate lock on microorganisms causing hemodialysis catheter-related infections. AB - PURPOSE: This in vitro study tested the effectiveness of a novel 30% ethanol/4% trisodium citrate (TSC) lock solution against the most common pathogens causing hemodialysis catheter-related infections. METHODS: Clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) (n = 4), methicillin-sensitive S. aureus (MSSA) (n = 8), methicillin-resistant Staphylococcus epidermidis (MRSE) (n = 8), Pseudomonas aeruginosa (n = 4) and Escherichia coli (n = 4) were tested in duplicate. Bacterial suspensions of each isolate were made in a control solution of normal saline and Mueller-Hinton broth (MHB), and in a lock solution of ethanol 30%, TSC 4% and MHB. Suspensions were incubated at 37 degrees C for 48 h. Colony counts were determined from samples collected at t = 0 h (before exposure to the ethanol/TSC lock), t = 1 h (one hour after exposure to the ethanol/TSC lock), t = 24 h and t = 48 h. To confirm the absence of viable organisms in the lock solution, the remaining volume at 48 h was filtered through a 0.45 microm filter. The filter was rinsed with 15 mL sterile water and plated on tryptic soy agar (TSA). RESULTS: All controls demonstrated significant growth over 48 h. In the lock solutions, initial inocula were reduced to 0 viable colonies by t = 1 h (6-log kill), and there was no growth at t = 24 and 48 h. Filtering of lock solutions also showed no growth. These results were consistent among duplicates of all isolates. CONCLUSIONS: The 30% ethanol/4% TSC lock solution consistently eradicated MRSA, MSSA, MRSE, P. aeruginosa and E. coli within 1 h of exposure. Experiments are currently underway to test this novel lock solution on preventing biofilm production by these pathogens. PMID- 18161673 TI - Stenosis at the area of transposition - an under-recognized complication of transposed brachiobasilic fistulas. AB - BACKGROUND: With an increased focus on native AV fistula creation in hemodialysis patients, a transposed brachiobasilic fistula (tBBF) is becoming an increasingly utilized option. This study describes the outcomes of tBBFs in a chronic hemodialysis population. In particular, we focus on the incidence and location of stenosis, and review the impact of angioplasty on these lesions. METHODS: A retrospective cohort study using all patients with a tBBF created between January 2001 and December 2004. RESULTS: Of the 543 fistulas created during the study period, 93 were tBBFs. The mean age of patients was 65 years, 56% were male and 55% were diabetic. Stenosis occurred in 54% (46/85) of fistulas; the location of stenosis in the majority (74%) was at or near the area of basilic vein transposition and 50% of fistulas with stenosis in this location required three or more angioplasties. Primary (unassisted) patency was 42% at one year in this cohort. Secondary patency was 68% at 1 year and 58% and 53% at 2 and 3 years respectively. CONCLUSION: In a cohort of hemodialysis patients who received a tBBF, we describe a reasonable primary and secondary patency rate and a high rate of stenosis at the point of transposition of the basilic vein. Such stenosis usually requires multiple percutaneous or surgical interventions to ensure or reestablish conduit patency. Further study is required regarding the optimal surgical technique, monitoring, and treatment of stenosis of this fistula type including the utility of repeat angioplasty. PMID- 18161674 TI - Functional patency of autogenous AV fistulas for hemodialysis. AB - BACKGROUND: Although AV fistulas are the preferred access for hemodialysis and have low complication rates, failure to function remains high and time to first dialysis may be several months. METHODS: Data from a Computerized Patient Record System of patients undergoing AV fistula from October 2000 to March 2006 were reviewed for type of fistula, interval from AV fistula construction to first hemodialysis, patency period, and complication rate. RESULTS: 129 patients were identified who underwent 155 autogenous AV fistula constructions. The average age was 62.1 (range 40-84) years old. 114 radiocephalic and 41 brachiocephalic fistulas were performed. 57 (50%) radiocephalic fistulas allowed successful hemodialysis after an average length of 13+/-5 weeks with a primary patency of 13+/-4 months. 24 (42%) fistulas subsequently thrombosed, 7 (12%) developed fistula stenosis, and 2 (4%) developed steal syndrome. 28 (68%) brachiocephalic fistulas reached successful hemodialysis after 6+/-2 weeks with a primary patency of 16+/-7 months. Eleven (42%) of the brachiocephalic fistulas that reached hemodialysis remained patent while four (15%) thrombosed. Two (8%) brachiocephalic fistulas thrombosed before reaching hemodialysis. There were two incidences (5%) of steal syndrome in the brachiocephalic group with one case being severe leading to tissue loss in the hand. CONCLUSION: Brachiocephalic fistulas were superior to radiocephalic in both time to maturity, primary patency, and functional primary patency. Brachiocephalic fistulas had a higher maturation rate and were less likely to fail once hemodialysis began. Vascular surgeons should develop better patient selection to predict which fistulas will function successfully rather than risk complications of prolonged central catheters. PMID- 18161675 TI - Accessory veins and radial-cephalic arteriovenous fistula non-maturation: a prospective analysis using contrast-enhanced magnetic resonance angiography. AB - PURPOSE: To determine if large caliber accessory veins are associated with radial cephalic arteriovenous fistula (RC-AVF) non-maturation. METHODS: RC-AVFs were created in 15 consecutive patients (radial artery and cephalic vein diameter > 2 mm, in the absence of arterial inflow or venous outflow stenoses or occlusions). Contrast-enhanced magnetic resonance angiography (CE-MRA) was performed preoperatively for the determination of vessel diameters, stenoses and occlusions. The location and caliber of accessory veins was determined. Vascular access (VA) function was monitored and all interventions required to obtain a functioning VA were recorded. Non-maturation was defined as a nonfunctional VA at 2 months after creation. The predictive value of accessory vein caliber for prediction of RC-AVF non-maturation was evaluated using receiver operating characteristic (ROC) analysis. RESULTS: Non-maturation occurred in 10 (67%) out of 15 RC-AVFs. Large caliber accessory veins (n = 4), venous stenosis (n = 3) or both (n = 2) were associated with RC-AVF non-maturation. The presence of large caliber accessory veins was the only significant predictor for RC-AVF non maturation (p = 0.01). Preoperatively detected accessory veins with a diameter > 70% of the cephalic vein diameter, had a sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of 80, 100, 100 and 91% for prediction of RC-AVF non-maturation on patient level. Accessory vein ligation and dilatation of venous stenosis resulted in an overall salvage success rate of 89% (8/9). CONCLUSION: Large caliber accessory veins are associated with RC-AVF non-maturation. Ligation of large caliber accessory veins is a successful salvage procedure in a substantial group of patients. Furthermore, ligation of these accessory veins during initial RC-AVF creation can potentially reduce non maturation rates; and therefore, preoperative assessment of accessory veins is recommended. PMID- 18161676 TI - Risk factors for the development of cephalic arch stenosis. AB - PURPOSE: The creation of a vascular access is necessary in hemodialysis patients, including those with marginal vessels. Upper arm fistulae are attractive due to the ease of creation and of achieving high access flow rates. Cephalic arch stenosis (CAS) can lead to failure of upper arm fistulae and is increasingly identified. We hypothesized that CAS is promoted by high blood flow rates, brachiocephalic fistulae, and an angle of cephalic vein insertion approaching 90 degrees. METHODS: All patients requiring a fistulogram between January 2004 and May 2006 had surveillance fluoroscopy of the central veins. Demographic, clinical and laboratory parameters were collected and the angle of the cephalic vein insertion measured by 3 blinded independent observers. RESULTS: Fifty-eight patients had fistulograms and CAS was detected in 18 subjects. Significant differences between the CAS and non-CAS groups were brachiocephalic fistula site (p = 0.046), access flow (mL/min) (p = 0.012), and absence of diabetes (p = 0.03). Univariate predictors of CAS include access flow (per 100 mL/min) (p = 0.042), platelet count (p = 0.031) and calcium-phosphate product (p = 0.026). The relationship of brachiocephalic site and CAS was confounded by access flow [(per 100 mL/min)*brachiocephalic fistula site (p = 0.016)] and fistula age [brachiocephalic fistula site*fistula age (p = 0.017)]. In multivariate analysis, renovascular disease, calcium-phosphate product, platelet count and access flow (per 100 mL/min)*brachiocephalic fistula predicted CAS (p < 0.001, Negelkerke's R Square = 0.55). The angle of insertion of the cephalic vein was not predictive for CAS. CONCLUSIONS: CAS may be a long-term consequence of high blood flow rates. The interaction of access flow and brachiocephalic fistula supports the hypothesis that high flow through a brachiocephalic fistula promotes CAS. The multiple factors influencing cephalic arch remodeling require further research. PMID- 18161677 TI - Effect of upper arm brachial basilic and prosthetic forearm arteriovenous fistula on left ventricular hypertrophy. AB - BACKGROUND: Creation of an arteriovenous fistula (AVF) may increase left ventricular hypertrophy in the hemodialysis population. Aim of this study was to compare the effects of a brachial-basilic (BB) AVF and the prosthetic brachial antecubital forearm loop access (PTFE) on cardiac performance. METHODS: Patients were randomized to receive BB-AVF or prosthetic brachial-antecubital forearm loop access. Before and three months after AVF creation patients underwent an echocardiographic examination. Mann-Whitney U-test was used to compare relative increase between the measured cardiac parameters for the two groups. RESULTS: Twenty-seven patients participated in the study. The relative increase in left ventricular parameters was not significantly different between the two groups. Only left ventricular end-diastolic diameter tended to be of significance. Mean blood flow through the brachial artery was 1680+/-156 and 1450+/-221 mL/min three months after surgery for the PTFE and the BB-AVF group, respectively. CONCLUSION: After three months of follow-up, changes in cardiac structure were comparable between patients with BB and PTFE AVFs. Also access flow was comparable at this time. In general, the effects of creation of a fistula on LV structure were limited. Longer follow up time may be needed to explore the long term effects of different vascular accesses on cardiac function. PMID- 18161678 TI - Pitfall in pediatric dialysis: malposition of a dialysis catheter mimicking azygos continuation syndrome. AB - Central venous catheters are established as vascular access in hemodialysis therapy. Vascular catheter misdirection may occur and is a well known problem. We present a rare catheter malposition in a young dialysis patient with consequent dilatation of the azygos vein system, simulating the appearance of an azygos continuation syndrome. PMID- 18161679 TI - Using arteriovenous fistulae as a dual access for hemodialysis and total parenteral nutrition administration is feasible with a good outcome: a case series. AB - End stage renal failure patients requiring long term total parenteral nutrition (TPN) often have multiple central line placements due to line infection or occlusion. Sometimes this can cause central venous stenosis or even occlusion. We present three cases in this consecutive series, in which we have successfully used arteriovenous fistulae for both hemodialysis and long term TPN administration as an alternative route without any complications. We therefore think that native AVF and grafts can be used as dual access for hemodialysis and TPN administration provided careful case selection, counselling and follow-up. PMID- 18161681 TI - Fundamental vibrational frequencies and dominant resonances in methylamine isotopologues by ab initio and density functional theory methods. AB - Ab initio and density functional theory (DFT) calculations were performed for obtaining fundamental vibrational frequencies of methylamine, CH3NH2, and its deuterated variants CH3ND2, CD3NH2, and CD3ND2. The calculations were carried out using the CCSD(T) coupled cluster approximation with cc-pVTZ and cc-pVQZ basis sets, and by the DFT method with the semiempirical hybrid functional B97-1 with polarization consistent pc-2 and pc-3 basis sets. Reasonable performance of the DFT harmonic and ab initio harmonic calculations was found, which improved considerably upon combination of the harmonic fundamental frequencies with anharmonic corrections from the smaller, pc-2, basis. The computed anharmonic fundamental frequencies of methylamine isotopologues agree very well with the experimental values and represent a useful tool for assignment and analysis of the dominant resonances. PMID- 18161682 TI - Structure and singly occupied molecular orbital analysis of anionic tautomers of guanine. AB - Recently, we reported the discovery of adiabatically bound anions of guanine that might be involved in the processes of DNA damage by low-energy electrons and in charge transfer through DNA. These anions correspond to some tautomers that have been ignored thus far. They were identified using a hybrid quantum mechanical combinatorial approach in which an energy-based screening was performed on the library of 499 tautomers with their relative energies calculated with quantum chemistry methods. In the current study, we analyze the adiabatically bound anions of guanine in two aspects: (1) the geometries and excess electron distributions are analyzed and compared with anions of the most stable neutrals to identify the sources of stability; (2) the chemical space of guanine tautomers is explored to verify if these new tautomers are contained in a particular subspace of the tautomeric space. The first task involves the development of novel approaches-the quantum chemical data like electron density, orbital, and information on its bonding/antibonding character are coded into holograms and analyzed using chemoinformatics techniques. The second task is completed using substructure analysis and clustering techniques performed on molecules represented by 2D fingerprints. The major conclusion is that the high stability of adiabatically bound anions originates from the bonding character of the pi orbital occupied by the excess electron. This compensates for the antibonding character that usually causes significant buckling of the ring. Also, the excess electron is more homogenously distributed over both rings than in the case of anions of the most stable neutral species. In terms of 2D substructure, the most stable anionic tautomers generally have additional hydrogen atoms at C8 and/or C2 and they do not have hydrogen atoms attached to C4, C5, and C6. They also form an "island of stability" in the tautomeric space of guanine. PMID- 18161683 TI - Molecular dynamics of a calix[4]arene-containing polymer in dichloromethane solution: ability of the solvent molecules to fill the cavity of the macrocycle. AB - The structure and dynamics of the dichloromethane solvent around the calix[4]arene units contained in the molecular actuator poly(calix[4]arene bis bithiophene) have been examined using a 1-micros molecular dynamics simulation. Results indicate that a solvent molecule fills the cavity associated to the cone conformation of the macrocycle during a significant period of time, especially when the actuator is not contracted. The position of such solvent molecule presents a fourfold symmetry with a maximum orientation toward the center of each ring contained in the calix[4]arene. Frequently, the solvent molecule located inside the cavity is rapidly exchanged for another molecule of the bulk. Thus, the number of dichloromethane molecules that reside more than 0.5 ns inside the cavity is relatively small. Finally, we detected that a significant number of solvent molecules are able to migrate from the cavity of one calix[4]arene to the cavity/ies of other/s, suggesting that the dynamics of the bulk solvent is important for the formation of these inclusion complexes. PMID- 18161684 TI - A dual-level state-specific time-dependent density-functional theory. AB - A highly efficient new algorithm for time-dependent density-functional theory (TDDFT) calculations is presented. In this algorithm, a dual-level approach to speed up DFT calculations (Nakajima and Hirao, J Chem Phys 2006, 124, 184108) is combined with a state-specific (SS) algorithm for TDDFT (Chiba et al., Chem Phys Lett 2006, 420, 391). The dual-level SS-TDDFT algorithm was applied to excitation energy calculations of typical small molecules, the Q bands of the chlorophyll A molecule, the charge-transfer energy of the zincbacteriochlorin-bacteriochlorin model system, and the lowest-lying excitation of the circumcoronene molecule. As a result, it was found that the dual-level SS-TDDFT gave correct excitation energies with errors of 0.2-0.3 eV from the standard TDDFT approach, with much lower CPU times for various types of excitation energies of large-scale molecules. PMID- 18161685 TI - Globally convergent computation of chemical equilibrium composition. AB - We report the Newton-Raphson based globally convergent computational method for determination of chemical equilibrium composition. In the computation of chemical equilibrium composition, an appearance of nonpositive value of number of moles of any component leads to discrepancy. The process of conditional backtracking and adaptive set of refining factors for Newton-Raphson steps are employed to resolve the problem. The mathematical formulation proposed by Heuze et al. (J Chem Phys 1985, 83, 4734) has been solved using proposed computational method, instead of empirical iterative formulation, as proposed by them. Results for the same numerical example, used by Heuze et al. (J Chem Phys 1985, 83, 4734) and White et al. (J Chem Phys 1958, 28, 751) are presented in addition to decomposition of Cyclotrimethylenetrinitramine for fixed temperature and pressure. It is observed that the proposed method is efficient and globally convergent. An even noteworthy finding is that the set of refining factors can be chosen from the range 0.1 to eta, where eta may be greater than one depending on how smoothly system of nonlinear equations is dependant on corresponding variable. Related analysis and results are discussed. PMID- 18161686 TI - Assessment of performance of the general purpose polarizable force field QMPFF3 in condensed phase. AB - The recently introduced force field (FF) QMPFF3 is thoroughly validated in gas, liquid, and solid phases. For the first time, it is demonstrated that a physically well-grounded general purpose FF fitted exclusively to a comprehensive set of high level vacuum quantum mechanical data applied as it is to simulation of condensed phase provides high transferability for a wide range of chemical compounds. QMPFF3 demonstrates accuracy comparable with that of the FFs explicitly fitted to condensed phase data, but due to high transferability it is expected to be successful in simulating large molecular complexes. PMID- 18161687 TI - Dynamic structures of phosphodiesterase-5 active site by combined molecular dynamics simulations and hybrid quantum mechanical/molecular mechanical calculations. AB - Various quantum mechanical/molecular mechanical (QM/MM) geometry optimizations starting from an x-ray crystal structure and from the snapshot structures of constrained molecular dynamics (MD) simulations have been performed to characterize two dynamically stable active site structures of phosphodiesterase-5 (PDE5) in solution. The only difference between the two PDE5 structures exists in the catalytic, second bridging ligand (BL2) which is HO- or H2O. It has been shown that, whereas BL2 (i.e. HO-) in the PDE5(BL2 = HO-) structure can really bridge the two positively charged metal ions (Zn2+ and Mg2+), BL2 (i.e. H2O) in the PDE5(BL2 = H2O) structure can only coordinate Mg2+. It has been demonstrated that the results of the QM/MM geometry optimizations are remarkably affected by the solvent water molecules, the dynamics of the protein environment, and the electronic embedding charges of the MM region in the QM part of the QMM/MM calculation. The PDE5(BL2 = H2O) geometries optimized by using the QM/MM method in different ways show strong couplings between these important factors. It is interesting to note that the PDE5(BL2 = HO-) and PDE5(BL2 = H2O) geometries determined by the QM/MM calculations neglecting these three factors are all consistent with the corresponding geometries determined by the QM/MM calculations that account for all of these three factors. These results suggest the overall effects of these three important factors on the optimized geometries can roughly cancel out. However, the QM/MM calculations that only account for some of these factors could lead to considerably different geometries. These results might be useful also in guiding future QM/MM geometry optimizations on other enzymes. PMID- 18161688 TI - Theoretical study on the hydrolysis mechanism of N,N-dimethyl-N'-(2-oxo-1, 2 dihydro-pyrimidinyl)formamidine: water-assisted mechanism and cluster-continuum model. AB - The hydrolysis reaction of N,N-dimethyl-N'-(2-oxo-1, 2-dihydro pyrimidinyl)formamidine (DMPFA), a model compound of the antivirus drug amidine 3TC (3TC = 2', 3'-dideoxy-3'-thiacytidine), is investigated by the hybrid density functional theory B3LYP/6-31+G (d,p) method. The hydrolysis reaction of the title compound is predicted to undergo via two pathways, each of which is a stepwise process. Path A is the addition of H2O to the C=N double bond in the amidine group to form a tetrahedral structure in its first step, and then the transfer of the H atom of hydroxyl leads to the corresponding products via four possible channels. Path B simultaneously involves the nucleophilic attack of H2O to the C atom of the C=N bond and the proton transfer to the N atom of amino group leading to the cleavage of the C-N single bond in the amidine group. The results indicate that path A is more favorable than path B in the gas phase. Moreover, to simulate the title reaction in aqueous solution, water-assisted mechanism and the cluster continuum model, based on the SCRF/CPCM model, are taken into account in our work. The results indicate that it is rational for two water molecules served as a bridge to assist in the first step of path A and that cytosine rather than the cytosine-substituted formamide should be released from the tetrahedral intermediate via s six-membered cycle transition state (channel 2). Our calculations exhibit that the process toward the tetrahedral intermediate is the rate-determining step both in the gas phase and in aqueous solution. PMID- 18161689 TI - Trends in CE-MS 2005-2006. AB - CE-MS has gained further attention as a multidimensional analytical approach. The number of publications dealing with this technique is still increasing on the level of application as well as method development-oriented approaches. Additionally, 15 reviews were published the last two years focusing on CE-MS. An overview of all papers found to have been published within 2005 and 2006 is given in tabulated form. Additionally, four promising technical trends are chosen to be presented in detail: (i) chip-based CE-MS, (ii) capillary coatings in CE-MS, (iii) new trends in CEC-MS and (iv) the application of 2-D CE-MS. PMID- 18161690 TI - Recent applications in CEC. AB - In this review, research papers on CEC are summarized that have been published between May 2005 and May 2007. Application-oriented research is discussed in which CEC is used in biochemical and pharmaceutical studies, in the analysis of food and natural products, and in industrial, environmental, and forensic analysis. Some trends and developments in separation science that may increase the applicability of the separation technique CEC are highlighted: 2-D separation systems and the application of nano- and microfluidic devices in separations. PMID- 18161691 TI - Sensitive chiral analysis by CE: an update. AB - A general view of the different strategies used in the last years to enhance the detection sensitivity in chiral analysis by CE is provided in this article. With this purpose and in order to update the previous review by Garcia-Ruiz et al., the articles appeared on this subject from January 2005 to March 2007 are considered. Three were the main strategies employed to increase the detection sensitivity in chiral analysis by CE: (i) the use of off-line sample treatment techniques, (ii) the employment of in-capillary preconcentration techniques based on electrophoretic principles, and (iii) the use of alternative detection systems to the widely employed on-column UV-Vis absorption detection. Combinations of two or three of the above-mentioned strategies gave rise to adequate concentration detection limits up to 10(-10) M enabling enantiomer analysis in a variety of real samples including complex biological matrices. PMID- 18161692 TI - Ionic liquids and CE combination. AB - This paper reviews and discusses the analytical combination of ionic liquids (IL) with CE. On the one hand, it shows CE as a powerful technique to separate impurities from IL as well as being capable to determine physical and chemical properties of IL. On the other hand, it also shows how IL are employed in CE separations to enhance resolution, peak efficiency and peak symmetry. Specifically, IL are used as additives in CZE, NACE, and MEKC and as support coatings of the capillary wall in electrochromatography. The integrity of the IL in the electrophoretic system as well as the roles of the cations and anions of the IL in the electrophoretic separation are also discussed. PMID- 18161693 TI - Rapid-on-chip determination of dielectric properties of biological cells using imaging techniques in a dielectrophoresis dot microsystem. AB - Dielectrophoresis is a technique whereby polarisable particles are manipulated by non-uniform alternating electric fields. A specific application of this technique is deducing the dielectric properties of cells from analysis of the dielectrophoretic spectrum of that particular cell population. We have developed a new microelectrode geometry consisting of two parallel electrode planes, one of which is patterned with arrays of circular apertures or 'dots'. The radial symmetry of the dots means that the polarisability of the particles within the dot can be directly related to change shifts in light transmission through the dot, and quantified from analysis of digital images. We have validated our system using well-characterised cell types and found a high degree of agreement to published data. Furthermore, we have observed that at high particle concentrations, electrostatic inter-particle repulsion causes spontaneous, rapid particle re-dispersion over the dot volume upon removal of an applied electric field. This allows the automated acquisition of a spectrum of 26 data points in approximately 15 min. PMID- 18161697 TI - Microemulsion electrokinetic chromatography hyphenated to atmospheric pressure photoionization mass spectrometry. AB - CZE is an appropriate technique for separating charged species, but lacks selectivity for neutral compounds. Alternative approaches such as microemulsion electrokinetic chromatography (MEEKC) have been developed to broaden its range of applications. Hyphenation of MEEKC with MS is an attractive perspective since it can enhance sensitivity and selectivity. The on-line coupling of MEEKC with MS, however, is not straightforward due to the low compatibility of non-volatile surfactant additives (e.g. SDS) and the commonly used API source, namely ESI. In order to hyphenate MEEKC with MS detection, the atmospheric pressure photoionization (APPI) source was investigated. Possibilities offered by the coupling of MEEKC with APPI-MS were highlighted for the complex separation of ionized and neutral compounds in both the positive and negative modes. MEEKC-APPI MS performance, in terms of selectivity, efficiency and sensitivity was compared to CZE-ESI-MS and MEEKC-ESI-MS for the screening of doping substances (beta blockers, central stimulants, diuretics, etc). Relevant selectivity and detectability, particularly for neutral, structurally related and isobaric compounds was demonstrated with the MEEKC-APPI-MS approach opening new avenues for CE-MS, in addition to the well-established CZE-ESI-MS technique. PMID- 18161698 TI - Sepsis-induced cholestasis. PMID- 18161699 TI - Dissecting the genetic susceptibility for cholangiocarcinoma in primary sclerosing cholangitis. PMID- 18161701 TI - Does a latte a day keep the hepatologist away? PMID- 18161700 TI - Platelet count is not a predictor of the presence or development of gastroesophageal varices in cirrhosis. AB - Current guidelines recommend esophagogastroduodenoscopy (EGD) in patients with cirrhosis to screen for gastroesophageal varices (GEV). Thrombocytopenia has been proposed as a noninvasive test to predict the presence of GEV. There is no agreement regarding a specific platelet count (PLT) that can reliably predict GEV. The present longitudinal study aims to (1) further investigate the relationship between varices and PLT at the time of endoscopy, (2) investigate whether changes in PLT from the baseline over time can predict the development of GEV, and (3) investigate whether changes in PLT correlate with the hepatic venous pressure gradient (HVPG). A secondary analysis was conducted for 213 subjects with compensated cirrhosis with portal hypertension but without GEV enrolled in a randomized, placebo-controlled, double-blind trial of a nonselective beta-blocker used to prevent GEV. PLTs were obtained every 3 months, and HVPG measurements and EGD were done annually. The PLTs were compared between subjects who did and did not develop GEV. In a median follow-up of 54.9 months, 84 patients developed GEV. PLT was greater than 150,000 in 15% of patients at the development of GEV. A receiver operating curve did not show any PLT with high sensitivity or specificity for the presence of GEV. Subjects with clinically insignificant portal hypertension (HVPG < 10 mm Hg) whose PLT remained greater than 100,000 had a 2-fold reduction in the occurrence of GEV (P = 0.0374). A significant correlation was found between HVPG and PLT at the baseline, year 1, and year 5 (P < 0.0001). CONCLUSION: Cross-sectional or longitudinal evaluations of PLTs are inadequate noninvasive markers for GEV. Patients with mild portal hypertension whose PLT remains greater than 100,000 have significantly less risk of GEV. Although HVPG correlates somewhat with PLT, changes in PLT cannot be used as a surrogate for HVPG changes. PMID- 18161702 TI - Infrared spectroscopy and ab initio theory of isolated H5O2+: from buckets of water to the Schrodinger equation and back. PMID- 18161703 TI - Li6PS5X: a class of crystalline Li-rich solids with an unusually high Li+ mobility. PMID- 18161704 TI - Living donor liver transplantation: The ethics and the practice. PMID- 18161705 TI - The neck-liver axis. Madelung disease as further evidence for an impact of body fat distribution on hepatic histology. PMID- 18161706 TI - Asymmetric counteranion-directed catalysis for the epoxidation of enals. PMID- 18161708 TI - Supramolecular gels formed by amphiphilic low-molecular-weight gelators of N alpha,N epsilon-diacyl-L-lysine derivatives. AB - Simple L-lysine derivatives, N(alpha)-hexanoyl-N(epsilon)-lauroyl-L-lysine (1), its alkali metal salts (2-4), and two-component compounds that consist of 1 with 2 to 4, were synthesized and their hydrogelation and organogelation properties were studied. Addition of hydrochloric acid to an aqueous solution of the alkali metal salt at room temperature produced a translucent hydrogel. This hydrogelation occurred as a result of a change in nanostructure from micelle-like aggregates to nanofibers, which was induced by partial protonation of the carboxylate to form a carboxylic acid. On the other hand, two-component low molecular-weight gelators exhibited amphiphilic gelation behavior and functioned as not only hydrogelators, but also as organogelators. FTIR studies revealed that lateral ionic interactions between the carboxylate, alkali metal cation, carboxylic acid, and protons, in addition to hydrogen-bonding and van der Waals interactions play a very important role in hydrogelation. Furthermore, it was found that the water-insoluble carboxylic acid compound underwent a precipitation dissolution transition with a thermally reversible sol-gel transition in the two component gelator systems. PMID- 18161707 TI - Acute hepatitis C virus infection: a chronic problem. PMID- 18161709 TI - Interleukin-22, interleukin-17, and other cytokines: a wall is coming down. PMID- 18161710 TI - Towards a tunable tautomeric switch in azobenzene biomimetics: implications for the binding affinity of 2-(4'-hydroxyphenylazo)benzoic acid to streptavidin. AB - The tautomeric equilibria of 2-(4'-hydroxyphenylazo)benzoic acid (HABA) and 2 (3',5'-dimethyl-4'-hydroxyphenylazo)benzoic acid (3',5'-dimethyl-HABA) have been studied by a combination of spectroscopic and computational methods. For neutral HABA in solvents of different polarity (toluene, chloroform, DMSO, DMF, butanol, and ethanol) the azo tautomer (AT) is largely predominant. For monoanionic HABA, the hydrazone tautomer (HT) is the only detected species in apolar solvents such as toluene and chloroform, while the AT is the only detected species in water and a mixture of both tautomers is detected in ethanol. Comparison of the results obtained for HABA and its 3',5'-dimethylated derivative shows that dimethylation of the hydroxybenzene ring shifts the tautomeric preferences towards the hydrazone species. These findings have been used to examine the differences in binding affinity to streptavidin, as the lower affinity of HABA can be explained in terms of the larger energetic cost associated with the tautomeric shift to the bioactive hydrazone species. Overall, these results suggest that a balanced choice of chemical substituents, embedding environment, and pH can be valuable for exploitation of the azo-hydrazone tautomerism of HABA biomimetics in biotechnological applications. PMID- 18161711 TI - FibroTest has better diagnostic and prognostic values than the aspartate aminotransferase-to-platelet ratio index in patients with chronic hepatitis C. PMID- 18161712 TI - A pattern recognition based fluorescence quenching assay for the detection and identification of nitrated explosive analytes. AB - Herein we report a differential array of micelle-solubilized fluorophores for the detection and identification of small nitrated analytes, such as the explosives TNT, tetryl, RDX and HMX. The quenching ability of the analytes can be used to correlate their analyte identity, wherein the quenching patterns generated from the differential array are used in linear discriminant analysis (LDA). LDA results in a well-clustered two-dimensional plot, and a jack-knife analysis of the data suggests that this system can be used to identify unknown samples of analyte with 96 % accuracy and with a detection limit of 19 muM. PMID- 18161713 TI - Predictors of response of U.S. veterans to treatment for the hepatitis C virus. PMID- 18161715 TI - Interactions with hydrophobic clusters in the urea-unfolded membrane protein OmpX. PMID- 18161716 TI - Comparative properties of insulin-like growth factor 1 (IGF-1) and [Gly7D-Ala]IGF 1 prepared by total chemical synthesis. PMID- 18161717 TI - Locoregional treatment for hepatocellular carcinoma: From clinical exploration to robust clinical data, changing standards of care. PMID- 18161718 TI - The largest 15N-15N coupling constant across an NHN hydrogen bond. PMID- 18161719 TI - Hepatitis C virus living off the fat of the land. PMID- 18161720 TI - Racial disparities in the management of hospitalized patients with cirrhosis and portal hypertension. PMID- 18161721 TI - Catching the rainbow: light harvesting in dye-sensitized solar cells. PMID- 18161722 TI - Catalytic asymmetric fluorination comes of age. PMID- 18161724 TI - Conflict of interest policy. PMID- 18161723 TI - Nonalcoholic fatty liver disease (NAFLD) and cardiac lipotoxicity: Another piece of the puzzle. PMID- 18161725 TI - Exploring the substrate specificity of a mycobacterial polyprenol monophosphomannose-dependent alpha-(1-->6)-mannosyltransferase. AB - A series of synthetic alpha-(1-->6)-linked octyl mannopyranoside oligomers was evaluated as potential acceptors of a polyprenol monophosphomannose-dependent alpha-(1-->6)-mannosyltransferase that is involved in the biosynthesis of the mannan core of mycobacterial lipoarabinomannan. Initial evaluation demonstrated that the enzyme recognizes di-, tri- and tetramannosides (5, 6 and 7) as substrates with different activities. While the highest mannosyltransferase activities were observed when the di- and trisaccharide were used as substrates, diminished enzymatic activity was seen with the tetramannoside. As octyl alpha-D mannopyranosyl-(1-->6)-alpha-D-mannopyranoside (5) appears to be the minimum structural element required for mannosyltransferase catalysis, a panel of methoxy and deoxy disaccharide analogues (8-21) were used to probe the substrate specificity of the enzyme further. In terms of the steric requirements at the active site, the enzyme does not recognize either C2'- and C2-methoxy analogues as substrates, a result that suggests that the alpha-(1-->2)-mannopyranosyl branches, which are present in the mannan core of LAM must be added on a larger alpha-(1-->6)-oligomannan intermediate. In contrast, the presence of a methoxy functionality at the C3', C3, C4' and C4 positions are somewhat tolerated by the enzyme, although diminished enzyme activities were observed with the C4'- and C4 methoxy analogues. Moreover, the 2'- and 4-hydroxyl groups appear not to be critical for substrate binding at the active site, as both 2'- and 4-deoxy analogues are substrates for the enzyme. In contrast, replacement of the hydroxyl groups at other positions essentially abolished enzymatic activity. Further kinetic characterization of the enzyme by using the effective acceptor substrates gave apparent K(M) values ranging from 111 to 437 microM, which are within two fold higher or lower than that for the parent dimannoside (5). Although the K(M) values indicate that the enzyme binds those acceptors with comparable affinities, the C4'-methoxy analogue (12) turns over more slowly than the others, as indicated by the apparent V(max) values. PMID- 18161726 TI - Can matrix metalloproteinases be targeted in hepatic ischemia/reperfusion injury? PMID- 18161727 TI - Parallel analysis of two analytes in solutions or on surfaces by using a bifunctional aptamer: applications for biosensing and logic gate operations. AB - A bifunctional aptamer that includes two aptamer units for cocaine and adenosine 5'-monophosphate (AMP) is blocked by a nucleic acid to form a hybrid structure with two duplex regions. The blocked bifunctional aptamer assembly is used as a functional structure for the simultaneous sensing of cocaine or AMP. The blocked bifunctional aptamer is dissociated by either of the two analytes, and the readout of the separation of the sensing structure is accomplished by a colorimetric detection, by a released DNAzyme, or by electronic means that use Faradaic impedance spectroscopy or field-effect transistors. In one configuration, the blocked bifunctional aptamer structure is separated by the substrates cocaine or AMP, and the displaced blocker units act as a horseradish peroxidase-mimicking DNAzyme that permits the colorimetric detection of the analytes. In the second system, the blocked bifunctional aptamer hybrid is associated with a Au electrode. The displacement of the aptamer by any of the substrates alters the interfacial electron transfer resistance at the electrode surface, thus providing an electronic signal for the sensing process. In the third configuration, the blocked aptamer hybrid is linked to the gate of a field effect transistor device. The separation of the complex by means of any of the analytes, cocaine, or AMP alters the gate potential, and this allows the electronic transduction of the sensing process by following the changes in the gate-to-source potentials. The different systems enable not only the simultaneous detection of the two analytes, but they provide a functional assembly that performs a logic gate "OR" operation. PMID- 18161729 TI - External validation of FIB-4: diagnostic accuracy is limited in elderly populations. PMID- 18161730 TI - The role of the conserved threonine in P450 BM3 oxygen activation: substrate determined hydroxylation activity of the Thr268Ala mutant. AB - The hydroxylation activity of the Thr268Ala mutant of P450(BM3) has been shown to occur to varying degrees with small alterations in the structure of a fatty-acid substrate. Ten substrates were investigated, including straight chain, branched chain and cis-cyclopropyl substituted fatty acids with a straight-chain length that varied between 12 and 16 carbon atoms. The efficacy of the hydroxylation activity appeared to be governed by the chain length of the substrate. Substrates possessing 14 to 15 carbons afforded the highest levels of activity, which were comparable with the wild-type enzyme. Outside of this window, straight-chain fatty acids showed reduced activity over the other substrate types. These results provide a cautionary tale concerning the loss of ferryl activity in such cytochrome P450 threonine to alanine mutants, as the nature of the substrate can determine the extent to which hydroxylation chemistry is abolished. PMID- 18161731 TI - H2O2/nitrite-induced post-translational modifications of human hemoglobin determined by mass spectrometry: redox regulation of tyrosine nitration and 3 nitrotyrosine reduction by antioxidants. AB - Covalent modifications of proteins by endogenous reactive nitrogen oxide species lead to cytotoxic effects that are implicated in diseases associated with chronic infections and inflammation. Tyrosine nitration is a major post-translational modification of proteins by reactive nitrogen oxide species. Recent studies suggest that nitrotyrosine is not a permanent protein modification. We previously demonstrated that lipoyl dehydrogenase is capable of converting 3-nitrotyrosine into 3-aminotyrosine in the presence of certain reducing agents. In this study, we compared the abilities of various hemoproteins, hemin, and the cobalt containing cofactor cyanocobalamin to mediate H(2)O(2)/nitrite-dependent tyrosine nitration and found that these hemoproteins and metal-containing cofactors also catalyzed the reduction of 3-nitrotyrosine to various extents in the presence of thiol reducing agents or ascorbate. The H(2)O(2)/nitrite-induced post translational modifications of human hemoglobin identified by nanoLC/nanospray ionization tandem mass spectrometric analysis of the tryptic digest include nitration of tyrosine and tryptophan, as well as oxidation of methionine and cysteine residues. Nitration of human hemoglobin by H(2)O(2)/nitrite was detected on Tyr24 and Tyr42 (alpha-chain) and on Tyr130 and Trp15 (beta-chain) in the alphabeta-dimer. Oxidation of methionine and cysteine residues was also observed. Furthermore, hemoglobin also catalyzed nitro reduction of 3-nitrotyrosine to form 3-aminotyrosine, at Tyr24 in the alpha-chain peptide of human Hb in the presence of ascorbate. The enhanced peroxidase activity of nitrated hemoglobin can be reversed by the antioxidant ascorbate. These results suggest a possible in vivo pathway for hemoglobin contributing to denitration of nitrated proteins through redox regulation. PMID- 18161732 TI - Hepatitis C virus genotype 1b, cirrhosis, and risk of hepatocellular carcinoma. PMID- 18161734 TI - Scalable and concise synthesis of dichlorofluorescein derivatives displaying tissue permeation in live zebrafish embryos. PMID- 18161737 TI - Solid-state NMR spectroscopy of amyloid proteins. PMID- 18161738 TI - Antiproliferative agents that interfere with the cell cycle at the G1-->S transition: further development and characterization of a small library of stilbene-derived compounds. AB - In this continuation of our research on derivatives containing the stilbene privileged structure or that are derived from it, we report the results of further studies carried out on the previously initiated collection of compounds. We used a parallel synthetic approach to rapidly obtain small sets of compounds and started the annotation of the library in progress by calculating some physicochemical properties to be eventually correlated with biological activities. A pharmacophore for the antiproliferative activity was also built to summarize the features of the library. We evaluated the antiproliferative and pro apoptotic activities of all compounds as well as the cell-cycle effects of some representative compounds. After in-depth investigations, 3'-phenyl [1,1';4',1'']terphenyl-4,3'',5''-triol showed the most interesting biological profile, as it interferes with cell-cycle progression at the G(1)-->S transition, acting on retinoblastoma phosphorylation and inducing cell differentiation. PMID- 18161739 TI - The calculation of polar surface area from first principles: an application of quantum chemical topology to drug design. AB - The calculation of polar surface areas (PSA) from the electron density using quantum chemical topology (QCT) and a newly developed algorithm to determine isodensity surface areas is described. PSA values were calculated from the atomic partitioning of B3LYP/6-311G* wavefunctions and the results described herein represent the first application of this new algorithm. PSA values were calculated for forty drugs and compared to the topological polar surface area (TPSA) and those calculated by the QikProp program. Oral bioavailabilities predicted from the QCT PSA values for a subset of twenty drugs (the Palm set) were similar to those predicted by the dynamic polar surface area (DPSA) and in general, are in agreement with the observed values. Overall, PSA values obtained from QCT were generally similar to the DPSA, TPSA, and QikProp values, though differences in fragment contributions were found, with nitrogen-bearing functional groups showing the largest variation between methods. Differences between methods showed how the calculation of the PSA is dependent on the method used and, therefore, judicious application of the upper limits used in the prediction of oral bioavailability is warranted. These results also indicate that, because of the differences in the way PSA values are calculated, values from the different methods should not be used interchangeably. PMID- 18161740 TI - Carbonic anhydrase inhibitors: binding of indanesulfonamides to the human isoform II. AB - Indanesulfonamides are interesting lead compounds for designing selective inhibitors of the different isoforms of the zinc enzyme Carbonic Anhydrase (CA). Herein, we report for the first time the X-ray crystal structure of two such derivatives, namely indane-5-sulfonamide and indane-2-valproylamido-5 sulfonamide, in complex with the physiologically dominant human isoform II. The structural analysis reveals that, although these two inhibitors have quite similar chemical structures, the arrangement of their indane ring within the enzyme active site is significantly diverse. Thus, our findings suggest that the introduction of bulky substituents on the indane-sulfonamide ring may alter the binding mode of this potent class of CA inhibitors, although retaining good inhibitory properties. Accordingly, the introduction of bulky tail moieties on the indane-sulfonamide scaffold may represent a powerful strategy to induce a desired physicochemical property to an aromatic sulfonamide or to obtain inhibitors with diverse inhibition profiles and selectivity for various mammalian CAs. PMID- 18161741 TI - Lanthanide coordination polymers constructed from infinite rod-shaped secondary building units and flexible ligands. AB - Three lanthanide coordination polymers constructed from infinite rod-shaped secondary building units (SBUs), [Nd(2)(H(2)O)(2)(cis-chdc)(2)(trans-chdc)]2H(2)O (1), Nd(2)(H(2)O)(4)(trans-chdc)(3) (2), and [Sm(2)(H(2)O)(2)(cis-chdc)(trans chdc)(2)]4H(2)O (3) (chdcH(2)=1,4-cyclohexanedicarboxylic acid), were hydrothermally synthesized and structurally characterized. The structures of 1-3 are modulated by different ratios of the cis and trans configurations of chdc(2-) ligands, which was achieved by temperature control in the hydrothermal reactions. Crystal-structure analysis revealed that 1 is a four-connected pcu-type rod packing network built from cross-linking of rod-shaped neodymium-oxygen SBUs by cis- and trans-chdc(2-) ligands in a 2:1 ratio, 2 displays a complicated six connected hex-type rod packing structure built by connection of rod-shaped neodymium-oxygen SBUs and trans-chdc(2-) ligands, and 3 features an unprecedented five-connected rod packing pattern constructed from rod-shaped samarium-oxygen SBUs and cis- and trans-chdc(2-) ligands in a 1:2 ratio. PMID- 18161742 TI - Chiral-catalyst-based convergent synthesis of HIV protease inhibitor GRL-06579A. AB - Catalytic asymmetric synthesis of GRL-06579A (1), an HIV-1 protease inhibitor effective against multi-protease-inhibitor-resistant viruses, is described. A convergent strategy that utilizes heterobimetallic multifunctional catalysts developed in our group is a key feature of the synthesis. The chirality of the bicyclic tetrahydrofuran unit of 1 was introduced through Al-Li bis(binaphthoxide) (ALB) catalyst-controlled Michael addition of dimethyl malonate to racemic 4-O-protected cyclopentenone. ALB afforded not only the trans adduct with up to 96% ee from a matched substrate through kinetic resolution, but also the cis adduct with 99% ee through a catalyst-controlled Michael addition to a mismatched substrate. The Michael addition to produce the unusual cis adduct is described in detail. The framework of the bicyclic tetrahydrofuran was constructed by an intramolecular oxy-Michael reaction. The amino alcohol unit was constructed by an La-Li3-tris(binaphthoxide) (LLB)-catalyzed diastereoselective nitroaldol reaction of N-Boc aldehyde (Boc = tert-butoxycarbonyl) derived from L phenylalanine. LLB promoted the nitroaldol reaction without racemization of the chiral aldehyde to give the nitroaldol adduct in 85% yield and with 93:7 diastereoselectivity and over 99% ee. PMID- 18161743 TI - Mechanisms of action of interferon and ribavirin in chronic hepatitis C: Summary of a workshop. PMID- 18161744 TI - Macrophage activation by endogenous danger signals. AB - Macrophages are cells that function as a first line of defence against invading microorganisms. One of the hallmarks of macrophages is their ability to become activated in response to exogenous 'danger signals'. Most microbes have molecular patterns (PAMPS) that are recognized by macrophages and trigger this activation response. There are many aspects of the activation response to PAMPS that are recapitulated when macrophages encounter endogenous danger signals. In response to damaged or stressed self, macrophages undergo physiological changes that include the initiation of signal transduction cascades from germline-encoded receptors, resulting in the elaboration of chemokines, cytokines and toxic mediators. This response to endogenous mediators can enhance inflammation, and thereby contribute to autoimmune pathologies. Often the overall inflammatory response is the result of cooperative activation signals from both exogenous and endogenous signals. Macrophage activation plays a critical role, not only in the initiation of the inflammatory response but also in the resolution of this response. The clearance of granulocytes and the elaboration of anti-inflammatory mediators by macrophages contribute to the dissolution of the inflammatory response. Thus, macrophages are a key player in the initiation, propagation and resolution of inflammation. This review summarizes our understanding of the role of macrophages in inflammation. We pay particular attention to the endogenous danger signals that macrophages may encounter and the responses that these signals induce. The molecular mechanisms responsible for these responses and the diseases that result from inappropriately controlled macrophage activation are also examined. PMID- 18161745 TI - Cellular and molecular mechanisms of fibrosis. AB - Fibrosis is defined by the overgrowth, hardening, and/or scarring of various tissues and is attributed to excess deposition of extracellular matrix components including collagen. Fibrosis is the end result of chronic inflammatory reactions induced by a variety of stimuli including persistent infections, autoimmune reactions, allergic responses, chemical insults, radiation, and tissue injury. Although current treatments for fibrotic diseases such as idiopathic pulmonary fibrosis, liver cirrhosis, systemic sclerosis, progressive kidney disease, and cardiovascular fibrosis typically target the inflammatory response, there is accumulating evidence that the mechanisms driving fibrogenesis are distinct from those regulating inflammation. In fact, some studies have suggested that ongoing inflammation is needed to reverse established and progressive fibrosis. The key cellular mediator of fibrosis is the myofibroblast, which when activated serves as the primary collagen-producing cell. Myofibroblasts are generated from a variety of sources including resident mesenchymal cells, epithelial and endothelial cells in processes termed epithelial/endothelial-mesenchymal (EMT/EndMT) transition, as well as from circulating fibroblast-like cells called fibrocytes that are derived from bone-marrow stem cells. Myofibroblasts are activated by a variety of mechanisms, including paracrine signals derived from lymphocytes and macrophages, autocrine factors secreted by myofibroblasts, and pathogen-associated molecular patterns (PAMPS) produced by pathogenic organisms that interact with pattern recognition receptors (i.e. TLRs) on fibroblasts. Cytokines (IL-13, IL-21, TGF-beta1), chemokines (MCP-1, MIP-1beta), angiogenic factors (VEGF), growth factors (PDGF), peroxisome proliferator-activated receptors (PPARs), acute phase proteins (SAP), caspases, and components of the renin-angiotensin-aldosterone system (ANG II) have been identified as important regulators of fibrosis and are being investigated as potential targets of antifibrotic drugs. This review explores our current understanding of the cellular and molecular mechanisms of fibrogenesis. PMID- 18161746 TI - Nod-like proteins in inflammation and disease. AB - The field of innate immunity has undergone an enormous upheaval during the last decade. The discovery of different groups of proteins, called pattern recognition molecules (PRMs), which detect microbial components, so-called pathogen associated molecular patterns (PAMPs) and trigger protective responses, had a huge impact on the understanding of innate immune responses. Among the PRMs, the intracellular Nod-like receptors (NLRs) have recently been identified as key mediators of inflammatory and immune responses. The NLR family is divided into subfamilies on the basis of their different signal transduction domains, and recent studies have highlighted the role of certain NLRs, including Nod1, Nod2, Nalp3, Ipaf and Naip5, in the detection of intracellular microbes and possibly 'danger signals'. In this review, we summarize the current knowledge on the function of these proteins in immunity and inflammation, with a focus on their participation in different disease pathologies. PMID- 18161747 TI - Innate immunity in inflammatory bowel disease: a disease hypothesis. AB - Crohn's disease arises from a defective interaction between the highly concentrated mass of bacteria in the gastrointestinal tract and the underlying tissues. It has generally been believed to result from an excessively exuberant inflammatory response or from 'autoimmunity'. Recent evidence has emerged that the problem is instead a failure of the way in which the body responds to the penetration of bacteria and other bowel contents through the intestinal mucosal barrier. Rather than Crohn's disease being caused by excessive inflammation, the primary mechanism is actually that of an immunodeficiency. Failure of inflammatory mediator production leads to insufficient recruitment of neutrophils, resulting in inadequate removal of bacteria and other debris. This impairment of acute inflammation can be compensated in some circumstances by signalling through NOD2. If not cleared, the foreign material in the bowel wall is taken up within macrophages, eliciting a granulomatous reaction and the local and systemic sequelae so characteristic of Crohn's disease. PMID- 18161748 TI - The role of TLR activation in inflammation. AB - The Toll-like receptor family was originally identified in Drosophila, where it provides important developmental and immunological signalling. In mammals, the developmental signal appears to have been lost, but the immunological defence role of these receptors has been expanded to provide broad recognition of bacterial, fungal, viral and parasitic pathogens. There is increasing evidence that these receptors go beyond the recognition of microbial molecules to sense host tissue damage. Recognition of host molecules and commensal microbes is also involved in the restoration of normal tissue architecture after injury and in maintenance of epithelial health. Recent developments in the TLR field highlight the importance of these molecules to human health and disease and demonstrate that their targeting, to boost immunity or inhibit inflammation, is both feasible and also potentially challenging. PMID- 18161749 TI - Molecular and cellular themes in inflammation and immunology. AB - This issue of the Journal of Pathology contains a series of cutting-edge review articles that deal with the broad issue of inflammatory and immunological disease mechanisms. Of necessity, these reviews deal with selected topics but the mixture of articles on specific signalling pathways and mediators with articles addressing individual organ systems provides a broad overview of the field. These contributions provide insight into current areas of debate in inflammation and immunology. In particular, they highlight current interest in the interface between innate and adaptive immunity and present intriguing prospects for future therapeutic developments in a variety of disease areas. PMID- 18161750 TI - Innate and adaptive mechanisms to control [corrected] pathological intestinal inflammation. AB - The intestine is the home of a tremendous number of commensal organisms that have a primary role in host metabolism. As a consequence, the gut mucosa has evolved multiple layers of protection. This review highlights both innate and adaptive mechanisms that prevent bacterial invasion and abnormal intestinal inflamamation, how a failure of these mechanisms may be involved in the pathogenesis of inflammatory bowel diseases, and discusses new findings implicating dendritic cells as central to the induction of active mucosal tolerance to commensal bacteria. PMID- 18161751 TI - The highway code of T cell trafficking. AB - Coordinated migratory events are required for the development of effective and regulated immunity. Naive T lymphocytes are programmed to recirculate predominantly in secondary lymphoid tissue by non-specific stimuli. In contrast, primed T cells must identify specific sites of antigen location in non-lymphoid tissue to exert targeted effector responses. Following priming, T cells acquire the ability to establish molecular interactions mediated by tissue-selective integrins and chemokine receptors (homing receptors) that allow their access to specific organs, such as the skin and the gut. Recent studies have shown that an additional level of specificity is provided by the induction of specific T cell migration into the tissue following recognition of antigen displayed by the endothelium. In addition, co-stimulatory signals (such as those induced by CD28 and CTLA-4 molecules) have been shown not only to regulate T cell activation and differentiation, but also to orchestrate the anatomy of the ensuing T cell response. PMID- 18161752 TI - TNF-mediated inflammatory disease. AB - TNF was originally described as a circulating factor that can cause necrosis of tumours, but has since been identified as a key regulator of the inflammatory response. This review describes the known signalling pathways and cell biological effects of TNF, and our understanding of the role of TNF in human disease. TNF interacts with two different receptors, designated TNFR1 and TNFR2, which are differentially expressed on cells and tissues and initiate both distinct and overlapping signal transduction pathways. These diverse signalling cascades lead to a range of cellular responses, which include cell death, survival, differentiation, proliferation and migration. Vascular endothelial cells respond to TNF by undergoing a number of pro-inflammatory changes, which increase leukocyte adhesion, transendothelial migration and vascular leak and promote thrombosis. The central role of TNF in inflammation has been demonstrated by the ability of agents that block the action of TNF to treat a range of inflammatory conditions, including rheumatoid arthritis, ankylosing spondylitis, inflammatory bowel disease and psoriasis. The increased incidence of infection in patients receiving anti-TNF treatment has highlighted the physiological role of TNF in infectious diseases. PMID- 18161753 TI - The role of natural killer T cells in lung inflammation. AB - Invariant NK T cells (iNKT) bridge the innate and adaptive immune response, being characterized by the ability to use invariant T cell receptors to recognize glycolipid antigens presented by CD1d, leading to an explosive cytokine effector response. As such it has been proposed that iNKT cells perform important roles as both effector and regulatory cells in a wide range of disease settings. These roles have been characterized in experiments depending on the use of iNKT-null mice, due to lack of either CD1d expression or Jalpha18 and the use of CD1d tetramers loaded with the model glycolipid antigen, alpha-galactosylceramide (alphaGalCer). Several studies have examined lung disease, infectious and allergic, in humans and mice. While the lung itself does not carry an exceptionally large population of iNKT cells (compared with, say, the liver), it does appear to be a site at which these cells can exert a profound effect. Several models of bacterial, fungal and viral murine lung infection have been investigated that have sometimes produced conflicting results. Abrogation of iNKT cell function in knockouts is often associated with disease exacerbation, indicating a regulatory role in lung infection. Studies in murine asthma models and in patients have similarly probed the role of iNKT cells in these settings. While there are again somewhat contradictory findings, evidence suggests a likely role for iNKT cells in mediating airway hyper-responsiveness (AHR), but probably not in Th2 polarization or lung eosinophilia. In marginally different models, administration of alphaGalCer has either ameliorated or exacerbated AHR. Different studies of BAL from human asthma patients show variously that there is either a very enlarged population of iNKT cells in the asthmatic lung, or that there is no significant difference from controls. Taken together, there are some observations that argue compellingly for an important role of iNKT cells in the lung, but resolution of some of the contradictory findings may await the development of reagents capable of providing alternative readouts of iNKT activation in these diverse disease settings. PMID- 18161754 TI - The immunology of sepsis. AB - Sepsis, the systemic inflammatory response to infection, is considered the major cause of death among critically ill patients in the developed world. While there is a general view that this reflects contributions from both the pathogen and the host with respect to an inappropriate inflammatory response, there is a lack of agreement as to the key immune mechanisms. This has been reflected in the diverse range of immunotherapies tested in clinical trials, often with rather marginal effects. The case has been made for a pathogenic role of excessive immunity, the so-called 'cytokine storm', and for a role of too little immunity through immune paralysis. Apoptosis is implicated as a key mechanism in both this immune paralysis and the multi-organ failure that is a feature of severe sepsis. A number of polymorphisms have been implicated in susceptibility to sepsis, including cytokine genes, HLA class II and caspase-12. In this review we focus in particular on the role of group A streptococci in severe sepsis. Here the effect of bacterial superantigens appears to be a correlate of inflammatory activation, although the precise evolutionary role of the superantigens remains unclear. PMID- 18161755 TI - Stress proteins in CNS inflammation. AB - Stress proteins or heat shock proteins (HSPs) are ubiquitous cellular components that have long been known to act as molecular chaperones. By assisting proper folding and transport of proteins, and by assisting in the degradation of aberrant proteins, they play key roles in cellular metabolism. The frequent accumulation of insoluble protein aggregates during chronic neurodegenerative disorders suggests failure of HSP functions to be a common denominator among such diseases. Recent developments have clarified that functions of HSPs extend well beyond their role in protein folding and degradation alone. Stress-inducible HSPs also regulate apoptosis, antigen presentation, inflammatory signalling pathways and, intriguingly, also serve as extracellular mediators of inflammation. Several receptors have been identified for extracellular HSPs, which control inflammatory pathways similar to those activated by cytokines and chemokines. In this review, both the traditional and the exciting novel functions of HSPs are discussed, with a focus on their relevance for neurodegeneration and neuroinflammation. Recent advances in this field suggest that HSPs represent attractive novel targets as well as therapeutic entities for CNS disorders. PMID- 18161756 TI - Control of apoptosis in autoimmunity. AB - Apoptosis and the subsequent removal of apoptotic cells underpin a healthy immune system. They are crucial for both the maintenance of self-tolerance and the contraction of clonally expanded lymphocytes at the conclusion of immune responses. Aberrant apoptosis and the disposal of apoptotic cells is implicated in the development of both systemic and organ-specific autoimmune disease and is a major contributing factor in disease susceptibility. Dissection of the molecular mechanisms involved in dysregulated apoptosis may reveal pathways which can be targeted for more effective therapeutic intervention. This review highlights the molecular events underlying programmed cell death and apoptotic cell uptake, and summarizes recent studies that link impaired apoptotic death to autoimmunity. PMID- 18161757 TI - IL-10 and the resolution of infections. AB - Chronic viral infections pose serious health concerns, as secondary complications such as immunodeficiencies and cancers are common. Treating such infections with conventional vaccine approaches has proved to be difficult. Studies in animals and humans suggest that vaccine failure is probably due to exhaustion of antiviral T cell responses, which occurs in a number of chronic infections. Attempts to elucidate the causes of impairment of antiviral immunity have pointed to a role for the immunomodulatory cytokine IL-10 in the ability of viruses to establish persistence. Induction of IL-10 production by the host during chronic infection appears to be one of the viral means to alter the class of the antiviral immune response and induce generalized immune suppression. Recent work by us and others suggests that it is possible to resuscitate antiviral immunity by interfering with the IL-10 signalling pathway. Targeting IL-10 thus constitutes a promising alternative to conventional vaccine strategies which have not proved to be successful in treating chronic infections. In addition, sterile cure may be achieved with minimal side-effects by combining agents that alter the IL-10 signalling pathway with other compounds, such as antiviral drugs or interferon, but also agents neutralizing other crucial elements of T cell exhaustion, such as PD-1. PMID- 18161758 TI - Immune activation and inflammation in HIV-1 infection: causes and consequences. AB - Thorough research on HIV is progressively enabling us to understand the intricate mechanisms that link HIV-1 infection to the onset of immunodeficiency. The infection and depletion of CD4(+) T cells represent the most fundamental events in HIV-1 infection. However, in recent years, the role played by chronic immune activation and inflammation in HIV pathogenesis has become increasingly apparent: quite paradoxically, immune activation levels are directly associated with HIV-1 disease progression. In addition, HIV-1-infected patients present intriguing similarities with individuals of old age: their immune systems are characterized by a loss of regenerative capacity and an accumulation of ageing T cells. In this review, we discuss the potential reasons for the establishment of sustained immune activation and inflammation from the early stages of HIV-1 infection, as well as the long-term consequences of this process on the host immune system and health. A simplified model of HIV pathogenesis is proposed, which links together the three major facets of HIV-1 infection: the massive depletion of CD4(+) T cells, the paradoxical immune activation and the exhaustion of regenerative capacity. PMID- 18161759 TI - A classification system for faecal incontinence based on anorectal investigations. AB - BACKGROUND: Faecal incontinence is a socially disabling condition that affects a heterogeneous population of patients. There is no standardization of investigations, and treatment outcomes are variable. The major limitation for comparing the results from different studies is the lack of a pretreatment classification of incontinence. The aim of this study was to review the anorectal investigation findings and propose a simple, repeatable classification for faecal incontinence. METHODS: Patients who had anorectal investigations for defaecatory disorders from February 2000 to September 2006 were analysed retrospectively. All patients had anorectal manometry, anal mucosal electrosensitivity testing and endoanal ultrasonography. RESULTS: Of a total of 1294 patients, 135 were excluded, leaving 1159 (460 continent and 699 incontinent) for analysis. The patients were divided into four groups: traumatic incontinence, neuropathic faecal incontinence, combined faecal incontinence and idiopathic faecal incontinence. The manometric variables and demographics were distinct in these groups. CONCLUSION: Patients with faecal incontinence can be classified into different groups with distinct pathophysiological variables. Such a classification system will enable comparison and interpretation of the outcomes of different studies and also help in the selection of patients for appropriate treatments. PMID- 18161760 TI - Randomized clinical trial evaluating elective laparoscopic appendicectomy for chronic right lower-quadrant pain. AB - BACKGROUND: It is questionable whether elective appendicectomy can effectively reduce persistent or recurrent right lower-quadrant abdominal pain due to chronic or recurrent appendicitis. METHODS: This single-centre double-blind randomized clinical trial studied the effects of elective laparoscopic appendicectomy on pain 6 months after operation in patients with persistent or recurrent lower quadrant pain. A secondary outcome evaluated was the relationship between clinical response and appendiceal histopathology. The analysis was performed on an intention-to-treat basis. RESULTS: Forty patients were randomized to laparoscopic appendicectomy (18) or laparoscopic inspection only (22). Postoperative pain scores differed significantly between the groups, favouring appendicectomy (P = 0.005). Relative risk calculations indicated that there was a 2.4 (95 per cent confidence interval (c.i.) 1.3 to 4.0) times greater chance of improvement in pain after laparoscopic appendicectomy. The number needed to treat was 2.2 (95 per cent c.i. 1.5 to 6.5). There was no association between postoperative pain scores and histopathology findings. CONCLUSION: Persistent or recurrent lower abdominal pain can be treated by elective appendicectomy with significant pain reduction in properly selected cases. Histopathology may not be abnormal. REGISTRATION NUMBER: ISRCTN48831122 (http://www.controlled-trials.com). PMID- 18161761 TI - Late morbidity after duodenum-preserving pancreatic head resection with bile duct reinsertion into the resection cavity. AB - BACKGROUND: Reinsertion of the distal common bile duct (CBD) into the pancreatic resection cavity during duodenum-preserving pancreatic head excision (DPPHE) may be an alternative option to Whipple resection or bilioenteric anastomosis when chronic pancreatitis is associated with CBD stenosis. METHODS: Outcome in 82 patients with chronic pancreatitis who underwent DPPHE with CBD reinsertion was compared with that in 432 who had DPPHE without reinsertion and 50 who had a Whipple procedure or pylorus-preserving pancreatoduodenectomy (PPPD). RESULTS: There were no deaths after DPPHE with CBD reinsertion, compared with four (0.9 per cent) after DPPHE without reinsertion and three (6 per cent) after classical resection. Overall morbidity rates were 30, 28.9 and 36 per cent respectively. Fifteen patients (18 per cent) who had DPPHE with CBD reinsertion developed a stricture at the reinsertion site, compared with a long-term stricture rate of 2.3 per cent (ten patients) after DPPHE without CBD reinsertion and 4 per cent (two patients) after PPPD/Whipple resection. CONCLUSION: Although associated with a high incidence of anastomotic stricture, reinsertion of the CBD into the resection cavity as part of DPPHE can be used to preserve duodenal passage and offers an alternative to extended resection for chronic pancreatitis. PMID- 18161762 TI - The impact of published recommendations on the management of penetrating abdominal injury. AB - BACKGROUND: In patients with penetrating abdominal injury (PAI), haemodynamic instability and peritonitis are indications for laparotomy, but it remains uncertain whether laparotomy is indicated for evisceration, retained foreign body and pneumoperitoneum. In 1989, a review of 107 patients with PAI revealed a 78.5 per cent laparotomy rate, with 35 per cent considered unnecessary. The aim of this study was to review current practice in the same hospitals. METHODS: A retrospective review included case notes from 224 patients with PAI presenting to three hospitals between 2001 and 2005. RESULTS: Some 206 patients (92.0 per cent) were male and the mean age was 30.5 years. Aetiologies were stabbing (96.4 per cent), impalement (2.7 per cent) and gunshot wound (0.9 per cent). Laparotomy was performed in 48 patients (21.4 per cent), and was positive in 33 and unnecessary or negative in 15. Haemodynamic instability and peritonitis were strong indicators of positive laparotomy; seven of 13 laparotomies for evisceration alone were negative, as were two of four for retained foreign bodies. CONCLUSION: The laparotomy rate fell from 78.5 to 21.4 per cent over 25 years. The rate of unnecessary or negative laparotomy did not change. Isolated evisceration and retained foreign body remain relative indications. PMID- 18161763 TI - Microbubble-enriched lavage fluid for treatment of experimental peritonitis. AB - BACKGROUND: Relaparotomies and closed postoperative peritoneal lavage (CPPL) are performed to treat persistent peritonitis. This experimental animal study compared open abdominal lavage with CPPL, and evaluated the potential of microbubble-enriched lavage fluids to improve the efficiency of CPPL and reduce clinical morbidity, mortality and cost. METHODS: Fluorescent polystyrene spheres were injected intraperitoneally into 22 male Wistar rats to simulate localized peritonitis. After 18 h the rats received open abdominal lavage and CPPL, with and without microbubbles. Microbubbles were obtained by adding ultrasound contrast agents to continuous ambulatory peritoneal dialysis fluid. RESULTS: Open abdominal lavage was 3.5 times more effective in particle removal than CPPL, owing to better fluid dynamics. The introduction of air-liquid interfaces in the form of microbubbles made CPPL up to 2.4 times more effective than lavage without bubbles. Best detachment results were obtained when microbubbles with a flexible surfactant shell and longer blood elimination half-life were used. CONCLUSION: Open abdominal and CPPL lavage techniques are not efficient beyond a certain duration and volume as they do not cause bacterial detachment from the peritoneal membrane. Using surface tension forces from microbubbles significantly enhanced polystyrene particle detachment. These findings may have great consequences for the treatment of patients with peritonitis. PMID- 18161764 TI - Another patient with an umbilical hernia and massive ascites: what to do? PMID- 18161765 TI - Extracorporeal hepatic resection for unresectable giant hepatic hemangiomas. PMID- 18161766 TI - Use of a metallic-wall stent in the magnet compression anastomosis technique for bile duct obstruction after liver transplantation. PMID- 18161767 TI - Prognostic impact of lactate in acute liver failure. PMID- 18161769 TI - In defense of the administration of perioperative steroids in liver transplantation. PMID- 18161770 TI - Human herpesvirus 6 infection in adult living related liver transplant recipients. AB - To analyze human herpesvirus 6 (HHV-6) infection in adult living related liver transplantation, we performed a virological analysis, including viral isolation, serological assay, and real-time polymerase chain reaction, of serially collected blood samples from 67 recipients. In addition, cytokine levels were measured to determine their role in viral reactivation. HHV-6 was isolated from only 4 recipients (6.0%), and viral DNA was detected in 15 (22.4%) of the 67 recipients. A significant increase in HHV-6 immunoglobulin G antibody titers was observed in 19 (28.4%) of the 67 recipients. Finally, 26 recipients (38.8%) had HHV-6 reactivation 2-6 weeks after transplantation. HHV-6 associated clinical features were analyzed in the 17 recipients presenting with either viremia or DNAemia. Two recipients with viremia and 3 recipients with DNAemia had unexplained fever at the time of viral infection. An increase in aminotransferase levels was observed in 2 recipients with viremia and 3 recipients with DNAemia. Recipients with liver cirrhosis caused by hepatitis B virus or hepatitis C virus infection as the underlying disease were more likely to have HHV-6 infection (P = 0.025). Mortality at the last follow-up in recipients with HHV-6 reactivation was significantly higher than in those without viral reactivation (P = 0.0118). Plasma interleukin-6 levels were significantly higher in the recipients with HHV 6 viremia than in the recipients without viremia at 4 weeks post-transplant (P = 0.0411). Moreover, tumor necrosis factor alpha levels were also higher in recipients with HHV-6 viremia (P < 0.0001) or reactivation (P = 0.0011) than in recipients without viremia or reactivation 4 weeks post-transplant. PMID- 18161771 TI - Early invasive Listeria monocytogenes infection after orthotopic liver transplantation: case report and review of the literature. AB - Infection with Listeria monocytogenes is rare, with a reported annual incidence of 4.4 cases per million individuals. It is caused by a gram-positive rod-shaped bacterium (Listeria monocytogenes) that can be found in soil, vegetation, water, sewage, and silage and in feces of humans and animals. It is a facultative intracellular pathogen with the ability to survive and multiply in phagocytic host cells, even in adverse environmental circumstances. Listeriosis has rarely been reported after orthotopic liver transplantation, and transplant physicians are often unfamiliar with the clinical presentation of this rare but virulent infection, which accounts for 20%-30% mortality in affected individuals. We present a case of invasive Listeria infection causing bacteremia and peritonitis in the early postoperative period after cadaveric liver transplantation in a previously asymptomatic patient. PMID- 18161772 TI - Olfactory and gustatory sensory changes to tobacco smoke in pregnant smokers. AB - Models of smoking behavior change include addiction, social, and behavioral concepts. The purpose of this study was to explore the prevalence of two biologic factors, olfactory and gustatory responses to tobacco smoke, as potentially powerful contributors to smoking behavior change among pregnant women. Data were obtained from 209 pregnant smokers. The majority of women reported olfactory (62%) and gustatory (53%) aversions to tobacco. Aversions first appeared during the first trimester of pregnancy. Women who experienced olfactory aversions were more likely also to experience gustatory aversions. Olfactory aversions were associated with women smoking less. Aversions to tobacco smoke are common among pregnant smokers, are associated with women smoking less, and could help explain pregnant women's smoking patterns. PMID- 18161773 TI - English-language acculturation predicts academic performance in nursing students who speak English as a second language. AB - Students who speak English as a second language (ESL) face considerable challenges in English language universities, but little is known about the relationship between English-language acculturation and academic performance. A prospective, correlational design was used to validate the English Language Acculturation Scale (ELAS), a measure of the linguistic aspect of acculturation, and to determine the relationship between English-language acculturation and academic achievement among 273 first-year nursing students. Exploratory factor analyses demonstrated that the ELAS was a valid and reliable measure (alpha = .89). When ELAS scores were examined in relation to students' grades, students with the lowest ELAS scores also had the lowest mean subject grades, highlighting the need to place greater emphasis on identifying English-language acculturation among ESL students. PMID- 18161774 TI - Arboreal nesting as anti-predator adaptation by savanna chimpanzees (Pan troglodytes verus) in southeastern Senegal. AB - Chimpanzees (Pan troglodytes) make nests for resting and sleeping, which is unusual for anthropoid primates but common to all great apes. Arboreal nesting has been linked to predation pressure, but few studies have tested the adaptive nature of this behavior. We collected data at two chimpanzee study sites in southeastern Senegal that differed in predator presence to test the hypothesis that elevated sleeping platforms are adaptations for predator defense. At Assirik in the Parc National du Niokolo-Koba, chimpanzees face four species of large carnivore, whereas at Fongoli, outside national park boundaries, humans have exterminated almost all natural predators. We quantified the availability of vegetation at the two sites to test the alternative hypothesis that differences in nesting reflect differences in habitat structure. We also examined possible sex differences in nesting behavior, community demographic differences, seasonality and nest age differences as variables also potentially affecting nest characteristics and nesting behavior between the two sites. Chimpanzees at Fongoli nested at lower heights and farther apart than did chimpanzees at Assirik and sometimes made nests on the ground. The absence of predators outside of the national park may account for the differences in nest characteristics at the two sites, given the similarities in habitat structure between Fongoli and Assirik. However, Fongoli chimpanzees regularly build arboreal nests for sleeping, even under minimal predation pressure, and this requires explanation. PMID- 18161776 TI - Spontaneous clearance of hepatitis C virus after liver transplantation in two patients coinfected with hepatitis C virus and human immunodeficiency virus. AB - Spontaneous resolution of chronic hepatitis C virus (HCV) infection is exceedingly rare and poorly understood. As HCV and human immunodeficiency virus (HIV) have shared routes of transmission, HCV coinfection is estimated to affect 15%-30% of the HIV-positive population. We report 2 patients with HCV-HIV coinfection who underwent orthotopic liver transplantation at our center and had spontaneous clearance of their chronic HCV infection after transplantation without any anti-HCV treatment. Both patients showed no evidence of HCV recurrence for more than 3 years despite long-term immunosuppressant therapy. Spontaneous clearance of chronic HCV infection can occur in HIV-HCV-coinfected patients after liver transplantation. The mechanism of this phenomenon remains unclear. PMID- 18161777 TI - Triple-phase computed tomography and intraoperative flow measurements improve the management of portosystemic shunts during liver transplantation. AB - Ligation of portosystemic shunts in patients with cirrhosis undergoing liver transplantation has been recommended to avoid insufficient portal vein (PV) flow. Shunts are not always recognized pretransplantation because intraoperative PV flow assessment is not routinely attempted. As a result of a posttransplantation PV thrombosis in a recipient with a large portosystemic shunt and a PV flow <1 L/minute, we employed triple-phase computed tomography with vascular reconstruction and intraoperative graft flow measurement to determine the need for inflow modification in our next 16 patients with large portosystemic shunts. Subsequently, 6 patients with large portosystemic shunts and PV flows 0.05) were observed among the two treatments throughout the implant intervals. However, by the end of the 8 weeks, the quantity of bone marrow was two times greater (p < 0.05) in the control group than in the XC group. In conclusion, the xenograft composite promotes formation of new bone in a similar fashion to autogenous bone and could therefore be considered a biomaterial with potential applications as a bone substitute in maxillary sinus floor augmentation. PMID- 18161781 TI - In vitro microTBS of one-bottle adhesive systems: sound versus artificially created caries-affected dentin. AB - This in vitro study aimed to evaluate a pH-cycling model for simulation of caries affected dentin (CAD) surfaces, by comparing the bond strength of etch-and-rinse adhesive systems on sound and artificially-created CAD. Dentin substrates with different mineral contents and morphological patterns were created by submitting buccal bovine dentin to the following treatments: (1) immersion in artificial saliva during the experimental period (sound dentin, SD), or (2) induction to a CAD condition by means of a dynamic pH-cycling model (8 cycles, demineralization for 3 h followed by mineralization for 45 h). The bond strength of Excite or Prime and Bond NT adhesive systems was assessed using the microtensile bond strength (microTBS) test. Dentin microhardness was determined by cross-sectional Knoop evaluations. Resin-dentin morphology after the treatments was examined by scanning electron microscopy. SD produced significantly higher microTBS than CAD for both adhesives evaluated, without differences between materials. CAD exhibited lower microhardness than SD. Morphological analysis showed marked distinctions between SD and CAD bonded interfaces. Under the conditions of this study, differences in morphological pattern and dentin mineral content may help to explain resin-dentin bond strengths. The proposed pH-cycling model may be a suitable method to simulate CAD surfaces for bonding evaluations. PMID- 18161782 TI - Polypeptide-catalyzed silica for dental applications. AB - Polypeptides such as polylysine have been shown to catalyze the condensation and direct the structure of silica from precursor solutions under ambient conditions. Several of the reaction parameters have been shown to mediate this activity. Specifically, mechanical perturbation seems to play a role in the formation of hierarchical structures. Most studies have been conducted in solution, but biomedical and particularly dental applications will likely require control of biosilicified coatings, films or particle formation on surfaces. Tetraethylorthosilicate was reacted with polylysine and then spin coated onto a surface. The process parameters catalyst structure, pH, buffer: ethanol ratio and percentage of cocatalyst polyethyleneimine were varied to determine their effects on the formed silica. The chemical nature and morphology of the silica were investigated with FTIR and SEM, respectively and reaction rates were monitored with a colorimetric assay. Our results show that these process parameters had only minor effects on composition, but the catalyst conformation influenced the degree of hydration while the pH, choice of solvent and cocatalyst strongly influenced morphology. We also found that perturbation from spin coating significantly influences the silicification dynamics. The ability to catalyze nano- to micron-sized mineral with different morphologies using polypeptides could have numerous dental applications including, sealing of dentin tubules, in situ reinforcement of resin interfaces or preparation of implant surfaces. PMID- 18161783 TI - Clinical trials: rising costs limit innovation. PMID- 18161784 TI - Danazol therapy for aplastic anemia refractory to immunosuppressive therapy. AB - Although there are anecdotal reports of the efficacy of danazol in the treatment of aplastic anemia (AA), there has been no systematic study to clarify its efficacy and toxicity. Therefore, we assessed the efficacy of danazol for treatment of patients with AA refractory to immunosuppressive therapy (IST) and those who relapsed after IST, in a prospective clinical trial. Sixteen patients (12 males and four females; six severe cases and 10 moderate cases) were treated with 300 mg of danazol daily for 12 weeks. All patients completed the treatment period without occurrence of severe toxicity. Three female patients achieved partial remission, whereas only two of the 12 male patients did so. None of the responders had shown a response to previous IST or an increase in the percentage of paroxysmal nocturnal hemoglobinuria (PNH)-type cells which are known to be a marker for a good response to IST. These findings indicate that danazol is effective for a subset of AA patients, and particularly for female patients with AA refractory to IST. PMID- 18161785 TI - Development and validation of SIMS: an instrument for measuring quality of life of adults with sickle cell disease. AB - In recent years, there has been a growing interest in the assessment of quality of life (QOL) issues, particularly in chronic debilitating conditions. Several instruments have been developed, tested, and validated in the general population and in other chronic diseases; however, few studies have examined QOL issues in adults with sickle cell disease (SCD). We developed Sickle Cell Impact Measurement Scale (SIMS), an instrument for measuring the QOL of adults with SCD. The 142-item multi-dimensional SIMS questionnaire was developed using 4 validated instruments and additional questions based upon recommendations of patient focus groups. The SIMS was self-administered to 106 SCD and 45 rheumatoid arthritis (RA) patients over 4 sites. SIMS was evaluated on measures of both internal consistency and construct validity. Item reduction was performed based on results of factor analysis. The SIMS achieved good internal consistency, with a Cronbach's alpha coefficient reported of 0.86, and distinguished between patients with SCD and RA. Overall, QOL did not differ significantly among SCD and RA patients. However, SCD patients scored higher in both physical and social domains, which was expected and reflected the differences in the pathophysiology of each disease. The SIMS is a reliable, valid, and responsive questionnaire, which functions well as a discriminative instrument for the measure of health related QOL (HRQOL) of adults with SCD. The SIMS is currently being administered to adults with SCD across several centers for further validation to become a disease-specific, global QOL instrument. PMID- 18161786 TI - WT1 protein expression in childhood acute leukemia. AB - In patients with acute leukemia, Wilms' tumor gene 1 (WT1) has been used as a target for the detection of minimal residual disease (MRD) by PCR techniques. The expression of WT1 protein, however, has not been extensively studied. To determine the relation between expression of WT1 transcripts and of the encoded protein, we examined leukemic cell lines and primary childhood leukemia samples using both real-time quantitative PCR (RQ-PCR) and flow cytometry. WT1 protein was highly expressed in the leukemic cell lines K562, HL-60, PLB 985, KG-1a and CEM. By contrast, 40 primary samples of acute lymphoblastic leukemia (ALL; B-ALL, n = 15 and T-ALL, n = 10) and acute myeloid leukemia (n = 15) expressed low levels of WT1 protein. RQ-PCR detected WT1 transcript levels in the same range as reported in earlier studies in childhood acute leukemia. The results of this study indicate the following: (i) there are considerable discrepancies between WT1 transcripts and protein expression; (ii) WT1 is not a suitable marker for flow cytometric MRD detection in childhood acute leukemia. PMID- 18161787 TI - Validation of a targeted DNA microarray for the clinical evaluation of recurrent abnormalities in chronic lymphocytic leukemia. AB - Recurrent genomic alterations, mainly losses and gains of specific chromosomes and/or regions, in chronic lymphocytic leukemia (CLL) are recognized as important independent predictors of prognosis and disease progression. The current standard clinical practice for identifying these alterations is chromosome analysis and in situ hybridization with probes targeting 4-5 chromosome regions. We sought to apply array comparative genomic hybridization (array-CGH) technology for the simultaneous detection of genomic imbalances of all loci implicated in CLL. DNA from enriched B-cells from CLL patients were analyzed by array-CGH on a customized CLL BAC array. Copy number changes were detected in 87% of samples with a sensitivity of 100% in samples with clonal abnormalities present in at least 23% of the cells. Furthermore, in nine cases genomic alterations were observed that were undetectable by standard cytogenetic and/or FISH analyses. One of these patients had a 13q14 deletion that was missed by the clinical CLL FISH panel probe set. Our results suggest that a subset of potentially significant genomic alterations in CLL is being missed by the current available techniques. Furthermore, this pilot study clearly shows the robustness, high sensitivity, and high specificity for the targeted CLL microarray analysis as well as the potential for use in routine screening in CLL. PMID- 18161790 TI - Influences of passivating elements on the corrosion and biocompatibility of super stainless steels. AB - Biometals need high corrosion resistance since metallic implants in the body should be biocompatible and metal ion release should be minimized. In this work, we designed three kinds of super stainless steel and adjusted the alloying elements to obtain different microstructures. Super stainless steels contain larger amounts of Cr, Mo, W, and N than commercial alloys. These elements play a very important role in localized corrosion and, thus, their effects can be represented by the "pitting resistance equivalent number (PREN)." This work focused on the behavior which can arise when the bare surface of an implant in the body is exposed during walking, heavy exercise, and so on. Among the experimental alloys examined herein, Alloy Al and 316L stainless steels were mildly cytotoxic, whereas the other super austenitic, duplex, and ferritic stainless steels were noncytotoxic. This behavior is primarily related to the passive current and pitting resistance of the alloys. When the PREN value was increased, the passivation behavior in simulated body solution was totally different from that in acidic chloride solution and, thus, the Cr(2)O(3)/Cr(OH)(3) and [Metal oxide]/[Metal + Metal oxide] ratios of the passive film in the simulated body solution were larger than those in acidic chloride solution. Also, the critical current density in simulated body solution increased and, thus, active dissolution may induce metal ion release into the body when the PREN value and Ni content are increased. This behavior was closely related to the presence of EDTA in the simulated body solution. PMID- 18161791 TI - Transmission electron microscopy and electron diffraction study of BSA-loaded quaternized chitosan nanoparticles. AB - Chitosan nanoparticles, O-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride (O-HTCC) nanoparticles and bovine serum albumin (BSA) loaded chitosan and O-HTCC nanoparticles of a size (about 200-600 nm) were obtained through the process of ionic gelation between chitosan or O-HTCC and sodium tripolyphosphate (TPP). The physicochemical properties of nanoparticles made from chitosan, O HTCC, BSA loaded chitosan, and BSA loaded O-HTCC were determined by transmission electron microscopy (TEM), polarized optical microscopy (POM), photon correlation spectroscopy (PCS), and X-ray diffraction (XRD) pattern. Zeta potential was also performed to understand the surface properties of nanoparticles and their ability to bind negatively charged BSA. TEM, POM, and XRD suggested that ionic-gelation process significantly influenced the crystallinity of BSA, and greater chain realignment in the BSA-loaded chitosan and O-HTCC nanoparticles. PCS revealed that BSA-loaded chitosan nanoparticles were bigger than chitosan nanoparticles in size and BSA-loaded O-HTCC nanoparticles were smaller than O-HTCC nanoparticles in size. PMID- 18161792 TI - What motivates hate crimes based on sexual orientation? Mediating effects of anger on antigay aggression. AB - The aim of this study was to examine the role of anger in response to gay men within three theoretical models of antigay aggression. Participants were 135 exclusively heterosexual men who completed a structured interview designed to assess sexual prejudice, anger in response to a vignette depicting a nonerotic male-male intimate relationship (i.e. partners saying "I love you", holding hands, kissing), and past perpetration of antigay aggression. Among identified antigay assailants, motivations for one earlier assault (i.e. sexual prejudice, peer dynamics, thrill seeking) were also assessed. Results indicated that anger fully mediated the relationship between sexual prejudice and antigay aggression, partially mediated the effect of peer dynamics on antigay aggression, and did not account for the relationship between thrill seeking and antigay aggression. These findings indicate that anger in response to gay men facilitates antigay aggression among some, but not all, antigay perpetrators. PMID- 18161793 TI - The cell cycle control protein cdc25C is present, and phosphorylated on serine 214 in the transition from germinal vesicle to metaphase II in human oocyte meiosis. AB - Cdc25C is a dual specificity phosphatase essential for dephosphorylation and activation of cyclin-dependent kinase 1 (cdk1), a prerequisite step for mitosis in all eucaryotes. Cdc25C activation requires phosphorylation on at least six sites including serine 214 (S214) which is essential for metaphase/anaphase transit. Here, we have investigated S214 phosphorylation during human meiosis with the objectives of determining if this mitotic phosphatase cdc25C participates in final meiotic divisions in human oocytes. One hundred forty-eight human oocytes from controlled ovarian stimulation protocols were stained for immunofluorescence: 33 germinal vesicle (GV), 37 metaphase stage I (MI), and 78 unfertilized metaphase stage II (MII). Results were stage dependent, identical, independent of infertility type, or stimulation protocol. During GV stages, phospho-cdc25C is localized at the oocyte periphery. During early meiosis I (MI), phosphorylated cdc25C is no longer detected until onset of meiosis I. Here, phospho-cdc25C localizes on interstitial microtubules and at the cell periphery corresponding to the point of polar body expulsion. As the first polar body reaches the periphery, phosphorylated cdc25C is localized at the junction corresponding to the mid body position. On polar body expulsion, the interior signal for phospho-cdc25C is lost, but remains clearly visible in the extruded polar body. In atresic or damaged oocytes, the polar body no longer stains for phospho-cdc25C. Human cdc25C is both present and phosphorylated during meiosis I and localizes in a fashion similar to that seen during human mitotic divisions implying that the involvement of cdc25C is conserved and functional in meiotic cells. PMID- 18161794 TI - Confirmation of the role of N-acetyltransferase 2 in teratogen-induced cleft palate using transgenics and knockouts. AB - Previous work on Dilantin- and hydrocortisone-induced cleft palate and cleft lip with or without cleft palate using congenics for the N-acetyltransferase loci (Nat1 and Nat2 are closely linked) and recombinant inbred lines implicated the Nat1,2 region in susceptibility to teratogen-induced orofacial clefting. Since Nat1 does not differ between the two strains, Nat2 appeared to be responsible. We have now tested this conclusion using transgenics and knockouts. Transgenics for human NAT1 (equivalent to mouse Nat2) and knockouts for Nat2 were tested for susceptibility to Dilantin, hydrocortisone, and 6-aminonicotinamide-induced orofacial clefting. We found that Nat2 greatly influences teratogen-induced orofacial clefting on the A/J background but not on the C57BL/6J background. The magnitude and direction of the effects depended on which teratogen was used. The Nat2 knockout did not make C57BL/6J susceptible or A/J (already with very low activity) more susceptible but significantly decreased sporadic clefting in the A/J strain. We conclude that only the A/J strain, with several loci affecting orofacial clefting, is influenced by Nat2. PMID- 18161795 TI - Caroli's disease and liver transplantation. PMID- 18161796 TI - Mission poorly accomplished: the protective role of natural killer cells in recurrent hepatitis C after liver transplantation. PMID- 18161797 TI - Is there a "NAC" to treating acute liver failure in children? PMID- 18161798 TI - Geography, transplant centers, and recipients: what can we learn? PMID- 18161799 TI - Caroli's disease and outcomes after liver transplantation. AB - Caroli's disease is a rare autosomal recessive disorder characterized by intrahepatic cystic dilatation of the bile ducts that, when progressive, leads to intrahepatic stones, recurrent cholangitis, portal hypertension, cholangiocarcinoma, and liver failure. Liver transplantation is a promising curative option for advanced Caroli's disease. The aim of this study was to determine the outcomes of liver transplantation in unselected patients with Caroli's disease and recommend an evidence-based therapeutic algorithm for the management of Caroli's disease. Of the 78,124 patients transplanted in the United States between 1987 and 2006, 104 had Caroli's disease; 96 of these underwent liver alone, and 8 underwent combined liver/kidney transplantation. The patient survival and graft survival were analyzed by Kaplan-Meier survival analysis, and risk of death and risk of graft loss were analyzed by Cox proportional hazards regression. The overall 1-, 3-, and 5-year graft (79.9%, 72.4%, and 72.4%) and patient (86.3%, 78.4%, and 77%) survival rates were excellent for patients after liver transplantation. For combined liver/kidney transplantation (n = 8), the 1 year patient survival and graft survival were 100%. Proportional hazards analysis identified Asian ethnicity, elevated bilirubin, requirement of life support or hospitalization prior to transplantation, and a cold ischemia time greater than 12 hours as associated with increased risk of both graft loss and death. A history of prior transplant or prior abdominal surgery was also associated with increased risk of graft loss. In conclusion, liver transplantation is an excellent treatment option for patients with advanced Caroli's disease and should be considered in a timely fashion to prevent worsening complications including refractory cholangitis and cholangiocarcinoma. PMID- 18161800 TI - Greater travel time to a liver transplant center does not adversely affect clinical outcomes. AB - The effect of patient travel time to a transplant center on outcomes is unknown. We compared outcomes between patients living >3 hours (Group A) vs. 90 days to list, listing, survival while listed, transplantation, and posttransplantation survival. Covariates included Model for End-Stage Liver Disease (MELD) score, hepatocellular carcinoma (HCC), alcoholic liver disease, insurance type, and psychosocial score. There were 38 (23%) patients in Group A and 128 (77%) in Group B. Median MELD scores were 14.5 (range, 6-36) for Group A and 14.0 (range, 7-32) for Group B (p = 0.20). Groups were similar for age, gender, diagnosis, psychosocial score, insurance, and HCC variables. Group A was not independently associated with >90 days to list (odds ratio, 0.98; 95% confidence interval [CI], 0.4-2.4). Kaplan-Meier cumulative probabilities for listing, transplantation, and 1-yr posttransplantation survival were similar (A vs. B: 0.77 vs. 0.83, 0.70 vs. 0.69, and 0.85 vs. 0.86, respectively; all p values >0.05). Being in Group A remained insignificant in terms of probability of listing, transplantation, and posttransplantation survival by Cox proportional hazard modeling. Survival on the list was significantly better for Group A (A: 1.0, B: 0.55; p = 0.02). Fewer patients at high MELD score in Group A and referral biases may explain this difference. In conclusion, after entering evaluation, patients living >3 hours away from a transplant center have comparable outcomes to those living closer. PMID- 18161801 TI - Phylogenetic relationships of South American alligatorids and the caiman of Madeira River. AB - We analyzed DNA sequences of the mitochondrial cytochrome b gene (cyt b), the nuclear Recombination Activating Gene 1 (RAG1) and the myelocytomatosis oncogene (MYC) to infer the phylogenetic relationship of Caiman crocodilus and Caiman yacare, and other South American alligatorid crocodilian species. Phylogenetic relationships were robustly supported with Paleosuchus sister to Melanosuchus and Caiman. Phylogenetic relationships of C. crocodilus and C. yacare were unclear as these two species share mitochondrial and nuclear haplotypes. Specifically this sharing occurs among specimens of C. yacare and C. crocodilus from the Madeira River drainage. Two potential explanations stand out: secondary contact followed by hybridization, and differentiation along a cline. Current data cannot resolve between these two competing hypotheses. In comparison with C. yacare and C. crocodilus, Paleosuchus trigonatus and Paleosuchus palpebrosus are very well differentiated and also show surprising haplotypic diversity in spite of their phenotypic similarity. PMID- 18161802 TI - In vitro degradation and in vivo biocompatibility study of a new linear poly(urethane urea). AB - Segmented poly(urethane urea)s (PUUs) with hard segments derived only from methyl 2,6-diisocyantohexanoate (LDI) without the use of a chain extender have previously been described. These materials, which contain hard segments with multiple urea linkages, show exceptionally high strain capability (1600-4700%). In the study reported here, the rate and effect of hydrolysis of these materials were determined for gamma-sterilized and nonsterilized samples. Materials investigated contained PCL, PTMC, P(TMC-co-CL), P(CL-co-DLLA), or P(TMC-co-DLLA) as soft segments and, as well as their mechanical properties, changes in mass, inherent viscosity (I.V.), and thermal properties were studied over 20 weeks. Results showed that the degradation rate was dependant on the soft segment structure, with a higher rate of degradation for the polyester-dominating PUUs exhibiting a substantial loss in I.V. A tendency of reduction of tensile strength and strain hardening was seen for all samples. Also, loss in elongation at break was detected, for PUU-P(CL-DLLA) it went from 1600% to 830% in 10 weeks. Gamma radiation caused an initial loss in I.V. and induced more rapid hydrolysis compared with nonsterilized samples, except for PUU-PTMC. A cytotoxicity test using human fibroblasts demonstrated that the material supports cell viability. In addition, an in vivo biocompatibility study showed a typical foreign body reaction after 1 and 6 weeks. PMID- 18161803 TI - Membrane permeability properties of dental adhesive films. AB - This study evaluated the permeability properties of five experimental resin membranes that ranged from relatively hydrophobic to relatively hydrophilic to seal acid-etched dentin saturated with water or ethanol. The experimental resins (R1, R2, R3, R4, and R5) were evaluated as neat bonding agents or as solutions solvated with ethanol (70% resin/30% ethanol). The quality of dentin sealing by these experimental resins was expressed in terms of reflection coefficients calculated as the ratio of the effective osmotic pressure to the theoretical osmotic pressure of test solutions. The effective osmotic pressure produced across resin-bonded dentin was induced in hypertonic solutions (CaCl(2) or albumin) at zero hydrostatic pressure. The outward fluid flow induced by these solutions was brought to zero by applying an opposing negative hydrostatic pressure. The least hydrophilic resins blends, R1 and R2, exhibited significantly (p < 0.05) higher reflection coefficients than the most hydrophilic resins (R4 and R5) in both conditions of dentin saturation (water and ethanol). The reflection coefficients of neat resins were, in general, significantly higher when compared with their corresponding solvated versions in both conditions of dentin saturation. In dentin saturated with ethanol, bonding with neat or solvated resins, resulted in reflection coefficients that were significantly higher when compared with the results obtained in dentin saturated with water. Reflection coefficients of CaCl(2) (ca. 1 x 10(-4)) were significantly lower (p < 0.05) than for albumin (ca. 3 x 10(-2)). Application of hydrophobic resins may provide better sealing of acid-etched dentin if the substrate is saturated with ethanol, instead of water. PMID- 18161804 TI - Metal ion release from fracture fixation devices: a potential marker of implant failure. AB - Stainless steel is the alloy most frequently used for fracture fixation devices (FFD). We aimed to verify if the ion release evaluation could be a surrogate marker of performance and could allow an early detection of implant failure in patients with stainless steel FFD. We measured the nickel (Ni) and chromium (Cr) serum content in patients undergoing the retrieval of stainless steel plates (group I) or intramedullary nails (group II), because of consolidation or failure. Forty-five healthy donors were recruited as controls. Furthermore, the diagnostic performance of these values was evaluated: analysis power, sensitivity, specificity, positive and negative predictive values, likelihood ratios, and diagnostic accuracy were calculated. A significant increase of ion values was demonstrated in patients with failed plates, compared with the values recorded in patients with well-fixed plates (p = 0.002 for Cr and p = 0.002 for Ni), and in healthy subjects (p = 0.0002 for Cr and p = 0.003 for Ni). No significant difference was found between stable implants and controls (p = 0.8 for Cr and p = 0.06 for Ni). A high specificity (0.92 for Cr and 1.00 for Ni), positive predictive value (0.87 for Cr and 1.00 for Ni), and positive likelihood ratio (9.10 for Cr) were calculated for ion testing in plates. The substantial metal content elevations in patients with plates and the positive likelihood ratio above 5 for chromium testing suggest that ion dosage may be a useful surrogate marker for the presence of malfunctioning of these devices, perhaps before the onset of clinical and radiographic changes. PMID- 18161805 TI - Cumulative effect of disinfection procedures on microhardness and tridimensional stability of a poly(methyl methacrylate) denture base resin. AB - Microwave irradiation has been used for disinfection of dentures instead of chemical solutions; yet, its effect on resin properties after repeated procedures still is unclear. This study evaluated the cumulative effect of two disinfection methods on Knoop microhardness and tridimensional stability of a poly(methyl methacrylate) denture base resin. For the microhardness measurement, 24-resin discs received mechanical polishing and were submitted to the following treatments: (1) control (no disinfection), (2) chemical disinfection (immersion in 100ppm chloride solution for 24h), or (3) microwave disinfection (irradiation at 690 W for 6 min). Disinfection procedures were performed twice (T1, T2) with a 7-day interval. Knoop microhardness was recorded after polishing (T0) and after T1 and T2. For the dimensional stability test (measured by the adaptation of the denture bases), 36-maxillary denture bases were obtained from type III dental stone casts duplicated from a metallic master model and submitted to the disinfection treatment. Adaptation of denture bases was measured at baseline (T0) and after T1 and T2 by weighing a vinyl polysiloxane film reproducing the gap between resin base and master model. Data were analyzed by ANOVA GLM for repeated measures and Bonferroni's test, alpha = 0.05. Knoop microhardness was not modified by any disinfection procedure but decreased over time. Denture resin bases submitted to microwave disinfection had gradual increase of distortion over time, while bases immersed in chloride solution did not differ from the control group and remained dimensionally stable from T1 to T2. PMID- 18161806 TI - Neural tissue engineering of the CNS using hydrogels. AB - Current therapies have limited capacity to curtail disease progression or damage of the central nervous system (CNS) of adult mammals and successful regeneration following injury or disease does not occur. Regeneration of the CNS is limited by physical and chemical inhibitory barriers within the injured environment and the absence of positive cues that elicit and guide repair. Neural tissue engineering strategies focus on developing scaffolds that artificially generate favourable cellular microenvironments that attempt to tip the balance in favour of regeneration. Some recent advances using scaffolds to promote regeneration within the CNS, particularly in conjunction with stem cells, has generated promising results. This review focuses on hydrogel scaffolds which have been used extensively in neural tissue engineering applications and addresses the physical and chemical modifications of these materials to promote nerve regeneration. PMID- 18161807 TI - Effect of cyclic loading on bonding of fiber posts to root canal dentin. AB - This study evaluated the effect of mechanical loading on microtensile bond strength (MTBS) when luting fiber posts to root canal dentin. Forty maxillary premolars were endodontically treated, and the roots were prepared for post cementation using the FRC Postec system. A light-cured composite (Clearfil Photo Core, Kuraray) was used in combination with four adhesive systems: Single Bond (3M ESPE), Multilink Primer (Vivadent), Clearfil Photo Bond (Kuraray), and Clearfil New Bond (Kuraray). A composite build-up was performed around the root to provide adequate gripping during testing. For each experimental group, half of the specimens were load cycled under 90 N (5000 cycles, 3 cycles/s). Specimens were cut to obtain beams with the post in the center and with the radicular dentin overlaid by the composite build-up on each side. Microtensile testing was performed with a universal testing machine at a cross-head speed of 0.5 mm/min. The failure mode was classified under a stereomicroscope. ANOVA and Student Newman-Keuls multiple comparison tests were performed. After 24 h, all adhesives performed similarly. When specimens were subjected to mechanical loading, decreases in MTBS were observed for Clearfil New Bond and Multilink Primer. PMID- 18161808 TI - Iodine inhibits antiadhesive effect of PEG: implications for tissue engineering. AB - BACKGROUND: The success of a biomaterial implant may be affected by the surface chemistry's impact on protein adsorption. We have developed a series of poly(ethylene glycol) (PEG) containing, tyrosine-derived polycarbonates, which have been rendered radio-opaque by the iodination of tyrosine units in the copolymer backbone for use in resorbable biomedical implants including vascular stents and grafts. We tested the hypothesis that protein adsorption along with seeding, growth, and migration of human aortic smooth muscle cells (SMC) and human aortic endothelial cells (EC) will be modified by the presence of iodine and PEG within the polymer composition. METHODS: Thin films of these polymers were prepared for the protein-material and cell-material interaction studies. Dot blot, SDS-PAGE, and XPS were used to evaluate relative protein adsorption. Cell adhesion and growth studies were performed using an MTS assay. Cell migration was evaluated using an injury model. RESULTS: The presence of PEG attenuated protein adsorption, cell adhesion, and growth. With the subsequent incorporation of iodine, protein adsorption markedly increased while the antiadhesive effect of PEG was counteracted by iodine for EC and SMC adhesion and SMC growth. CONCLUSION: Iodine incorporation into the polymer resulted in increased protein adsorption thus counteracting the effect of PEG. PMID- 18161809 TI - Adhesion to chondroitinase ABC treated dentin. AB - Dentin bonding relies on complete resin impregnation throughout the demineralised hydrophilic collagen mesh. Chondroitin sulphate-glycosaminoglycans are claimed to regulate the three-dimensional arrangement of the dentin organic matrix and its hydrophilicity. The aim of this study was to investigate bond strength of two etch-and-rinse adhesives to chondroitinase ABC treated dentin. Human extracted molars were treated with chondroitinase ABC and a double labeling immunohistochemical technique was applied to reveal type I collagen and chondroitin 4/6 sulphate distribution under field emission in-lens scanning electron microscope. The immunohistochemical technique confirmed the effective removal of chondroitin 4/6 sulphate after the enzymatic treatment. Dentin surfaces exposed to chondroitinase ABC and untreated specimens prepared on untreated acid-etched dentin were bonded with Adper Scotchbond Multi-Purpose or Prime and Bond NT. Bonded specimens were submitted to microtensile testing and nanoleakage interfacial analysis under transmission electron microscope. Increased mean values of microtensile bond strength and reduced nanoleakage expression were found for both adhesives after chondroitinase ABC treatment of the dentin surface. Adper Scotchbond Multi-Purpose increased its bond strength about 28%, while bonding made with Prime and Bond NT almost doubled (92% increase) compared to untreated specimens. This study supports the hypothesis that adhesion can be enhanced by removal of chondroitin 4/6 sulphate and dermatan sulphate, probably due to a reduced amount of water content and enlarged interfibrillar spaces. Further studies should validate this hypothesis investigating the stability of chondroitin 4/6 and dermatan sulphate-depleted dentin bonded interface over time. PMID- 18161810 TI - The electrochemical behavior and surface analysis of Ti49.6Ni45.1Cu5Cr0.3 alloy for orthodontic usage. AB - The aim of this study was to investigate the electrochemical behavior of Ti(49.6)Ni(45.1)Cu(5)Cr(0.3) (TiNiCuCr) alloy in artificial saliva solutions with a wide rage of pH values and to characterize the surface passive film after polarization tests. This article represents the ideal, static environment and associated electrochemical response and comparison values. The corrosion behavior of TiNiCuCr alloy was systematically studied by open circuit potential, potentiodynamic, potentiostatic, and electrochemical impedance techniques. Potentiodynamic and potentiostatic test results showed that the corrosion behavior of TiNiCuCr was similar to that of NiTi alloy. Both corrosion potential (E(corr)) and pitting corrosion potential (E(b)) showed a pH-dependent tendency that E(corr) and E(b) decreased with the increase of the pH value. X-ray photoelectron spectroscopy results revealed the composition of the passive film consisted mainly of TiO(2) with a little amount of Ni oxides (NiO/Ni(2)O(3)) that was identical with NiTi alloy. Besides Ni, a Cu enriched sub-layer was also found. The nickel ion release rate showed a typical time-related decrease as examined by ICP/OES. In conclusion, the addition of Cu and Cr had little effect on the corrosion behavior of NiTi or on the composition and the structure of the passive film. PMID- 18161811 TI - In vivo response of porous hydroxyapatite and beta-tricalcium phosphate prepared by aqueous solution combustion method and comparison with bioglass scaffolds. AB - Pure hydroxyapatite (HAp) and a biphasic calcium phosphate [containing 90% of beta-tri-calcium phosphate (beta-TCP) and 10% HAp] were tailored through an aqueous solution combustion synthesis. Porous struts were prepared using all the powders along with bioglass, a known bioactive material, and subsequently characterized. Sterilized struts were implanted to the lateral side of radius bone of 24 black Bengal goats of either sex, in which a blank hole was left unfilled in a group of six specimens to act as control. The bone formation response of the three implanting materials in vivo has been studied using scanning electron microscope and histological analysis in contrast with positive controls. Push-out tests were used to assess the mechanical strength at the bone biomaterial interface. It was observed that interfacial response was strongly dependent on combinations of different physical and chemical parameters. The surface of beta-TCP exhibited similar characteristics of bone and was distinct from those of intervening apatite layer of bioglass. Lower bone ingrowth and reduced strength was observed with HAp compared to beta-TCP/bioglass-based implants. Bone formation response of the Ca-P material varied according to the composition of the implanting material, which could be tailored through this novel synthesis. PMID- 18161812 TI - In vitro degradation and in vivo biocompatibility of poly(lactic acid) mesh for soft tissue reinforcement in vaginal surgery. AB - This study was aimed at evaluating the in vitro degradation, the in vivo biocompatibility and at comparing the effects of two methods of sterilization on poly(L-lactic acid) (PLA(94)) resorbable mesh. The mesh was manufactured to be used as surgical soft tissue reinforcement in the vaginal area. Samples of 100 mg of PLA(94) mesh (10 x 10 mm(2)) were immersed in isoosmolar 0.13M, pH 7.4 phosphate buffer solution at 37 degrees C, during 12 months. The hydrolytic degradation up to 12 months after immersion was monitored by measuring weight loss, mesh area changes, and by various analytical techniques namely Differential scanning calorimetry (DSC), capillary zone electrophoresis (CZE), size exclusion chromatography (SEC), and environmental scanning electron microscopy (ESEM). Specimens of nonsterilized, ethylene-oxide (ETO) sterilized, and gamma-ray sterilized PLA(94) mesh were compared. Fifteen samples were implanted in an incisional hernia Wistar rat model. Histopathology was performed up to 90 days after implantation to evaluate the inflammatory response and the collagen deposition. Although the decrease of molecular weight due to polymer chain scissions started 6 weeks after in vitro immersion, water-soluble degradation products and decrease of tensile strength appeared after 8 months only. Analyses showed also that ETO sterilization did not affect the degradation of the PLA(94) mesh. In contrast, gamma-ray sterilization increased very much the sensitivity of the mesh to the hydrolytic degradation. In vivo, the PLA(94) mesh exhibited good biocompatibility over the investigated time period. PMID- 18161813 TI - Fatigue crack propagation rates in PMMA bone cement cannot be reduced to a single power law. AB - Cement mantles around metallic implants have pre-existing flaws (shrinkage induced cracks, laminations, and endosteal surface features) and their fatigue failure is related to the fatigue crack propagation (FCP) rate of bone cement. We estimated the relevant in vivo range of cyclic stress intensity factor (DeltaK) around a generic femoral stem (0-1 MPa square root(m)) and determined that previous FCP data did not adequately cover this range of DeltaK. Vacuum-mixed standard bone cement was machined into ASTM E647 standard compact notched tension specimens. These were subject to sinusoidal loading (R = 0.1) at 5 Hz in 37 degrees C DI water, covering a DeltaK range of 0.25-1.5 MPa square root(m) (including a decreasing DeltaK protocol). FCP-rate data is normally reduced to a power-law fit relating crack growth rate (da/dn) to DeltaK. However, a substantial discontinuity was observed in our data at around DeltaK = 1, so two power-law fits were used. Over the physiologically plausible range of DeltaK, cracks grew at a rate of 2.9 E -8 x DeltaK(2.6) m/cycle. Our data indicated that FCP-rates for 0.5 > DeltaK > 0.3 MPa square root(m) are between 10 E -8 and 10 E 8 m/cycle, 1 or 2 orders of magnitude greater than predicted by extrapolating from previous models based on higher DeltaK data. PMID- 18161814 TI - Assessment of tissue repair in full thickness chondral defects in the rabbit using magnetic resonance imaging transverse relaxation measurements. AB - The purpose of this study was to determine if the noninvasive and nondestructive technique of magnetic resonance imaging could be used to quantify the amount of repair tissue that fills surgically-induced chondral defects in the rabbit. Sixteen 4-mm diameter full-thickness chondral defects were created. A photopolymerizable hydrogel was used to seal the defects as a treatment modality. At 5 weeks, the animals were sacrificed and the distal femur was subjected to MRI analyses at high field (9.4 T). The transverse relaxation time (T(2)) in each defect was measured. Histology and histomorphometric analysis were used to quantify the amount of repair tissue that filled each defect. The relationship between T(2) and percent tissue fill was found to fit well to a negatively sloped, linear model. The linear (Pearson's product-moment) correlation coefficient was found to be r = -0.82 and the associated coefficient of determination was r(2) = 0.67. This correlation suggests that the MRI parameter T(2) can be used to track changes in the amount of repair tissue that fills cartilage defects. This would be especially useful in in vivo cartilage tissue engineering studies that attempt to determine optimal biomaterials for scaffold design. PMID- 18161815 TI - Changes in stiffness of demineralized dentin following application of collagen crosslinkers. AB - It is thought that increasing the strength of the dentin matrix using crosslinking agents may improve both the strength and the durability of resin dentin bonds. The purpose of this study was to evaluate the effect of two collagen crosslinking agents (glutaraldehyde, GD and grape seed extract, GSE) on the modulus of elasticity of demineralized dentin. Sound molar fragments were fully demineralized and divided into five groups according to the type and concentration of crosslinking agents: 2.5% GD; 5% GD, 25% GD; 0.65% GSE; 6.5% GSE. Specimens were immersed in their respective solution and tested at baseline, 10 min, 30 min, 1 h, 2 h, 4 h. The elastic modulus of dentin was significantly affected by the treatment (p < 0.01) and exposure time (p < 0.01). There was a statistically significant interaction between the two factors evaluated (treatment vs. time p < 0.01). Mean baselines values varied between 4.8 and 6.2 MPa in water; after 4 h of treatment the values increased between 34.9 and 242.5 MPa, that were treatment time and agent dependent. The use of these collagen crosslinkers to increase the stiffness of demineralized dentin, was both concentration and time dependent. PMID- 18161816 TI - The influence of the acidic component of the gas-foaming porogen used in preparing an injectable porous calcium phosphate cement on its properties: acetic acid versus citric acid. AB - In the present study, macroporous calcium phosphate cements (CPCs) were prepared using a porogen; that is, the gas-foaming technique. The objective was to investigate the influence of the acidic component of the porogen (acetic acid versus citric acid) on several properties of a specified CPC. In all of the cements prepared, the basic component of the porogen was the same, namely, NaHCO(3), and it was added to the powder phase of the cement, while the acidic component of the porogen was dissolved in the liquid phase of the cement. The cements were characterized in terms of initial setting time, porosity, crystallinity, injectability and compressive strength. Also, XRD, FTIR, and SEM techniques were employed to evaluate the phase composition, the chemical groups and the morphological aspects of the porous cements during setting. It was found that the presence of a porogen in a CPC led to significant decreases in both its initial setting time and compressive strength. A CPC made using acetic acid contained a larger amount of the apatite phase but was significantly less injectable and less porous than when citric acid was used. PMID- 18161817 TI - Polypeptide nanocoatings for preventing dental and orthopaedic device-associated infection: pH-induced antibiotic capture, release, and antibiotic efficacy. AB - Implant-associated infection is one of the most common and problematic complications for dental and orthopaedic patients. Modification of currently used implant surfaces aimed at bestowing them with antibacterial properties is a promising approach in the development of new biomaterials. In this study, a novel nanotechnology, that is, electrostatic self-assembly, was developed to construct biomimetic polypeptide nanocoatings on commonly used metal implants. A model antibacterial drug, cefazolin, was captured in the polypeptide nanocoating and its release was studied. We have shown that the capture and release of cefazolin was pH-induced and could be controlled, and the developed antibiotic-incorporated polypeptide multilayer nanocoatings could prevent Staphylococcus aureus colonization thus showing great potential for preventing implant-associated infection. PMID- 18161818 TI - Clinical evaluations of OSTEON as a new alloplastic material in sinus bone grafting and its effect on bone healing. AB - PURPOSE: The objective of this study was to clinically evaluate the use of OSTEON as a sinus graft material and to measure the effect of healing at 4 and 6 months after surgery. MATERIALS AND METHODS: After sinus graft using OSTEON in 17 patients, bone specimens were collected from lateral sinus using 2.0-mm trephine bur at the time of 4 or 6 months after surgery. Histology of the bone specimens was prepared and the percentage of newly formed bone fraction, lamellar bone/woven bone ratio (LB/WB), and newly formed bone/graft material ratio (NB/GM) were measured to indicate the suitability of the materials and the successful healing of the graft. RESULTS: The morphology of OSTEON was observed to be interconnected, with 77% porosity and a pore size of 300-500 microm. After implantation, the mean percentage of newly formed bone fraction after 4 months and 6 months surgery was 40.6 and 51.9%, respectively. Statistical analysis indicated no significant difference (p = 0.135) in the newly formed bone fraction between the two postoperative periods. The mean LB/WB ratio after 4 months and 6 months surgery was 0.14 and 0.45, respectively, with significant difference observed between the two postoperative periods (p = 0.027). Additionally, the mean NB/GM ratio after 4 months and 6 months surgery was 1.95 and 7.72, respectively, with significant difference observed between the two postoperative periods (p = 0.046). CONCLUSION: It was concluded that OSTEON is suitable for use in sinus graft application since desirable time-dependent healing was demonstrated. PMID- 18161819 TI - Solvent/non-solvent sintering: a novel route to create porous microsphere scaffolds for tissue regeneration. AB - Solvent/non-solvent sintering creates porous polymeric microsphere scaffolds suitable for tissue engineering purposes with control over the resulting porosity, average pore diameter, and mechanical properties. Five different biodegradable biocompatible polyphosphazenes exhibiting glass transition temperatures from -8 to 41 degrees C and poly (lactide-co-glycolide), (PLAGA) a degradable polymer used in a number of biomedical settings, were examined to study the versatility of the process and benchmark the process to heat sintering. Parameters such as: solvent/non-solvent sintering solution composition and submersion time effect the sintering process. PLAGA microsphere scaffolds fabricated with solvent/non-solvent sintering exhibited an interconnected porosity and pore size of 31.9% and 179.1 mum, respectively which was analogous to that of conventional heat sintered PLAGA microsphere scaffolds. Biodegradable polyphosphazene microsphere scaffolds exhibited a maximum interconnected porosity of 37.6% and a maximum compressive modulus of 94.3 MPa. Solvent/non-solvent sintering is an effective strategy for sintering polymeric microspheres, with a broad spectrum of glass transition temperatures, under ambient conditions making it an excellent fabrication route for developing tissue engineering scaffolds and drug delivery vehicles. PMID- 18161820 TI - Bone regeneration by using scaffold based on mineralized recombinant collagen. AB - Bone regeneration was achieved in the 15-mm segmental defect model in the radius of rabbit by using the scaffold based on mineralized recombinant collagen for the first time. The recombinant collagen was recombinant human-like type I collagen, which was produced by cloning a partial cDNA that was reversed by mRNA from human collagen alpha1(I) and transferred to E. coli. The scaffold material nano hydroxyapatite/recombinant human-like collagen/poly(lactic acid) (nHA/RHLC/PLA) was developed by biomimetic synthesis. Thermo gravimetric analysis, X-ray diffraction and scanning electron microscopy were applied to exhibit that the scaffold showed some features of natural bone both in main component and hierarchical microstructure. The percentages of organic phase and inorganic phase of nHA/RHLC were similar to that of natural bone. The three-dimensional porous scaffold materials mimic the microstructure of cancellous bone. In the implantation experiment, the segmental defect was healed 24 weeks after surgery, and the implanted composite was completely substituted by new bone tissue. The results of the implantation experiment were very comparable with that of the scaffold based on mineralized animal-sourced collagen. It is concluded that the scaffold based on mineralized recombinant collagen maintains the advantages of mineralized animal-sourced collagen, while avoids potential virus-dangers. The scaffold is a promising material for bone tissue engineering. PMID- 18161821 TI - Study of the micro-structural properties of RISUG--a newly developed male contraceptive. AB - A new male contraceptive given the name RISUG (an acronym for reversible inhibition of sperm under guidance) and presently undergoing advanced clinical trials has been developed. When injected into the lumen of the vas deferens, its polyelectrolytic nature induces a surface charge imbalance on sperm membrane system leading to the leakage of enzymes essential for fertilization. Contact mode atomic force microscopy (AFM) has been used to analyze quantitatively the micro-structural properties of RISUG and its precipitate in various systems. Hydrolysis of the contraceptive gel resulted in the formation of pores of varying dimensions. RISUG being a highly charged molecule, as evident from zeta potential measurements, has a tendency to form a complex with ionic biomolecules present in the seminal plasma. This is supported by the experimental observations using AFM. This RISUG-biomolecule complex possibly acts as an ionic trap for spermatozoa passing through the vas deferens. Micro-structural properties of RISUG including amplitude (root mean square, peak-to-valley distance, skewness and kurtosis) and spatial roughness have been studied to understand its response to various physiological conditions. Significant alterations in the surface charge distribution of the sperm cell is observed on exposure to RISUG. PMID- 18161822 TI - Effect of temporary filling materials on repair bond strengths of composite resins. AB - Endodontic access cavities sometimes can be prepared through a permanent composite restoration. Between the appointments, temporary cements are used to seal access cavities and may have negative effect on bonding of further composite restoration. The purpose of this study was to compare shear bond strength of composite to composite which had been in contact with various temporary filling materials. Standard cavities were prepared on 160 acrylic resin blocks, obturated with composite resin (Clearfil AP-X, Kuraray, Japan) and randomly divided into eight groups (n = 20). Group 1 received no treatment. From group 2-8, composite surfaces were covered with the following cements temporarily: Zinc-oxide/calcium sulphate (Cavit-G, ESPE, Germany), two different Zinc-Oxide-Eugenol materials (ZnOE, Cavex, Holland and IRM, Dentsply, USA), Zinc-phosphate cement (Adhesor, Spofa-Dental, Germany), Zinc-polycarboxylate cement (Adhesor-Carbofine, Spofa Dental, Germany), Glass-Ionomer-Cement (Argion-Molar, Voco, Germany), or light curing temporary material (Clip, Voco, Germany). The cements were removed mechanically after 1 week storage in distilled water at 37 degrees C and composite surfaces were treated with a self-etch adhesive system (SE-Bond, Kuraray, Japan). Composite resin build-ups were created on composite surfaces. Shear bond strength values were measured using universal testing machine at crosshead speed of 1 mm/min. The data was calculated in MPa and statistically analyzed using one-way ANOVA and Tukey tests. Eugenol-containing cements significantly reduced shear bond strengths of composite to composite (p < 0.05), while the other temporary materials had no adverse effect on shear bond strength (p > 0.05). These findings suggested that temporary filling materials except eugenol-containing materials have no negative effect on composite repair bond strengths. PMID- 18161823 TI - Hybrid coating of hydroxyapatite and collagen within poly(D,L-lactic-co-glycolic acid) scaffold. AB - To improve the cell-affinity of biodegradable polymer scaffold, coating hydroxyapatite (HA) or collagen on the surface of polymer materials seemed to be a strategy to combine both advantages of them. The objective of this study was to develop a novel method to introduce HA and collagen inside polymer scaffold uniformly. HA and collagen suspension was mixed with paraffin microspheres, and molded to form a composite sample. After the sample was dried, HA/collagen composite was left among and on the surface of paraffin microspheres. Poly(D,L lactic-co-glycolic acid) (PLGA) (50/50) solution was cast into the inter-space of the paraffin microspheres and dried. Afterwards, the paraffin was dissolved and removed, HA/collagen was transferred to the surface and even inside of the pore wall of PLGA scaffolds. Collagen fibers and HA particles which were inlaid inside the PLGA pore wall could help to enhance the coating strength between HA/Col coating and the pore wall surface of the PLGA scaffold. The scaffolds with HA/Col coating were expected to exhibit desirable properties in bone tissue engineering. PMID- 18161824 TI - Y-TZP zirconia run against highly crosslinked UHMWPE tibial inserts: knee simulator wear and phase-transformation studies. AB - BACKGROUND: Zirconia (ZrO(2)) ceramics combined with highly cross-linked polyethylene appears to be a promising approach to minimize wear in artificial knee joints. The wear performance of yttria-stabilized zirconia (YZr) femoral condyles on 7-Mrad tibial inserts was compared in a knee simulator to CoCr bearing on 3.5-Mrad inserts. METHODS: The knee design was the Bi-Surface type with a 9-year clinical history in Japan (JMM, Japan). A displacement-controlled knee simulator was used with kinematics that included 20 degrees flexion, +/-5 degrees rotation, and 6 mm anterior/posterior translation. Lubricant was alpha calf serum, test duration was 10 million cycles (10 Mc), and wear was measured by weight-loss techniques. The wear zones were studied by laser interferometry, scanning electron microscopy, and Raman microprobe spectroscopy. RESULTS: At 10 Mc the wear rates of the CoCr controls averaged 4.5 mm(3)/Mc. This was within 7% of the prior estimate at 5-Mc duration and comparable to Bi-Surface wear data from another laboratory. The CoCr condyles increased in roughness (R(a)) from <50 nm to average R(a) = 250 nm due to linear scratching. The ceramic condyles remained pristine throughout the wear study (R(a) <7 nm). With the YZr/7-Mrad combination, the weight change had a positive slope over at 10 Mc, which meant that the actual polyethylene wear was unmeasurable. Microscopic examinations at 10 Mc showed that the zirconia surfaces were intact and there was no detectable change from tetragonal to monoclinic phase. INTERPRETATION: Our laboratory knee wear simulation appeared very supportive of the 9-year YZr/PE clinical results with Bi-Surface total knee replacements in Japan. PMID- 18161825 TI - Physiological and psychological symptoms of grief in widows. AB - In this cross-sectional descriptive survey of 173 widows, we describe their grief symptoms using an open-ended questionnaire and the Revised Grief Experience Inventory. Self-reported physical symptoms included pain, gastro-intestinal problems, medical/surgical conditions, sleep disturbances, and neurological/circulatory issues. Psychological symptoms were reported as depression, anxiety, and loneliness. The mean total grief score was 71.4 +/- 30.2 (possible range 22-132). Physical symptoms were significantly correlated with the total mean grief score, and subscales of physical distress, depression, existential tension, and guilt. When the subjects were grouped by years since loss, (range 1-5 years), there was no significant decrease found in overall grief scores, suggesting symptoms of grief experienced by widows may continue up to at least 5 years. PMID- 18161826 TI - Occupational hypersensitivity pneumonitis due to molds in an onion and potato sorter. AB - A 42-year-old female non-smoking onion and potato sorter developed work-related shortness of breath, cough, fatigue and flu-like symptoms. The diagnosis of hypersensitivity pneumonitis was based on patchy infiltrates in both lungs on high resolution computed tomography and lymphocytosis of 71% in a bronchoalveolar lavage sample with a CD4/CD8 ratio of 0.4. Exposure cessation and initial corticosteroid therapy resulted in complete recovery. IgG antibodies to Penicillium species and Fusarium solani cultivated from samples from the patient's workplace were detected in the patient's serum and cross-reactivity was demonstrated within Penicillium species, but also between Penicillium species and Aspergillus fumigatus. We conclude that occupational hypersensitivity pneumonitis due to molds may develop in onion and potato sorters. PMID- 18161827 TI - Traditional nutritional analyses of figs overestimates intake of most nutrient fractions: a study of ficus perforata consumed by howler monkeys (Alouatta palliata mexicana). AB - There continue to remain uncertainty concerning the nutritional importance of figs (Ficus spp.) in the primate diet. Although studies have been performed on the nutritional analysis of fig pulp and seed fractions separately, there has not been an attempt to quantify the contribution of animal matter within figs. Here we report nutritional values of figs (Ficus perforata) (Urostigma) consumed by a troop of howler monkeys (Alouatta palliata mexicana). Separate nutritional assays were performed on the pulp, seed, and animal fraction. Whole-fig analyses significantly exaggerate the concentrations of all nutrients (lipids especially) as seeds, which represent a large proportion of the fig (45%), cannot be digested. Animal matter only represents 1% of the fig, and augments fig protein and lipid content by 0.44 and 0.30%, respectively. This represents the 11 and 9% of the fraction available for digestion. Differences in fig consumption were observed between age and sex classes (P=0.04) and periods of the day (P=0.001); females consumed more figs than males and the highest consumption of figs was observed in the afternoon. F. perforata figs may be an important component of the howler monkeys diet owing to their high content of water and calcium. PMID- 18161828 TI - Safety and efficacy of N-acetylcysteine in children with non-acetaminophen induced acute liver failure. AB - Acute liver failure (ALF) carries a high mortality in children. N-acetylcysteine (NAC), an antioxidant agent that replenishes mitochondrial and cytosolic glutathione stores, has been used in the treatment of late acetaminophen-induced ALF and non-acetaminophen-induced ALF. In our unit, NAC was introduced as additional treatment for non-acetaminophen-induced ALF in 1995. The aim of this study was to evaluate the safety and efficacy of NAC in children with ALF not caused by acetaminophen poisoning. A retrospective review of medical records of 170 children presenting with nonacetaminophen-induced ALF between 1989 and 2004 was undertaken. ALF was defined as either international normalized ratio of prothrombin time (INR) > 2 and abnormal liver function or INR >1.5 with encephalopathy and abnormal liver function. Children were divided into the following groups: Group 1 (1989-1994), standard care (n = 59; 34 [58%] male; median age 2.03 yr, range 0.003-15.8 yr); and Group 2 (1995-2004), standard care and NAC administration (n = 111; 57 [51%] male; median age 3.51 yr, range 0.005 17.4 yr). NAC was administered as a continuous infusion (100 mg/kg/24 hours) until INR < 1.4, death, or liver transplantation (LT). The median duration of NAC administration in Group 2 was 5 (range, 1-77) days. Complications were noted in 8 (10.8%) children: rash in 3, arrhythmia in 3, and dizziness and peripheral edema in 1. One child had an allergic reaction (bronchospasm) and NAC was stopped. A total of 41 (71%) children in Group 1 vs. 85 (77%) in Group 2 required admission to intensive care, P = not significant (ns). The length of intensive care stay was 6 (range, 1-58) days in Group 1 vs. 5 (range, 1-68) days in Group 2, P = ns and length of hospital stay was 25 (range, 1-264) days vs. 19 (range, 1-201) days, P = 0.05. The 10-yr actuarial survival was 50% in Group 1 compared to 75% in Group 2, P = 0.009. Survival with native liver occurred in 13 (22%) in Group 1 vs. 48 (43%) in Group 2, P = 0.005; 15 (25%) in Group 1 died without transplant vs. 21 (19%) in Group 2, P = ns; and LT was performed in 32 (54%) vs. 42 (38%), P = ns. Death after transplantation occurred in 15 (39%) in Group 1 vs. 8 (16%) in Group 2, P = 0.02. In conclusion, NAC is safe in non-acetaminophen-induced ALF. In this retrospective study NAC was associated with a shorter length of hospital stay, higher incidence of native liver recovery without transplantation, and better survival after transplantation. PMID- 18161829 TI - Pretransplantation CD56(+) innate lymphocyte populations associated with severity of hepatitis C virus recurrence. AB - Cluster of differentiation (CD)56(+) lymphocytes are believed to play important roles in the innate immune response to viral infections by production of interferon (IFN)-gamma and/or the recognition of virally infected cells, but their role in liver transplantation (LT) has not been characterized. Here, for the first time, we examine the phenotypic and functional features of these cells in patients undergoing LT for hepatitis C virus (HCV)-related liver failure. The study was comprised of four patient groups: patients with mild HCV recurrence (n = 9), severe HCV recurrence (n = 10), patients with non-HCV-related liver failure (n =10), and normal healthy subjects (n = 10). Pre-LT, the frequency of circulating CD56(+) lymphocytes was significantly lower in patients who subsequently developed severe HCV recurrence, relative to those patients who developed mild histologic recurrence, as well as non-HCV controls. HCV was associated with impaired lymphokine-activated killing and natural cytotoxicity. We found that natural T (NT) cells that coexpressed CD4/CD8 or expressed CD8 alone were more frequent in patients who subsequently developed severe recurrence. In contrast, NT cells that expressed only CD4 appeared to be depleted in HCV infection relative to controls. A significantly higher percentage of NTs in both HCV groups expressed the inhibitory receptor NKG2A relative to HCV negative controls with liver disease. In conclusion, these results demonstrate a previously unappreciated association between pretransplantation CD56(+) lymphocytes and outcome of HCV recurrence and provide novel mechanistic insights into the immunopathogenesis of HCV recurrence, as well as potential targets for therapeutic manipulation. PMID- 18161830 TI - Liver transplantation for argininosuccinic aciduria: clinical, biochemical, and metabolic outcome. AB - We report successful liver transplantation in a young adult with argininosuccinic aciduria but without cirrhosis. Plasma amino acid profile normalized and brain magnetic resonance spectroscopy indicated improved metabolism after transplantation. The general well-being of the patient and his quality of life improved. We suggest that orthotopic liver transplantation should be considered for patients with argininosuccinic aciduria even in the absence of cirrhosis, with the aim of correcting (at least in part) central nervous system metabolism, thereby preventing further neurological deterioration. PMID- 18161831 TI - Effect of in vitro degradation of poly(D,L-lactide)/beta-tricalcium composite on its shape-memory properties. AB - The in vitro degradation characteristic and shape-memory properties of poly(D,L lactide) (PDLLA)/beta-tricalcium phosphate (beta-TCP) composites were investigated because of their wide application in biomedical fields. In this article, PDLLA and crystalline beta-TCP were compounded and interesting shape memory behaviors of the composite were first investigated. Then, in vitro degradation of the PDLLA/beta-TCP composites with weight ratios of 1:1, 2:1, and 3:1 was performed in phosphate buffer saline solution (PBS) (154 mM, pH 7.4) at 37 degrees C. The effect of in vitro degradation time for PDLLA/beta-TCP composites on shape-memory properties was studied by scanning electron microscopy, differential scanning calorimetry, gel permeation chromatography, X ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The changes of structural morphology, glass transition temperature (T(g)), molecular weight, and weight loss of composites matrix and pH change of degradation medium indicated that shape-memory effects at different degradation time were nonlinearly influenced because of the breaking down of polymer chain and the formation of degradation products. Furthermore, the results from XRD and FTIR implied that the degradation products, for example, hydroxyapatite (HA), calcium hydrogen phosphate (CaHPO(4)), and calcium pyrophosphate (Ca(2)P(2)O(7)) phases also had some effects on shape-memory properties during the degradation. PMID- 18161832 TI - Effect of keratin-gelatin and bFGF-gelatin composite film as a sandwich layer for full-thickness skin mesh graft in experimental dogs. AB - Skin grafts are indicated when there is a major loss of skin. Full-thickness skin graft is an ideal choice to reconstruct defect of irregular surface that is difficult to immobilize. Full-thickness mesh grafts can be applied to patch large skin defect when there is less donor site in extensively traumatized and burned surgical patients. The concept of using natural biomaterials such as keratin, basic fibroblast growth factor is slowly gaining popularity in the field of medical research to achieve early healing. The main objective of this study is to evaluate the efficacy of gelatin conjoined with keratin processed from the poultry feather and commercially available basic fibroblast growth factor (bFGF) as a sandwich layer in promoting the viability of full-thickness skin mesh grafts. The efficacy was assessed from the observation of clinical, bacteriological, and histopathological findings in three groups of experimental dogs. The clinical observations such as color, appearance and discharge, and hair growth were selected as criteria which indicated good and early acceptance of graft in keratin-gelatin (group II). On bacteriological examination, Staphylococcus aureus and Proteus was identified in few animals. Histopathological study of the patched graft revealed early presences of hair follicles; sebaceous gland, and normal thickness of the epidermis in keratin gelatin in group II treated animals compared with other group (group I-control, group III-bFGF-gelatin). PMID- 18161833 TI - Prevention of postoperative adhesions by a novel honeycomb-patterned poly(lactide) film in a rat experimental model. AB - Intraperitoneal adhesion is a serious problem concerning abdominal surgery. This study evaluated the performance of a honeycomb-patterned poly(lactide) (HCPLA) film as a physical barrier for preventing postoperative adhesion. HCPLA films were prepared using dioleoylphosphatidylethanolamine (DOPE) or a copolymer of dodecylacrylamide and omega-carboxyhexylacrylamide (CAP) as a surfactant (HCPLA DOPE and HCPLA-CAP, respectively). In an in vivo adhesion prevention experiment, male Sprague-Dawley rats underwent standard cecum abrasion before midline laparotomy. We placed 2 cm x 2 cm HCPLA and flat films on the gliding interfaces; untreated rats formed the control group. After 1 week, adhesion was scored from 0 to 4. No significant difference was observed in the scores among groups, but macroscopic differences in adhesion prevention were observed. The adhesive strength of HCPLA-DOPE (18.1 +/- 1.2 g) to skinless chicken breast was significantly higher than that of the flat film (15.2 +/- 0.8 g, p < 0.05). Further, the adhesion score after 1 week for the HCPLA-DOPE group (1.6 +/- 0.2) was significantly lower than that for the control group (3.0 +/- 0.3, p < 0.05) but comparable to that for the Seprafilm group (1.4 +/- 0.3). These results demonstrated the potential of HCPLA-DOPE as a physical barrier for preventing postoperative adhesion. PMID- 18161834 TI - Preparation of a bonelike apatite-polymer fiber composite using a simple biomimetic process. AB - A bonelike apatite-polymer fiber composite may be useful as an implant material to replace bone, the enthesis of a tendon, and the joint part of a ligament. We treated an ethylene-vinyl alcohol copolymer (EVOH) plate and knitted EVOH fibers with an oxygen plasma to produce oxygen-containing functional groups on their surfaces. The plasma-treated samples were alternately dipped in alcoholic calcium and phosphate ion solutions three times to deposit apatite precursors onto their surfaces. The surface-modified samples formed a dense and uniform bonelike surface apatite layer after immersion for 24 h in a simulated body fluid with ion concentrations approximately equal to those of human blood plasma. The adhesive strength between the apatite layer and the sample's surface increased with increasing power density of the oxygen plasma. The apatite-EVOH fiber composite obtained by our process has similarities to natural bone in that apatite crystals are deposited on organic polymer fibers. The resulting composite would possess osteoconductivity due to the apatite phase. With proper polymer selection and optimized synthesis techniques, a composite could be made that would have bonelike mechanical properties. Hence, the present surface modification and coating process would be a promising route to obtain new implant materials with bonelike mechanical properties and osteoconductivity. PMID- 18161835 TI - The synthesis and characterization of polymerizable and biocompatible N-maleic acyl-chitosan. AB - Biocompatible and polymerizable natural macromolecules have been found to provide great advantages in the preparation of hydrogels, which have wide applications in the fields of tissue engineering and polymeric drug delivery systems. To develop a new biocompatible polymerizable chitosan derivative, N-maleic acyl-chitosan (NMCS) was synthesized in this study. This novel biomaterial was designed from the N-acylation of chitosan with maleic anhydride introducing functional carboxyl and vinylated (--C[double bond]C--) groups. The structure of NMCS was characterized by FTIR, (1)H NMR, element analysis, and X-ray diffraction (XRD). NMCS can be dissolved into water because of its decreased crystallinity compared with chitosan. The NMCS's multiporous and microgel morphology was revealed by transmission electron microscope (TEM). Crosslinked hydrogel films can be successfully obtain through the macromolecular polymerization of NMCS. Subsequently, 3T3 fibroblasts were cultured onto the surface of the polymerized NMCS (P-NMCS) films to examine the capability of cell attachment and proliferation. Results from the cell culture demonstrate that P-NMCS films provide significant improvement in cell attachment and proliferation over unmodified chitosan. The improved P-NMCS cytocompatibility is expected to provide substantial contributions to tissue engineering in the future. PMID- 18161836 TI - Adhesion of a self-etching system to dental substrate prepared by Er:YAG laser or air abrasion. AB - The purpose of this study was to assess the microtensile bond strength of a self etching adhesive system to enamel and dentin prepared by Er:YAG laser irradiation or air abrasion, as well as to evaluate the adhesive interfaces by scanning electron microscopy (SEM). For microtensile bond strength test, 80 third molars were randomly assigned to five groups: Group I, carbide bur, control (CB); II, air abrasion with standard tip (ST); III, air abrasion with supersonic tip (SP); IV, Er:YAG laser 250 mJ/4 Hz (L250); V, Er:YAG laser 300 mJ/4 Hz (L300). Each group was divided into two subgroups (n = 8) (enamel, E and dentin, D). E and D surfaces were treated with the self-etching system Adper Prompt L-Pop and composite buildups were done with Filtek Z-250. Sticks with a cross-sectional area of 0.8 mm(2) (+/-0.2 mm(2)) were obtained and the bond strength tests were performed. Data were submitted to ANOVA and Tukey's test. For morphological analysis, disks of 30 third molars were restored, sectioned and prepared for SEM. Dentin presented the highest values of adhesion, differing from enamel. Laser and air-abrasion preparations were similar to enamel. Dentin air-abrasion with standard tip group showed higher bond strength results than Er:YAG-laser groups, however, air-abrasion and Er:YAG laser groups were similar to control group. SEM micrographs revealed that, for both enamel and dentin, the air-abrasion and laser preparations presented irregular adhesive interfaces, different from the ones prepared by rotary instrument. It was concluded that cavity preparations accomplished by both Er:YAG laser energies and air abrasion tips did not positively influence the adhesion to enamel and dentin. PMID- 18161837 TI - Novel extracellular matrix for cell sheet recovery using genetically engineered elastin-like protein. AB - Elastin-like peptides (ELPs) sequences are repeats of the pentapeptide GVGVP, and they have the ability to coaggregate reversibly, depending on the temperature. By exploiting this characteristic, a novel extracellular matrix protein (ECM) containing ELP was developed genetically to harvest a cell sheet from a culture dish. One of the ELP constructs, G288, consisted of 288 repeats of the sequence GVGVGP (G); it was attached to a hydrophobic dish surface. Next, cells with the sequence His-G36RG36, which has a His tag and an RGD sequence (R) that promotes attachment of the cell between the G36 sequences, consisted of 36 repeats of the sequence GVGVP, were added to the dish. After these cells became confluent, the temperature was changed to 20 degrees C in order to reverse the coaggregation. At this temperature, cells could be detached from the dish as a cell sheet. This genetically engineering method for construction of thermoresponsive ECM would be suitable to modify ECM with further functional domains. PMID- 18161838 TI - Predicting immunosuppressant dosing in the early postoperative period with noninvasive indocyanine green elimination following orthotopic liver transplantation. AB - Twenty adult patients undergoing orthotopic liver transplantation (OLT) were enrolled in this study, with the noninvasive indocyanine green plasma disappearance rate (ICG-PDR) measured both during and after OLT to assess the relationship between ICG-PDR and the ability of patients to achieve therapeutic postoperative tacrolimus immunosuppressant blood levels. Liver function was determined at both 2 and 18 hours post reperfusion with the ICG-PDR k value (1/min). Postoperative standard serum measures of liver function as well as liver biopsies were also collected and analyzed. The median ICG-PDR k value for the study group at 2 hours post reperfusion was 0.20 (0.16, 0.27), whereas at 18 hours post reperfusion, it was 0.22 (0.18, 0.35). The median change in the k value between the two ICG-PDR measurements was 0.05 (-0.02, 0.07) with P = 0.02. There was an interaction between the postoperative day 1 (18 hours post reperfusion) ICG-PDR k value and the linear increase in the tacrolimus blood level, such that the greater the k value was, the more gradual the observed rise was in tacrolimus over time [that is, the longer it took to achieve a therapeutic blood level (>12 ng/mL), P = 0.003]. Of the 16 patients that received tacrolimus, comparable dosing on a per kilogram body weight basis was observed. Also, no significant association between ICG-PDR k values and postoperative liver biopsy results was seen. This study demonstrates that the ICG-PDR measurement is a modality with the potential to assist in achieving adequate blood levels of tacrolimus following OLT. PMID- 18161839 TI - Recurrent hepatitis C after liver transplantation: on-treatment prediction of response to peginterferon/ribavirin therapy. AB - Sustained virologic response (SVR) in the treatment of recurrent hepatitis C virus (HCV) infection after liver transplantation (LT) remains suboptimal. We evaluated efficacy of pegylated interferon alfa (PEG) and ribavirin (RBV) (PEG/RBV) combination therapy in LT recipients with recurrent HCV and predictive values of rapid virological response (RVR) and early virologic response (EVR). Between January 2001 and October 2005, LT recipients with recurrent HCV were intended to be treated for 48 weeks with PEG/RBV combination therapy independent of genotype or virologic response [53 patients (79% genotype 1)]. On-treatment predictor of response at week 4 (RVR) was defined as undetectable HCV RNA, and at week 12 (EVR) as undetectable HCV RNA or a >2 log(10) drop from pretreatment viral load. SVR was seen in 19 (35%) patients. Patients with genotype 2/3 were more likely to achieve SVR than those with genotype 1 (87% versus 23%; P = 0.001). The highest rate of SVR was seen in patients with RVR [specificity and positive predictive value (PPV) = 100%] while the highest rate of treatment failure was seen in those who did not have EVR [sensitivity and negative predictive value (NPV) = 100%]. The NPV of RVR to identify those who will not achieve SVR was also very high (88%). EVR had low PPV (63%) to identify those with SVR. In conclusion, PEG/RBV combination therapy is effective in the treatment of post-LT recurrent HCV. On-treatment virologic monitoring is highly predictive of SVR and may optimize the virologic response and minimize toxicity. Given its high PPV and NPV, RVR appears to be the most appropriate decision time point for continuation of therapy. PMID- 18161840 TI - Association between donor-recipient serum sodium differences and orthotopic liver transplant graft function. AB - Previous studies have shown that donor hypernatremia and possibly recipient hyponatremia negatively impact graft function after orthotopic liver transplant (OLT). The purpose of this retrospective investigation was to determine whether measured differences in serum sodium values between cadaveric donors and OLT recipients (DeltaNa(+)) influence immediate postoperative allograft function and short-term patient outcomes. Two hundred and fifty patients that underwent OLT from January 2001 to December 2005 were included in this study. The DeltaNa(+) for each donor recipient pair was correlated with standard postoperative liver function tests as well as recipient length of intensive care unit stay (LOICUS), length of hospital stay (LOHS) and recipient survival. The relationship between donor hypernatremia (serum sodium >or= 155 mEq/mL), recipient hyponatremia (serum sodium level 64 years were independent predictive factors of osteoporosis in multivariate analysis. In females, back pain was an independent factor for osteoporosis. CONCLUSION: The results of this study revealed that prevalence of osteoporosis and vertebral bone deformity rate were high in gastric cancer patients, regardless of post-gastrectomy duration and operation type. Early diagnosis and a proper management plan must be established in these patients. PMID- 18161917 TI - JAK-STAT pathway in carcinogenesis: is it relevant to cholangiocarcinoma progression? AB - The features of JAK-STAT signaling in liver cells are discussed in the current review. The role of this signaling cascade in carcinogenesis is accentuated. The possible involvement of this pathway and alteration of its elements are compared for normal cholangiocytes, cholangiocarcinoma predisposition and development. Prolactin and interleukin-6 are described in detail as the best studied examples. In addition, the non-classical nuclear translocation of cytokine receptors is discussed in terms of its possible implication to cholangiocarcinoma development. PMID- 18161919 TI - Effect of arsenic trioxide on vascular endothelial cell proliferation and expression of vascular endothelial growth factor receptors Flt-1 and KDR in gastric cancer in nude mice. AB - AIM: To investigate the effect of arsenic trioxide (As2O3) on expression of vascular endothelial growth factor receptor-1 (VEGFR-1, Flt-1) and VEGFR-2 (KDR) in human gastric tumor cells and proliferation of vascular endothelial cells. METHODS: The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were treated with As2O3. Microvessel density (MVD) and expression of Flt-1 and KDR were detected by immunofluorescence laser confocal microscopy. SGC-7901 cells were treated respectively by exogenous recombinant human VEGF165 or VEGF165 + As2O3. Cell viability was measured by MTT assay. Cell viability of ECV304 cells was measured by MTT assay, and cell cycle and apoptosis were analyzed using flow cytometry. RESULTS: The tumor growth inhibition was 30.33% and 50.85%, respectively, in mice treated with As2O3 2.5 and 5 mg/kg. MVD was significantly lower in arsenic-treated mice than in the control group. The fluorescence intensity levels of Flt-1 and KDR were significantly less in the arsenic-treated mice than in the control group. VEGF165 may accelerate growth of SGC7901 cells, but As2O3 may disturb the stimulating effect of VEGF165. ECV304 cell growth was suppressed by 76.51%, 71.09% and 61.49% after 48 h treatment with As2O3 at 0.5, 2.5 and 5 micromol/L, respectively. Early apoptosis in the As2O3-treated mice was 2.88-5.1 times higher than that in the controls, and late apoptosis was 1.17-1.67 times higher than that in the controls. CONCLUSION: Our results showed that As2O3 delays tumor growth, inhibits MVD, down-regulates Flt-1 and KDR expression, and disturbs the stimulating effect of VEGF165 on the growth of SGC7901 cells. These results suggest that As2O3 might delay growth of gastric tumors through inhibiting the paracrine and autocrine pathways of VEGF/VEGFRs. PMID- 18161920 TI - Inhibitory effect of schisandrin B on gastric cancer cells in vitro. AB - AIM: To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro. METHODS: SC-B consisted of schisandrin B, aloe-emodin, and Astragalus polysaccharides. Exponentially growing human gastric cancer SGC-7901 cells were divided into six treatment groups: (1) control group (RPMI 1640 medium); (2) negative control group (2% DMSO); (3) positive control group (50 mg/L 5-Fluorouracil, 5-FU); (4) low-dose group (LSC, final concentration of schisandrin B, 25 mg/L); (5) moderate-dose group (MSC, final concentration of schisandrin B, 50 mg/L); (6) high-dose group (HSC, final concentration of schisandrin B, 100 mg/L). Follow-up was done at 12 48 h. An MTT (Methylthiazolyldiphenyl-tetrazolium bromide) assay was used to examine the inhibitory effect of SC-B on gastric cancer cells. The mitosis index was assessed using an inverted microscope. Flow cytometry was used to visualize the cell cycle. An RT-PCR (Reverse transcription-Polymerase chain reaction) based assay was used to detect mRNA expression for cyclin D1 and glyceraldehyde-3 phosphate dehydrogenase (GAPDH). RESULTS: The MTT assay showed that the number of living cells in the LSC, MSC and HSC groups was significantly smaller than that in the DMSO-treated group (P < 0.05) at 12-48 h. The inhibitory rate (IR) of the LSC group was 41.15% +/- 3.86%, 59.24% +/- 5.34% and 69.93% +/- 7.81% at 12, 24 and 48 h, respectively. The IR of the MSC group was 42.82% +/- 4.94%, 62.68% +/- 7.58% and 71.79% +/- 8.12% at 12, 24 and 48 h, respectively. The IR of the HSC group was 37.50% +/- 3.21%, 40.34% +/- 2.98% and 61.99% +/- 4.88% at 12, 24 and 48 h, respectively. These results suggested that a moderate dosage had the most obvious inhibitory efficacy at 48 h. Compared to the DMSO group, the mitosis index of the LSC, MSC, HSC groups was greatly decreased (P < 0.05) at all time points. Any dose of SC-B suppressed mitosis within 12-48 h. Compared to the DMSO group, the percentage of cells in the G0/G1 phase of the MSC group was greatly increased, and that of the S + G2M phase was greatly decreased, while the percentage of cell inhibition (PCI) in the MSC group was greatly increased (P < 0.05). This suggested that SC-B could exclusively arrest cells in the G0/G1 phase. Cyclin D1 mRNA expression was lower in the MSC group than that in the DMSO group (P < 0.05). CONCLUSION: SC-B can inhibit the proliferation and aberrant mitosis of human gastric cancer SCG-7901 cells in vitro. This inhibitory effect may be due to the down-regulation of cyclin D1 mRNA expression, which causes cell cycle arrest of gastric cancer cells. PMID- 18161921 TI - New anti-proliferative agent, MK615, from Japanese apricot "Prunus mume" induces striking autophagy in colon cancer cells in vitro. AB - AIM: To investigate the anti-neoplastic effects of MK615, an extract from the Japanese apricot (Prunus mume), against colon cancer cells. METHODS: Three colon cancer cell lines, SW480, COLO, and WiDr, were cultured with MK615. Growth inhibition was evaluated by cell proliferation assay and killing activity was determined by lactate dehydrogenase assay. Induction of apoptosis was evaluated by annexin V flow cytometry. Morphological changes were studied by light and electron microscopy, and immunofluorescence staining with Atg8. RESULTS: MK615 inhibited growth and lysed SW480, COLO and WiDr cells in a dose-dependent manner. Annexin V flow cytometry showed that MK615 induced apoptosis after 6 h incubation, at which point the occurrence of apoptotic cells was 68.0%, 65.7% and 64.7% for SW480, COLO, and WiDr cells, respectively. Light and electron microscopy, and immunofluorescence staining with Atg8 revealed that MK615 induced massive cytoplasmic vacuoles (autophagosomes) in all three cell lines. CONCLUSION: MK615 has an anti-neoplastic effect against colon cancer cells. The effect may be exerted by induction of apoptosis and autophagy. PMID- 18161922 TI - Chronic hepatitis C virus infection: prevalence of extrahepatic manifestations and association with cryoglobulinemia in Bulgarian patients. AB - AIM: To assess the prevalence of extrahepatic manifestations in Bulgarian patients with chronic hepatitis C virus (HCV) infection and identify the clinical and biological manifestations associated with cryoglobulinemia. METHODS: The medical records of 136 chronically infected HCV patients were reviewed to assess the prevalence of extrahepatic manifestations. Association between cryoglobulin positivity and other manifestations were identified using chi2 and Fisher's exact test. Risk factors for the presence of extrahepatic manifestations were assessed by logistic regression analysis. RESULTS: Seventy six percent (104/136) of the patients had at least one extrahepatic manifestation. Clinical manifestations included fatigue (59.6%), kidney impairment (25.0%), type 2 diabetes (22.8%), paresthesia (19.9%), arthralgia (18.4%), palpable purpura (17.6%), lymphadenopathy (16.2%), pulmonary fibrosis (15.4%), thyroid dysfunction (14.7%), Raynaud's phenomenon (11.8%), B-cell lymphoma (8.8%), sicca syndrome (6.6%), and lichen planus (5.9%). The biological manifestations included cryoglobulin production (37.5%), thrombocytopenia (31.6%), and autoantibodies: anti-nuclear (18.4%), anti-smooth muscle (16.9%), anti-neutrophil cytoplasm (13.2%) and anti cardiolipin (8.8%). All extrahepatic manifestations showed an association with cryoglobulin-positivity, with the exception of thyroid dysfunction, sicca syndrome, and lichen planus. Risks factors for the presence of extrahepatic manifestations (univariate analysis) were: age > or = 60 years, female gender, virus transmission by blood transfusions, longstanding infection (> or = 20 years), and extensive liver fibrosis. The most significant risks factors (multivariate analysis) were longstanding infection and extensive liver fibrosis. CONCLUSION: We observed a high prevalence of extrahepatic manifestations in patients with chronic HCV infection. Most of these manifestations were associated with impaired lymphoproliferation and cryoglobulin production. Longstanding infection and extensive liver fibrosis were significant risk factors for the presence of extrahepatic manifestations in HCV patients. PMID- 18161923 TI - Calcitriol analog ZK191784 ameliorates acute and chronic dextran sodium sulfate induced colitis by modulation of intestinal dendritic cell numbers and phenotype. AB - AIM: To investigate the effects of ZK1916784, a low calcemic analog of calcitriol on intestinal inflammation. METHODS: Acute and chronic colitis was induced by dextran sodium sulfate (DSS) according to standard procedures. Mice were treated intraperitoneally with ZK1916784 or placebo and colonic inflammation was evaluated. Cytokine production by mesenterial lymph node (MLN) cells was measured by ELISA. Immunohistochemistry was performed to detect intestinal dendritic cells (DCs) within the colonic tissue, and the effect of the calcitriol analog on DCs was investigated. RESULTS: Treatment with ZK191784 resulted in significant amelioration of disease with a reduced histological score in acute and chronic intestinal inflammation. In animals with acute DSS colitis, down-regulation of colonic inflammation was associated with a dramatic reduction in the secretion of the proinflammatory cytokine interferon (IFN)-gamma and a significant increase in intereleukin (IL)-10 by MLN cells. Similarly, in chronic colitis, IL-10 expression in colonic tissue increased 1.4-fold when mice were treated with ZK191784, whereas expression of the Th1-specific transcription factor T-beta decreased by 81.6%. Lower numbers of infiltrating activated CD11c+ DCs were found in the colon in ZK191784-treated mice with acute DSS colitis, and secretion of proinflammatory cytokines by primary mucosal DCs was inhibited in the presence of the calcitriol analog. CONCLUSION: The calcitriol analog ZK191784 demonstrated significant anti-inflammatory properties in experimental colitis that were at least partially mediated by the immunosuppressive effects of the derivate on mucosal DCs. PMID- 18161924 TI - Acanthus ilicifolius plant extract prevents DNA alterations in a transplantable Ehrlich ascites carcinoma-bearing murine model. AB - AIM: To investigate the chemopreventive efficacy of the Indian medicinal plant Acanthus ilicifolius L Acanthaceae in a transplantable Ehrlich ascites carcinoma (EAC)-bearing murine model. METHODS: Male Swiss albino mice were divided into four groups: Group A was the untreated normal control; Group B was the EAC control mice group that received serial, intraperitoneal (ip) inoculations of rapidly proliferating 2 x 10(5) viable EAC cells in 0.2 mL of sterile phosphate buffered saline; Group C was the plant extract-treated group that received the aqueous leaf extract (ALE) of the plant at a dose of 2.5 mg/kg body weight by single ip injections, once daily for 10, 20 and 30 consecutive days following tumour inoculation (ALE control); and Group D was the EAC + ALE-treatment group. The chemopreventive potential of the ALE was evaluated in a murine model by studying various biological parameters and genotoxic markers, such as tumour cell count, mean survival of the animals, haematological indices, hepatocellular histology, immunohistochemical expression of liver metallothionein (MT) protein, sister-chromatid exchanges (SCEs), and DNA alterations. RESULTS: Treatment of the EAC-bearing mice with the ALE significantly (P < 0.001) reduced viable tumour cell count by 68.34% (228.7 x 10(6) +/- 0.53) when compared to EAC control mice (72.4 x 10(6) +/- 0.49), and restored body and organ weights almost to the normal values. ALE administration also increased (P < 0.001) mean survival of the hosts from 35 +/- 3.46 d in EAC control mice to 83 +/- 2.69 d in EAC + ALE-treated mice. Haematological indices also showed marked improvement with administration of ALE in EAC-bearing animals. There was a significant increase in RBC count (P < 0.001), hemoglobin percent (P < 0.001), and haematocrit value (P < 0.001) from 4.3 +/- 0.12, 6.4 +/- 0.93, and 17.63 +/- 0.72 respectively in EAC control mice to 7.1 +/- 0.13, 12.1 +/- 0.77, and 30.23 +/- 0.57 respectively in EAC + ALE treated group, along with concurrent decrement (P < 0.001) in WBC count from 18.8 +/- 0.54 in EAC control to 8.4 +/- 0.71 in EAC + ALE. Furthermore, treatment with ALE substantially improved hepatocellular architecture and no noticeable neoplastic lesions or foci of cellular alteration were observed. Daily administration of the ALE was found to limit liver MT expression, an important marker of cell proliferation with concomitant reduction in MT immunoreactivity (62.25 +/- 2.58 vs 86.24 +/- 5.69, P < 0.01). ALE was also potentially effective in reducing (P < 0.001) the frequency of SCEs from 14.94 +/- 2.14 in EAC control to 5.12 +/- 1.16 in EAC + ALE-treated group. Finally, in comparison to the EAC control, ALE was able to suppress in vivo DNA damage by abating the generations of 'tailed' DNA by 53.59% (98.65 +/- 2.31 vs 45.06 +/- 1.14, P < 0.001), and DNA single-strand breaks (SSBs) by 38.53% (3.14 +/- 0.31 vs 1.93 +/- 0.23, P < 0.01) in EAC-bearing murine liver. CONCLUSION: Our data indicate that, ALE is beneficial in restoring haematological and hepatic histological profiles and in lengthening the survival of the animals against the proliferation of ascites tumour in vivo. Finally, the chemopreventive efficacy of the ALE is manifested in limiting MT expression and in preventing DNA alterations in murine liver. The promising results of this study suggest further investigation into the chemopreventive mechanisms of the medicinal plant A. ilicifolius in vivo and in vitro. PMID- 18161925 TI - Choice of laxatives and colonoscopic preparation in pregnant patients from the viewpoint of obstetricians and gastroenterologists. AB - AIM: To elucidate the preferences of gastroenterologists at our institution and compare them to those of obstetricians when making decisions in the pregnant patient, including which type of bowel preparations to use for flexible sigmoidoscopy or colonoscopy, as well as which laxatives can be used safely. METHODS: Surveys were mailed to all attending gastroenterologists (n = 53) and obstetricians (n = 99) at our institution. Each survey consisted of the 14 most common laxative or motility agents used in pregnancy and inquired about the physician's prescribing habits in the past as well as their willingness to prescribe each medication in the future. The survey also listed four common bowel preparations used prior to colonoscopy and sigmoidoscopy and asked the physician to rank the order of the preferred agent in each case. RESULTS: With regard to common laxatives, both gastroenterologists and obstetricians favor the use of Metamucil, Colace, and Citrucel. Both groups appear to refrain from using Fleets Phosphosoda and Castor oil. Of note, obstetricians are less inclined to use PEG solution and Miralax, which is not the case with gastroenterologists. In terms of comparing bowel preparations for colonoscopy, 50% of gastroenterologists prefer to use PEG solution and 50% avoid the use of Fleets Phosphosoda. Obstetricians seem to prefer Fleets Phosphosoda (20%) and tend to avoid the use of PEG solution (26%). With regard to bowel preparation for sigmoidoscopy, both groups prefer Fleets enema the most (51%), while magnesium citrate is used least often (38%). CONCLUSION: It is clear that preferences in the use of bowel cleansing preparations between the two groups exist, but there have not been many case controlled human studies in the pregnant patient that give clear cut indications for using one versus another drug. In light of the challenge of performing controlled trials in pregnant women, more extensive surveys should be undertaken to gather a larger amount of data on physician's experiences and individual preferences. PMID- 18161926 TI - Activity and safety of pegylated liposomal doxorubicin, 5-fluorouracil and folinic acid in inoperable hepatocellular carcinoma: a phase II study. AB - AIM: To improve the results of New therapeutic strategies in hepatocellular carcinoma (HCC). We have conducted a phase II study with pegylated liposomal doxorubicin (PLD), 5-fluorouracil (5FU) and folinic acid (FA). METHODS: Thirty one patients with hystologically-confirmed, inoperable HCC, received combination chemotherapy with PLD 25 mg/mq on d 1, 5FU 1200 mg/mq in 48 h continuous infusion, and oral FA 30 mg on d 1 and 2 every 3 wk until disease progression or intolerable toxicity. RESULTS: The median age was 65 years (range 41-82) and 28 patients were hepatitis C virus seropositive (90%). The majority of patients were Child-Pugh Class B (55%). Two patients showed a partial response (PR), and 16 had stable disease (SD). With a median follow-up of 14 mo, the median time to progression of all evaluable patients was 4 mo (95% CI 1.7-7). Median overall survival was 9 mo (95% CI 3-24 mo). After 1 year, 9 of 18 PR/SD patients were alive. Chemotherapy was well tolerated. CONCLUSION: PLD/FU/FA combination seems capable of achieving durable stabilization of HCC. The manageable toxicity supports a role for combination with other anticancer agents. PMID- 18161927 TI - PlexinA1 expression in gastric carcinoma and its relationship with tumor angiogenesis and proliferation. AB - AIM: To explore the expression of PlexinA1 in gastric carcinoma and its relationship with tumor angiogenesis and proliferation. METHODS: PlexinA1 mRNA and protein expressions of Semaphorin6D were measured using semi-quantity reverse transcription PCR and Western blotting in 20 cases of gastric carcinoma and corresponding normal gastric mucosa. PlexinA1, Ki-67 expression and microvessel density (MVD) were detected by immunohistochemistry in 50 cases of gastric carcinoma and 20 cases of normal gastric mucosa. RESULTS: The mRNA and protein expressions of PlexinA1 in gastric carcinoma were significantly higher than that in normal gastric mucosa (0.71 +/- 0.37 vs 0.60 +/- 0.25, P = 0.0299 < 0.05, and 0.47 +/- 0.16 vs 0.21 +/- 0.08, P = 0.0000 < 0.01), and MVD within tumor tissues increased significantly with PlexinA1 mRNA expression (r =0.8736, P < 0.01) and PlexinA1 protein expression (r = 0.7286, P < 0.01), and MVD of the PlexinA1 positive staining group (25.25 +/- 3.93) was significantly higher than that of the negative group (19.56 +/- 1.75), (P < 0.01). Proliferation index of tumor cells within tumor tissues were positively correlated with PlexinA1 mRNA expression (r = 0.5420, P = 0.014 < 0.01) and PlexinA1 protein expression (r = 0.5024, P = 0.024 < 0.05). The proliferation index of the PlexinA1 positive staining group (567.69 +/- 125.61) was significantly higher than that of the negative group (369.58 +/- 116.88), (P < 0.01). CONCLUSION: PlexinA1 may play an important role in the occurrence and development of gastric carcinoma, and be related to tumor angiogenesis and proliferation. PMID- 18161928 TI - Serum pepsinogen levels and their influencing factors: a population-based study in 6990 Chinese from North China. AB - AIM: To explore the essential characteristics of serum pepsinogen (PG) levels in Chinese people, by analyzing the population-based data on the serum levels of PG I and II and the PG I/II ratio, and their influencing factors in Chinese from North China. METHODS: A total of 6990 subjects, who underwent a gastric cancer screening in North China from 1997 to 2002, were collected in this study. Serum pepsinogen levels were measured by enzyme-linked immunosorbent assay (ELISA). H pylori status was determined by histological examination and H pylori-IgG ELISA. The cut-off point was calculated by using receiving operator characteristics (ROC) curves. Factors linked to serum PG I/II ratio were identified using a multivariate logistic regression. RESULTS: The serum PG I and PG II levels were significantly higher in males than in females (95.2 microg/L vs 79.7 microg/L, P < 0.01; 12.1 microg/L vs 9.4 microg/L, P < 0.01), PG I/II ratio was significantly lower in males than in females (7.9 vs 8.3, P < 0.01). The PG I/II ratio decreased significantly in the aged groups following the progression of gastric mucosa from normal to non-atrophic and atrophic lesions (10.4, 8.8, and 6.6, respectively). The serum PG I and II levels were significantly higher in patients with H pylori infection than in those without H pylori infection (88.7 microg/L vs 81.4 microg/L, P < 0.01; 11.4 microg/L vs 8.4 microg/L, P < 0.01), while the PG I/II ratio was significantly lower in patients with H pylori infection than in those without H pylori infection (7.7 vs 9.6, P < 0.01). For patients with atrophic lesions, the area under the PG I/II ROC curve was 0.622. The best cut off point for PG I/II was 6.9, with a sensitivity of 53.2%, and a specificity of 67.5%. Factors linked to PG I/II were sensitive to identified PG using a multinomial logistic regression relying on the following inputs: males (OR: 1.151, 95% CI: 1.042-1.272, P = 0.006), age > or = 61 years (OR: 1.358, 95% CI: 1.188-1.553, P = 0.000), atrophic lesion (OR: 2.075, 95% CI: 1.870-2.302, P = 0.000), and H pylori infection (OR: 1.546, 95% CI: 1.368-1.748, P = 0.000). CONCLUSION: The essential characteristics of serum PG levels in Chinese are significantly skewed from the normal distribution, and influenced by age, sex, gastric mucosa lesions and H pylori infection. PG I/II ratio is more suitable for identifying subgroups with different influence factors compared with PG I or PG II alone. PMID- 18161929 TI - Gastrointestinal tract distribution of Salmonella enteritidis in orally infected mice with a species-specific fluorescent quantitative polymerase chain reaction. AB - AIM: To identify and understand the regular distribution pattern and primary penetration site for Salmonella enteritidis (S. enteritidis) in the gastrointestinal tract. METHODS: Based on the species-specific DNA sequence of S. enteritidis from GenBank, a species-specific real-time, fluorescence-based quantitative polymerase chain reaction (FQ-PCR) was developed for the detection of S. enteritidis. We used this assay to detect genomic DNA of S. enteritidis in the gastrointestinal tract, including duodenum, jejunum, ileum, cecum, colon, rectum, esophagus and stomach, from mice after oral infection. RESULTS: S. enteritidis was consistently detected in all segments of the gastrointestinal tract. The jejunum and ileum were positive at 8 h post inoculation, and the final organ to show a positive result was the stomach at 18 h post inoculation. The copy number of S. enteritidis DNA in each tissue reached a peak at 24-36 h post inoculation, with the jejunum, ileum and cecum containing high concentrations of S. enteritidis, whereas the duodenum, colon, rectum, stomach and esophagus had low concentrations. S. enteritidis began to decrease and vanished at 2 d post inoculation, but it was still present up to 5 d post inoculation in the jejunum, ileum and cecum, without causing apparent symptoms. By 5 d post inoculation, the cecum had significantly higher numbers of S. enteritidis than any of the other areas (P < 0.01), and this appeared to reflect its function as a repository for S. enteritidis. CONCLUSION: The results provided significant data for clarifying the pathogenic mechanism of S. enteritidis in the gastrointestinal tract, and showed that the jejunum, ileum and cecum are the primary sites of invasion in normal mice after oral infection. This study will help to further understanding of the mechanisms of action of S. enteritidis. PMID- 18161930 TI - Effects of a Shuangling Fuzheng anticancer preparation on the proliferation of SGC-7901 cells and immune function in a cyclophosphamide-treated murine model. AB - AIM: To study the inhibitory effects of a Shuangling Fuzheng anticancer preparation (SFAP) on the human gastric cancer cell line SGC-7901 in vitro as well as its immune-modulated effects in a cyclophosphamide-treated murine model. METHODS: MTT experiments and immunocytochemistry ABC experiments were performed for detecting the proliferation of SGC-7901 cells in vitro and protein expression of c-myc. The staphylococcal protein A (SPA) rosette test was utilized for measuring the ratio of T-lymphocyte subsets from peripheral blood in a cyclophosphamide-treated murine model. Enzyme-linked immunosorbant assay (ELISA) was performed for measuring the levels of serum sIL-2R in treated mice, while immunoturbidimetry was used for measuring the levels of immunoglobulins (Ig). RESULTS: SFAP (40-640 mg/L, 48 h) inhibited the proliferation of SGC-7901 cells, and a positive correlation was noted between inhibitory effects and dosage. At a dosage of 160-320 mg/L in cultured cells, the expression of c-myc was decreased. SFAP (50-200 mg/kg) increased the percentage of CD3+ and CD4+ T-lymphocytes, the ratio of CD4/CD8, and the contents of Ig such as IgM, IgG or IgA, but decreased the levels of serum sIL-2R in peripheral blood from cyclophosphamide-treated mice. CONCLUSION: SFAP can inhibit the proliferation of SGC-7901 cells via the c myc gene. In addition, SFAP can modulat the cellular and humoral immunity in cyclophosphamide-induced immunosuppressed mice. PMID- 18161931 TI - ShRNA-mediated gene silencing of beta-catenin inhibits growth of human colon cancer cells. AB - AIM: To observe the gene silencing mediated by the specific shRNA targeted against beta-catenin and its effect on cell proliferation and cycle distribution in the human colon cancer cell line Colo205. METHODS: Two shRNA plasmid vectors against beta-catenin were constructed and transfected into Colo205 cells with Lipofectamine2000. The down-regulations of beta-catenin, c-myc and cyclinD1 expressions were detected by RT-PCR and western blot analysis. The cell proliferation inhibitions were determined by MTT assay and soft agar colony formation assay. The effect of these two beta-catenin shRNAs on cell cycle distribution and apoptosis was examined by flow cytometry. RESULTS: These two shRNA vectors targeted against beta-catenin efficiently suppressed the expression of beta-catenin and its down stream genes, c-myc and cyclinD1. The expression inhibition rates were around 40%-50% either at the mRNA or at the protein level. The shRNA-mediated gene silencing of beta-catenin resulted in significant inhibition of cell growth both on the culture plates and in the soft agar. Moreover, the cancer cells showed significant G0/G1 arrest and increased apoptosis at 72 h post transfection due to gene silencing. CONCLUSION: These specific shRNAs targeted against beta-catenin could have a gene silencing effect and block the WNT signaling pathway. They could inhibit cell growth, increase apoptosis, and induce cell cycle arrest in Colo205 cells. ShRNA interference against beta-catenin is of potential value in gene therapy of colon cancer. PMID- 18161932 TI - Predictive value of D-dimer for portal vein thrombosis after portal hypertension surgery in hepatitis B virus-related cirrhosis. AB - AIM: To evaluate the predictive value of D-dimer as a predictive indicator of portal vein thrombosis (PVT) after portal hypertension surgery in hepatitis B virus-related cirrhosis. METHODS: A prospective study was carried out in 52 patients who had undergone surgery for portal hypertension in hepatitis B virus related cirrhosis. Changes in perioperative dynamic D-dimer were observed. The sensitivity, specificity, positive predictive values and negative predictive values of D-dimer were calculated, and ROC curves were analyzed. RESULTS: The D dimer levels in the group developing postoperative PVT was significantly higher than those in the group not developing PVT (P = 0.001), and the ROC semiquantitative and qualitative analysis of D-dimer showed a moderate predictive value in PVT (semi-quantitative value Az = 0.794, P = 0.000; qualitative analysis: Az = 0.739, P = 0.001). CONCLUSION: Dynamic monitoring of D-dimer levels in patients with portal hypertension after surgery can help early diagnosis of PVT, as in cases where the D-dimer levels steadily increase and exceed 16 microg/mL, the possibility of PVT is very high. PMID- 18161933 TI - Combined treatment of hepatocellular carcinoma with partial splenic embolization and transcatheter hepatic arterial chemoembolization. AB - AIM: To prospectively evaluate the efficacy and safety of partial splenic embolization (PSE) combined with transcatheter hepatic arterial chemoembolization (TACE) in treatment of hepatocellular carcinoma (HCC). METHODS: Fifty patients suffering from primary HCC associated with hypersplenism caused by cirrhosis were randomly assigned to 2 groups: group A receiving PSE combined with TACE (n = 26) and group B receiving TACE alone (n = 24). Follow-up examinations included calculation of peripheral blood cells (leukcytes, platelets and red blood cells) and treatment-associated complications. RESULTS: Prior to treatment, there was no significant difference in sex, age, Child-Pugh grade, tumor diameter, mass pathology type and peripheral blood cell counts between the 2 groups. After treatment, leukocyte and platelet counts were significantly higher in group A during the 3-mo follow-up period (P < 0.05), but lower in group B (P < 0.05). Severe complications occurred in 3 patients (11.5%) of group A and in 19 patients (79.2%) of group B (P < 0.05), and there was no significant difference in symptoms of post-embolization syndrome, including abdominal pain, fever, mild nausea and vomiting between the 2 groups (P > 0.05). CONCLUSION: PSE combined with TACE is more effective and safe than TACE alone for patients with HCC associated with hypersplenism caused by cirrhosis. PMID- 18161934 TI - Surgical management in biliary restricture after Roux-en-Y hepaticojejunostomy for bile duct injury. AB - AIM: To discuss the surgical method and skill of biliary restricture after Roux en-Y hepaticojejunostomy for bile duct injury. METHODS: From November 2005 to December 2006, eight patients with biliary restricture after Roux-en-Y hepaticojejunostomy for bile duct injury were admitted to our hospital. Their clinical data were analyzed retrospectively. RESULTS: Bile duct injury was caused by cholecystectomy in the eight cases, including seven cases with laparoscopic cholecystectomy and one with mini-incision choleystectomy. According to the classification of Strasberg, type E1 injury was found in one patient, type E2 injury in three, type E3 injury in two and type E4 injury in two patients. Both of the type E4 injury patients also had a vascular lesion of the hepatic artery. Six patients received Roux-en-Y hepaticojejunostomy for the second time, and one of them who had type E4 injury with the right hepatic artery disruption received right hepatectomy afterward. One patient who had type E4 injury with the proper hepatic artery lesion underwent liver transplantation, and the remaining one with type E3 injury received external biliary drainage. All the patients recovered fairly well postoperatively. CONCLUSION: Roux-en-Y hepaticojejunostomy is still the main approach for such failed surgical cases with bile duct injury. Special attention should be paid to concomitant vascular injury in these cases. The optimal timing and meticulous and excellent skills are essential to the success in this surgery. PMID- 18161935 TI - Littoral-cell angioma of the spleen: a case report. AB - Littoral-cell angioma (LCA) is a primary splenic vascular tumor that arises from the normal littoral cells lining the sinus channels of the splenic red pulp. We report a case of LCA of the spleen, which has been infrequently communicated in the literature. A 76-year-old man with a 2-wk history of weight loss, abdominal pain and changes in bowel habits was admitted to our hospital. Imaging studies (CT and MRI) showed multiple lesions in the spleen. Splenectomy was performed. Lining cells were positive for CD31/CD68 markers. Our case was associated with a serrated colonic adenoma. LCA is a benign vascular tumor of the spleen that needs to be included in the differential diagnosis of multiple splenic nodules. PMID- 18161936 TI - Surgical intervention may not always be required in gossypiboma with intraluminal migration. AB - Gossypiboma is the technical term for a retained surgical sponge. Because of legal-ethical concerns, there have not been many publications on this topic. Delays in diagnosis and treatment might increase mortality and morbidity. Radiological imaging is used in diagnosis. We present a case of gossypiboma that had fistulized to bulbous following hydatic cyst surgery. We established the diagnosis with endoscopy and followed its migration endoscopically. PMID- 18161937 TI - Imatinib-induced fatal acute liver failure. AB - Imatinib mesylate is a drug that has been approved for treatment of chronic myeloid leukemia (CML) in blast crisis, accelerated or chronic phase, and also for advanced gastrointestinal stromal tumors. Severe hepatic toxicity and three deaths from hepatic failure have been reported. We report the case of a 51-year old woman who was admitted to our institution with severe acute hepatitis. She was diagnosed with CML and began treatment with imatinib mesylate at a dose of 400 mg/d. Five months after beginning treatment, she developed severe hepatitis associated with coagulopathy, and was admitted to our institution. She had been consuming acetaminophen 500-1000 mg/d after the onset of symptoms. She had a progressive increase in bilirubin level and a marked decrease of clotting factor V. Five days after admission, grade II encephalopathy developed and she was referred for liver transplantation. Her clinical condition progressively deteriorated, and 48 h after being referred for transplantation she suffered a cardiac arrest and died. This report adds concern about the possibility of imatinib-mesylate-induced hepatotoxicity and liver failure, particularly in the case of concomitant use with acetaminophen. Liver function tests should be carefully monitored during treatment and, with the appearance of any elevation of liver function tests, treatment should be discontinued. PMID- 18161938 TI - Snake venomics of Central American pitvipers: clues for rationalizing the distinct envenomation profiles of Atropoides nummifer and Atropoides picadoi. AB - We report the proteomic characterization of the Central American pitvipers Atropoides nummifer and Atropoides picadoi. The crude venoms were fractionated by reverse-phase high-performance liquid chromatography (HPLC), followed by analysis of each chromatographic fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequencing, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass fingerprinting, and collision-induced dissociation-tandem mass spectrometry (CID-MS/MS) of tryptic peptides. Each venom contained a number of bradykinin-potentiating peptides and around 25-27 proteins of molecular masses in the range of 7-112 kDa, belonging to only nine different toxin families (disintegrin, DC fragment, snake venom vascular endothelial growth factor, phospholipases A2, serine protease, cysteine rich secretory proteins, C-type lectins, L-amino acid oxidase, and Zn2+-dependent metalloproteases), albeit distinctly distributed among the two Atropoides species. In addition, A. nummifer expresses low amounts of a three-finger toxin not detected in the venom of A. picadoi. The major toxins of A. nummifer belong to the PLA2 (relative abundance, 36.5%) and the serine proteinase (22%) families, whereas the most abundant A. picadoi toxins are Zn2+-dependent metalloproteinases (66.4%). We estimate that the similarity of venom proteins between the two Atropoides taxa may be around 14-16%. The high degree of differentiation in the venom proteome among congeneric taxa emphasizes unique aspects of venom composition of related species of Atropoides snakes and points to a strong role for adaptive diversification via natural selection as a cause of this distinctiveness. On the other hand, their distinct venom toxin compositions provide clues for rationalizing the low hemorrhagic, coagulant, and defibrinating activities and the high myotoxic and proteolytic effects evoked by A. nummifer snakebite in comparison to other crotaline snake venoms and the high hemorrhagic activity of A. picadoi. PMID- 18161939 TI - Membrane protein identification: N-terminal labeling of nontryptic membrane protein peptides facilitates database searching. AB - Membrane proteins are fairly refractory to digestion especially by trypsin, and less specific proteases, such as elastase and pepsin, are much more effective. However, database searching using nontryptic peptides is much less effective because of the lack of charge localization at the N and C termini and the absence of sequence specificity. We describe a method for N-terminal-specific labeling of peptides from nontryptic digestions of membrane proteins, which facilitates Mascot database searching and can be used for relative quantitation. The conditions for digestion have been optimized to obtain peptides of a suitable length for mass spectrometry (MS) fragmentation. We show the effectiveness of the method using a plasma membrane preparation from a leukemia cell line and demonstrate a large increase in the number of membrane proteins, with small extra membranar domains being identified in comparison to previous published methods. PMID- 18161940 TI - Identification of hepatocellular-carcinoma-associated antigens and autoantibodies by serological proteome analysis combined with protein microarray. AB - To comprehensively study autoantibodies in patients with hepatocellular carcinoma (HCC), we used an approach-based serology and proteomics technologies. Total proteins extracted from HepG2 cells and HepG2.2.15 cells were separated by two dimensional gel electrophoresis (2DE) and then transferred onto polyvinylidene difluoride (PVDF) membranes, which were subsequently incubated with sera from HCC patients or from normal controls. As a result, 13 HCC-associated antigens were identified. Antigenicity of eight proteins was further confirmed using recombinant proteins by Western blotting (WB) and protein microarray. The results of antigen microarray analysis showed strong signals of keratin 8 and lamin A/C in chronic hepatitis controls; therefore, the autoantibodies to keratin 8 and lamin A/C may not be HCC-specific. These two antigens were removed from subsequent analyses. The frequencies of positive reactions to DEAD (Asp-Glu-Ala Asp) box polypeptide 3, eukaryotic translation elongation factor 2 (eEF2), apoptosis-inducing factor (AIF), heterogeneous nuclear ribonucleoprotein A2 (hnRNP A2), prostatic binding protein, and triosephosphate isomerase (TIM) were significantly higher in HCC than in chronic hepatitis and normal individuals. Positive reactions to DEAD box polypeptide 3, eEF2, AIF, and prostatic binding protein were significantly more frequent in HCC than in any other cancer. The sensitivity of any individual antigen in HCC at stage I ranged from 50 to 85%. When the combinations of six antigens were analyzed, the sensitivity increased to 90%. We conclude that the detection of autoantibodies against the six antigens may have value on early diagnosis of HCC. PMID- 18161941 TI - Diterpenoid glycosides from the bitter fern Gleichenia quadripartita. AB - Fifteen new diterpenoid glycosides (1a-n, 2) were isolated from an Argentine collection of the bitter fern Gleichenia quadripartita along with the known flavonoid glycoside afzelin. Structure elucidation was accomplished by 1D and 2D NMR spectroscopy and by high-resolution MS analyses. In addition, X-ray crystallographic analysis of a monocrystal of 1a as well as chemical derivatization of 1h and 1m were performed to confirm their structures. PMID- 18161942 TI - Crotonkinins A and B and related diterpenoids from Croton tonkinensis as anti inflammatory and antitumor agents. AB - Cytotoxicity-guided phytochemical investigation of a methanolic extract of Croton tonkinensis afforded two new kaurane diterpenoids (1, 2) and 10 known ent-kaurane type diterpenoids (3- 12). The structures of 1 and 2 were based on analysis of spectroscopic and mass spectral data. Compounds 3- 12 were identified by comparison of their spectroscopic and physical data with those reported in the literature. Selected compounds from this plant were examined for cytotoxic and anti-inflammatory activities. Compounds 4 and 9 showed the highest cytotoxic activity against the tested tumor cell lines. Compounds 3, 4, 6, 8, 9, and 11 had IC 50 values less than 5 microM and were more potent than the nonspecific NOS inhibitor L-NAME in inhibiting LPS-induced NO production. PMID- 18161943 TI - Double-stranded RNA homopolymer poly(rC).poly(rG) for a new pH-sensitive drug carrier. AB - Double-stranded RNA homopolymer poly(rC).poly(rG) has been used as a new pH sensitive drug carrier. The poly(rC).poly(rG) had proton buffering capacity around pH 6, owing to protonation of cytosine, as determined by acid-base titration. By circular dichroism measurement, the protonation caused conformational change of the RNA. The poly(rC).poly(rG) and doxorubicin (Dox), as an anticancer drug, formed the complexes which released the drugs at endosomal pH. The resulting complex exhibited higher anticancer activity than the Dox alone. These results result suggest that the poly(rC).poly(rG) is a promising biopolymer for a new class of pH-sensitive drug carriers. PMID- 18161944 TI - Biomarkers in toxicology and risk assessment: informing critical dose-response relationships. AB - Tremendous advances have been made in the study of biomarkers related to carcinogenesis during the past 20 years. This perspective will briefly review improvements in methodology and instrumentation that have increased our abilities to measure the formation, repair, and consequences of DNA adducts. These biomarkers of exposure, along with surrogates such as protein adducts, have greatly improved our understanding of species differences in metabolism and effects of chemical stability and DNA repair on tissue differences in molecular dose. During this same time frame, improvements in assays for biomarkers of effect have provided better data and an improved understanding of the dose responses for both gene and chromosomal mutations. A framework analysis approach was used to examine the mode of action of genotoxic chemicals and the default assumption that cancer can be expected to be linear at very low doses. This analysis showed that biomarkers of exposure are usually linear at low doses, with the exception being when identical adducts are formed endogenously. Whereas biomarkers of exposure extrapolate down to zero, biomarkers of effect can only be interpolated back to the spontaneous or background number of mutations. The likely explanation for this major difference is that at high exposures, the biology that results in mutagenesis is driven by DNA damage resulting from the chemical exposure. In contrast, at very low exposures, the biology that results in mutagenesis is driven by endogenous DNA damage. The shapes of the dose response curves for biomarkers of exposure and effect can be very different, with biomarkers of effect better informing quantitative estimates of risk for cancer, a disease that results from multiple mutations. It is also clear, however, that low dose data on mutagenesis are needed for many more chemicals. PMID- 18161946 TI - APCI interface for LC- and SEC-MS analysis of synthetic polymers: advantages and limits. AB - The main advantage of the APCI interface for the LC-MS analysis of synthetic polymers resides in its compatibility with the main chromatographic modes: reversed-phase liquid chromatography, normal-phase liquid chromatography, and size exclusion chromatography in organic phase, with the usual flow rates. Moreover, APCI can be used in positive or negative modes. Representative applications are described to highlight benefits and limitations of the LC-APCI MS technique with the analysis of industrial polymers up to molecular masses of 5 kDa: polyethers; polysiloxanes; and copolymers of siloxanes. Results are discussed in regard to those obtained by more classical techniques: SEC and MALDI MS. The use of an APCI interface in LC-MS and SEC-MS coupling applied to synthetic polymers is efficient up to 2000-4500 Da. The main drawback of the APCI interface is the in-source decomposition that is observed above m/z = 2000-3000 and can induce an underestimation of average molecular weights. However, APCI allows detection on a wide range of polarity of sample/solvent and appears to be complementary to ESI. PMID- 18161947 TI - Mechanism of inhibition of the GluR2 AMPA receptor channel opening by 2,3 benzodiazepine derivatives. AB - 2,3-Benzodiazepine derivatives are drug candidates synthesized for potential treatment of various neurodegenerative diseases involving the excessive activity of AMPA receptors. Here we describe a rapid kinetic investigation of the mechanism of inhibition of the GluR2Qflip AMPA receptor channel opening by two 2,3-benzodiazepine derivatives that are structurally similar (BDZ-2 and BDZ-3). Using a laser-pulse photolysis technique with a time resolution of approximately 60 mus, we measured the effects of these inhibitors on both the channel opening rate and the whole-cell current amplitude. We found that both compounds preferably inhibit the open-channel state, although BDZ-2 is a more potent inhibitor in that it inhibits the open-channel state approximately 5-fold stronger than BDZ-3 does. Both compounds bind to the same noncompetitive site. Binding of an inhibitor to the receptor involves the formation of a loose, partially conducting channel intermediate, which rapidly isomerizes to a tighter complex. The isomerization reaction is identified as the main step at which the receptor distinguishes the structural difference between the two compounds. These results suggest that addition of a bulky group at the N-3 position on the diazepine ring, as in BDZ-3, does not alter the mechanism of action, or the site of binding, but does lower the inhibitory potency, possibly due to an unfavorable interaction of a bulky group at the N-3 position with the receptor site. The new mechanistic revelation about the structure-reactivity relationship is useful in designing conformation-specific, more potent noncompetitive inhibitors for the GluR2 AMPA receptor. PMID- 18161948 TI - Pyridoxamine protects proteins from functional damage by 3-deoxyglucosone: mechanism of action of pyridoxamine. AB - Pyridoxamine (PM) is a promising drug candidate for treatment of diabetic nephropathy. The therapeutic effect of PM has been demonstrated in multiple animal models of diabetes and in phase II clinical trials. However, the mechanism of PM therapeutic action is poorly understood. One potential mechanism is scavenging of pathogenic reactive carbonyl species (RCS) found to be elevated in diabetes. We have suggested previously that the pathogenicity of RCS methylglyoxal (MGO) may be due to modification of critical arginine residues in matrix proteins and interference with renal cell-matrix interactions. We have also shown that this MGO effect can be inhibited by PM (Pedchenko et al. (2005) Diabetes 54, 2952-2960). These findings raised the questions of whether the effect is specific to MGO, whether other structurally different physiological RCS can act via the same mechanism, and whether their action is amenable to PM protection. In the present study, we have shown that the important physiological RCS 3-deoxyglucosone (3-DG) can damage protein functionality, including the ability of collagen IV to interact with glomerular mesangial cells. We have also demonstrated that PM can protect against 3-DG-induced protein damage via a novel mechanism that includes transient adduction of 3-DG by PM followed by irreversible PM-mediated oxidative cleavage of 3-DG. Our results suggest that, in diabetic nephropathy, the therapeutic effect of PM is achieved, in part, via protection of renal cell-matrix interactions from damage by a variety of RCS. Our data emphasize the potential importance of the contribution by 3-DG, along with other more reactive RCS, to this pathogenic mechanism. PMID- 18161949 TI - Cooperation between a salt bridge and the hydrophobic core triggers fold stabilization in a Trp-cage miniprotein. AB - Miniproteins are adequate models to study various protein-structure modifying effects such as temperature, pH, point mutation(s), H-bonds, salt bridges, molecular packing, etc. Tc5b, a 20-residue Trp-cage protein is one of the smallest of such models with a stable 3D fold (Neidigh J. W. et al. (2002) Nat. Struct. Biol. 9, 425-430). However, Tc5b exhibits considerable heat-sensitivity and is only stable at relatively low temperatures. Here we report a systematic investigation of structural factors influencing the stability of Tc5b by solving its solution structure in different environments, varying temperature, and pH. The key interactions identified are the hydrophobic stacking of the aromatic rings of Tyr3 and Trp6 and the salt bridge formed between Asp9 and Arg18. To verify the importance of these interactions, selected variants (mutated, glycosylated and truncated) of Tc5b were designed, prepared, and investigated by NMR. Indeed, elimination of either of the key interactions highly destabilizes the structure. These observations enabled us to design a new variant, Tc6b, differing only by a methylene group from Tc5b, in which both key interactions are optimized simultaneously. Tc6b exhibits enhanced heat stability and adopts a stable fold at physiological temperature. PMID- 18161950 TI - DNA polymerase beta fidelity: halomethylene-modified leaving groups in pre-steady state kinetic analysis reveal differences at the chemical transition state. AB - The mechanism of DNA polymerase beta-catalyzed nucleotidyl transfer consists of chemical steps involving primer 3' OH deprotonation, nucleophilic attack, and pyrophosphate leaving-group elimination, preceded by dNTP binding which induces a large-amplitude conformational change for Watson-Crick nascent base pairs. Ambiguity in the nature of the rate-limiting step and active-site structural differences between correct and incorrect base-paired transition states remain obstacles to understanding DNA replication fidelity. Analogues of dGTP where the beta-gamma bridging oxygen is replaced with fluorine-substituted methylene groups have been shown to probe the contribution of leaving-group elimination to the overall catalytic rate (Biochemistry 46, 461-471). Here, the analysis is expanded substantially to include a broad range of halogen substituents with disparate steric and electronic properties. Evaluation of linear free energy relationships for incorporation of dGTP analogues opposite either template base C or T reveals a strong correlation of log(kpol) to leaving group pKa. Significantly different kpol behavior is observed with a subset of the analogues, with magnitude dependent on the identity of the nascent base pair. This observation, and the absence of an analogous effect on ground state analogue binding (Kd values), points to active-site structural differences at the chemical transition state. Reduced catalysis with bulky halo-containing substrates is manifested in the fidelity of T-G incorporation, where the CCl2-bridging analogue shows a 27-fold increase in fidelity over the natural dGTP. Solvent pH and deuterium isotope effect data are also used to evaluate mechanistic differences between correct and mispaired incorporation. PMID- 18161951 TI - Theoretical survey of the potential energy surface of methyl nitrite + Cu+ reaction. AB - The gas-phase reaction of methyl nitrite with Cu+ has been investigated using density functional theory. The geometries and energies of all the stationary points involved in the reaction have been investigated at the B3LYP/6 311+G(2df,2pd) level. Seven different structures of the encounter complexes could be formed when Cu+ attacking at different electronegative heteroatoms of trans and cis conformational isomers of methyl nitrite, in which the inner oxygen attacks account for the most stable complexes. Extensive conversions could take place for these complexes converting into each other. Various mechanisms leading to the loss of NO and HNO are analyzed in terms of the topology of the potential energy surface. The reaction proceeds exclusively from the inner oxygen attachments, followed by four different mechanisms, i.e., direct dissociation, direct H abstraction, N-O activation, and C-H activation, where the former two provide direct channels for the respective losses of NO and HNO, the third one accounts for both of the losses, and C-H activation is unlikely to be important due to the energetics. PMID- 18161953 TI - Unimolecular reactions of chemically activated CF2BrCF2CH3 and CF2BrCF2CD3: evidence for 1,2-FBr interchange. AB - Vibrationally excited CF2BrCF2CH3 and CF2BrCF2CD3 molecules were prepared with 96 kcal mol-1 energy at room temperature by the recombination of CF2BrCF2 and CH3 (CD3) radicals. The observed unimolecular reactions are 1,2-BrF interchange to give CF3CFBrCH3 (CD3) molecules and 2,3-FH (FD) elimination; the rate constants are 2.2 x 10(5) (1.5 x 10(5)) s(-1) and 2.0 x 105 (0.75 x 10(5)) s(-1), respectively. The CF3CFBrCH3 (CD3) molecules rapidly, relative to the reverse reaction, eliminate HBr or DBr to give the observed product CF3CF=CH2 (CD2). Density functional theory at the B3PW91/6-311+G(2d,p) level was used to obtain vibrational frequencies and moments of inertia of the molecule and transition states for subsequent calculations of statistical rate constants for CF2BrCF2CH3 and CF2BrCF2CD3. Matching experimental and calculated rate constants gave threshold energies of 62 and 66 kcal mol-1 for 1,2-BrF interchange and 2,3-FH elimination, respectively. The BrF interchange reaction is compared to ClF interchange from CF2ClCF2CH3 and CF2ClCHFCH3. PMID- 18161954 TI - Viscosity dependence of the kinetic parameters of the radical ion pair in homogeneous solution determined by optically detected X- and ku-band electron spin resonance. AB - We investigated the dynamics of the radical ion pairs formed by photoinduced electron-transfer reaction from zinc tetraphenyl porphyrin to 2-methyl-1,4 naphthoquinone in mixtures of 2-propanol and cyclohexanol. By the irradiation of a resonant X- (9.16 GHz) or Ku-band (17.41 GHz) microwave pulse, the time profiles of the transient absorptions was modified and the yields of escaping radical ions decreased. From these experiments, we determined the kinetic parameters of the radical ion pairs at various solvent viscosities. The recombination rates of the singlet pairs were (4 +/- 4), (8 +/- 3), and (16 +/- 3) x 10(6) s(-1) at 5, 10, and 15 cP, respectively. The escape rates were (1.7 +/ 0.2), (1.4 +/- 0.1), and (0.9 +/- 0.2) x 10(6) s(-1) at 5, 10, and 15 cP, respectively. The viscosity dependence of the kinetic parameters was followed by the simple continuum diffusion model. PMID- 18161955 TI - Dynamical optimization for partition theory. AB - How to partition a chemical system into its constituent parts is a classic problem of theoretical chemistry. A formally exact solution has recently been developed, partition theory (PT), based on density functional theory [Cohen, M. H.; Wasserman, A. J. Phys. Chem. A 2007, 111, 2229]. PT presents a constrained optimization problem to which the Car-Parrinello (CP) method of electronic structure theory is well suited. We propose here a generalization of the CP method suitable for PT and thereby make way for its practical numerical implementation. We demonstrate that this CP implementation of PT need not increase the complexity of the computation of the system's electronic structure. The scheme provides an exact DFT formulation of, e.g., atoms in molecules theory that is amenable to numerical implementation. PMID- 18161956 TI - Thermostability of salt bridges versus hydrophobic interactions in proteins probed by statistical potentials. AB - The temperature dependence of the interactions that stabilize protein structures is a long-standing issue, the elucidation of which would enable the prediction and the rational modification of the thermostability of a target protein. It is tackled here by deriving distance-dependent amino acid pair potentials from four datasets of proteins with increasing melting temperatures (Tm). The temperature dependence of the interactions is determined from the differences in the shape of the potentials derived from the four datasets. Note that, here, we use an unusual dataset definition, which is based on the Tm values, rather than on the living temperature of the host organisms. Our results show that the stabilizing weight of hydrophobic interactions (between Ile, Leu, and Val) remains constant as the temperature increases, compared to the other interactions. In contrast, the two minima of the Arg--Glu and Arg--Asp salt bridge potentials show a significant Tm dependence. These two minima correspond to two geometries: the fork--fork geometry, where the side chains point toward each other, and the fork--stick geometry, which involves the N(epsilon) side chain atom of Arg. These two types of salt bridges were determined to be significantly more stabilizing at high temperature. Moreover, a preference for more-compact salt bridges is noticeable in heat-resistant proteins, especially for the fork--fork geometry. The Tm dependent potentials that have been defined here should be useful for predicting thermal stability changes upon mutation. PMID- 18161957 TI - Ordinal classification using Comparative Molecular Field Analysis. AB - Comparative Molecular Field Analysis (CoMFA) is most widely used as one of the 3 dimensional QSAR (3D-QSAR) methods to identify the relationship between chemical structure and biological activity. Conventional CoMFA requires at least 3 orders of experimental data, such as IC50 and Ki, to obtain a good model, although practically there are many screening assays where biological activity is measured only by a rating scale. Hence, rating classification-oriented CoMFA coupled with ordinal logistic regression has been developed, and its predictive ability and 3D graphical analysis ability have been investigated. As a result, this novel CoMFA (Logistic CoMFA) has been found to be more robust than conventional CoMFAs in both predictive and 3D graphical analysis abilities. Furthermore, Logistic CoMFA is useful since it can provide the probability of each rank. PMID- 18161958 TI - Last step of the para route of the Kolbe-Schmitt reaction. AB - This work is an extension of our investigations of the Kolbe--Schmitt reaction mechanism. The last step in the para route of the carboxylation reaction of alkali metal phenoxides is investigated at the B3LYP/LANL2DZ level of theory. Among several examined pathways, two mechanisms are proposed: the one involving a successive rearrangement of hydrogen and the other one based on the formation of free radicals. The former pathway is energetically comparable to the last step of the carboxylation reaction in the ortho position, whereas the latter pathway requires higher activation energy. Bearing in mind that the Kolbe--Schmitt reaction is performed at high temperatures, we assume that both reaction paths are plausible. PMID- 18161959 TI - The trouble with QSAR (or how I learned to stop worrying and embrace fallacy). AB - A general feeling of disillusionment with QSAR has settled across the modeling community in recent years. Most practitioners seem to agree that QSAR has not fulfilled the expectations set for its ability to predict biological activity. Among the possible reasons that have been proposed recently for this disappointment are chance correlation, rough response surfaces, incorrect functional forms, and overtraining. Undoubtedly, each of these plays an important role in the lack of predictivity seen in most QSAR models. Likely to be just as important is the role of the fallacy cum hoc ergo propter hoc in the poor prediction seen with many QSAR models. By embracing fallacy along with an over reliance on statistical inference, it may well be that the manner in which QSAR is practiced is more responsible for its lack of success than any other innate cause. PMID- 18161960 TI - Excited-state absorption and sign tuning of nonlinear refraction in porphyrin derivatives. AB - The nonlinear absorptions and nonlinear refractions of free-base porphyrin (P2) and Zn-porphyrin (ZnP2) were studied using the Z-scan technique at 532 nm with different pulse durations. Both P2 and ZnP2 exhibit reverse saturated absorption attributed to excited-state absorption. The coordination of P2 by Zn ion can alter the nonlinear refraction sign from negative to positive at 4 ns pulse. The results indicate that the sign of self-lensing can be tuned by the coordination of Zn in the porphyrin derivatives. In the case of longer pulse duration, the thermal effect was enhanced to dominate the nonlinear refraction sign, leading to the negative nonlinear refraction repeated appearance. PMID- 18161961 TI - Sonoluminescence of Na atom from NaCl solutions doped with ethanol. AB - Sonoluminescence spectra from argon-saturated NaCl solution were measured in the concentration range of 0.5-4 M at the frequency of 138 kHz. The line broadening of sodium atom emission was observed at various acoustic powers in the range from 1.8 to 16.2 W. The sodium D line showed a maximum intensity at a NaCl concentration of 2 M, which corresponded to the maximum production of OH radicals estimated by KI dosimetry. The effects of the addition of a small amount of ethanol on the line width and intensity were closely investigated at various acoustic powers. The sodium line width increases with ethanol concentration and also with power, whereas the line intensity is strongly quenched with increasing ethanol concentration. The results conclusively show that the sodium emission occurs in the gas phase within bubbles. The line broadening is due to interactions with high-pressure argon, and the maximum relative density of gas at bubble collapse was estimated to be 59.5 from the comparison with spectroscopic data. Further line broadening and quenching upon the addition of ethanol arise from collisions with gaseous products obtained from the decomposition of ethanol. The mechanism of sodium excitation is inferred to be as follows. Sodium ions enter bubbles as droplets, and salts are formed because of the high temperature within bubbles. Sodium atoms are generated by the dissociation of salts and then undergo electronic excitation by OH and H radicals. PMID- 18161962 TI - Synthesis and characterization of the cycloheptatrienyl tantalum "mixed-sandwich" compounds (c5r5)ta(c7h7). AB - The molecule (cycloheptatrienyl)(cyclopentadienyl)tantalum, (C(5)H(5))Ta(C(7)H(7)) (1), and its methylcyclopentadienyl and pentamethylcyclopentadienyl analogues (C(5)H(4)Me)Ta(C(7)H(7)) (2), and (C(5)Me(5))Ta(C(7)H(7)) (3) have been synthesized by magnesium reduction of the corresponding (C(5)R(5))TaCl(4) species in the presence of cycloheptatriene. The crystal structures of 2 and 3 show that the two rings are planar and essentially parallel to each other. Interestingly, the Ta-C distances to the C(7)H(7) ring are significantly shorter (by about 0.1 A) than those to the cyclopentadienyl ring; the difference reflects stronger bonding to the C(7)H(7) ring. A comparison with the structures of other (C(5)R(5))M(C(7)H(7)) shows that the M-C distances to the seven-membered ring are especially sensitive to the d-orbital energies of the metal center and its ability to engage in delta bonding with the ring. For 1 3, the EPR spectra at room temperature consist of octets due to the tantalum nuclear spin. Both A(iso) and g(iso) increase as the number of methyl groups on the cyclopentadienyl ring increases. EPR spectra of 1-3 as frozen glasses correspond to axial symmetry, and the hyperfine couplings and g factors are deduced from simulations. PMID- 18161963 TI - RhCl3-assisted C-H and C-S bond scissions: isomeric self-association of organorhodium(iii) thiolato complex. synthesis, structure, and electrochemistry. AB - The ligating properties of alkyl 2-(phenylazo)phenyl thioether 1 (HL(R); R = Me, CH(2)Ph) toward Rh(III) have been examined. A novel hexacoordinated orthometalated rhodium(III) thiolato complex trans-[Rh(L)Cl(PPh3)2] 5 has been synthesized from 1 and RhCl(3).3H(2)O in the presence of excess PPh(3) via in situ C(sp(2))-H and C(sp(3))-S bond scissions, which is the first example for a coordination compound of [L](2-). We were also able to isolate the intermediate organothioether rhodium(III) compound trans-[Rh(L(R))Cl(2)(PPh(3))] 6 with 1 equiv of PPh(3) relative to both 1 and RhCl(3).3H2O in the course of the synthesis of the S-dealkylated product. PPh(3) plays a crucial role in the C(sp(3))-S cleavage process. A plausible mechanistic pathway is presented for C-S bond cleavage, and reductive cleavage by single-electron transfer mechanism is likely to be operative. The electronically and coordinatively saturated thiolato complex 5, indefinitely stable in the solid state, undergoes spontaneous self dimerization in solution via dissociation of one coordinated PPh3 molecule to afford edge-shared bioctahedral anti-[Rh(L)Cl(PPh(3))]2 7 and syn [Rh(L)Cl(PPh(3))]2 8 isomers. All the synthesized organosulfur rhodium(III) compounds were isolated as both air- and moisture-stable solids and spectroscopically characterized in both solution and solid states. In addition, all the representative members have been authenticated by single-crystal X-ray structure analyses. Availability of the isomeric dimers provides an opportunity to recognize the presence of noncovalent intramolecular "metallochelate metallochelate" interaction in the sterically encumbered syn isomer. Unlike other organosulfur rhodium complexes, the monomeric thiolato complex 5 exhibits a fully reversible oxidative wave at 0.82 V vs Ag/AgCl, which is supposed to be primarily centered on the thiolato sulfur atom, and such perception is consistent with the DFT study. Formation of rhodium-bound thiyl radical cation 5(*+) by electrochemical oxidation was scrutinized by EPR spectroscopy. PMID- 18161964 TI - Tetra- and dinuclear nickel(II)-vanadium(IV/V) heterometal complexes of a phenol based N2O2 ligand: synthesis, structures, and magnetic and redox properties. AB - The tetra- and binuclear heterometallic complexes of nickel(II)-vanadium(IV/V) combinations involving a phenol-based primary ligand, viz., N,N'-dimethyl-N,N' bis(2-hydroxy-3,5-dimethylbenzyl)ethylenediamine (H2L1), are reported in this work. Carboxylates and beta-diketonates have been used as ancillary ligands to obtain the tetranuclear complexes [Ni(II)(2)V(V)(2)(RCOO)(2)(L(1))(2)O(4)] (R = Ph, 1; R = Me(3)C, 2) and the binuclear types [(beta-diket)Ni(II)L(1)V(IV)O(beta diket)] (3 and 4), respectively. X-ray crystallography shows that the tetranuclear complexes are constructed about an unprecedented heterometallic eight-membered Ni(2)V(2)O(4) core in which the (L(1))(2)- ligands are bound to the Ni center in a N(2)O(2) mode and simultaneously bridge a V atom via the phenoxide O atoms. The cis-N(2)O(4) coordination geometry for Ni is completed by an O atom derived from the bridging carboxylate ligand and an oxo O atom. The latter two atoms, along with a terminal oxide group, complete the O5 square pyramidal coordination geometry for V. Each of the dinuclear compounds, [(acac)Ni(II)L(1)V(IV)O(acac)] (3) and [(dbm)Ni(II)L(1)V(IV)O(dbm)] (4) [Hdbm = dibenzoylmethane], also features a tetradentate (L(1))(2)- ligand, Ni in an octahedral cis-N(2)O(4) coordination geometry, and V in an O(5) square-pyramidal geometry. In 3 and 4, the bridges between the Ni and V atoms are provided by the (L(1))(2)- ligand. The Ni...V separations in the structures lie in the narrow range of 2.9222(4) A (3) to 2.9637(5) A (4). The paramagnetic Ni centers (S = 1) in 1 and 2 are widely separated (Ni...Ni separations are 5.423 and 5.403 A) by the double V(V)O(4) bridge that leads to weak antiferromagnetic interactions (J = -3.6 and -3.9 cm-1) and thus an ST = 0 ground state for these systems. In 3 and 4, the interactions between paramagnetic centers (Ni(II) and V(IV)) are also antiferromagnetic (J = -8.9 and -10.0 cm-1), leading to an S(T) = 1/2 ground state. Compound 4 undergoes two one-electron redox processes at E(1/2) = +0.66 and -1.34 V vs Ag/AgCl reference due to a V(IV/V) oxidation and a Ni(II)/I reduction, respectively, as indicated by cyclic and differential pulse voltammetry. PMID- 18161965 TI - Unusual structural types in manganese cluster chemistry from the use of N,N,N',N' tetrakis(2-hydroxyethyl)ethylenediamine: Mn8, Mn12, and Mn20 Clusters. AB - The syntheses, crystal structures, and magnetochemical characterization are reported for three new mixed-valent Mn clusters [Mn(8)O(3)(OH)(OMe)(O(2)CPh)7(edte)(edteH(2))](2)CPh) (1), [Mn(12)O(4)(OH)(2)(edte)(4)C(l6)(H(2)O)(2)] (2), and [Mn(20)O(8)(OH)(4)(O(2)CMe)(6)(edte)(6)](ClO(4))(2) (3) (edteH(4) = (HOCH(2)CH(2))(2)NCH(2)CH(2)N(CH(2)CH(2)OH)(2) = N,N,N',N'-tetrakis(2 hydroxyethyl)ethylenediamine). The reaction of edteH(4) with Mn(O(2)CPh)(2), MnCl(2), or Mn(O(2)CMe)(2) gives 1, 2, and 3, respectively, which all possess unprecedented core topologies. The core of 1 comprises two edge-sharing [Mn(4)O(4)] cubanes connected to an additional Mn ion by a micro(3)-OH- ion and two alkoxide arms of edteH(22-). The core of 2 consists of a [Mn(12)(micro(4 )O)(4)](24+) unit with S4 symmetry. The core of 3 consists of six fused [Mn(4)O(4)] cubanes in a 3 x 2 arrangement and linked to three additional Mn atoms at both ends. Variable-temperature, solid-state dc and ac magnetization (M) studies were carried out on complexes 1-3 in the 5.0-300 K range. Fitting of the obtained M/Nmicro(B) vs H/T data by matrix diagonalization and including only axial zero-field splitting (ZFS) gave ground-state spin (S) and axial ZFS parameter (D) of S = 8, D = -0.30 cm-1 for 1, S = 7, D = -0.16 cm-1 for 2, and S = 8, D = -0.16 cm-1 for 3. The combined work demonstrates that four hydroxyethyl arms on an ethylenediamine backbone can generate novel Mn structural types not accessible with other alcohol-based ligands. PMID- 18161966 TI - Nickel(II) chain with alternating end-on/end-to-end single azido bridges: a combined structural, magnetic, and theoretical study. AB - The reaction of a tridentate Schiff base LH (L-: 1,1,1-trifluoro-7 (dimethylamino)-4-methyl-5-aza-3-hepten-2-onato) with a Ni(II) salt in the presence of azide salt has led to a new alternating end-on (EO)/end-to-end (EE) azido-bridged Ni(II) chain of formula {[Ni2(micro1,1-N3)(micro1,3 N3)(L)2(MeOH)2]}n. Its originality lies in the presence of single EE and EO coordination modes for the azide. It crystallizes in the C2/c space group, a=21.570(7) A, b=10.79(1) A, c=16.154(5) A, beta=120.81(2) degrees, Z=4. The chain can be viewed as {Ni2(N3)(L)2(MeOH)2}+ dimeric units linked to each other in a zigzag pattern by the other azide. Magnetic susceptibility and magnetization measurements have been performed and revealed that the chain can magnetically be depicted as isolated {Ni2(N3)} units exhibiting antiferromagnetic interaction (JAF approximately -37 cm(-1)). Ab initio calculations confirmed the efficient magnetic coupling through the EE bridge and vanishingly small EO {Ni2(micro1,1 N3)} interactions. PMID- 18161967 TI - Coupling of CpCr(CO)3 and heterocyclic dithiadiazolyl radicals. synthetic, x-ray diffraction, dynamic NMR, EPR, CV, and DFT studies. AB - The reaction of the 1,2,3,5-dithiadiazolyls (4-R-C(6)H(4)CN(2)S(2))(2) (R = Me, 2a; Cl, 2b; OMe, 2c; and CF3, 2d) and (3-NC-5-tBu-C(6)H(3)CN(2)S(2))(2) (2e) with [CpCr(CO)(3)](2) (Cp = eta(5)-C(5)H(5)) (1) at ambient temperature respectively yielded the complexes CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(4)R) (R = 4-Me, 3a; 4-Cl, 3b; 4-OMe, 3c; and 4-CF(3), 3d) and CpCr(CO)(2)(eta(2)-S(2)N(2)CC(6)H(3)-3-(CN)-5 (tBu)) (3e) in 35-72% yields. The complexes 3c and 3d were also synthesized via a salt metathesis method from the reaction of NaCpCr(CO)(3) (1B) and the 1,2,3,5 dithiadiazolium chlorides 4-R-C(60H(4)CN(2)S(2)Cl (R = OMe, 8c; CF(3), 8d) with much lower yields of 6 and 20%, respectively. The complexes were characterized spectroscopically and also by single-crystal X-ray diffraction analysis. Cyclic voltammetry experiments were conducted on 3a-e, EPR spectra were obtained of one electron-reduced forms of 3a-e, and variable temperature 1H NMR studies were carried out on complex 3d. Hybrid DFT calculations were performed on the model system [CpCr(CO)(2)S(2)N(2)CH] and comparisons are made with the reported CpCr(CO)(2)(pi-allyl) complexes. PMID- 18161968 TI - Intermolecular interaction-induced hierarchical transformation in 1D nanohybrids: analysis of conformational changes by 2D correlation spectroscopy. AB - We have examined both self-assembly and confinement effect in room-temperature ionic liquid (RTIL)-aluminum hydroxide hybrids (RAHs) to attain a fundamental understanding of special phenomena in nanoscale spaces as well as to design functional nanomaterials for practical applications. Phase-controlled one dimensional (1D) RAHs were synthesized through a simple ionothermal process. The RAHs were hierarchically transformed in terms of the molecular structures, morphologies, and phases of the materials during the ionothermal process with respect to the concentration of RTIL. In addition to the hierarchical transformation, the RTIL/aluminum hydroxide nanohybrids revealed unexpected physical behaviors, including thermal transition variation of the RTIL in confined environments and a phase transition from nanosolid to nanoliquid affected by changes of the melting points. More importantly, intermolecular interaction induced-self-assembly and confinement effect of RTILs inside an integrated hybrid system, which have not been clearly explained to date, were analyzed by 2D infrared correlation spectroscopy (2D IR COS); dynamic behaviors of RTILs, i.e., sequentially spatial reorientation and kinetically conformational changes, were attributed to the interactions between RTILs and aluminum hydroxides. 2D IR COS offers a new way to interpret highly complex, veiled systems such as the formation mechanism of nanoparticles, biomineralization, self/supramolecular assembly, and nanoconfinement. PMID- 18161969 TI - [Te2As2]2-: a planar motif with "conflicting" aromaticity. AB - [K(18-crown-6)]2[Te2As2] is the first four-membered ring Zintl anion of elements from groups XV and XVI. The anion has an unexpected triplet aromatic ground state. PMID- 18161970 TI - Probing mode and site of substrate water binding to the oxygen-evolving complex in the S2 state of photosystem II by 17O-HYSCORE spectroscopy. AB - In the oxygen-evolving complex (OEC) of photosystem II (PSII) molecular oxygen is formed from two substrate water molecules that are ligated to a mu-oxo bridged cluster containing four Mn ions and one Ca ion (Mn4OxCa cluster; Ox symbolizes the unknown number of mu-oxo bridges; x >or= 5). There is a long-standing enigma as to when, where, and how the two substrate water molecules bind to the Mn4OxCa cluster during the cyclic water-splitting reaction, which involves five distinct redox intermediates (Si-states; i = 0,...,4). To address this question we employed hyperfine sublevel correlation (HYSCORE) spectroscopy on H217O-enriched PSII samples poised in the paramagnetic S2 state. This approach allowed us to resolve the magnetic interaction between one solvent exchangeable 17O that is directly ligated to one or more Mn ions of the Mn4OxCa cluster in the S2 state of PSII. Direct coordination of 17O to Mn is supported by the strong (A approximately 10 MHz) hyperfine coupling. Because these are properties expected from a substrate water molecule, this spectroscopic signature holds the potential for gaining long-sought information about the binding mode and site of one of the two substrate water molecules in the S2 state of PSII. PMID- 18161971 TI - Syntheses, structures, and coordination chemistry of phosphole-containing hybrid calixphyrins: promising macrocyclic P,N2,X-mixed donor ligands for designing reactive transition-metal complexes. AB - The syntheses, structures, and coordination chemistry of phosphole-containing hybrid calixphyrins (P,N2,X-hybrid calixphyrins) and the catalytic activities of their transition-metal complexes are reported. The 5,10-porphodimethene type 14pi P,(NH)2,X- and 16pi-P,N2,X-hybrid calixphyrins (X = O, S, NH) are prepared via acid-promoted dehydrative condensation between a sigma4-phosphatripyrrane and the corresponding 2,5-bis[hydroxy(phenyl)methyl]heteroles followed by DDQ oxidation. Both spectroscopic and crystallographic data of the hybrid calixphyrins have revealed that the conformation and size of the macrocyclic platforms as well as the oxidation state of the -conjugated pyrrole-heterole-pyrrole (N-X-N) units vary considerably depending on the combination of heteroles. The sigma3-P,(NH)2,S and sigma3-P,N2,S-hybrids react with Pd(OAc)2 and Pd(dba)2, respectively, to afford the same Pd(II)-P,N2,S-hybrid complex, in which the calixphyrin platform is regarded as a dianionic ligand. In the complexation with [RhCl(CO)2]2 in dichloromethane, the sigma3-P,N2,S-hybrid behaves as a neutral ligand to afford an ionic Rh(I)-P,N2,S-hybrid complex, whereas the sigma3-P,N2,NH-hybrid behaves as an anionic ligand to produce Rh(III)-P,N3-hybrid complexes. In the latter reaction, it is likely that a neutral Rh(I)-P,N3-hybrid complex, generated as a highly nucleophilic intermediate, undergoes C-Cl bond activation of the solvent. The complexation of AuCl(SMe2) with the sigma3-P,N2,X-hybrids (X = S, NH) leads to the formation of the corresponding Au(I)-monophosphine complexes. The spectral data and crystal structures of these metal complexes exhibit the hemilabile nature of the phosphole-containing hybrid calixphyrin platforms derived from the flexible phosphole unit and the redox active N-X-N units. The hybrid calixphyrin palladium and -rhodium complexes catalyze the Heck reaction and hydrosilylations, respectively, implying that the metal center in the core is capable of activating the substrates under appropriate reaction conditions. The present results demonstrate the potential utility of the phosphole-containing hybrid calixphyrins as a new class of macrocyclic P,N2,X-mixed donor ligands for designing highly reactive transition-metal complexes. PMID- 18161972 TI - Trading templates: supramolecular transformations between {CoII13} and {CoII12} nanoclusters. AB - "Nuclearity switching" from a {Co13} supercluster to a {Co12} species via the addition of CO(3)2- anions is reported and can be traced in solution using electrospray MS techniques. In addition, cryospray MS can be used to identify the entire cluster in solution despite the relative lability of its constituents. PMID- 18161973 TI - Time-programmed peptide helix inversion of a synthetic metal complex triggered by an achiral NO3- anion. AB - Dynamic and efficient inversion of peptide helices by an achiral NO3- anion was programmed in terms of time scale from milliseconds to hours. PMID- 18161974 TI - Polarizing the Nazarov cyclization: the impact of dienone substitution pattern on reactivity and selectivity. AB - The impact of dienone substitution on the Nazarov cyclization has been examined in detail. Substrates bearing different substituents at each of four positions on the dienone backbone were systematically probed in order to identify trends leading to higher reactivity and better selectivity. Desymmetrization of the pentadienyl cation and oxyallyl cation intermediates through placement of polarizing groups at both the C-2 and C-4 positions was found to be particularly effective. These modifications allowed cyclizations to occur in the presence of catalytic amounts of mild Lewis acids. It was also found that stereoconvergent cyclization of mixtures of E and Z isomers of alkylidene beta-ketoesters occurred via an efficient isomerization process that occurred under the reaction conditions. PMID- 18161975 TI - Designed amino acid ATRP initiators for the synthesis of biohybrid materials. AB - A synthetic strategy to prepare peptide-polymer conjugates with precise sites of attachment is described. Amino acids modified with atom transfer radical polymerization (ATRP) initiators for the polymerization of styrenes and methacrylates were prepared. Fmoc-4-(1-chloroethyl)-phenylalanine (5) was synthesized in four steps from Fmoc-tyrosine. HATU-mediated amidation with glycine-OMe resulted in dipeptide (6). The initiator was effective for Cu(I)/bipyridine mediated bulk polymerization of styrene. Kinetic studies indicated a controlled polymerization, with high conversion (97%), and a polydispersity index (PDI) of 1.25. Fmoc-O-(2-bromoisobutyryl)-serine tert-butyl ester (10) was synthesized from Fmoc-Ser(OTrt)-OH in three steps. This initiator was employed in the ATRP of 2-hydroxyethyl methacrylate (HEMA), and kinetic studies indicated a controlled polymerization. Different monomer to initiator ratios resulted in poly(HEMA) of different molecular weights and narrow PDIs (1.14-1.25). Conversions were between 70 and 99%. HEMA modified with N-acetyl-D glucosamine (GlcNAc) was also polymerized to 84% conversion and the resulting PDI was 1.19. The t-butyl ester protecting group of 10 was removed, and the resulting amino acid (11) was incorporated into VM(11)VVQTK by standard solid-phase peptide synthesis. Polymerization resulted in the glycopolymer-peptide conjugate in 93% conversion and a PDI of 1.14. PMID- 18161976 TI - The chemical bond between Au(I) and the noble gases. Comparative study of NgAuF and NgAu+ (Ng = Ar, Kr, Xe) by density functional and coupled cluster methods. AB - The nature of the chemical bond between gold and the noble gases in the simplest prototype of Au(I) complexes (NgAuF and NgAu+, where Ng = Ar, Kr, Xe), has been theoretically investigated by state of art all-electron fully relativistic DC CCSD(T) and DFT calculations with extended basis sets. The main properties of the molecules, including dipole moments and polarizabilities, have been computed and a detailed study of the electron density changes upon formation of the Ng-Au bond has been made. The Ar-Au dissociation energy is found to be nearly the same in both Argon compounds. It almost doubles along the NgAuF series and nearly triples in the corresponding NgAu+ series. The formation of the Ng-Au(I) bonds is accompanied by a large and very complex charge redistribution pattern which not only affects the outer valence region but reaches deep into the core-electron region. The charge transfer from the noble gas to Au taking place in the NgAu+ systems is largely reduced in the fluorides but the Ng-Au chemical bond in the latter systems is found to be tighter near the equilibrium distance. The density difference analysis shows, for all three noble gases, a qualitatively identical nature of the Ng-Au bond, characterized by the pronounced charge accumulation in the middle of the Ng-Au internuclear region which is typical of a covalent bond. This bonding density accumulation is more pronounced in the fluorides, where the Au-F bond is found to become more ionic, while the overall density deformation is more evident and less localized in the NgAu+ systems. Accurate density difference maps and charge-transfer curves help explain very subtle features of the chemistry of Au(I), including its peculiar preference for tight linear bicordination. PMID- 18161977 TI - Detection and mechanistic relevance of transient ligand radicals formed during [Ru(bpy)2(OH2)]2O4+-catalyzed water oxidation. AB - Mechanistic proposals to account for the reactivity of water-oxidizing ruthenium diimine complexes have often invoked participation of covalently hydrated or pseudobase intermediates formed by reaction of solvent with the polypyridyl ligands. Probing for these intermediates has proven difficult because the concentrations of detectable reactive species are very low under commonly used experimental conditions. However, we have recently found that these transients accumulate in photocatalytic oxidation systems at neutral pH. In this work, we show that the reaction rates of these transient species correlate with catalytic activity and, therefore, that they meet minimal kinetic criteria to be true reaction intermediates. PMID- 18161978 TI - Formation of Pd/Au nanostructures from Pd nanowires via galvanic replacement reaction. AB - Bimetallic nanostructures with non-random metal atoms distribution are very important for various applications. To synthesize such structures via benign wet chemistry approach remains challenging. This paper reports a synthesis of a Au/Pd alloy nanostructure through the galvanic replacement reaction between Pd ultrathin nanowires (2.4 +/- 0.2 nm in width, over 30 nm in length) and AuCl3 in toluene. Both morphological and structural changes were monitored during the reaction up to 10 h. Continuous changes of chemical composition and crystalline structure from Pd nanowires to Pd68Au32 and Pd45Au55 alloys, and to Au nanoparticles were observed. More interestingly, by using combined techniques such as high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), energy dispersive X-ray spectrometry (EDS), UV-vis absorption, and extended X-ray absorption fine structure (EXAFS) spectroscopy, we found the formation of Pd68Au32 non-random alloy with Au-rich core and Pd-rich shell, and random Pd45Au55 alloy with uniformly mixed Pd and Au atom inside the nanoparticles, respectively. Density functional theory (DFT) calculations indicated that alkylamine will strongly stabilize Pd to the surface, resulting in diffusion of Au atoms into the core region to form a non-random alloy. We believe such benign synthetic techniques can also enable the large scale preparation of various types of non-random alloys for several technically important catalysis applications. PMID- 18161979 TI - Total synthesis and structural revision of the marine macrolide neopeltolide. AB - The total synthesis and structural revision of the marine natural product neopeltolide is reported. The key bond-forming step involves a Lewis acid catalyzed intramolecular cyclization to install the tetrahydropyran ring and the macrocycle simultaneously. This type of cyclization is the first of its kind and assembles the carbon backbone of the natural product efficiently. The synthesis of the reported structure revealed differences in the data between the natural and synthetic material. After significant investigation, the diastereomeric molecule with the C11 and C13 configurations inverted was synthesized using the initial route. This compound matches the data reported for neopeltolide (1H, 13C, HRMS, IR, NOESY, [alpha]), thereby establishing the correct overall structure for this potent macrolide natural product, including the relative and absolute stereochemistry. PMID- 18161980 TI - A highly conjunctive beta-keto phosphonate: application to the synthesis of pyridine alkaloids xestamines C, E, and H. AB - The synthesis of a novel beta-keto gamma-xanthyl phosphonate has been achieved. This highly conjunctive reagent has been utilized in a combination of radical and ionic reactions to create new carbon-carbon bonds. Its usefulness was demonstrated by realizing the first total synthesis of naturally occurring pyridine alkaloids xestamines C, E, and H. PMID- 18161981 TI - A non-Diels-Alder approach to the cis-decalin core of branimycin. AB - The synthesis of the highly substituted cis-decalin core of branimycin has been accomplished. A catalytic copper mediated SN2' opening of oxabicycle 7 with Grignard reagent and ring-closing metathesis served as key transformations. PMID- 18161982 TI - A facile and efficient synthesis of polyfunctionalized pyridin-2(1H)-ones from beta-oxo amides under Vilsmeier conditions. AB - A facile and efficient one-pot synthesis of polysubstituted pyridin-2(1H)-ones from a variety of beta-oxo amides under Vilsmeier conditions is described, and a mechanism involving sequential halogenation, formylation and intramolecular nucleophilic cyclization is proposed. PMID- 18161983 TI - A DFT study on the mechanism of hydrosilylation of unsaturated compounds with neutral hydrido(hydrosilylene)tungsten complex. AB - Recently, Tobita et al. reported stoichiometric hydrosilylation reactions of acetone and acetonitrile with neutral hydrido(hydrosilylene)tungsten complexes Cp'(CO)2(H)W=Si(H)[C(SiMe(3))(3)] (Cp' = Cp*, C(5)Me(4)Et). The mechanisms of the hydrosilylation reactions of unsaturated compounds (ketone and nitrile) with the tungsten complexes have been investigated with the B(3)LYP density functional theory method. Four possible reaction mechanisms were studied. The results of the calculations indicate that the hydrosilylation of acetone proceeds via a metal hydride migration mechanism proposed by Tobita et al., while the hydrosilylation of nitrile occurs through a silyl migration mechanism, analogous to the modified Chalk-Harrod mechanism. The [2(sigma)+2(pi)] additions of various CX (CX = C=O or CN) multiple bonds with the Si-H bonds in the neutral complexes have very high barriers although similar additions were found feasible in other related cationic complexes. All the hydrosilylation reactions studied here give stable tungsten silylene or tungsten-silyl products, which are not easily converted into the starting hydrido(hydrosilylene)tungsten complexes when reacting with a hydrosilane substrate molecule. Therefore, we predict that hydrosilylation of acetonitrile and acetone catalyzed by these tungsten complexes is difficult to achieve. PMID- 18161984 TI - Intermolecular hydroaminations via strained (E)-cycloalkenes. AB - A photoinduced procedure for the intermolecular hydroamination of alkenes using azoles is described. This reaction occurs in modest to good yield for 6- and 7 membered cyclic alkenes. Upon irradiation at 254 nm in the presence of methyl benzoate and a small amount of triflic acid as an additive (20 mol %), imidazoles, pyrazoles, triazoles, and tetrazole can react with the alkene to afford complex Markovnikov adducts. The proposed mechanism involves photoisomerization to generate highly strained (E)-cycloalkene intermediates and (E)-cycloalkene protonation followed by reaction with the azole nucleophile. Alkene isomerization was found to be a competing side reaction under these conditions. PMID- 18161985 TI - A novel synthesis of chiral guanidinium receptors and their use in unfolding the energetics of enantiorecognition of chiral carboxylates. AB - A novel synthetic route to the versatile chiral bicyclic guanidinium building block is described making use of l-methionine as a starting material from the natural chiral pool. Furthermore, the synthetic elaboration of this building block is shown in the construction of macrocyclic and open chain hosts, respectively. The host design employs urea functions as the connecting units and supplementary anchor groups for the complexation of anions. The binding studies of these hosts with various chiral and achiral oxoanions are performed by isothermal titration calorimetry. A trend analysis of the binding energetics in an ensemble of structurally similar guests discloses the importance of geometrical confinement of the guest. Association entropy rather than free energy (affinity) is identified as an indicator of structural uniqueness needed to distinguish configurational isomers in the recognition of enantiomeric carboxylates by the chiral guanidinium hosts. PMID- 18161986 TI - A new solvent-dependent mechanism for a triazolinedione ene reaction. AB - The ene reaction between 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD) and tetramethylethylene has been investigated using QM/MM calculations in water, methanol, DMSO, and acetonitrile. The effects of solvation on the mechanism and rates of reaction are elucidated using two-dimensional potentials of mean force (PMF) simulations utilizing free-energy perturbation theory and Monte Carlo statistical mechanics. A new mechanism is proposed where direct formation of an open dipolar intermediate following the addition of PTAD to the alkene is rate limiting and the pathway toward ene product is significantly dependent on the reaction medium. In protic solvents, the open dipolar intermediate may proceed directly to the ene product or reversibly form an aziridinium imide (AI) intermediate that does not participate in the reaction. However, in aprotic solvents the open intermediate is short-lived (<10-11 s) and the ene product forms via the AI intermediate. The calculated free energies of activation are in close agreement with those derived from experiment, e.g., DeltaG of 14.9 kcal/mol compared to 15.0 kcal/mol in acetonitrile. Density functional theory calculations at the (U)B3LYP/6-311++G(2d,p) level using the CPCM continuum solvent model were also carried out and confirmed a zwitterionic, and not diradical, open intermediate present in the reaction. Only the QM/MM methodology was able to accurately reproduce the experimental rates and differentiate between the protic and aprotic solvents. Solute-solvent interaction energies, radial distribution functions, and charges are analyzed and show that the major factor dictating the changes in reaction path is hydrogen bond stabilization of the charge separations spanning 2 to 4 atoms in the intermediates and transition states. PMID- 18161987 TI - Total synthesis of (+)-achalensolide based on the rh(i)-catalyzed allenic Pauson Khand-type reaction. AB - The first total synthesis of (+)-achalensolide was achieved from a commercially available d-(-)-isoascorbic acid. The known epoxide, derived from d-(-) isoascorbic acid, was converted into the allenyne, the Rh(I)-catalyzed Pauson Khand-type reaction of which directly provided the bicyclo[5.3.0]decane system, a core framework of the title natural product. The construction of the gamma lactone moiety and some chemical modifications resulted in the completion of the total synthesis of (+)-achalensolide. PMID- 18161988 TI - Threonine-509 is a determinant of apparent affinity for both substrate and cations in the human Na+/dicarboxylate cotransporter. AB - The Na+/dicarboxylate cotransporter (NaDC1) is involved in the absorption of citric acid cycle intermediates from the lumen of the renal proximal tubule and small intestine. The NaDC1 orthologues from human (h) and rabbit (rb) exhibit differences in citrate and cation transport properties. The citrate Km and sodium KNa values are much larger in human than rabbit NaDC1. Our previous study showed that transmembrane helices (TM) 7, 10, and 11 and associated loop regions contain the amino acids that are important in determining the differences in apparent citrate affinity, whereas TM10 and 11 determine differences in apparent sodium affinity. Chimera R10 (hNaDC1 with a substitution of TM10 and associated loop from rbNaDC1) contains only four amino acid differences between rb and hNaDC1. This chimera has similar apparent affinity for succinate and sodium as the wild type rbNaDC1, and an intermediate Km for citrate. To identify individual residues in the TM10 region that determine functional differences between rb and hNaDC1, four mutants were made in which the rabbit sequence was substituted for that of the hNaDC1. Mutants with a serine or threonine at position 509 (or 512 in rbNaDC1) in TM10 have partial changes in Km for citrate and succinate but larger changes in apparent affinity for cations and substrate specificity for four carbon dicarboxylates. The results show that the serine or threonine at position 509 (h) or 512 (rb) is the most important determinant of functional differences in apparent affinity for substrate and cations. Furthermore, the results suggest that the cation and substrate binding sites are located in close proximity to one another in NaDC1. PMID- 18161989 TI - Disulfide bond-dependent mechanism of protection against oxidative stress in pyruvate-ferredoxin oxidoreductase of anaerobic Desulfovibrio bacteria. AB - Oxidative decarboxylation of pyruvate forming acetyl-coenzyme A is a crucial step in many metabolic pathways. In most anaerobes, this reaction is carried out by pyruvate-ferredoxin oxidoreductase (PFOR), an enzyme normally oxygen sensitive except in Desulfovibrio africanus (Da), where it shows an abnormally high oxygen stability. Using site-directed mutagenesis, we have specified a disulfide bond dependent protective mechanism against oxidative conditions in Da PFOR. Our data demonstrated that the two cysteine residues forming the only disulfide bond in the as-isolated PFOR are crucial for the stability of the enzyme in oxidative conditions. A methionine residue located in the environment of the proximal [4Fe 4S] cluster was also found to be essential for this protective mechanism. In vivo analysis demonstrated unambiguously that PFOR in Da cells as well as two other Desulfovibrio species was efficiently protected against oxidative stress. Importantly, a less active but stable Da PFOR in oxidized cells rapidly reactivated when returned to anaerobic medium. Our work demonstrates the existence of an elegant disulfide bond-dependent reversible mechanism, found in the Desulfovibrio species to protect one of the key enzymes implicated in the central metabolism of these strict anaerobes. This new mechanism could be considered as an adaptation strategy used by sulfate-reducing bacteria to cope with temporary oxidative conditions and to maintain an active dormancy. PMID- 18161990 TI - Phosphorylation of c-Abl by protein kinase Pak2 regulates differential binding of ABI2 and CRK. AB - The tyrosine kinase c-Abl is implicated in a variety of cellular processes that are tightly regulated by c-Abl kinase activity and/or by interactions between c Abl and other signaling molecules. The interaction of c-Abl with the Abl interactor protein Abi2 is shown to be negatively regulated by phosphorylation of serines 637 and 638. These serines are adjacent to the PxxP motif (PTPPKRS637S638SFR) that binds the SH3 domain of Abi. Phosphorylation of the Abl 593-730 fragment by Pak2 dramatically reduces Abi2 binding ( approximately 90%). Mutation of serines 637-639 to alanine (3A) or aspartate (3D) results in an increased tyrosine kinase activity of c-Abl 3D, and a slight reduction of the activity of the 3A mutant, as compared to wild-type (WT) c-Abl. The interaction between Abi2 and c-Abl 3D is inhibited by 80%, as compared to WT c-Abl or c-Abl 3A. This is accompanied by a 2-fold increase in binding of Crk to c-Abl 3D. The data indicate a molecular mechanism whereby phosphorylation of c-Abl by Pak2 inhibits the interaction between the SH3 domain of Abi2 and the PxxP motif of c Abl. This phosphorylation enhances the association of c-Abl with the substrate Crk and increases c-Abl-mediated phosphorylation of Crk, thus altering the association of Crk with other signaling molecules. PMID- 18161991 TI - Conformational changes below the Tm: molecular dynamics studies of the thermal pretransition of ribonuclease A. AB - Recent work suggests that some native conformations of proteins can vary with temperature. To obtain an atomic-level description of this structural and conformational variation, we have performed all-atom, explicit-solvent molecular dynamics simulations of bovine pancreatic ribonuclease A (RNase A) up to its melting temperature (Tm approximately 337 K). RNase A has a thermal pretransition near 320 K [Stelea, S. D., Pancoska, P., Benight, A. S., and Keiderling, T. A. (2001) Protein Sci. 10, 970-978]. Our simulations identify a conformational change that coincides with this pretransition. Between 310 and 320 K, there is a small but significant decrease in the number of native contacts, beta-sheet hydrogen bonding, and deviation of backbone conformation from the starting structure, and an increase in the number of nonnative contacts. Native contacts are lost in beta-sheet regions and in alpha1, partially due to movement of alpha1 away from the beta-sheet core. At 330 and 340 K, a nonnative helical segment of residues 15-20 forms, corresponding to a helix observed in the N-terminal domain swapped dimer [Liu, Y. S., Hart, P. J., Schulnegger, M. P., and Eisenberg, D. (1998) Proc. Natl. Acad. Sci. U.S.A. 95, 3437-3432]. The conformations observed at the higher temperatures possess nativelike topology and overall conformation, with many native contacts, but they have a disrupted active site. We propose that these conformations may represent the native state at elevated temperature, or the N' state. These simulations show that subtle, functionally important changes in protein conformation can occur below the Tm. PMID- 18161992 TI - Structural dynamics of myoglobin: FTIR-TDS study of NO migration and binding. AB - By using Fourier transform infrared photolysis difference spectroscopy combined with temperature derivative spectroscopy at cryogenic temperatures, we have measured infrared spectra of the stretching absorption on nitric oxide (NO) in the heme-bound and photodissociated states of ferrous and ferric nitrosyl myoglobin (MbNO) and a few site-specific Mb mutants. The NO absorption was utilized as a sensitive local probe of ligand interactions with active-site residues and movements within the protein. By comparison with results obtained in previous spectroscopic and structural studies of carbonmonoxy myoglobin (MbCO), the MbNO data were interpreted in structural terms. In the NO-bound state, conformational heterogeneity was inferred from the appearance of multiple bands, arising from different electrostatic interactions with active site residues, most importantly, His-64. In ferrous MbNO, a primary photoproduct site similar to site B of MbCO was found, as indicated by a characteristic NO stretching spectrum. In ferric MbNO, the His-64 side chain appears to interfere with trapping of NO in this site; only a very weak photoproduct spectrum was observed in Mb variants in which His-64 was present. Upon extended illumination, the photoproduct spectrum changed in a characteristic way, indicating that NO readily migrates to a secondary docking site C, the Xe4 cavity, in which the ligand performs librational motions on the picosecond time scale. This docking site may play a role in the physiological NO scavenging reaction. Surprisingly, NO cannot be trapped at all in secondary docking site D, the Xe1 cavity. PMID- 18161993 TI - On the contribution of water-mediated interactions to protein-complex stability. AB - Protein-water interactions have long been recognized as a major determinant of chain folding, conformational stability, binding specificity and catalysis. However, the detailed effects of water on stabilizing protein-protein interactions remain elusive. A way to test experimentally the contribution of water-mediated interactions is by applying double mutant cycle analysis on pairs of residues that do not form direct interactions, but are bridged by water. Seven such interactions within the interface between TEM1 and BLIP proteins were evaluated. No significant interaction free energy was found between either of them. Water can bridge interactions, but also stabilize the structure of the monomer. To distinguish between these, we performed a bioinformatic analysis using AQUAPROT (http://bioinfo.weizmann.ac.il/aquaprot) to determine the degree of water conservation between the bound and unbound states. 29 structures of twelve complexes and 20 related monomers were analyzed. Of the 262 water molecules located within the interfaces, 145 were conserved between the unbound and bound structures. Strikingly, all 50 buried or partially buried waters in the monomer structures were conserved at the same location in the bound structures. Thus, buried waters have an important role in stabilizing the monomer fold rather than contributing to protein-protein binding, and are not replaced by residues from the incoming protein. Taking together the experimental and bioinformatics evidence suggests that exposed waters within the interface may be good sites for protein engineering, while buried or mostly buried waters should be left unchanged. PMID- 18161995 TI - Pentacene nanorails on Au(110). AB - We studied the molecular orientation of pentacene monolayer phases on the Au(110) surface by means of near-edge X-ray absorption spectroscopy at the carbon K-shell and scanning tunneling microscopy. The highest coverage phase, displaying a (6 x 8) symmetry, is found to be formed by two types of differently oriented molecules mimicking regular arrays of nanorails. Flat-lying molecules, aligned side-by-side with the long molecular axis along the [001] direction, form long crosstie chains extending in the [110] direction. In between the adjacent flat chains, additional molecules, tilted by 90 degrees around their molecular axis, line up head-to-tail into rails extending along [110]. These molecules are very weakly hybridized with the substrate, as indicated by their lowest unoccupied molecular orbitals, which closely resemble those of the free molecule. The nanorail structure is found to be stable up to 420 K in vacuum and to also remain in place after exposure to air, thus being a template well suited for further self-assembly of organic heterostructures. The tilted quasi-free molecules open the possibility for an optimal lateral pi-coupling to other molecules or molecular assemblies. PMID- 18161994 TI - Opposing effects of inositol hexakisphosphate on rod arrestin and arrestin2 self association. AB - The robust cooperative formation of rod arrestin tetramers has been well established, whereas the ability of other members of the arrestin family to self associate remains controversial. Here, we used purified arrestins and multi-angle light scattering to quantitatively compare the propensity of the four mammalian arrestin subtypes to self-associate. Both non-visual and cone arrestins only form oligomers at very high non-physiological concentrations. However, inositol hexakisphosphate (IP6), a fairly abundant form of inositol in the cytoplasm, greatly facilitates self-association of arrestin2. Arrestin2 self-association equilibrium constants in the presence of 100 microM IP6 suggest that an appreciable proportion could exist in an oligomeric state but only in intracellular compartments where its concentration is 5-10-fold higher than average. In contrast to arrestin2, IP6 inhibits self-association of rod arrestin, indicating that the structure of these two tetramers in solution is likely different. PMID- 18161996 TI - Formation of gold branched plates in diluted solutions of poly(vinylpyrrolidone) and their use for the fabrication of near-infrared-absorbing films and coatings. AB - Ribbon-like and branched gold nano- and microstructures were produced by simple heating of diluted aqueous solutions of poly(vinylpyrrolidone) (PVP) and HAuCl4. The reaction was carried out in a one-pot, one-step process at mild temperatures. Modification of the synthesis variables allowed the obtaining of structures with different sizes and branching degrees which formed stable hydrosols with characteristic colors. A mechanism for the growth of the crystals was proposed, based on the aggregation of metal units followed by reorientation and attachment processes facilitated by the presence of low concentrations of the polymer. These anisotropic structures were used to obtain large-area porous coatings on metallic, plastic, and glass substrates and to synthesize homogeneous polymer composites. The resulting gold-modified materials showed an important increase of absorption in the near-infrared (NIR) region of the electromagnetic spectrum, which could find interesting applications in the development of NIR-absorbing filters and coatings. PMID- 18161997 TI - Porous conducting polymer/heteropolyoxometalate hybrid material for electrochemical supercapacitor applications. AB - A porous conducting polymer/heteropolyoxometalate hybrid material that displays high specific capacitance and low ionic resistance has been prepared for electrochemical supercapacitor applications. Polypyrrole/phosphomolybdate composite films were chemically synthesized in tetrahydrofuran in the presence of sodium sulfate, which acts as a porogen. While the phosphomolydic acid could be removed from the film upon rinsing with pure tetrahydrofuran or acetone, rinsing with water or methanol resulted in retention of the heteropolyoxometalate at a level high enough to easily observe its electrochemistry. The retained phosphomolybdate exhibits fast and reversible redox behavior, adding a significant amount of pseudocapacitance to the polymer. Porous films were obtained by leaching out the sodium sulfate porogen from the films using water. The morphology obtained using this method is altered by varying the monomer-to porogen ratio. Increasing the porosity increases the rate at which the hybrid material can be charged/discharged (i.e., oxidized/reduced) by increasing the ionic conductivity and in turn lowering the resistor-capacitor time constant of the material. The ability to tune the porosity of the material allows the optimization of performance characteristics for use in supercapacitor applications. Impedance measurements indicate that the ionic conductivity of these porous structures can be increased more than an order of magnitude over that observed for standard conducting polymer films and that the hybrid material displays peak specific capacitance of around 700 F/g as well as excellent reversibility and cyclability. PMID- 18161998 TI - Relation between the lower consolute boundary and the structure of mesoporous silica materials. AB - In this study, we have shed some light on the relation between the position of the lower consolute boundary of various nonionic surfactants in water and the structure of the mesoporous silica materials synthesized from these surfactants based systems. In the first part, the lower consolute boundary was shifted by adding salts. Depending on the features of the phase diagram, we have looked for either a salting out or a salting in effect. Mesoporous materials were prepared from a micellar solution of the investigated surfactants. Results clearly evidenced that the cooperative self-assembly mechanism is not favored if the lower consolute boundary is not shifted toward high temperatures. Moreover, the higher the difference between the phase separation temperature and the temperature at which the silica precursor is added to the surfactant solution, the better the mesopore ordering is. In the second part, this tendency has been confirmed by using a hydrogenated surfactant as additive. PMID- 18161999 TI - Effect of hyperbranched surface-migrating additives on the electrospinning behavior of poly(methyl methacrylate). AB - Poly(methyl methacrylate) (PMMA) was electrospun in the presence of a low molecular weight, hyperbranched poly(ethylene imine) additive partially functionalized with perfluorinated and aliphatic end-groups (M(n) approximately 1600 g/mol). The additive exhibited surface segregation with an insignificant influence on the rheological behavior of PMMA solutions. A morphological transition from beaded electrospun fibers to uniform fibers was observed upon introduction of additive at low PMMA concentrations. XPS revealed a surface enrichment of fluorine and nitrogen, which are both present in the hyperbranched additive. Surface fluorine content depended primarily on the amount of additive in solution, and a dependency on the PMMA/additive weight ratio was not observed. PMID- 18162000 TI - Toward reliable algorithmic self-assembly of DNA tiles: a fixed-width cellular automaton pattern. AB - Bottom-up fabrication of nanoscale structures relies on chemical processes to direct self-assembly. The complexity, precision, and yield achievable by a one pot reaction are limited by our ability to encode assembly instructions into the molecules themselves. Nucleic acids provide a platform for investigating these issues, as molecular structure and intramolecular interactions can encode growth rules. Here, we use DNA tiles and DNA origami to grow crystals containing a cellular automaton pattern. In a one-pot annealing reaction, 250 DNA strands first assemble into a set of 10 free tile types and a seed structure, then the free tiles grow algorithmically from the seed according to the automaton rules. In our experiments, crystals grew to approximately 300 nm long, containing approximately 300 tiles with an initial assembly error rate of approximately 1.4% per tile. This work provides evidence that programmable molecular self-assembly may be sufficient to create a wide range of complex objects in one-pot reactions. PMID- 18162001 TI - The importance of strong carbon-metal adhesion for catalytic nucleation of single walled carbon nanotubes. AB - Density functional theory is used to show that the adhesion between single-walled carbon nanotubes (SWNTs) and the catalyst particles from which they grow needs to be strong to support nanotube growth. It is found that Fe, Co, and Ni, commonly used to catalyze SWNT growth, have larger adhesion strengths to SWNTs than Cu, Pd, and Au and are therefore likely to be more efficient for supporting growth. The calculations also show that to maintain an open end of the SWNT it is necessary that the SWNT adhesion strength to the metal particle is comparable to the cap formation energy of the SWNT end. This implies that the difference between continued and discontinued SWNT growth to a large extent depends on the carbon-metal binding strength, which we demonstrate by molecular dynamics (MD) simulations. The results highlight that first principles computations are vital for the understanding of the binding strength's role in the SWNT growth mechanism and are needed to get accurate force field parameters for MD. PMID- 18162002 TI - Identifying the mechanism of biosensing with carbon nanotube transistors. AB - Carbon nanotube transistors have outstanding potential for electronic detection of biomolecules in solution. The physical mechanism underlying sensing however remains controversial, which hampers full exploitation of these promising nanosensors. Previously suggested mechanisms are electrostatic gating, changes in gate coupling, carrier mobility changes, and Schottky barrier effects. We argue that each mechanism has its characteristic effect on the liquid gate potential dependence of the device conductance. By studying both the electron and hole conduction, the sensing mechanisms can be unambiguously identified. From extensive protein-adsorption experiments on such devices, we find that electrostatic gating and Schottky barrier effects are the two relevant mechanisms, with electrostatic gating being most reproducible. If the contact region is passivated, sensing is shown to be dominated by electrostatic gating, which demonstrates that the sensitive part of a nanotube transistor is not limited to the contact region, as previously suggested. Such a layout provides a reliable platform for biosensing with nanotubes. PMID- 18162003 TI - Dysmenorrhea in adolescents: diagnosis and treatment. AB - Dysmenorrhea occurs in the majority of adolescent girls and is the leading cause of recurrent short-term school absence in this group. In the vast majority of cases, a presumptive diagnosis of primary dysmenorrhea can be made based on a typical history of low anterior pelvic pain coinciding with the onset of menses and lasting 1-3 days with a negative physical examination. Risk factors for primary dysmenorrhea include nulliparity, heavy menstrual flow, and smoking. Poor mental health and social supports are other associations. Empiric therapy for primary dysmenorrhea can be initiated without diagnostic testing. Effective therapies include NSAIDs, oral contraceptives, and pharmacologic suppression of menstrual cycles. In atypical, severe, or refractory cases, imaging and/or laparoscopy should be performed to investigate secondary causes of dysmenorrhea. The most common cause of secondary dysmenorrhea is endometriosis, the treatment of which may include medical and surgical approaches. Pharmacologic treatment of young women with pain related to endometriosis is similar to treatment of primary dysmenorrhea but may infrequently include gonadotropin-releasing hormone agonists in severe refractory cases. PMID- 18162004 TI - Corticosteroid-resistant nephrotic syndrome with focal and segmental glomerulosclerosis : an update of treatment options for children. AB - Corticosteroid-resistant nephrotic syndrome (CRNS) with focal and segmental glomerulosclerosis (FSGS) is a heterogeneous disorder and the most severe and frequent type of all glomerulopathies in children leading to end-stage renal failure. The podocyte is at the center of development and progress of FSGS; this unique cell type plays a major role in the integrity of glomerular structure and permeability. The rate of complete remission of CRNS after induction therapy using different immunosuppressant agents is reported to range between 30% and 84%, depending on the treatment schedule and on the underlying defects of FSGS. Children with genetic types of FSGS barely respond to immunosuppressant therapies and over-treatment prior to transplantation should be avoided. The response of children with an idiopathic type of FSGS to immunosuppressants is superior to those with genetic FSGS. However, many children with idiopathic FSGS do not enter complete remission if they are under-treated, for example, with short-term immunosuppressant monotherapies. If immunosuppressant treatment fails, these patients will have to undergo renal transplantation. However, as unknown pathogenetic mechanisms may persist, more than one-third of these patients with idiopathic FSGS develop a rapid recurrence of CRNS that responds poorly to further long-term therapeutic attempts. In contrast with previously published data, this review takes into account recently identified genetic etiologies of CRNS, and superior results with long-term combination therapy in idiopathic forms to avoid over- and under-treatment. PMID- 18162005 TI - Management of chronic daily headache in children and adolescents. AB - Chronic daily headache (CDH) occurs in 1-2% of children and adolescents. It can evolve from either episodic tension-type headache or episodic migraine, or can appear with no previous headache history. As with other primary headache disorders, treatment is based on the level of disability. There are children and adolescents who cope well, but there are others who are markedly disabled by their chronic headaches. As in adults, children and adolescents with CDH are at risk for medication overuse. CDH is a diagnosis of exclusion, based on a thorough history, normal physical examination, and negative neuroimaging findings. Along with the chronic headaches, children with this condition may have co-morbid sleep problems, autonomic dysfunction, anxiety, and/or depression. Principles of treatment include identifying migrainous components, stopping medication overuse, stressing normalcy, using rational pharmacotherapy, and addressing co-morbid conditions. Successful outcomes often involve identifying an appropriate headache preventative, reintegration into school, and family participation in resetting realistic expectations. PMID- 18162006 TI - Managing Crohn disease in children and adolescents : focus on tumor necrosis factor antagonists. AB - Crohn disease (CD) is a chronic inflammatory disorder of the gastrointestinal tract characterized by a relapsing course and variable presentation that often includes abdominal pain, diarrhea, and fatigue. CD frequently presents during childhood, resulting in pediatric-specific complications, such as growth failure and delayed puberty. Conventional drug therapy for moderate to severe pediatric CD includes induction of remission with corticosteroids, and maintenance of remission with immunomodulators. Patients who have an inadequate response to standard therapy are being increasingly treated with anti-tumor necrosis factor alpha (TNFalpha) agents. Infliximab has been the most widely studied anti TNFalpha agent in pediatric CD, and has been shown to be efficacious in this condition. Adalimumab has been proven to be efficacious in adults with CD, but there has been only a single case report in children. CDP571 has been tested in 20 children with CD, showing some efficacy. Finally, thalidomide therapy has been associated with improvement in two small case series. Toxicities of these agents include infusion reactions, infections, malignancies, neurologic disorders, and hematologic derangements. PMID- 18162007 TI - Switching from neurostimulant therapy to atomoxetine in children and adolescents with attention-deficit hyperactivity disorder : clinical approaches and review of current available evidence. AB - This review provides practical information on and clinical reasons for switching children and young people with attention-deficit hyperactivity disorder (ADHD) from neurostimulants to atomoxetine, detailing currently available evidence, and switching options. The issue is of particular relevance following recent guidance from the National Institute for Health and Clinical Excellence and European ADHD guidelines endorsing the use of atomoxetine, along with the stimulants methylphenidate and dexamphetamine, in the management of ADHD in children and adolescents in the UK. The selective norepinephrine (noradrenaline) reuptake inhibitor, atomoxetine, is a non-stimulant drug licensed for the treatment of ADHD in children and adolescents, and in adults who have shown a response in childhood. Following the once-daily morning dose, its therapeutic effects extend through the waking hours, into late evening, and in some patients, through to early the next morning. Atomoxetine may be considered for patients who are unresponsive or incompletely responsive to stimulant treatment, have co-morbid conditions (e.g. tics, anxiety, depression), and have sleep disturbances or eating problems, for patients in whom stimulants are poorly tolerated, and for situations where there is potential for drug abuse or diversion. Atomoxetine has been shown to be effective in relapse prevention and there is suggestion that atomoxetine may have a positive effect on global functioning; specifically health related quality of life, self-esteem, and social and family functioning. According to one study, approximately 50% of non-responders to methylphenidate will respond to atomoxetine therapy and approximately 75% of responders to methylphenidate will also respond to atomoxetine. Atomoxetine may be initiated by a schedule of dose increases and cross-tapering with methylphenidate. A slow titration schedule with divided doses minimizes the impact of adverse events within the first several weeks of treatment. Atomoxetine may be co-administered with methylphenidate during the switching period without undue concern for adverse events, such as cardiovascular effects (although monitoring of blood pressure and heart rate is necessary). Atomoxetine may be discontinued abruptly and patients may miss the occasional dose without rebound effects or discontinuation syndrome. A trial period of at least 6-8 weeks, perhaps longer, is recommended before evaluation of the overall tolerability and efficacy of atomoxetine. We conclude that patients with ADHD can be switched from neurostimulants, specifically methylphenidate, to atomoxetine, and may benefit from symptom improvement. PMID- 18162009 TI - What clinicians need to know about neuroprotective issues in bipolar disorder. AB - Neuroimaging and pathohistologic research may show whether bipolar disorder is a degenerative and progressive condition, if detectable changes in the brain record past occurrences, and whether these changes are reversible. Interpreting neuroimaging studies has been complicated by a lack of identification of patient mood states and differences in patients' medication status, but several observations have been made. Pathohistologic research suggests that bipolar disorder is associated with significant cell pathology. PMID- 18162008 TI - Clinical uses of dexmedetomidine in pediatric patients. AB - Dexmedetomidine is being used off-label as an adjunctive agent for sedation and analgesia in pediatric patients in the critical care unit and for sedation during non-invasive procedures in radiology. It also has a potential role as part of anesthesia care to prevent emergence delirium and postanesthesia shivering. Dexmedetomidine is currently approved by the US FDA for sedation only in adults undergoing mechanical ventilation for <24 hours. Pediatric experiences in the literature are in the form of small studies and case reports. In patients sedated for mechanical ventilation and/or opioid/benzodiazepine withdrawal, the loading dose ranged from 0.5 to 1 microg/kg and was usually administered over 10 minutes, although not all patients received loading doses. This patient group also received a continuous infusion at rates ranging from 0.2 to 2 microg/kg/h, with higher rates used in burn patients and those with withdrawal following > or =24 hours of opioid/benzodiazepine infusion. The dexmedetomidine dosage used for anesthesia and sedation during non-invasive procedures, such as radiologic studies, ranged from a loading dose of 1-2 microg/kg followed by a continuous infusion at 0.5-1.14 microg/kg/h, with most patients spontaneously breathing. For invasive procedures, such as awake craniotomy or cardiac catheterization, dosage ranged from a loading dose of 0.15 to 1 microg/kg followed by a continuous infusion at 0.1-2 microg/kg/h. Adverse hemodynamic and respiratory effects were minimal; the agent was well tolerated in most patients. The efficacy of dexmedetomidine varied depending on the clinical situation: efficacy was greatest during non-invasive procedures, such as magnetic resonance imaging (MRI), and lowest during invasive procedures, such as cardiac catheterization. Dexmedetomidine may be useful in pediatric patients for sedation in a variety of clinical situations. The literature suggests potential use of dexmedetomidine as an adjunctive agent to other sedatives during mechanical ventilation and opioid/benzodiazepine withdrawal. In addition, because of its minimal respiratory effects, dexmedetomidine has also been used as a single agent for sedation during non-invasive procedures such as MRI. However, additional studies in pediatric patients are warranted to further evaluate its safety and efficacy in all age ranges. PMID- 18162010 TI - Recognition and treatment of obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is prevalent, chronic, and potentially disabling. It is characterized by recurrent, unwanted, and distressing thoughts (obsessions) and repetitive, irresistible, behaviors (compulsions). Individuals with OCD recognize that the obsessions and compulsions are senseless or excessive yet they are unable to stop these behaviors. Some etiologic theories of OCD suggest a biological origin, including hypotheses involving the serotonergic system, the glutamatergic system, the orbital cortex and the basal ganglia, and streptococcal throat infections in children. Standard treatments for OCD include selective serotonin reuptake inhibitors (SSRIs) and cognitive-behavior therapy. Combining SSRIs with other medications has relatively little empirical support; however, the adjunctive use of antipsychotics has been shown to be effective. Neurosurgery, such as deep brain stimulation, has also been shown to be effective in select patients with debilitating and refractory OCD. PMID- 18162011 TI - Identifying and treating cognitive impairments in elderly patients. AB - Elderly patients with depression are at risk for developing mild cognitive impairment (MCI), and some patients with MCI may go on to develop Alzheimer's disease. This interactive Case and Comment follows Mr. P through the initial diagnosis and treatment of MCI and introduces measurements to predict his likelihood of converting to Alzheimer's disease. PMID- 18162012 TI - Mortality risk in patients with schizophrenia participating in premarketing atypical antipsychotic clinical trials. AB - OBJECTIVE: Given the concern that mortality rates may be increased in geriatric patients exposed to atypical antipsychotic agents, we assessed mortality rates for adult patients with schizophrenia assigned to an investigational antipsychotic (olanzapine, quetiapine, risperidone, or ziprasidone), a control antipsychotic (haloperidol or chlorpromazine), or placebo in preapproval clinical development programs to assess relative risk with atypical antipsychotics as compared to typical antipsychotics or placebo. METHOD: We reviewed safety data (from clinical trials conducted from approximately 1982 to 2002) for 16,791 adult patients with schizophrenia (DSM-III or DSM-IV criteria) in U.S. Food and Drug Administration (FDA) Summary Basis of Approval (SBA) reports for 6 antipsychotic drugs. Mortality rates were calculated for each treatment group (investigational agent, active control, or placebo) on the basis of patient exposure years (PEY) and gross mortality. We compared the differences in mortality rates between placebo and investigational agents, active controls, and all antipsychotic drugs combined using odds ratios. RESULTS: By PEY analysis, the mortality rate for patients assigned to placebo treatment was significantly higher (p < .05) than for either the investigational antipsychotic (OR = 0.23, 95% CI = 0.13 to 0.45) or the active control group (OR = 0.19, 95% CI = 0.08 to 0.45). Although rates based on gross mortality were also higher with placebo treatment, statistical significance was only seen when comparing patients assigned to placebo with those assigned to the active control antipsychotic group (OR = 0.35, 95% CI = 0.15 to 0.82). CONCLUSIONS: Despite reported excess mortality with antipsychotic use in elderly patients with dementia, SBA data did not reveal a similar increased risk of antipsychotics in adult patients with schizophrenia. However, methodological limitations of the FDA SBA reports may affect the generalizability of these findings. PMID- 18162013 TI - Adiponectin as a potential biomarker for the metabolic syndrome in Chinese patients taking clozapine for schizophrenia. AB - OBJECTIVE: As the metabolic syndrome is an important side effect of some antipsychotics, use of a biomarker will enable clinicians to identify metabolic changes more effectively than anthropometry and biochemistry. Adiponectin, an adipocyte-derived hormone, serves as a central regulatory protein in many of the physiologic pathways controlling lipid and carbohydrate metabolism. The aim of this study is to determine the possible relationship between adiponectin and the metabolic syndrome among Chinese patients taking clozapine for schizophrenia. METHOD: The study sample consisted of 188 hospitalized Chinese patients with schizophrenia (DSM-IV criteria) who had been receiving clozapine for at least 3 months. Cross-sectional anthropometric measurements, biochemical analysis, and serum adiponectin levels were assessed to determine the prevalence of metabolic syndrome. Retrospective chart reviews were conducted to obtain demographic data, age at which clozapine treatment was initiated, and weight change after the initiation of clozapine treatment. The study was conducted from March to September of 2005. RESULTS: The prevalence of the metabolic syndrome was 28.4%. Adiponectin levels were negatively associated with weight change after the initiation of clozapine treatment, systolic blood pressure, diastolic blood pressure, body weight, body mass index (BMI), waist circumference, serum triglycerides, and insulin and were positively associated with high-density lipoprotein cholesterol. Multiple logistic regression analysis showed that age (OR = 1.083, p = .009), BMI (OR = 1.423, p < .001), and serum adiponectin (OR = 0.847, p = .01) each correlated significantly with the presence of the metabolic syndrome. CONCLUSION: Independent of age and BMI, hypoadiponectinemia is a potential biomarker of the metabolic syndrome in patients taking clozapine for schizophrenia. PMID- 18162014 TI - Divalproex in the treatment of acute bipolar depression: a preliminary double blind, randomized, placebo-controlled pilot study. AB - OBJECTIVE: To determine the efficacy of divalproex (extended release) in the treatment of acute nonrefractory bipolar depression. METHOD: In a stratified, double-blind, randomized, placebo-controlled trial, 18 acutely depressed bipolar outpatients (DSM-IV criteria) received either divalproex monotherapy (target dose level, 70-90 ng/dL) (N = 9) or placebo (N = 9) for 6 weeks. Patients were recruited between January 2004 and May 2005. Clinical assessment on the Montgomery-Asberg Depression Rating Scale (MADRS) determined primary efficacy. RESULTS: The divalproex treatment group showed significantly greater reduction in MADRS scores compared to placebo (group x time interaction, p = .0078). Absolute effect size of estimated MADRS total score reduction over time was 13.6 points with divalproex versus 1.4 points with placebo (p = .003, linear growth curve model). Standardized effect size was large (Cohen d = 0.81). MADRS item analyses demonstrated improvement in core mood symptoms more than in anxiety or insomnia symptoms. There was also a modest but significant association between MADRS and Mania Rating Scale scores in the divalproex group (r = 0.29, df = 51, p = .03), but not in the placebo group (r = -0.15, df = 35, p = .36). CONCLUSIONS: Divalproex appeared to be an effective treatment for acute nonrefractory bipolar depression, which is consistent with previous small randomized studies. Some evidence of benefit in the depressive mixed state was observed. Confirmation or refutation with larger randomized clinical trials is warranted. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT00226343. PMID- 18162015 TI - Paroxetine versus placebo and other agents for depressive disorders: a systematic review and meta-analysis. AB - OBJECTIVE: To compare paroxetine with placebo and other antidepressants across multiple efficacy and tolerability outcomes. DATA SOURCES: Searches were conducted in MEDLINE (1966-2004), EMBASE (1980-2004), CINAHL (1982-2004), all Evidence-Based Medicine Reviews (1991-2004), HealthSTAR (1975-2004), BIOSIS (1980 2004), and PsycINFO (1840-2004). Medical Subject Headings (MeSH) included "paroxetine" OR "Paxil" exploded. The searches were not restricted by language, publication type, or study design. STUDY SELECTION: A study report was included if it described a randomized trial of paroxetine versus placebo or other antidepressants for patients with depressive disorders. Records were screened independently by 2 reviewers under the supervision of another reviewer. DATA EXTRACTION: Three investigators abstracted data, including study design, trial characteristics, and psychiatric assessment tools, using a prespecified form. Two investigators assessed quality of reporting using Jadad's scale. DATA SYNTHESIS: We included 62 unique randomized controlled trials. Paroxetine yielded consistently and significantly better remission (rate difference [RD]: 10% [95% CI = 6 to 14]), clinical response (RD: 17% [95% CI = 7 to 27]), and symptom reduction (effect size: 0.2 [95% CI = 0.1 to 0.3]) than placebo. Such consistency in the evidence base was not observed between paroxetine and other antidepressants. Pairwise comparisons of paroxetine and venlafaxine, mirtazapine, mianserin, or fluoxetine yielded inconsistent results across efficacy outcomes. Controlled-release paroxetine was the only antidepressant with significantly fewer dropouts due to adverse events than immediate-release paroxetine (RD: 5% [95% CI = 0.1 to 11]). CONCLUSIONS: There were no significant and valid differences between paroxetine and other antidepressants to suggest that multiple modes of action improve clinical outcomes. PMID- 18162016 TI - Symptoms of sexual dysfunction in patients treated for major depressive disorder: a meta-analysis comparing selegiline transdermal system and placebo using a patient-rated scale. AB - OBJECTIVE: Spontaneous reports of sexual side effects were infrequent during placebo-controlled clinical trials of selegiline transdermal system (STS). The objective of this study was to examine the impact of STS 6 mg/24 hours on various domains of sexual function in patients with major depressive disorder (MDD), using a patient-rated questionnaire. METHOD: Data from 4 short-term (6 to 8 weeks), randomized, double-blind, placebo-controlled trials of STS in patients with MDD (DSM-IV criteria) were included in the meta-analysis (STS, N = 389; placebo, N = 400). The Medex Sexual Dysfunction Subscale was used to assess sexual interest, arousal, maintenance of interest, orgasm, and satisfaction. Estimates of the average effect of study drug on each item of sexual function and 95% confidence intervals were calculated using a fixed-effects model due to homogeneity of study means. The direct effect of STS versus placebo was estimated using multivariate regression models, with baseline item score as a covariate and controlling for improvement in depression. Analyses were performed on the total population and by gender. Data were collected between January 1997 and April 2000. RESULTS: Estimates of difference between STS and placebo demonstrated a nonsignificant trend toward a positive treatment effect of STS on most sexual function items and significant improvement in sexual satisfaction. For women, there was a significant positive effect on interest, maintaining interest during sex, and satisfaction. The direct effect of STS on changes in individual item scores was minimal in men and showed a trend for improvement in women. CONCLUSION: This meta-analysis suggests that short-term therapy with STS 6 mg/24 hours does not impair any aspect of sexual function in MDD patients as measured using a patient-rated questionnaire. PMID- 18162017 TI - Ethyl-eicosapentaenoic acid in first-episode psychosis: a randomized, placebo controlled trial. AB - OBJECTIVE: To investigate if ethyl-eicosapentaenoic acid (E-EPA) augmentation improves antipsychotic efficacy and tolerability in first-episode psychosis (FEP). METHOD: We performed a 12-week, randomized, double-blind, placebo controlled trial of 2-g E-EPA augmentation in 80 FEP patients. Sixty-nine patients were eligible for analysis; a post hoc analysis was computed for a subgroup of nonaffective FEP patients (N = 53). The first participant was included in November 2000 and the last participant completed the trial in August 2003. Primary outcome measures were symptom change scores and time to first response, while tolerability measures and cumulative antipsychotic dose were secondary outcome measures. RESULTS: Analysis of covariance controlling for baseline symptoms found no significant mean difference between E-EPA and placebo at week 12 for symptom change scores. Cox regression analysis revealed a significant treatment by diagnosis interaction (p = .024) for time to first response favoring E-EPA in nonaffective psychosis. Post hoc analysis for cumulative response rates further confirmed a higher response rate at week 6 (42.9% [15/35] vs. 17.6% [6/34] for all participants, p = .036; 54.2% [13/24] vs. 17.2% [5/29] for the nonaffective psychosis subset, p = .008); however, the difference at week 12 was no longer significant. Analysis of secondary outcome measures revealed that E-EPA-augmented participants needed 20% less antipsychotic medication between weeks 4 through 6 (p = .03), had less extrapyramidal side effects in the initial 9 weeks (p < .05 for all participants and for all timepoints), and reported less constipation (p = .011) and fewer sexual side effects (p = .016) than those treated with antipsychotic medication alone. CONCLUSION: The findings suggest that E-EPA may accelerate treatment response and improve the tolerability of antipsychotic medications. However, it was not possible to demonstrate a sustained symptomatic benefit of E-EPA in early psychosis, possibly due to a ceiling effect, since a high proportion of first episode patients already achieve symptomatic remission with antipsychotic medication alone. Further controlled trials in nonaffective early psychosis seem warranted. TRIAL REGISTRATION: Australian Clinical Trials Registry identifier 12605000267651 (http://actr.org.au). PMID- 18162018 TI - Comparison of intramuscular ziprasidone, olanzapine, or aripiprazole for agitation: a quantitative review of efficacy and safety. AB - OBJECTIVE: To compare the efficacy and safety of the intramuscular formulations of ziprasidone, olanzapine, and aripiprazole in treating agitation. DATA SOURCES: The pivotal registration trials were accessed by querying on-line literature and clinical trial databases. Pharmacovigilance data and posters were requested from the manufacturers. No date or language constraints were applied. STUDY SELECTION: Nine double-blind, randomized, controlled clinical trials were identified. DATA EXTRACTION: Number needed to treat (NNT) for response to treatment for agitation and number needed to harm (NNH) for extrapyramidal effects were calculated from the study reports. Additional safety outcomes subject to NNH analysis were obtained from product labeling. DATA SYNTHESIS: Using the a priori definitions of response at 2 hours after the first injection, NNT for response versus placebo (or placebo equivalent) in treating agitation for the pooled data at the recommended dose of ziprasidone 10-20 mg was 3 (95% CI = 2 to 4), for olanzapine 10 mg was 3 (95% CI = 2 to 3), and for aripiprazole 9.75 mg was 5 (95% CI = 4 to 8). Treatment-emergent adverse events occurring during the pivotal trials revealed statistically significant NNH versus placebo (or placebo equivalent) for aripiprazole for headache (NNH = 20, 95% CI = 11 to 170) and nausea (NNH = 17, 95% CI = 11 to 38), for ziprasidone in the treatment of headache (NNH = 15, 95% CI = 8 to 703), and for olanzapine in treatment-emergent hypotension (NNH = 50, 95% CI = 30 to 154). Olanzapine and aripiprazole had a more favorable extrapyramidal side effect profile compared to haloperidol. (There was no haloperidol treatment arm in the ziprasidone studies.) CONCLUSIONS: Although the lowest NNT, and hence strongest therapeutic effect, was seen for the studies of ziprasidone and olanzapine as opposed to aripiprazole, head-to-head controlled studies directly comparing these 3 agents are needed. PMID- 18162019 TI - Quality of treatment and disability compensation in depression: comparison of 2 nationally representative samples with a 10-year interval in Finland. AB - OBJECTIVE: Depressive disorders cause substantial work impairment that can lead to disability compensation. The authors compared treatment received for depression preceding disability pension between 2 nationally representative samples with a 10-year interval. METHOD: The medical statements for 2 random samples drawn from the Finnish national disability pension registers, representing populations granted a disability pension for DSM-III-R major depression during a 12-month period from October 1993 through September 1994 (N = 277) and for ICD-10 depressive disorders (F32-F33) from October 2003 through September 2004 (N = 265) were examined. The proportions of persons receiving weekly psychotherapy, antidepressants, adequate antidepressant dosage, sequential antidepressant trials, lithium augmentation, and electroconvulsive therapy (ECT) were compared. RESULTS: No significant differences emerged between the 2 samples, except for the adequacy of antidepressant dosage. Few subjects in either of the samples (8.7% for 1993-1994 vs. 10.6% for 2003-2004, p = .45) had received weekly psychotherapy. Most had received antidepressants (87.4% vs. 85.6%, p = .55) with increasingly adequate dosage (75.6% vs. 85.0%, p = .02), but only a minority had received sequential antidepressant trials (39.5% vs. 44.5%, p = .24). Lithium augmentation and ECT were rare (1.1% vs. 1.5%, p = .66 and 4.0% vs. 1.5%, p = .08, respectively). Even in 2003-2004, over half of the subjects were granted a disability pension without sequential antidepressant trials. CONCLUSION: This nationally representative study indicates that, despite an increased antidepressant use and improved practice guidelines for depression, a considerable proportion of the people granted long-term compensation for depression seem to be suboptimally treated. Given the enormous costs of the disability, attention to the quality of treatment provided for depression is warranted before long-term disability compensations are granted. PMID- 18162020 TI - The role of craving in relapse after discontinuation of long-term benzodiazepine use. AB - OBJECTIVE: Craving for benzodiazepines has never been examined as a factor of relapse after successful benzodiazepine discontinuation. In this study, we examined the predictive value of craving on benzodiazepine relapse. METHOD: A stepped-care intervention trial aimed to discontinue long-term benzodiazepine use in general practice. The first step was the sending of a letter to users advising them to gradually quit their use by themselves (i.e., minimal intervention). The second step, a supervised tapering-off program, was offered to those unable to discontinue by themselves. Craving was assessed by means of the Benzodiazepine Craving Questionnaire (BCQ). Multiple Cox proportional hazards regression analyses were performed to examine the effect of craving on subsequent relapse during a 15-month follow-up period in patients who had successfully quit their benzodiazepine use by themselves after the minimal intervention (N = 79) and in those patients who had successfully quit after the supervised tapering-off program (N = 45). Data were collected from August 1998 to December 2001. RESULTS: Thirty-five (44%) and 24 (53%) patients had relapsed after the minimal intervention and tapering-off program, respectively. Patients able to quit by themselves experienced very little craving. In this sample, craving was not related to relapse (p = .82). In patients who needed an additional supervised tapering-off program, higher craving scores were significantly related to relapse (hazard ratio = 1.26, 95% CI = 1.02 to 1.54, p = .029), when corrected for benzodiazepine characteristics, psychopathology, and personality characteristics. CONCLUSION: Craving is an independent factor of subsequent relapse after successful benzodiazepine discontinuation in long-term benzodiazepine users who are not able to quit their usage of their own accord. PMID- 18162021 TI - Comparative assessment of the incidence and severity of tardive dyskinesia in patients receiving aripiprazole or haloperidol for the treatment of schizophrenia: a post hoc analysis. AB - OBJECTIVE: To comparatively assess the incidence of tardive dyskinesia in patients with schizophrenia receiving either aripiprazole or haloperidol. METHOD: Data from 2 controlled, long-term trials of subjects meeting DSM-IV criteria for schizophrenia treated with aripiprazole (N = 861, 20-30 mg/day) or haloperidol (N = 433, 5-10 mg/day) were analyzed. The primary outcome measure was the rate of new-onset tardive dyskinesia. The analysis was limited to patients without baseline tardive dyskinesia. There were no significant differences between treatment groups in demographic or disease characteristics. The Abnormal Involuntary Movement Scale (AIMS) and Research Diagnostic Criteria for tardive dyskinesia were used to define the comparative incidence rates of long-term treatment-emergent tardive dyskinesia. RESULTS: A significantly lower percentage of aripiprazole-treated patients developed new-onset tardive dyskinesia compared with haloperidol-treated patients (p < .0001). The annualized rate of treatment emergent tardive dyskinesia was significantly lower in aripiprazole-treated versus haloperidol-treated patients. In patients without a baseline diagnosis of tardive dyskinesia, aripiprazole significantly improved AIMS scores compared with haloperidol (p .05, all 4 analyses). CONCLUSION: There does not appear to be any statistically detectable difference in the rapidity of antidepressant effect between bupropion and the SSRIs overall or escitalopram specifically. PMID- 18162023 TI - The fundamental structure of axis II personality disorders assessed in the National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: Little is known about the fundamental structure of core personality disorder psychopathology in the general population. The current study employed confirmatory factor analysis to evaluate competing models of patterns of personality disorder diagnoses in a nationally representative sample. METHOD: DSM IV and alternate models of the structure of personality disorder psychopathology were evaluated using data from the National Epidemiologic Survey on Alcohol and Related Conditions conducted between 2001 and 2002 (N = 43,093). Dimensional versus categorical representations of DSM-IV personality disorder structure were also tested. Face-to-face interviews were conducted in the United States. Participants were community-based respondents aged 18 years and older. Diagnoses and dimensional scores were made for antisocial, avoidant, dependent, histrionic, obsessive-compulsive, paranoid, and schizoid personality disorders. RESULTS: Multiple goodness-of-fit indicators provided support for a DSM-IV-based hierarchical model of personality disorders. In this model, the individual personality disorders were viewed as belonging to 1 of 3 latent factors or clusters (A, B, or C). In all of the models, the individual personality disorders were allowed to be an indicator for only a single latent cluster, and errors were not allowed to correlate with each other. In turn, these 3 clusters were viewed as comprising a single higher-order "Axis II personality disorder factor." The DSM-IV model was largely invariant across gender, Axis I comorbidity, and treatment-seeking status. A dimensionally based form of assessment of the DSM-IV personality disorders produced excellent goodness-of-fit indicators and produced low Akaike information criterion values (which are indicative of better-fitting models). CONCLUSIONS: The results from this confirmatory factor analysis in a large, nationally representative mental health survey supported the DSM-IV hierarchical organization of Axis II personality disorders. This model was significantly superior to viable alternative models of Axis II personality psychopathology. There was also evidence to suggest this model could obtain even stronger support if a dimensionally based form of diagnostic assessment was adopted in place of the dichotomous form of assessment (presence/absence) of personality disorders currently in use in the DSM-IV. PMID- 18162024 TI - A comparison of initial duloxetine dosing strategies in patients with major depressive disorder. AB - OBJECTIVE: To compare the effects of starting doses of duloxetine taken with or without food on tolerability and efficacy in patients with major depressive disorder (MDD). METHOD: This double-blind, concurrent-dose-controlled, parallel design trial contained a variable expected-duration placebo lead-in period and was conducted in adult outpatients with DSM-IV-TR-defined MDD at psychiatric outpatient sites between October 2004 and January 2006. In actuality, patients received placebo for 1 week and then were randomly assigned to duloxetine 30 mg once daily in the morning (q.a.m.) (N = 219), 30 mg twice daily (b.i.d.) (N = 213), or 60 mg q.a.m. (N = 215) for 1 week along with 1 of 2 instructions about food: take study drug with food or do not take within 1 hour of eating. For the remaining 5 weeks of acute treatment, all patients received 60 mg once daily. The primary objective was to compare incidence of treatment-emergent nausea at 30 mg q.a.m. versus 60 mg q.a.m. using item 112 (nausea) of the Association for Methodology and Documentation in Psychiatry adverse event scale (AMDP-5). Secondary outcome measures included mean change on AMDP-5 item 112, discontinuations due to adverse events, mean changes in AMDP-5 items and subscales, spontaneously reported treatment-emergent adverse events, and vital signs. Efficacy was evaluated by the 17-item Hamilton Rating Scale for Depression (HAM-D-17). RESULTS: The primary analysis, which combined data from both food groups, showed no significant difference in the incidence of nausea between starting doses of 30 mg q.a.m. and 60 mg q.a.m. (23% vs. 29%, respectively; p = .207). However, mean changes on the AMDP-5 nausea item revealed a significant main effect of food (p = .010) and a significant interaction between food and starting dose (p = .033). The food-by-dose interaction indicated that the benefit from taking drug with food was greatest in patients started at 60 mg q.a.m., and the benefit of starting at 30 mg q.a.m. was greatest in patients taking drug without food. In patients who took study drug without food, there was a significant difference across initial-dose groups for discontinuation due to adverse events (30 mg q.a.m. = 3.6%, 30 mg b.i.d. = 14.0%, 60 mg q.a.m. = 10.2%; 30 mg q.a.m. vs. 30 mg b.i.d., p = .008; 30 mg q.a.m. vs. 60 mg q.a.m., p = .066); however, in patients who took study drug with food, discontinuations due to adverse events did not significantly differ (30 mg q.a.m. = 5.4%, 30 mg b.i.d. = 7.5%, 60 mg q.a.m. = 7.4%; all p values > .50). Patients who started at 30 mg b.i.d. or 60 mg q.a.m. without food did not differ regarding mean changes (i.e., increases) in the common adverse events score after 1 week of treatment but had significantly greater mean changes than patients who started at 30 mg q.a.m. without food (0.87, 0.82, and 0, respectively; p < .05 vs. 30 mg b.i.d. and 60 mg q.a.m.). No significant differences were found between initial-dose groups in vital signs. CONCLUSIONS: These data imply that starting dulox-etine at 30 mg q.a.m. for 1 week with or without food or starting duloxetine at the therapeutic dose of 60 mg q.a.m. with food can improve the initial tolerability of the medication. Adding this information to existing knowledge of duloxetine will enable the clinician to tailor therapy most appropriately for the individual patient. CLINICAL TRIALS REGISTRATION: ClinicalTrials.gov identifier NCT 00191061. PMID- 18162025 TI - Are depressed outpatients with and without a family history of substance use disorder different? A baseline analysis of the STAR*D cohort. AB - OBJECTIVE: This report compares the baseline demographic and clinical characteristics of outpatients with nonpsychotic major depressive disorder (MDD) and a family history of substance use disorder (SUD) versus those with MDD and no family history of SUD. METHOD: Using data from the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study, we grouped participants with MDD (DSM-IV criteria) according to presence or absence of family history of SUD based on participant report. Between-group comparisons were made of demographic and clinical characteristics, depressive symptoms, and psychiatric co morbidities. Patients were enrolled from July 2001 until August 2004. RESULTS: Of 4010 participants, 46% had a positive family history of SUD. Those with a positive family history were less likely to be Hispanic (p = .0029) and more likely to be female (p = .0013). They were less educated (p = .0120), less likely to be married (p < .01), and more likely to be divorced (p < .01). They also reported an earlier age at onset of MDD, greater length of illness, and more major depressive episodes (all p < .001). They had an increased likelihood of recurrent MDD, more prior suicide attempts, and more concurrent psychiatric comorbidities, including posttraumatic stress disorder, SUD, and generalized anxiety disorder (all p < .0001). CONCLUSION: Depressed patients with a family history of SUD had a more severe previous course of depression, were more likely to have attempted suicide, and had a greater burden of psychiatric comorbid conditions than patients without such a family history. These findings represent important clinical features to be considered in the evaluation and treatment planning of patients with MDD. PMID- 18162026 TI - Alcohol and cannabis use in Urban, African American, first-episode schizophrenia spectrum patients: associations with positive and negative symptoms. AB - OBJECTIVE: On the basis of limited prior research on associations between symptoms and substance use in first-episode psychosis, a retrospective chart review was conducted to test 2 hypotheses: (1) the presence of positive symptoms is associated with alcohol use prior to admission and (2) the absence of prominent negative symptoms is associated with cannabis use prior to admission. METHOD: Eligible patients included those admitted for a first episode of psychosis in a public-sector, university-affiliated hospital that serves a predominantly African American, socially disadvantaged, urban population. The 72 patients included in the analysis were 18 to 40 years of age, and all were African American. Using a structured data collection instrument, discharge summaries of consecutively admitted patients from January 2002 to March 2005 were reviewed to extract data on basic demographic and clinical characteristics, the presence of 11 symptoms, and alcohol and cannabis use within 6 months prior to hospitalization. RESULTS: Alcohol use in the 6 months prior to hospitalization was associated with a higher frequency of positive psychotic symptoms among first episode patients. Cannabis use was associated with a lower likelihood of having prominent negative symptoms. These associations remained even after controlling for relevant covariates in logistic regression models. CONCLUSION: Although the direction of causality cannot be established, the association between positive psychotic symptoms and alcohol use may represent a self-medication effect, whereas the association between lesser negative symptoms and cannabis use may result from the fact that interpersonal deficits and reduced hedonic capacity minimize drug-seeking activities. PMID- 18162027 TI - Screening and treatment for women with mood disorders associated with reproductive events. PMID- 18162028 TI - Family and partner psychopathology and the risk of postpartum mental disorders. AB - OBJECTIVE: Family and partner psychopathology characterizes 2 different types of potential risk factors for mental disorders linked to both biological and psychosocial processes, and no studies have included both variables in a study of risk of postpartum mental disorders (PPMD). The aim was to assess how a history of mental disorders in either a first-degree family member or partner affects the risk of admission or outpatient contact with PPMD. METHOD: A population-based cohort study using Danish registers was conducted, and survival analyses were performed. A total of 1,188,822 men and women became first-time parents during the study period from 1973 to 2005. The main outcome measure was incident admission or outpatient contact for any mental disorder (according to ICD-8 or ICD-10 criteria) 0 to 12 months after the birth of a first live-born child. RESULTS: A total of 2174 mothers and 1175 fathers experienced an incident admission or outpatient contact during the 12 months after the birth of the child. Mothers with no family or partner psychopathology had an increased risk of admission/outpatient contact 0 to 30 days postpartum; relative risk (RR) = 3.49 (95% CI = 3.01 to 4.04) compared to the reference group. During the same period, mothers with observed psychopathology in relatives and/or partners were at higher risk of PPMD: for family psychopathology, RR = 6.47 (95% CI = 5.25 to 7.97); for partner psychopathology, RR = 6.86 (95% CI = 3.95 to 11.90); and for both family and partner psychopathology, RR = 10.94 (95% CI = 5.18 to 23.09). Additionally, a 24-fold increased risk of PPMD 0 to 30 days postpartum was found in women with a first-degree relative with bipolar affective disorder compared to the reference group. CONCLUSION: Results indicated that family psychopathology represents a particular risk in the immediate postpartum period, especially if a family member suffers from bipolar affective disorder compared to other diagnostic groups. However, additional studies are needed to establish if partner psychopathology is a risk factor for PPMD specifically or has a more general influence on risk of mental disorders throughout pregnancy and postpartum. PMID- 18162029 TI - Treatment of premenstrual worsening of depression with adjunctive oral contraceptive pills: a preliminary report. AB - OBJECTIVE: Despite the efficacy of antidepres-sants, depression can break through premenstrually. Oral contraceptive pills (OCPs) stabilize reproductive hormones and treat premenstrual dysphoric disorder. Management of depression that breaks through premenstrually has not been studied. METHOD: Women taking antidepressants with successfully treated depression, except during the late luteal phase (Montgomery-Asberg Depression Rating Scale [MADRS] score >or= 15) and high late luteal phase (Daily Rating of Severity of Problems scores) were randomly assigned to open-label ethinyl estradiol (EE) 30 mug/day plus drospirenone 3 mg/day (EE/DRSP) for 21 days and double-blinded treatment with EE 30 mug/day or placebo for days 22 through 28 of 2 cycles. Participants were recruited from community and psychiatry outpatient clinics and enrolled into this study in 2004-2005. RESULTS: Of 25 subjects who received EE/DRSP (N = 12 with EE and N = 13 with placebo), 21 completed treatment. For study completers, premenstrual MADRS (p = .0019) and Daily Rating of Severity of Problems scores (p = .0001) improved significantly in both groups. Outcome did not differ between groups. CONCLUSION: This study provides preliminary evidence that addition of EE/DRSP (+/- EE) to antidepressants may treat premenstrual breakthrough of depression. Stabilizing hormone levels with EE/DRSP may provide an important therapeutic option for women taking antidepressants whose symptoms break through premenstrually. PMID- 18162030 TI - Diagnosing and managing complicated ADHD. PMID- 18162031 TI - Diagnosis and treatment strategies for mixed episodes in bipolar disorder. PMID- 18162032 TI - What constitutes evidence-based pharmacotherapy for bipolar disorder? Part 1: first-line treatments. PMID- 18162034 TI - More on placebo response. PMID- 18162035 TI - Corpus callosum dysplasia in adult attention-deficit/ hyperactivity disorder: a case report. PMID- 18162036 TI - Treatment of cognitive deficits in schizophrenia with atomoxetine: a case report. PMID- 18162037 TI - A case of olanzapine-induced gestational diabetes mellitus in the absence of weight gain. PMID- 18162038 TI - Perspectives for the treatment of brucellosis in the 21st century: the Ioannina recommendations. PMID- 18162039 TI - The challenges of Chagas Disease-- grim outlook or glimmer of hope. PMID- 18162040 TI - Targeting of interferon-beta to produce a specific, multi-mechanistic oncolytic vaccinia virus. AB - BACKGROUND: Oncolytic viruses hold much promise for clinical treatment of many cancers, but a lack of systemic delivery and insufficient tumor cell killing have limited their usefulness. We have previously demonstrated that vaccinia virus strains are capable of systemic delivery to tumors in mouse models, but infection of normal tissues remains an issue. We hypothesized that interferon-beta (IFN beta) expression from an oncolytic vaccinia strain incapable of responding to this cytokine would have dual benefits as a cancer therapeutic: increased anticancer effects and enhanced virus inactivation in normal tissues. We report the construction and preclinical testing of this virus. METHODS AND FINDINGS: In vitro screening of viral strains by cytotoxicity and replication assay was coupled to cellular characterization by phospho-flow cytometry in order to select a novel oncolytic vaccinia virus. This virus was then examined in vivo in mouse models by non-invasive imaging techniques. A vaccinia B18R deletion mutant was selected as the backbone for IFN-beta expression, because the B18R gene product neutralizes secreted type-I IFNs. The oncolytic B18R deletion mutant demonstrated IFN-dependent cancer selectivity and efficacy in vitro, and tumor targeting and efficacy in mouse models in vivo. Both tumor cells and tumor-associated vascular endothelial cells were targeted. Complete tumor responses in preclinical models were accompanied by immune-mediated protection against tumor rechallenge. Cancer selectivity was also demonstrated in primary human tumor explant tissues and adjacent normal tissues. The IFN-beta gene was then cloned into the thymidine kinase (TK) region of this virus to create JX-795 (TK-/B18R-/IFN-beta+). JX-795 had superior tumor selectivity and systemic intravenous efficacy when compared with the TK-/B18R- control or wild-type vaccinia in preclinical models. CONCLUSIONS: By combining IFN-dependent cancer selectivity with IFN-beta expression to optimize both anticancer effects and normal tissue antiviral effects, we were able to achieve, to our knowledge for the first time, tumor specific replication, IFN-beta gene expression, and efficacy following systemic delivery in preclinical models. PMID- 18162041 TI - Neovascular age-related macular degeneration risk based on CFH, LOC387715/HTRA1, and smoking. AB - BACKGROUND: Age-related macular degeneration (AMD) is the major cause of blindness in the elderly. Those with the neovascular end-stage of disease have irreversible loss of central vision. AMD is a complex disorder in which genetic and environmental factors play a role. Polymorphisms in the complement factor H (CFH) gene, LOC387715, and the HTRA1 promoter are strongly associated with AMD. Smoking also contributes to the etiology. We aimed to provide a model of disease risk based on these factors. METHODS AND FINDINGS: We genotyped polymorphisms in CFH and LOC387715/HTRA1 in a case-control study of 401 patients with neovascular AMD and 266 controls without signs of disease, and used the data to produce genetic risk scores for the European-descent population based on haplotypes at these loci and smoking history. CFH and LOC387715/HTRA1 haplotypes and smoking status exerted large effects on AMD susceptibility, enabling risk scores to be generated with appropriate weighting of these three factors. Five common haplotypes of CFH conferred a range of odds ratios (ORs) per copy from 1 to 4.17. Most of the effect of LOC387715/HTRA1 was mediated through one detrimental haplotype (carriage of one copy: OR 2.83; 95% confidence interval [CI] 1.91 4.20), with homozygotes being at particularly high risk (OR 32.83; 95% CI 12.53 86.07). Patients with neovascular macular degeneration had considerably higher scores than those without disease, and risk of blinding AMD rose to 15.5% in the tenth of the population with highest predicted risk. CONCLUSIONS: An individual's risk of developing AMD in old age can be predicted by combining haplotype data with smoking status. Until there is effective treatment for AMD, encouragement to avoid smoking in those at high genetic risk may be the best option. We estimate that total absence of smoking would have reduced the prevalence of severe AMD by 33%. Unless smoking habits change or preventative treatment becomes available, the prevalence of AMD will rise as a consequence of the increasing longevity of the population. PMID- 18162042 TI - GATA3-driven Th2 responses inhibit TGF-beta1-induced FOXP3 expression and the formation of regulatory T cells. AB - Transcription factors act in concert to induce lineage commitment towards Th1, Th2, or T regulatory (Treg) cells, and their counter-regulatory mechanisms were shown to be critical for polarization between Th1 and Th2 phenotypes. FOXP3 is an essential transcription factor for natural, thymus-derived (nTreg) and inducible Treg (iTreg) commitment; however, the mechanisms regulating its expression are as yet unknown. We describe a mechanism controlling iTreg polarization, which is overruled by the Th2 differentiation pathway. We demonstrated that interleukin 4 (IL-4) present at the time of T cell priming inhibits FOXP3. This inhibitory mechanism was also confirmed in Th2 cells and in T cells of transgenic mice overexpressing GATA-3 in T cells, which are shown to be deficient in transforming growth factor (TGF)-beta-mediated FOXP3 induction. This inhibition is mediated by direct binding of GATA3 to the FOXP3 promoter, which represses its transactivation process. Therefore, this study provides a new understanding of tolerance development, controlled by a type 2 immune response. IL-4 treatment in mice reduces iTreg cell frequency, highlighting that therapeutic approaches that target IL-4 or GATA3 might provide new preventive strategies facilitating tolerance induction particularly in Th2-mediated diseases, such as allergy. PMID- 18162043 TI - A neural computation for visual acuity in the presence of eye movements. AB - Humans can distinguish visual stimuli that differ by features the size of only a few photoreceptors. This is possible despite the incessant image motion due to fixational eye movements, which can be many times larger than the features to be distinguished. To perform well, the brain must identify the retinal firing patterns induced by the stimulus while discounting similar patterns caused by spontaneous retinal activity. This is a challenge since the trajectory of the eye movements, and consequently, the stimulus position, are unknown. We derive a decision rule for using retinal spike trains to discriminate between two stimuli, given that their retinal image moves with an unknown random walk trajectory. This algorithm dynamically estimates the probability of the stimulus at different retinal locations, and uses this to modulate the influence of retinal spikes acquired later. Applied to a simple orientation-discrimination task, the algorithm performance is consistent with human acuity, whereas naive strategies that neglect eye movements perform much worse. We then show how a simple, biologically plausible neural network could implement this algorithm using a local, activity-dependent gain and lateral interactions approximately matched to the statistics of eye movements. Finally, we discuss evidence that such a network could be operating in the primary visual cortex. PMID- 18162044 TI - Indirect effects of ploidy suggest X chromosome dose, not the X:A ratio, signals sex in Drosophila. AB - In the textbook view, the ratio of X chromosomes to autosome sets, X:A, is the primary signal specifying sexual fate in Drosophila. An alternative idea is that X chromosome number signals sex through the direct actions of several X-encoded signal element (XSE) proteins. In this alternative, the influence of autosome dose on X chromosome counting is largely indirect. Haploids (1X;1A), which possess the male number of X chromosomes but the female X:A of 1.0, and triploid intersexes (XX;AAA), which possess a female dose of two X chromosomes and the ambiguous X:A ratio of 0.67, represent critical tests of these hypotheses. To directly address the effects of ploidy in primary sex determination, we compared the responses of the signal target, the female-specific SxlPe promoter of the switch gene Sex-lethal, in haploid, diploid, and triploid embryos. We found that haploids activate SxlPe because an extra precellular nuclear division elevates total X chromosome numbers and XSE levels beyond those in diploid males. Conversely, triploid embryos cellularize one cycle earlier than diploids, causing premature cessation of SxlPe expression. This prevents XX;AAA embryos from fully engaging the autoregulatory mechanism that maintains subsequent Sxl expression, causing them to develop as sexual mosaics. We conclude that the X:A ratio predicts sexual fate, but does not actively specify it. Instead, the instructive X chromosome signal is more appropriately seen as collective XSE dose in the early embryo. Our findings reiterate that correlations between X:A ratios and cell fates in other organisms need not implicate the value of the ratio as an active signal. PMID- 18162045 TI - Male or female? The answer depends on when you ask. PMID- 18162046 TI - A tertiary twist to the transglutaminase tale. PMID- 18162047 TI - Going against the grain. PMID- 18162048 TI - Cognitive dimensions of predator responses to imperfect mimicry. PMID- 18162049 TI - Sex, dose, and equality. PMID- 18162053 TI - Aberrant insulin receptor signaling and amino acid homeostasis as a major cause of oxidative stress in aging. AB - The mechanisms leading to the increase in free radical-derived oxidative stress in "normal aging" remains obscure. Here we present our perspective on studies from different fields that reveal a previously unnoticed vicious cycle of oxidative stress. The plasma cysteine concentrations during starvation in the night and early morning hours (the postabsorptive state) decreases with age. This decrease is associated with a decrease in tissue concentrations of the cysteine derivative and quantitatively important antioxidant glutathione. The decrease in cysteine reflects changes in the autophagic protein catabolism that normally ensures free amino acid homeostasis during starvation. Autophagy is negatively regulated by the insulin receptor signaling cascade that is enhanced by oxidative stress in the absence of insulin. This synopsis of seemingly unrelated processes reveals a novel mechanism of progressive oxidative stress in which decreasing antioxidant concentrations and increasing basal (postabsorptive) insulin receptor signaling activity compromise not only the autophagic protein catabolism but also the activity of FOXO transcription factors (i.e., two functions that were found to have an impact on lifespan in several animal models of aging). In addition, the aging-related decrease in glutathione levels is likely to facilitate certain "secondary" disease-related mechanisms of oxidative stress. Studies on cysteine supplementation show therapeutic promise. PMID- 18162054 TI - Rac and PI3 kinase mediate endothelial cell-reactive oxygen species generation during normoxic lung ischemia. AB - Abrupt reduction of flow (ischemia) leads to endothelial cell membrane depolarization, NADPH oxidase activation, and reactive oxygen species (ROS) generation in isolated rat and mouse lungs and in flow-adapted endothelial cells in vitro. Here we evaluated the role of PI-3-kinase and rac in activation of endothelial NADPH oxidase. Endothelium of isolated perfused mouse lungs labeled with 2',7'-dichlorodihydrofluorescein (H(2)DCF) or hydroethidine (HE) showed increased ROS generation with ischemia; these results were supported by TBARS measurement in whole-lung homogenate and by in vitro studies using flow-adapted mouse pulmonary microvascular endothelial cells. Ischemia-induced ROS generation in intact lung or isolated cells was blocked by pretreatment with Clostridium difficile toxin B, a rac inhibitor, and by wortmannin or LY294002, PI3 kinase inhibitors. In cells, immunofluorescence and immunoblot after subcellular fractionation showed ischemia-induced translocation of rac, p47(phox), and p67(phox) to the plasma membrane. Increased extracellular K(+) also resulted in rac translocation, providing evidence that this pathway is sensitive to alterations of endothelial cell membrane potential. These results indicate that PI-3-kinase and the small G protein rac are involved in the activation of endothelial cell NADPH oxidase that is associated with the acute loss of shear stress. PMID- 18162055 TI - BRCA1-mediated ubiquitination inhibits topoisomerase II alpha activity in response to oxidative stress. AB - Topoisomerase IIalpha is known to be critically involved in both cell proliferation and cell death. The mechanisms responsible for stress-dependent topoisomerase IIalpha alterations, however, remain unclear. This study focused on the behavior of topoisomerase IIalpha in response to oxidative stress induced by hydrogen peroxide (H(2)O(2)). The catalytic activity of topoisomerase IIalpha in MOLT-4 cells treated with H(2)O(2) decreased in parallel with the alteration of topoisomerase IIalpha expression. The ubiquitination of topoisomerase IIalpha was dependent on oxidative stress. BRCA1, a tumor-suppressor gene, appeared to be involved in these alterations in topoisomerase IIalpha. Furthermore, the retinoblastoma protein (pRb) was required for the ubiquitination of topoisomerase IIalpha by BRCA1. We conclude that the functions of topoisomerase IIalpha are regulated by ubiquitination on exposure to oxidative stress. PMID- 18162056 TI - Alexithymia in multiple sclerosis: relationship with fatigue and depression. AB - OBJECTIVE: The aim of this study was to investigate the prevalence of alexithymia in a sample of patients with multiple sclerosis (MS) and to further evaluate the association between alexithymia and the occurrence of common disabling MS-related symptoms such as fatigue and depression. METHODS: Fifty-eight relapsing-remitting MS patients treated with interferon (IFN)-beta-1a underwent a complete neurological evaluation, including Expanded Disability Status Scale score assessment. Alexithymia, depressive symptoms and fatigue were assessed using the 20-item Toronto Alexithymia Scale, Beck Depression Inventory and Fatigue Severity Scale. RESULTS: Prevalence of alexithymia was 13.8%, with 27.6% of patients presenting borderline alexithymia. Sixty-seven per cent of the patients complained of fatigue while 29.3% of them were depressed. Higher levels of fatigue and depression were found in alexithymic patients when compared with non alexithymic patients. Results from logistic regressions showed that alexithymia significantly contributes to the severity of fatigue and depression. CONCLUSIONS: Alexithymia was associated with increased severity of fatigue and depression. PMID- 18162057 TI - Near-infrared spectroscopy provides an index of blood flow and vasoconstriction in calf skeletal muscle during lower body negative pressure. AB - AIM: Near-infrared spectroscopy (NIRS) has been used previously for forearm blood flow estimation at rest and during exercise. In this study we applied NIRS to selectively monitor deep calf oxygenated haemoglobin (Hb) responses in order to estimate blood flow changes in the calf muscle during lower body negative pressure (LBNP). The purpose of this study was to test the hypothesis that changes in calf skeletal muscle oxygenated-Hb, after the removal of superficial tissue responses, were related to blood flow changes during orthostatic stress, and to determine the efficacy of using NIRS measurements as an index of vasoconstriction. METHODS: Twenty-nine subjects participated in this study. All attempted a graded LBNP trial from baseline (0 mmHg) to -60 mmHg LBNP in 10 mmHg steps at 5-min intervals. Calf blood flow changes were estimated by oxygenated-Hb responses in relation to changes in mercury strain gauge plethysmography and muscle sympathetic nerve activity (MSNA). RESULTS: Calf selective deep oxygenated Hb decreased continuously from -10 mmHg LBNP. Regression analysis showed that oxygenated-Hb was significantly related to declines in plethysmography evaluations of blood flow [oxygenated-Hb = (-1.57 +/- 0.26) + (1.86 +/- 0.49) plethysmography, r(2) = 0.87 +/- 0.09]. Changes in MSNA (total activity) were also inversely related to oxygenated-Hb (slope < 0, P = 0.037; r(2) = 0.52 +/- 0.15). CONCLUSION: These results suggest that changes in selective deep calf oxygenated-Hb can be utilized to estimate calf muscle blood flow changes that are most likely caused by vasoconstriction during graded LBNP. PMID- 18162058 TI - Ocular characteristics in 10 children with long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency: a cross-sectional study with long-term follow-up. AB - PURPOSE: To present long-term ocular complications and electroretinographic (ERG) findings in children with long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency - a life-threatening metabolic disease - and the relation to age at diagnosis, treatment and other clinical parameters. METHODS: Ten children with LCHAD deficiency underwent repeated ophthalmological evaluations including ERG. RESULTS: All 10 children developed chorioretinal pathology. Regardless of age at diagnosis, initiation of treatment and age at examination, inter-individual differences were present. Profound chorioretinal atrophy, severe visual impairment and progressive myopia had developed in two teenagers. Milder chorioretinopathy with or without subnormal visual acuity was present in all other children. ERG was pathological in seven children. The chorioretinopathy often started in the peripapillary or perimacular areas. In one patient, unilateral visual impairment was associated with fibrosis. CONCLUSION: Early diagnosis and adequate therapy might delay but not prevent the progression of retinal complications. Late diagnosis with severe symptoms at diagnosis, neonatal hypoglycaemia and frequent decompensations may increase the progression rate of the chorioretinopathy. LCHAD deficiency, a potentially lethal disease, is sometimes difficult to diagnose. Unusual chorioretinal findings should alert the ophthalmologist to the long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency, especially if there is a history of neonatal hypoglycaemia or failure to thrive. PMID- 18162059 TI - Tilted disc syndrome may mimic false visual field deterioration. AB - PURPOSE: Tilted disc syndrome is a congenital anomaly of the eye characterized by mostly upper temporal visual field defects. The aim of the present study was to evaluate the effect of gradual myopic correction in the improvement of visual field defects associated with tilted disc syndrome. METHODS: The visual field was examined in 38 eyes of 24 patients using standard Goldmann perimetry. The isoptres IV-4e, I-4e, I-3e and I-2e were plotted. The defective isoptres were tested again with gradually increasing myopic correction until no further change was noted. RESULTS: The most common type of defect was a relative upper temporal defect (19 eyes). Temporal relative defects were found in five eyes, upper altitudinal field defects in six eyes, an enlarged blind spot in four eyes, and an inferior field defect in one eye. The visual field defect partly or totally disappeared with increased myopic correction in 18 (50%) eyes. The mean improvement was 17.0 +/- 6.2 degrees and the mean additional myopic correction was 3.1 +/- 1.5 D. CONCLUSIONS: Even a small change in near correction during visual field examination may imply worsened or improved visual field defects in tilted disc syndrome. To prevent a false interpretation of field deterioration in a patient with tilted disc syndrome and glaucoma, visual field assessment should include examination with the myopic correction that provides the maximal improvement of the defective visual field. PMID- 18162060 TI - Visual field loss after tadalafil: a case report. PMID- 18162061 TI - Intracameral triamcinolone acetonide to control postoperative inflammation following cataract surgery with phacoemulsification. AB - PURPOSE: To explore the efficacy, safety and tolerability of 1 mg intracameral triamcinolone acetonide (TA) in controlling ocular inflammation in patients undergoing cataract surgery. METHODS: Sixty eyes of 60 patients undergoing cataract extraction with phacoemulsification at the Department of Ophthalmology, Baskent University School of Medicine were randomized into two groups. After surgery, eyes in group A were injected with 1 mg/0.1 ml TA into the anterior chamber, but eyes in group B were not. Postoperatively; in group B, topical prednisolone acetate 1% eyedrops were administered six times per day for 7 days, then four times per day for 15 days, to control postoperative inflammation. In group A, topical corticosteroids were not used. To evaluate the efficacy of intracameral TA, anterior chamber cells, anterior chamber flare and conjunctival hyperaemia were measured on postoperative days 1, 7 and 30 by slit-lamp biomicroscopy. The safety of intracameral TA was evaluated by visual acuity measurements, intraocular pressure values and fundus examination. Tolerance variables were assessed by the degree of burning, stinging and blurred vision. RESULTS: Both treatments were equally effective in controlling postoperative inflammation following phacoemulsification. No statistically significant differences between groups were observed for the efficacy, safety and tolerance variables, and no serious adverse events were observed. CONCLUSIONS: Intracameral TA of 1 mg can effectively be used to control postoperative inflammation after uncomplicated cataract surgery with phacoemulsification. This makes it possible to decrease the dosage and duration of topical prednisolone acetate. PMID- 18162062 TI - A prospective study on intravitreal bevacizumab (Avastin) for neovascular age related macular degeneration of different durations. AB - PURPOSE: Choroidal neovascularization (CNV) accounts for 85-90% of severe visual impairment in age-related macular degeneration (AMD). Vascular endothelial growth factor (VEGF) is a major factor mediating angiogenesis, and VEGF inhibitors have become a new treatment modality. In this prospective study, we used bevacizumab (Avastin), a recombinant monoclonal antibody to VEGF, to treat neovascular AMD. METHODS: The case material comprised 36 subjects (26 females, 10 males) aged 65 88 years with subfoveal neovascular AMD with all subtypes of CNV. There were two categories of patients: category I, long-standing CNV (12 months or more), preoperative visual acuity (VA) 0.16 (mean); category II, CNV (duration < 12 months), preoperative VA 0.25 (mean). Evaluation protocol included the Early Treatment Diabetic Retinopathy Study (ETDRS) VA, clinical ophthalmological examination, fluorescein angiography and optical coherence tomography (OCT). Intravitreal injections of bevacizumab (Avastin) (IVB), 1.25 mg (0.05 ml), were given under an operating microscope and aseptic conditions in a theatre for surgery with intervals of 4 or 6 weeks during the first 3 months and subsequently according to clinical assessment. The follow-up was 6 months in all cases. RESULTS: At 6 months, mean VA had improved by 4.6 ETDRS letters in the entire case material (P = 0.001), by 3.9 letters in category I (duration 12 months or more) and by 6.0 letters in category II (duration < 12 months). A total of 148 IVB (mean 4.1 injections/eye) were delivered during 6 months, the first 3 months comprising 3.1 IVB (mean) and the last 3 months 1.0 IVB (mean). No eyes suffered visual decline of 15 ETDRS letters. Fluorescein angiograms displayed stabilization or regression of CNV activity; OCT showed resorption of intraretinal oedema and subretinal fluid. No severe complications occurred but recurrence was common, and repeated IVBs were necessary in most cases during the 6-month period. CONCLUSION: When addressing the issue of frequency of IBV, we observed that 6-week intervals were sufficient because VA and CNV lesions generally stabilized at 4 weeks. The gain in VA was promising in eyes with < 12 months CNV duration. Even in eyes with a longer CNV duration, a slight visual improvement was observed when retinal oedema resorbed, although subretinal fibrosis and general cellular damage certainly limited recovery. PMID- 18162063 TI - Correlation between retinal nerve fibre layer thickness and retinal sensitivity. AB - PURPOSE: To evaluate the relationship between retinal nerve fibre layer (RNFL) thickness measured by optical coherence tomography (OCT) and light threshold values obtained with the Micro Perimeter 1 (MP1). METHODS: Forty-nine normal subjects and 37 glaucoma patients were included. Thickness of the RNFL around the optic disc was measured with Stratus OCT scans, and static threshold perimetry was performed with the MP1 at the same visit. Total average and mean 12-clock hour RNFL thickness were measured with Stratus OCT. An automated 4-2-1 staircase strategy with Goldmann III stimuli was used for MP1. The correspondence between Stratus OCT and MP1 RNFL measurements in each clock-hour was best fit by a second order polynomial. RESULTS: Average RNFL thickness in the normal group was significantly higher than that in the glaucoma group in each clock hour. Comparing the normal and glaucomatous groups, retinal sensitivity differed significantly at 6, 7, 8 and 11 o'clock. In these regions, structure-function relationships were generally stronger than in other regions. CONCLUSIONS: Structure-function associations may be strong when loss of nerve fibres is severe. However, significant reduction in retinal sensitivity does not emerge until the atrophy of nerve fibres is severe. PMID- 18162064 TI - Impaired eyeblink conditioning in children with fetal alcohol syndrome. AB - BACKGROUND: Eyeblink conditioning (EBC) is a Pavlovian paradigm that involves contingent temporal pairing of a conditioned stimulus (e.g., tone) with an unconditioned stimulus (e.g., air puff). Animal studies have shown that binge consumption of alcohol during pregnancy impairs EBC and that this impairment is likely mediated by a loss of neurons in the inferior olive and the cerebellar cortex and deep nuclei, as well as by a reduction in neural plasticity in the cerebellar deep nuclei. METHODS: Short delay EBC was examined in 98 5-year-old children born to women from the Coloured (mixed ancestry) community in Cape Town, South Africa, who were recruited prenatally and are participating in the first prospective longitudinal study of children with fetal alcohol syndrome (FAS). FAS status was assessed at 5 years by expert dysmorphologists. Two sessions of 50 trials each were administered to the children; a third session was administered the following day to those children who did not meet criterion of 40% conditioned responses in session 2. RESULTS: Not a single child with FAS met criterion for conditioning as contrasted with 75.0% of the controls. Whereas 86.7% of the controls who were conditioned met criterion by the end of Session 2, a large proportion of the relatively few alcohol-exposed nonsyndromal children who conditioned did not do so until Session 3. These alcohol effects on EBC persisted after controlling for IQ. Three of 4 microcephalic children who were not exposed to alcohol were successfully conditioned. CONCLUSIONS: This is the first prospective study to demonstrate impaired EBC in children diagnosed with FAS. Successful EBC in a microcephalic group supports the inference that the EBC deficit is specific to prenatal alcohol exposure and a potential biomarker for diagnosis of exposed children lacking the distinctive FAS dysmorphology. Delay EBC has a high sensitivity for identifying individuals with a diagnosis of probable FAS. PMID- 18162065 TI - Effect of alcohol on miR-212 expression in intestinal epithelial cells and its potential role in alcoholic liver disease. AB - BACKGROUND AND AIMS: Alcohol-induced gut leakiness is a key factor in alcoholic liver disease (ALD); it allows endotoxin to enter the circulation and initiate liver damage. Zonula occludens 1 (ZO-1) protein is a major component of tight junctions that regulates intestinal permeability. microRNAs (miRNAs) are recently discovered regulatory molecules that inhibit expression of their target genes. THE AIMS OF OUR STUDY WERE: (i) to investigate the effect of alcohol on miRNA-212 (miR-212) and on expression of its predicted target gene, ZO-1, (ii) to study the potential role of miR-212 in the pathophysiology of ALD in man. METHODS: Using a TaqMan miRNA assay system, we measured miR-212 expression levels in colon biopsy samples from patients with ALD and in Caco-2 cells (a human intestinal epithelial cell line) treated with or without EtOH. We measured ZO-1 protein levels using western blots. ZO-1 mRNA was assayed using real-time PCR. Intestinal barrier integrity was measured using fluorescein sulfonic acid clearance and immunofluorescent staining for ZO-1. RESULTS: Ethanol increased miR-212 expression, decreased ZO-1 protein levels, disrupted tight junctions, and increased the permeability of monolayers of Caco-2 cells. An miR-212 over expression is correlated with hyperpermeability of the monolayer barrier. miR-212 levels were higher in colon biopsy samples in patients with ALD than in healthy controls; ZO-1 protein levels were lower. CONCLUSION: These data suggest a novel mechanism for alcohol-induced gut leakiness, one in which EtOH induces miR-212 over-expression which causes gut leakiness by down-regulating ZO-1 translation. This mechanism is a potential therapeutic target for leaky gut in patients with or at risk for ALD. PMID- 18162066 TI - Parenting mechanisms in links between parents' and adolescents' alcohol use behaviors. AB - BACKGROUND: Adolescence has been identified as a critical period with regard to the initiation and early escalation of alcohol use. Moreover, research on familial risk and protective processes provides independent support for multiple domains of parental influence on adolescent drinking; including parents' own drinking behaviors, as well as the practices they employ to socialize their children. Despite this prevalence of findings, whether and how these distinct associations are related to one another is still not entirely clear. METHODS: The present study used data from 4,731 adolescents and their parents to test the nature of associations between (a) parents' frequencies of alcohol use and intoxication, and lifetime alcohol-related problems, (b) adolescents' perceptions of the parenting that they receive, and (c) adolescents' prevalence of alcohol use and intoxication at 14 and 17(1/2) years of age. As such, multiple mediation modeling was used to assess whether parental alcohol use behaviors influence adolescent alcohol use directly, or if they operate through indirect associations with various aspects of parenting that subsequently influence adolescent use. RESULTS: Examination of simple associations demonstrated that maternal and paternal alcohol use behaviors were positively linked with adolescent use behaviors at 14 and 17(1/2) years of age. Likewise, several parenting behaviors were independently associated with both parental and adolescent drinking. Examined collectively, multivariate path analyses indicated that associations between parents' and adolescents' alcohol-related behaviors were mediated, in part, by adolescents' perceptions of the parenting that they received, especially at 14 years of age. Furthermore, perceived parental monitoring and discipline had unique mediating capabilities, net the effects of all other parenting behaviors. CONCLUSIONS: This study demonstrates that parenting is an important mediator of the association between parental and adolescent drinking practices. An important area for future research will be to study how adolescents can avoid alcohol related problems despite being reared within a risk laden parenting environment and/or having parents who drink frequently. PMID- 18162067 TI - DSM-IV criteria endorsement patterns in alcohol dependence: relationship to severity. AB - BACKGROUND: In DSM-IV, the diagnostic threshold for alcohol dependence (AD) is met when a patient presents with at least 3 of 7 criteria. We have computed the predictive value for each individual DSM-IV AD criterion, and examined subtypes of AD criteria endorsement patterns and their associated severity indicators for community-dwelling AD individuals. METHODS: We utilized data from the 2001 to 2002 National Epidemiological Survey on Alcohol and Related Conditions (NESARC). Positive predictive values (PPV) for DSM-IV AD were computed for each of the individual criteria. Patterns of criteria endorsements were identified by latent class analysis (LCA). Sociodemographic status, age of onset and duration of AD, patterns of drinking, and drinking treatment history, were conditional on DSM criteria endorsement clusters, as indicators of the respondents' clinical severity. RESULTS: At the individual criterion level, the single criterion with the greatest PPV was D7--"Activities given up" with approximately 95% of drinking individuals who endorsed this DSM criterion correctly diagnosed as having DSM-IV AD. In addition to D7, only D5--"Physical/Psychological problems", and D6--"Time spent" had a PPV for AD substantially >50%. The LCA of AD endorsement patterns yielded a 6-cluster solution. The most common response pattern (34.5% of those with AD) was endorsement of 5 criteria: D1--"Quit/Control," D2--"Larger/Longer," D3--"Tolerance," D4--"Withdrawal," and D5--"Physical/Psychological problems." The most severe cluster (14%) was comprised of those who were likely to endorse 7/7 criteria. Cluster 1 (8.3%) did not include an endorsement of withdrawal, despite a heavy pattern of alcohol consumption. Unmarried status was associated with more severe criteria endorsement patterns. CONCLUSIONS: The present findings indicate a Guttman-like scaling of endorsement which yielded associations with severity for some of the concurrent indicators included in the analysis. However, severity measures did not always increase with DSM-IV AD criterion endorsement counts. Although endorsement of 6/7 or 7/7 criteria was associated with greater severity across a variety of indicators, fewer criteria were randomly associated with these measures. These data do not support the use of AD symptom counts as a phenotypic dependent variable. At least 2 extant diagnostic criteria showed relatively low PPV for AD, indicating a need for further assessment of these criteria with new symptoms or re-wording of the current symptom items. PMID- 18162068 TI - Ethanol, saccharin, and quinine: early ontogeny of taste responsiveness and intake. AB - BACKGROUND: Rat pups demonstrate high levels of immediate acceptance of ethanol during the first 2 weeks of postnatal life. Given that the taste of ethanol is most likely perceived by infant rats as a combination of sweet and bitter, high intake of ethanol early in ontogeny may be associated with age-related enhanced responsiveness to the sweet component of ethanol taste, as well as with ontogenetic decreases in sensitivity to its bitter component. Therefore, the present study compared responsiveness to ethanol and solutions with bitter (quinine) and sweet (saccharin) taste in terms of intake and palatability across the first 2 weeks of postnatal life. METHODS: Characteristic patterns of responsiveness to 10% (v/v) ethanol, 0.1% saccharin, 0.2% quinine, and water in terms of taste reactivity and fluid intake were assessed in rat pups tested on postnatal day (P) 4, 9, or 12 using a new technique of on-line monitoring of fluid flow through a two-channel intraoral cannula. Taste reactivity included analysis of ingestive and aversive responses following six intraoral infusions of the test fluids. This taste reactivity probe was followed by the intake test, in which animals were allowed to voluntarily ingest fluids from an intraoral cannula. RESULTS: Pups of all ages showed more appetitive responses to saccharin and ethanol than to water or quinine. No age-related differences were apparent in taste responsiveness to saccharin and ethanol. However, the age-related pattern of ethanol intake drastically differed from that of saccharin. Intake of saccharin increased from P4 to P9 and decreased substantially by P12, whereas intake of ethanol gradually increased from P4 to P12. Intake of ethanol was significantly lower than intake of saccharin on P9, whereas P12 pups took in more ethanol than saccharin. CONCLUSION: The findings of the present study indicate ontogenetic dissociations between taste reactivity to ethanol and saccharin and intake of these solutions, and suggest that high acceptance of ethanol early in ontogeny may not be associated with its orosensory properties but rather with the pharmacological effects of ethanol. PMID- 18162069 TI - Dose-dependent deficits in dual interstimulus interval classical eyeblink conditioning tasks following neonatal binge alcohol exposure in rats. AB - BACKGROUND: Neonatal alcohol consumption in rats is widely used to model cerebellar injury arising from 3rd-trimester human fetal alcohol exposure. Binge alcohol exposure of 5 g/kg/day or more over postnatal days (PD) 4 to 9 in rats damages the cerebellum and consequently impairs classical eyeblink conditioning (EBC). The present study sought to identify deficits in EBC using doses lower than those that have been reported previously following alcohol exposure limited to PD4-9. Complex conditioned response (CR) timing tasks utilizing 2 interstimulus intervals (ISIs) were used to test the hypothesis that 3 g/kg/day of alcohol would produce early onset and early peaked CRs, whereas 4 and 5 g/kg/day would impair CR acquisition. METHODS: Five neonatal treatment groups were used: (1) undisturbed controls, (2) sham intubation controls, (3) 3 g/kg/day of alcohol, (4) 4 g/kg/day of alcohol, or (5) 5 g/kg/day of alcohol. Intubations occurred over PD4-9. In adulthood, rats were trained using ISI discrimination (Experiment 1) or temporal uncertainty (Experiment 2) EBC tasks. In ISI discrimination, 2 distinct conditioned stimuli (CSs; tone and light) are reinforced with a periocular shock unconditioned stimulus (US) at 2 different CS US intervals. Temporal uncertainty is identical in design with the exception that the same CS is presented at both CS-US intervals. RESULTS: Alcohol-exposed subjects were impaired in CR acquisition in a task- and dose-dependent fashion. CR deficits were most salient in the peak amplitude measure and occurred in both tasks following alcohol exposure at 4 and 5 g/kg/day. Alcohol at a dosage of 3 g/kg/day impaired CR acquisition only in ISI discrimination. All alcohol doses failed to produce short latency CRs in either task. Alcohol-exposed subjects displayed later-onset and later-peaked CRs to the long-ISI CS in ISI discrimination relative to controls. CONCLUSIONS: ISI discrimination training may be ideal to identify CR deficits resulting from neonatal exposure to moderate alcohol doses. Applications of this EBC task to humans may enable reliable early identification and diagnosis of individuals with fetal alcohol spectrum disorders. PMID- 18162070 TI - Decreased immunoreactivity of the melanocortin neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH) after chronic ethanol exposure in Sprague-Dawley rats. AB - BACKGROUND: The melanocortin (MC) system is composed of peptides that are cleaved from the polypeptide precursor proopiomelanocortin (POMC). Recent pharmacologic and genetic evidence suggests that MC receptor (MCR) signaling modulates neurobiologic responses to ethanol and ethanol intake. Because ethanol decreases POMC mRNA levels, we determined if exposure to an ethanol-containing diet (ED) would significantly reduce central immunoreactivity of the MC peptide alpha-MSH in rats. We also determined if ethanol exposure would alter the immunoreactivity of agouti-related protein (AgRP), an endogenous MCR antagonist. METHODS: Male Sprague-Dawley rats were given 18 days of access to normal rodent chow or a control diet (CD), or short-term (4 days) or long-term (18 days) access to an ED. At the end of the study, rats were perfused with 4% paraformaldehyde and their brains were sectioned into two sets for processing with alpha-MSH or AgRP immunohistochemistry. RESULTS: Rats exposed to an ED showed significant reductions of central alpha-MSH immunoreactivity relative to rats exposed to a control diet (CD) or normal rodent chow. Ethanol-induced reductions of alpha-MSH immunoreactivity were site-specific and were noted in regions of the hypothalamus and extended amygdala, as well as the paraventricular nucleus of the thalamus. Because there were no differences in body weights or caloric intake between the CD and ED groups, reductions of alpha-MSH immunoreactivity in ED-treated rats are best explained by ethanol exposure rather than altered energy balance. No significant ethanol-induced alterations in hypothalamic AgRP immunoreactivity were detected. CONCLUSIONS: The present study shows that ethanol site specifically reduces alpha-MSH immunoreactivity in rat brain. These observations, in tandem with recent pharmacologic and genetic studies, suggest that the endogenous MC system modulates neurobiologic responses to ethanol. Thus, compounds which target MCRs may prove to have therapeutic value in the treatment of excessive ethanol consumption and/or the symptoms associated with ethanol withdrawal. PMID- 18162071 TI - Acculturation, drinking, and alcohol abuse and dependence among Hispanics in the Texas-Mexico border. AB - BACKGROUND: Acculturation has been linked to an increased prevalence of alcohol related problems. However, most of the research has been conducted with Hispanic populations in metropolitan areas of the United States, none of which is on the U.S.-Mexico border. This study examines the association between acculturation, heavy episodic drinking, and DSM-IV alcohol abuse and dependence among Hispanics in the Texas-Mexico border. METHODS: The study used data from a survey conducted (2002 to 2003) along the Texas-Mexico border and included 472 male and 484 female Hispanic adults from El Paso, the Rio Grande Valley, and colonias. Based on the Acculturation Rating Scale for Mexican Americans-II scale, respondents were coded into 4 acculturation categories: "very Mexican oriented,""Mexican bicultural,""Anglo bicultural," or "very Anglo/Anglicized.". RESULTS: Acculturation was related to lower rates of alcohol use disorders among men and a higher frequency of heavy episodic drinking among women. Multivariate analyses indicate that men who report heavy episodic drinking and those who are "very Mexican,""bicultural Mexican," or "bicultural Anglo" are more at higher risk for alcohol abuse and/or dependence compared with "very Anglo/Anglicized" men. For women, acculturation level did not predict alcohol disorders. Statistical analyses included testing for bivariate associations and multivariate logistic regression predicting heavy episodic drinking alcohol abuse or dependence. CONCLUSIONS: This study suggests that acculturation has different effects on drinking for men and women. This finding needs some attention as literature also indicates that women drink more and may develop more alcohol-related problems as they acculturate. This increase in women's drinking is probably because of U.S. society's more liberal norms governing female drinking. The "bimodal" distribution of risk, in which only men in "very Anglo" group are at a lower risk than the others, may be unique to the Border. The association between acculturation and alcohol use disorders does not appear to be linear and the effect of acculturation is not uniform on individuals' drinking behavior. PMID- 18162072 TI - Blockade of the corticotropin releasing factor type 1 receptor attenuates elevated ethanol drinking associated with drinking in the dark procedures. AB - BACKGROUND: Drinking in the dark (DID) procedures have recently been developed to induce high levels of ethanol drinking in C57BL/6J mice, which result in blood ethanol concentrations (BECs) reaching levels that have measurable affects on physiology and/or behavior. The present experiments determined whether the increased ethanol drinking caused by DID procedures can be attenuated by pretreatment with CP-154,526; a corticotropin releasing factor type-1 (CRF1) receptor antagonist. METHODS: In Experiment 1, male C57BL/6J mice received ethanol (20% v/v) in place of water for 4 hours, beginning with 3 hours into the dark cycle. On the fourth day, mice were given an intraperitoneal injection of one of the 4 doses of CP-154,526 (0, 1, 3, 10 mg/kg) 30 minutes before receiving their ethanol bottle. In Experiment 2, C57BL/6J mice had 2 hours of access to the 20% ethanol solution, beginning with 3 hours into the dark cycle on days 1 to 3, and 4 hours of access to the ethanol bottle on day 4 of DID procedures. Mice were given an intraperitoneal injection of one of the 4 doses of CP-154,526 (0, 1, 3, 10 mg/kg) 30 minutes before receiving their ethanol bottle on day 4. Tail blood samples were collected immediately after the 4-hour ethanol access period on the fourth day of each experiment. Additional control experiments assessed the effects of CP-154,526 on 4-hour consumption of a 10% (w/v) sucrose solution and open-field locomotor activity. RESULTS: In Experiment 1, the vehicle-treated group consumed approximately 4.0 g/kg/4 h of ethanol and achieved BECs of approximately 30 mg%. Furthermore, pretreatment with the CRF1 receptor antagonist did not alter ethanol consumption. On the other hand, procedures used in Experiment 2 resulted in vehicle-treated mice consuming approximately 6.0 g/kg/4 h of ethanol with BECs of about 80 mg%. Additionally, the 10 mg/kg dose of CP 154,526 significantly reduced ethanol consumption and BECs to approximately 3.0 g/kg/4 h and 27 mg%, respectively, relative to vehicle-treated mice. Importantly, the 10 mg/kg dose of the CRF1R antagonist did not significantly alter 4-hour sucrose consumption or locomotor activity. CONCLUSIONS: These data indicate that CRF1R signaling modulates high, but not moderate, levels of ethanol drinking associated with DID procedures. PMID- 18162073 TI - Key role of ethanol-derived acetaldehyde in the motivational properties induced by intragastric ethanol: a conditioned place preference study in the rat. AB - BACKGROUND: Acetaldehyde (ACD), the first metabolite of ethanol (EtOH), is produced peripherally by gastric and hepatic alcohol dehydrogenase (ADH) and centrally by brain catalase. In spite of the aversive properties classically ascribed to ACD, it has recently been suggested that ACD might mediate some of the motivational effects of EtOH. Accordingly, the relative role of ACD in the positive motivational properties of EtOH ingested is increasingly becoming the matter of debate. Thus, we studied the ability of intragastrically administered EtOH, ACD and EtOH-derived ACD to induce conditioned place preference (cpp) in rats. METHODS: Wistar rats were pretreated intraperitoneally with saline, the peripheral competitive inhibitor of ADH, 4-methylpyrazole (4-MP, 22.5, 45 or 67.5 mg/kg) or with the selective ACD-sequestrating agent, d-penicillamine (DP, 25 or 50 mg/kg), before the intragastric administration of saline, EtOH (0.5, 1 or 2 g/kg) or ACD (10, 20, or 40 mg/kg). The specificity of 4-MP and DP effects was addressed using morphine-induced cpp (2.5 mg/kg). RESULTS: Both, EtOH and ACD dose-dependently induced cpp; further, while EtOH-induced cpp was prevented by the administration of 4-MP and by DP, ACD-induced cpp was unaltered by 4-MP administration and prevented by DP. Both pretreatments did not interfere with morphine-induced cpp indicating that 4-MP and DP specifically modulate the motivational properties of EtOH and ACD. CONCLUSION: The ability of 4-MP and DP to decrease EtOH-induced cpp suggests that a reduction of ACD levels is crucial in depriving EtOH from its motivational properties as indexed by the cpp procedure. In addition, this conclusion is supported by the inefficacy of 4-MP in preventing ACD-induced cpp, and by its blockade observed after administration of the selective ACD sequestrating agent DP. The present results underscore the role of EtOH-derived ACD in EtOH-induced motivational properties as well as its abuse liability. PMID- 18162074 TI - The CRF-1 receptor antagonist, CP-154,526, attenuates stress-induced increases in ethanol consumption by BALB/cJ mice. AB - BACKGROUND: Corticotropin-releasing factor (CRF) signaling modulates neurobiological responses to stress and ethanol, and may modulate observed increases in ethanol consumption following exposure to stressful events. The current experiment was conducted to further characterize the role of CRF1 receptor (CRF1R) signaling in stress-induced increases in ethanol consumption in BALB/cJ and C57BL/6N mice. METHODS: Male BALB/cJ and C57BL/6N mice were given continuous access to 8% (v/v) ethanol and water for the duration of the experiment. When a baseline of ethanol consumption was established, animals were exposed to 5 minutes of forced swim stress on each of 5 consecutive days. Thirty minutes before each forced swim session, animals were given an intraperitoneal injection of a 10 mg/kg dose of CP-154,526, a selective CRF1R antagonist, or an equal volume of vehicle. The effect of forced swim stress exposure on consumption of a 1% (w/v) sucrose solution was also investigated in an ethanol-naive group of BALB/cJ mice. RESULTS: Exposure to forced swim stress significantly increased ethanol consumption by the BALB/cJ, but not of the C57BL/6N, mice. Stress-induced increases in ethanol consumption were delayed and became evident approximately 3 weeks after the first stressor. Additionally, forced swim stress did not cause increases of food or water intake and did not promote delayed increases of sucrose consumption. Importantly, BALB/cJ mice pretreated with the CRF1R antagonist showed blunted stress-induced increases in ethanol intake, and the CRF1R antagonist did not influence the ethanol drinking of non-stressed mice. CONCLUSIONS: The present results provide evidence that CRF1R signaling modulates the delayed increase of ethanol consumption stemming from repeated exposure to a stressful event in BALB/cJ mice. PMID- 18162075 TI - The reinforcing properties of salsolinol in the ventral tegmental area: evidence for regional heterogeneity and the involvement of serotonin and dopamine. AB - BACKGROUND: Salsolinol (SAL), the condensation product of acetaldehyde and dopamine, may be a factor contributing to alcohol abuse. Previous research indicated that both ethanol and acetaldehyde are self-administered into the posterior ventral tegmental area (VTA). The current study examined SAL self infusions into the VTA, and determined the involvement of dopamine neurons and 5 HT3 receptors in this process. METHODS: The intracranial self-administration technique was used to determine the self-infusion of SAL into the VTA of adult, male Wistar rats. The rats were placed in 2-lever (active and inactive) experimental chambers, and allowed to respond for the self-infusion of 0, 0.03, 0.1, 0.3, 1.0 or 3.0 microM SAL into the posterior or anterior VTA. In a second experiment, rats self-administered 0.3 microM SAL for the initial 4 sessions, co administered SAL with ICS-205,930 (a 5-HT3 receptor antagonist) or quinpirole (a D(2,3) receptor agonist) for sessions 5 and 6, and then only 0.3 microM SAL for session 7. RESULTS: Wistar rats, given 0.03 to 0.3 microM SAL, received more infusions per session than did the group given artificial cerebrospinal fluid (aCSF) alone (e.g., 41 infusions for 0.1 microM SAL versus 9 infusions for the aCSF group), and responded more on the active than inactive lever. These effects were observed in the posterior but not in anterior VTA. Co-infusion of 100 microM ICS-205,930, or quinpirole significantly reduced self-infusions and active lever responding. CONCLUSIONS: SAL produces reinforcing effects in the posterior VTA of Wistar rats, and these effects are mediated by activation of DA neurons and local 5-HT3 receptors. PMID- 18162076 TI - A comparison of results from an alcohol survey of a prerecruited Internet panel and the National Epidemiologic Survey on Alcohol and Related Conditions. AB - BACKGROUND: Given today's telecommunications environment, random digit dial (RDD) telephone surveys face declining response rates and coverage, and increasing costs. As an alternative to RDD, we surveyed participants in a randomly recruited standing Internet panel supplemented with a randomly sampled telephone survey of nonpanel members for a study of associations between onset of alcohol use and later alcohol-related problems. The purpose of this paper was to compare results from our survey with results from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a face-to-face probability sample survey of 43,093 adults, with a focus on associations between demographics, age of drinking onset, and alcohol dependence. METHODS: Demographic and drinking characteristics from our survey of 4,021 ever-drinkers between the ages of 18 and 39 years were compared with the characteristics of 11,549 similarly aged ever-drinkers from the NESARC. Weighted analyses accounting for sampling design compared these 2 samples on drinking characteristics over the past year and during a respondent's heaviest period of drinking, and in multivariate models examining associations between demographics, age of drinking onset, and lifetime alcohol dependence. RESULTS: Participants in the supplemented Internet panel were similar to the national population of 18- to 39-year-old ever drinkers on gender, education, and race/ethnicity, while adults who were aged 18 to 25 years were under-represented in the Internet panel. The supplemented Internet panel reported higher rates of moderate risk drinking over the past 12 months, lifetime high-risk drinking, and lifetime (ever) alcohol dependence. Estimates of the associations between alcohol dependence and age of drinking onset, risky drinking, and family history of alcohol problems did not significantly differ between the supplemented Internet sample and the NESARC survey. CONCLUSIONS: Randomly recruited Internet-based panels may provide an alternative to random digit dial telephone surveys and in person surveys for some studies of factors associated with alcohol-related problems. PMID- 18162077 TI - Regulation of motivation to self-administer ethanol by mGluR5 in alcohol preferring (P) rats. AB - BACKGROUND: Emerging evidence indicates that Group I metabotropic glutamate receptors (mGluR1 and mGluR5) differentially regulates ethanol self administration in several rodent behavioral models. The purpose of this work was to further characterize involvement of Group I mGluRs in the reinforcing effects of ethanol using a progressive ratio schedule of reinforcement. METHODS: Alcohol preferring (P) rats were trained to self-administer ethanol (15% v/v) versus water on a concurrent schedule of reinforcement, and the effects of the Group I mGluR antagonists were evaluated on progressive ratio performance. The rats were then trained to self-administer sucrose (0.4% w/v) versus water, and the effects of the antagonists were tested on progressive ratio performance. RESULTS: The mGluR1 antagonist, 3,4-dihydro-2H-pyrano[2,3]b quinolin-7-yl (cis-4 methoxycyclohexyl) methanone (JNJ 16259685; 0 to 1 mg/kg) and the mGluR5 antagonist, 6-methyl-2-(phenylethynyl) pyridine (MPEP; 0 to 10 mg/kg) dose dependently reduced ethanol break point. In separate locomotor activity assessments, the lowest effective dose of JNJ 16259685 (0.3 mg/kg) produced a motor impairment, whereas the lowest effective dose of MPEP (3 mg/kg) did not. Thus, the reduction in ethanol break point by mGluR1 antagonism was probably a result of a motor impairment. JNJ 16259685 (0.3 mg/kg) and MPEP (10 mg/kg) reduced sucrose break point and produced motor impairments. Thus, the reductions in sucrose break point produced by both Group I antagonists were probably because of nonspecific effects on motor activity. CONCLUSIONS: Together, these results suggest that glutamate activity at mGluR5 regulates motivation to self-administer ethanol. PMID- 18162078 TI - Ethanol increases fetal human neurosphere size and alters adhesion molecule gene expression. AB - BACKGROUND: Ethanol (ETOH) consumption by pregnant women can result in Fetal Alcohol Spectrum Disorder (FASD). To date, the cellular targets and mechanisms responsible for FASD are not fully characterized. Our aim was to determine if ETOH can affect fetal human brain-derived neural progenitor cells (NPC). METHODS: Neural progenitor cells were isolated by positive selection from normal second trimester fetal human brains (n = 4) and cultured, for up to 72 hours, in mitogenic media containing 0, 1, 10, or 100 mM ETOH. From 48 to 72 hours in culture, neurospheres generated in these conditions were filmed using time-lapse video microscopy. At the end of 72 hours, neurosphere diameter and roundness were measured using videographic software. Mitotic phase analysis of cell-cycle activity and apoptotic cell count were also performed at this time, by flow cytometry using propidium iodide (PI) staining. Real-time PCR was used to estimate expression of genes associated with cell adhesion pathways. RESULTS: Neurosphere diameter correlated positively (r = 0.87) with increasing ETOH concentrations. There was no significant difference in cell-cycle activity and no significant increase in apoptosis with increasing ETOH concentrations. Time-lapse video microscopy showed that ETOH (100 mM) reduced the time for neurosphere coalescence. Real-time PCR analysis showed that ETOH significantly altered the expression of genes involved in cell adhesion. There was an increase in the expression of alpha and beta Laminins 1, beta Integrins 3 and 5, Secreted phosphoprotein1 and Sarcoglycan epsilon. No change in the expression of beta Actin was observed while the expression of beta Integrin 2 was significantly suppressed. CONCLUSIONS: ETOH had no effect on NPC apoptosis but, resulted in more rapid coalescence and increased volume of neurospheres. Additionally, the expression of genes associated with cell adhesion was significantly altered. ETOH induced changes in NPC surface adhesion interactions may underlie aspects of neurodevelopmental abnormalities in FASD. PMID- 18162079 TI - Effects of ethanol on cytokine production after surgery in a murine model of gram negative pneumonia. AB - BACKGROUND: Both alcohol abuse and surgery have been shown to impair immune function. The frequency of postoperative infectious complications is 2- to 5-fold increased in long-term alcoholic patients, leading to prolonged hospital stay. Following surgery, an increase in interleukin (IL)-6 has been shown to be associated with increased tissue injury and interleukin 1-(IL-10) is known to represent an anti-inflammatory signal. The purpose of this study was to test the hypothesis that several days of excess alcohol consumption results in more pronounced immunosuppression. We assume that alcoholic animals show increased levels of IL-10 in response to infection and increased IL-6 due to a more pronounced lung pathology. METHODS: Thirty-two female Balb/c mice were pretreated with ethanol (EtOH) at a dose of (3.8 mg/g body weight) or saline (NaCl) for 8 days. At day 8 of the experiment all mice underwent a median laparotomy. Two days postsurgery mice were either applicated 10(4) CFU Klebsiella pneumoniae or received sham-infection with saline. A total number of 4 groups (EtOH/K. pneumoniae; NaCl/K. pneumoniae; EtOH/Sham-infection, NaCl/Sham-infection) was investigated and a clinical score evaluated. Twenty-four hours later mice were killed; lung, spleen, and liver were excised for protein isolation and histological assessment. IL-6 and IL-10 levels were detected by ELISA. RESULTS: Alcohol-exposed mice exhibited a worsened clinical appearance. The histological assessment demonstrated a distinct deterioration of the pulmonary structure in alcohol-treated animals. In the lung, IL-6 and IL-10 was significantly increased in alcohol-exposed infected mice compared to saline-treated infected mice. The clinical score correlated significantly with IL-6 (r = 0.71; p < 0.01) and IL-10 levels (r = 0.64; p < 0.01) in the lung. CONCLUSIONS: Ethanol treatment in this surgical model led to a more severe pulmonary infection with K. pneumoniae which was associated with more tissue destruction and increased levels of IL-6 and IL 10 and a worsened clinical score. PMID- 18162080 TI - Lesions of the extended amygdala in C57BL/6J mice do not block the intermittent ethanol vapor-induced increase in ethanol consumption. AB - BACKGROUND: The central extended amygdala (cEA) which includes the central nucleus of the amygdala (CeA) and the lateral posterior bed nucleus of the stria terminalis (BNSTLP), has been proposed to play a key role in excessive ethanol consumption in humans (Koob and Le Moal, 2005 Nat Neurosci 8:1442). To examine this relationship, we used a murine model of ethanol dependence (Becker and Lopez, 2004 Alcohol Clin Exp Res 28:1829; Lopez and Becker, 2005 Psychopharmacology (Berl) 181:688) and compared animals with sham lesions and electrolytic lesions of the CeA and BNSTLP. METHODS: Male C57BL/6J (B6) mice were first acclimated to a limited-access 2-bottle-choice preference procedure. The access period began 3 hours into the dark phase of the light-dark cycle and continued for 2 hours. Once acclimated (1 week), mice underwent chronic exposure to and intermittent withdrawal from ethanol vapor. The animals were then retested in the limited-access 2-bottle-choice preference procedure. In some experiments, electrolytic and sham lesions of the CeA or BNSTLP were performed prior to initiating the 2-bottle choice procedure. RESULTS: In a series of 5 preliminary experiments, mice were randomly assigned either to the standard intermittent ethanol vapor procedure or to the standard procedure but with air in the vapor chamber (control). The air-control procedure produced no change in ethanol intake when compared to baseline consumption. In contrast, intermittent ethanol vapor exposure increased ethanol consumption by almost 50%. The increase in consumption was associated with an increase in total fluid volume consumed and no change in ethanol preference. Lesions of both the BNSTLP and CeA significantly decreased baseline ethanol consumption, the former by decreasing fluid consumption and the latter by decreasing ethanol preference. Intermittent ethanol vapor exposure significantly increased consumption in both the BNSTLP- and CeA-lesioned animals, largely by increasing the total volume of fluid consumed. CONCLUSIONS: The results obtained clearly demonstrate that the cEA has a role in the regulation of ethanol consumption in the limited-access procedure. However, neither lesions of the CeA nor BNSTLP prevented the intermittent ethanol vapor-induced increase in consumption. These data do not preclude some role of the cEA in the increased ethanol consumption following intermittent ethanol vapor exposure, but would suggest that other brain regions also must have a significant influence. PMID- 18162081 TI - The association between coeliac disease and cardiovascular disease. AB - BACKGROUND: Coeliac disease is more prevalent than was previously thought. The association between coeliac disease and cardiovascular outcome is not clear. AIM: To investigate whether coeliac disease patients have an increased risk of cardiovascular events. METHODS: A community-based cohort study using a record linkage database. Three hundred and sixty-seven coeliac patients identified by a positive antiendomysial antibody test or a diagnosis with small bowel biopsy, and 5537 subjects who were tested and had a negative coeliac immunology, were included in the study. RESULTS: The crude rates of cardiovascular events were 9.5 per 1000 person-years (95% CI: 4.4-14.6) in the coeliac cohort and 8.9 per 1000 person-years (95% CI: 7.6-10.3) in the antiendomysial antibody-negative cohort. Compared with the antiendomysial antibody-negative cohort, the adjusted relative risk of cardiovascular events for coeliac cohort was 1.9 (95% CI: 1.00-3.60). When we excluded patients who had previous hospitalization for cardiovascular disease, the adjusted relative risk was 2.5 (95% CI: 1.22-5.01). The use of any cardiovascular drugs prior to and after entry to the study were 36% and 29% for the coeliac cohort (P = 0.05), and 34% and 26% for the antiendomysial antibody negative cohort (P < 0.01). CONCLUSION: Our findings suggest that coeliac disease seems to be associated with an increased risk of cardiovascular outcome. PMID- 18162082 TI - Review article: endoscopic antireflux procedures - an unfulfilled promise? AB - BACKGROUND: Most published reviews concerning the endoscopic treatment of gastro oesophageal reflux disease date back to 2005. AIM: To provide an updated review that includes all papers published up to 2007. METHODS: A Medline search from January 2005 to June 2007 was performed regarding endoscopic procedures aiming at treating gastro-oesophageal reflux disease. In addition, we retrieved the abstracts presented at Digestive Disease Week during the last 3 years. We included in the review both 'mechanistic' studies - that is, papers exploring the potential mechanism of action of the procedure/device - and studies trying to assess its clinical efficacy. RESULTS: During the last 3 years, the number of published papers has declined, and some devices are not available any more. The alleged mechanism(s) of action of the various devices or procedures is (are) still not completely elucidated; however, some concerns have arisen as far as durability and potential detrimental effects. Moreover, all the aspects of endoscopic therapy, except for its safety, are either insufficiently explored or not investigated at all, or assessed only in particularly selected patient subgroups. CONCLUSIONS: None of the proposed antireflux therapies has fulfilled the criteria of efficacy, safety, cost, durability and, possibly, of reversibility. There is at present no definite indication for endoscopic therapy of gastro-oesophageal reflux disease. We suggest a list of recommendations to be followed when a new endoscopic therapeutic procedure is to be assessed for use in clinical practice. PMID- 18162083 TI - Intravenous esomeprazole 40 mg vs. intravenous lansoprazole 30 mg for controlling intragastric acidity in healthy adults. AB - BACKGROUND: Intravenous (IV) formulations of proton pump inhibitors are effective for patients in whom oral therapy is not appropriate. AIM: To compare IV esomeprazole and IV lansoprazole for the control of intragastric pH. METHODS: In this open-label crossover study, healthy, Helicobacter pylori-negative adults were randomized to one of two treatment sequences, each consisting of two 5-day dosing periods of IV esomeprazole 40 mg or IV lansoprazole 30 mg. Twenty-four hour intragastric pH monitoring was conducted on days 1 and 5 of each dosing period. RESULTS: On days 1 and 5, intragastric pH was >4.0 significantly longer with esomeprazole than lansoprazole (least-squares means: day 1, 40.0% vs. 33.6%; day 5, 61.9% vs. 45.4%; both P < 0.0001). During the first 4 h of pH monitoring, intragastric pH was >4.0 significantly longer on days 1 and 5 with esomeprazole than lansoprazole (P < 0.0001). Kaplan-Meier estimates of median hours to stable pH >4.0 were 4.92 for esomeprazole and 5.75 for lansoprazole (P = 0.0014 for test on Gehan scores). CONCLUSION: In healthy adults, IV esomeprazole 40 mg controlled intragastric acidity faster and more effectively than IV lansoprazole 30 mg. PMID- 18162084 TI - Patterns of quantitative food-specific IgE-antibodies and reported food hypersensitivity in 4-year-old children. AB - BACKGROUND: Diagnosis of food hypersensitivity (FHS) is difficult and interpretation of food allergy tests is complicated. OBJECTIVE: To investigate the probability of reported FHS in relation to levels of food-specific IgE antibodies (AB) in a population-based setting of 4-year-old children (n = 2336). METHODS: Information on FHS was obtained from a questionnaire and specific IgE-AB to milk, egg, fish, peanut, soy and wheat were analysed. RESULTS: Thirty-one per cent of the children with reported FHS (n = 284) were sensitized (> or =0.35 kU(A)/l) to at least one of the tested foods compared with 11% of children without FHS (n = 2052). Furthermore, the probability of reported symptoms to milk, egg and fish increased with increasing levels of food-specific IgE-AB to the same food allergens. A similar trend was seen for peanut and wheat, but not for soy. Increasing levels of specific IgE-AB to milk or egg were also associated with an increasing risk of reported symptoms caused by other foods. CONCLUSIONS: Quantitative measurements of IgE-AB to milk, egg and fish are useful to evaluate IgE-associated FHS in preschool children also in a population based sample. Such measurements appear to be of limited value for soy bean and wheat, in particular as a screening method. PMID- 18162085 TI - The ATG16L1 gene variants rs2241879 and rs2241880 (T300A) are strongly associated with susceptibility to Crohn's disease in the German population. AB - OBJECTIVES: We analyzed ATG16L1, a recently identified Crohn's disease (CD) susceptibility gene, in a large cohort with inflammatory bowel disease (IBD) including potential interactions with other IBD genes as well as factors regulating its gene expression. METHODS: Genomic DNA from 2,890 Caucasians including 768 patients with CD, 507 patients with ulcerative colitis (UC), and 1,615 healthy controls was analyzed for 9 different ATG16L1 single nucleotide polymorphisms (SNPs). Genotyping included CARD15/NOD2 variants p.Arg702Trp, p.Gly908Arg, and p.Leu1007fsX1008 and polymorphisms in SLC22A4/OCTN1 (1672 C-->T) and SLC22A5/OCTN2 (-207 G-->C) as well as 10 CD-associated IL23R variants. The transcriptional regulation of ATG16L1 was studied in intestinal epithelial cells following stimulation with Toll-like receptor (TLR) ligands and proinflammatory cytokines and in a murine ileitis model and CD biopsies. RESULTS: All nine ATG16L1 gene variants analyzed displayed highly significant associations with CD demonstrating a CD-protective effect for the minor allele. The strongest associations were found for rs2241879 and the coding SNP rs2241880 (T300A); P= 3.6 x 10(-6) and 3.7 x 10(-6), respectively (OR 0.74, 95% CI 0.65-0.84 for both variants). The genotype-phenotype analysis revealed no significant associations. In UC, only rs6431660 was weakly disease-associated. There was no evidence for epistasis between the ATG16L1 gene and other susceptibility genes (IL23R, CARD15, SLC22A4/5). ATG16L1 mRNA expression was not upregulated in CD and murine ileitis, and was less than threefold increased in cells stimulated with proinflammatory cytokines and TLR ligands. CONCLUSION: ATG16L1 is a CD susceptibility gene without epistatic interaction with other CD susceptibility genes and is not upregulated in intestinal inflammation. PMID- 18162086 TI - Clinical relevance of preformed HLA donor-specific antibodies in kidney transplantation. AB - This study analyzes the influence of preformed DSA, identified by HLA-specific ELISA assays, on graft survival and evaluates the incidence of antibody-mediated rejection (AMR) in patients with and without pregraft desensitization. Kidney graft survival at 8 years was significantly worse in patients with DSA (n = 43) than in those without DSA (n = 194)(p = 0.03). The incidence of AMR in patients with DSA is 9-fold higher than in patients without DSA (p < 0.001) and their graft survival is significantly worse than in DSA patients without AMR and in non DSA patients (p = 0.005). The prevalence for AMR in patients with DSA detected on historic serum is 32.3% in nondesensitized patients and 41.7% in desensitized patients. The risk for AMR is significantly more elevated in patients with strongly positive DSA (score 6-8) compared to those with DSA score 4 (p < 0.001), and in patients with historic DSA+/CXM+ compared to those with DSA+/CXM- (p = 0.01). The presence of preformed DSA is strongly associated with graft loss in kidney transplants, related to an increased risk of AMR. Our findings demonstrate the importance of detection and characterization of DSA before transplantation. Stratification of this risk could be used to determine kidney allocation and to devise specific strategies for these patients. PMID- 18162087 TI - Why are nearly half of expanded criteria donor (ECD) kidneys not transplanted? PMID- 18162088 TI - Working together toward a national kidney paired donation program. PMID- 18162089 TI - Reducing bile leak following segmental liver transplantation: understanding biliary anatomy of the caudate lobe. AB - Bile leak in split and living donor liver transplantation is not an uncommon postoperative complication with significant morbidity to both donor and recipients. Nonanastomotic bile leaks in these transplants are less well characterized and generally described as cut-surface leaks. A proportion of these leaks may derive from biliary radicles draining the caudate lobe. Based on the caudate lobe biliary anatomy the authors describe measures that may help to reduce such complications after segmental liver transplantation. PMID- 18162090 TI - Risk factors for rejection and infection in pediatric liver transplantation. AB - Rejection and infection are important adverse events after pediatric liver transplantation, not previously subject to concurrent risk analysis. Of 2291 children (<18 years), rejection occurred at least once in 46%, serious bacterial/fungal or viral infections in 52%. Infection caused more deaths than rejection (5.5% vs. 0.6% of patients, p < 0.001). Early rejection (<6 month) did not contribute to mortality or graft failure. Recurrent/chronic rejection was a risk in graft failure, but led to retransplant in only 1.6% of first grafts. Multivariate predictors of bacterial/fungal infection included recipient age (highest in infants), race, donor organ variants, bilirubin, anhepatic time, cyclosporin (vs. tacrolimus) and era of transplant (before 2002 vs. after 2002); serious viral infection predictors included donor organ variants, rejection, Epstein-Barr Virus (EBV) naivety and era; for rejection, predictors included age (lowest in infants), primary diagnosis, donor-recipient blood type mismatch, the use of cyclosporin (vs. tacrolimus), no induction and era. In pediatric liver transplantation, infection risk far exceeds that of rejection, which causes limited harm to the patient or graft, particularly in infants. Aggressive infection control, attention to modifiable factors such as pretransplant nutrition and donor organ options and rigorous age-specific review of the risk/benefit of choice and intensity of immunosuppressive regimes is warranted. PMID- 18162091 TI - Do we know enough to mandate donor evaluation testing? PMID- 18162092 TI - Influenza vaccination is efficacious and safe in renal transplant recipients. AB - Whether influenza vaccination in solid-organ transplant recipients is efficacious remains a controversial issue. Furthermore, theoretical concerns have been raised regarding the safety of vaccination as it might trigger rejection of the allograft. The present prospective trial is aimed at investigating the antibody response and safety of influenza vaccination in renal transplant recipients (RTR). A total of 165 RTR and 41 healthy volunteers were vaccinated with a standard trivalent inactivated influenza vaccine. Hemagglutination-inhibiting (HI) antibodies were quantified before and 1 month after vaccination. Seroprotection (SP) and seroresponse (SR) were defined as a titer > or =40 and a 4-fold rise in HI titer, respectively. Similar SR rates were observed in both groups. Postvaccination SP rates in RTR amounted to 92.7%, 78.7% and 82.9% for A/H1N1, A/H3N2 and B, respectively. High baseline SP rates, most probably reflecting frequent preimmunizations, explain partly the high postvaccination SP rates. SR rate was independently and inversely associated with baseline SP rate. Mycophenolate mofetil (MMF) usage was associated with a 2.6-5-fold lower SR. Nonetheless, these patients showed good postvaccination SP rates. A booster dose did not enhance SP or SR rates. Influenza vaccination neither affected allograft function nor caused rejection episodes. In conclusion, influenza vaccination is efficacious and safe in renal transplantation. PMID- 18162093 TI - Comparison of histidine-tryptophan-ketoglutarate and University of Wisconsin preservation in renal transplantation. AB - Histidine-tryptophan-ketoglutarate (HTK) is replacing University of Wisconsin (UW) solution as the preservation fluid for renal allografts in many centers, but recent large-scale data to support this transition are lacking. We conducted a retrospective analysis of patient and graft outcomes after renal transplantation at our center, comparing 475 consecutive living donor and 317 deceased donor transplants since the adoption of HTK with equal numbers of grafts preserved using UW solution. Data collected included donor and recipient age, race, sex, comorbidities and graft ischemia time. Graft and patient survival, as well as the incidence of delayed graft function (DGF), were studied by Kaplan-Meier and Cox regression analysis. No significant difference was seen in either patient or graft survival. Deceased donor kidneys in the HTK group had a higher incidence of DGF than the UW cohort, whereas this trend was reversed in the case of living donor organs. In multivariate analysis, HTK was associated with a significant risk reduction on the incidence of DGF. Prolonged preservation with HTK compared to UW was not associated with excess risk to the graft or patient. In summary, HTK demonstrated efficacy similar to UW in terms of patient and graft survival. PMID- 18162094 TI - Histological evaluation of the sheep's cervical spine segment 3/4--a histomorphological and histomorphometrical study. AB - The 2-year-old sheep is a frequently used model in spinal orthopaedic research; however, so far no investigations have been performed on physiological tissue of untreated animals. In this study, the cervical spine segment 3/4 of eight 2-year old female Merino sheep was evaluated histologically and histomorphometrically. Specimens were stained with Masson-Goldner-Trichrome, Safranin-Orange/Lightgreen, Safranin-Orange/von Kossa and Astrablue. Analysis of bone mass, mass of cartilage and mineralized cartilage in a defined region of interest was performed. A new finding of the study was that--although 2-year-old sheep is regarded as adult- tissue of the growth plate was still detected. Moreover, this study revealed that in the nucleus pulposus of the 2-year-old sheep notochordal cells are still existent, suggesting a complex remodelling of the nucleus pulposus in sheep. PMID- 18162095 TI - Immunohistochemical localization of haptoglobin in porcine lungs. AB - The extravasation of erythrocytes into the lower respiratory tract occurs in numerous lung injuries and may lead to oxidative damages in lung tissues. Haptoglobin (Hp), the major haemoglobin-binding protein, is known to reduce lung injury associated with exposure to blood in mice. In pigs, Hp is a major acute phase protein and its serum concentrations are elevated in various infections of the respiratory tract. However, information on the porcine Hp response towards inflammatory stimuli is restricted to blood. We herein investigated the presence of Hp in lung tissues from pigs with acute and chronic bronchopneumonia via immunohistochemistry. Hp was localized in airway epithelial cells and immigrated leucocytes whereas in alveolar epithelial cells there was no distinct signal. Unaltered lungs showed less Hp-positive cells compared with lungs from pigs with acute or chronic bronchopneumonia. PMID- 18162096 TI - Bovine meroanencephaly and gastroschisis: a macro and microscopic study. AB - Congenital malformations correspond to one of the main causes of embryonic loss during the gestational process. They result from interaction of several factors such as multifactor heredity, chromosomal and genetic alterations and environmental agents; however, unknown aetiology also can be present. In this article, we have used 10 embryos, from a frigorific area of Dracena, SP, Brazil, which were fixed in Bouin solution for a macro- and micro-scopic description. We could verify the presence of an encephalic tissue mass on the embryo's dorsal cranial area, resulting from the non-formation of part of the cranial cap and from the non-closing of cephalic neuropore and consequent neuroepithelial cells disorganization. In the abdominal area, the embryos did not show the complete fusion of the body lateral pleats during the abdominal wall formation, and the liver extruded into the amniotic cavity without involvement of the intestine. PMID- 18162097 TI - Normal cross-sectional anatomy of the bovine digit: comparison of computed tomography and limb anatomy. AB - The purpose of this study was to define the structures of the digits and hoof in Holstein dairy cattle by using computed tomography scan (CT scan). Transverse, sagittal and dorsoplantar CT images of two isolated cattle cadaver digits were obtained using a Siemens ARTX2 Somatom. The CT images were compared to corresponding frozen cross-sections. Relevant anatomical structures were identified and labelled at each level. The CT images provided anatomical detail of the digits and hoof in Holstein dairy cattle. Transversal images provided excellent depiction of anatomical structures when compared to corresponding frozen cross-sections. The information presented in this paper would serve as an initial reference to the evaluation of CT images of the digits and hoof in Holstein dairy cattle. PMID- 18162098 TI - Arterial vascularization of the uropygial glands (gl. uropygialis) in the Japanese quail (Coturnix coturnix japonica) and silver polish (Gallus gallus domesticus). AB - This study focused on the morphological characteristics of the uropygial glands of the Japanese quail and silver polish, particularly the arterial nourishment. With this respect, coloured-latex injected animals were dissected and their glands and related arteries were observed. The gland in the Japanese quail was relatively longer than that in the silver polish. The first caudal segmental arteries in the Japanese quail were specialized as the uropygial gland artery while the fourth caudal segmental ones were continued as the uropygial gland artery in the silver polish. Branches from the second and third caudal segmental arteries and skin also contributed bilaterally to the vascularization of the gland in the Japanese quail. Observing the more complex arterial structure in the Japanese quail suggests more active uropygial gland in this species, when compared with the silver polish. PMID- 18162099 TI - Mapping of six miRNAs expressed in porcine skeletal muscle. PMID- 18162100 TI - Evaluation of six candidate genes for dilated cardiomyopathy in Irish wolfhounds. PMID- 18162101 TI - Linkage and association analyses of intragenic SNPs in the canine beta-crystallin genes CRYBB1, CRYBB2, CRYBB3, CRYBA1 and CRYBA4 with primary cataracts in wire haired Dachshunds. PMID- 18162102 TI - Mapping of the porcine FBN2, YWHAQ, CNN3, DCN, POSTN, SPARC, RBM39 and GNAS genes, expressed in foetal skeletal muscles. PMID- 18162103 TI - QTL for meat tenderness in the M. longissimus lumborum of cattle. AB - Meat tenderness has been difficult to improve using standard genetic selection. Marker assisted selection holds great promise if markers for meat tenderness can be identified. Here, we report quantitative trait loci (QTL) for beef tenderness identified in 598 animals of three Charolais-Brahman x Belmont Red pedigrees after screening the whole genome using 183 DNA markers. In addition to the usual Warner-Bratzler peak force measurements, tenderness was also measured using compression, adhesion and pressure-heat-treated peak force. Three QTL for meat tenderness in the M. longissimus lumborum muscle were found, two of which have not been reported before. One is located in the HEL9-CSSM47 interval on bovine chromosome 8 with a LOD of 3.1 and an effect of 1.02 phenotypic standard deviations for tensile strength of cooked muscle as measured by adhesion. A second QTL is located near CSRM60 on bovine chromosome 10 with a LOD of 2.4 and an effect of 0.48 phenotypic standard deviations for compression. The third QTL is in a region of bovine chromosome 7 that has previously been reported to have a QTL affecting peak force. This region also shows effects on compression and a combined tenderness index. These QTL are all for the myofibrillar component of meat tenderness. No QTL were found for pressure-treated peak force, which is an estimate of the connective tissue component muscle of meat tenderness. PMID- 18162104 TI - Fine-mapping of a marbling trait to a 2.9-cM region on bovine chromosome 7 in Japanese Black cattle. AB - To locate quantitative trait loci (QTL) for intramuscular fat deposition (marbling) in a local population of Japanese Black cattle, we performed a genome scan using a paternal half-sib family of Bull A. A marbling QTL was mapped in the region flanked by DIK0079 (20.7 cM) and TGLA303 (39.3 cM) on bovine chromosome (BTA) 7, affecting 5.0% of the total family variance. Haplotype analysis of the QTL region revealed that the marbling-increasing Q allele was transmitted from the dam. On the other hand, Bull B, a maternal half-sib of Bull A, did not receive the Q allele from its dam, based on the following findings: (i) a marbling QTL on BTA7 was not detected in the Bull B paternal half-sib family; (ii) recombination between DIK0079 (20.7 cM) and RM006 (25.4 cM) in the QTL region was observed in the maternal chromosome of Bull B; and (iii) the Q harbouring steers from Bull A exhibited significantly higher marbling than the steers from Bull B and the remaining steers from Bull A. To precisely compare the maternal chromosomes of both bulls, we constructed a bacterial artificial chromosome contig covering the region between DIK0079 and RM006 and developed DNA markers. The recombination occurred between DIK8042 and DIK8044, indicating that the marbling QTL was in a 2.9-cM region flanked by DIK0079 and DIK8044. PMID- 18162105 TI - The effect of a c.-8G>T polymorphism on the expression of cytochrome b5A and boar taint in pigs. AB - The level of cytochrome b5A (CYB5A) in pig testis is correlated with boar taint from androstenone and an AF016388:c.-8G>T polymorphism in CYB5A has been linked with low androstenone levels in the fat of pigs. In this study, we developed a polymerase chain reaction-based assay to genotype 1242 boars from eight lines for the c.-8G>T SNP. The c.-8T allele was found in all eight lines at a frequency ranging from 1.8% to 20.3% with an overall frequency of 8.6%. Significant deviations from Hardy-Weinberg equilibrium were found in the Hampshire, Landrace and Yorkshire breeds. The homozygous mutant c.-8TT occurred infrequently and was not found in some lines, but was consistently associated with low androstenone levels in fat. Both CYB5A mRNA and CYB5A protein levels were decreased in the c. 8TT genotype in a subset of Yorkshire boars, suggesting that low levels of CYB5A protein in the c.-8TT mutant were not due to inefficient translation of CYB5A mRNA. There were significant but modest marker effects on fat androstenone levels in Landrace, Yorkshire and a Large White/Duroc cross and fat skatole in Duroc and Sire Line breeds. There was no effect of CYB5A genotype on bulbourethral gland length, suggesting that this SNP will not affect reproductive traits. We conclude that the c.-8G>T SNP in the CYB5A gene has a significant but modest effect on boar taint in male pigs and could be useful in some breeds as part of a panel of SNP markers in a marker-assisted selection programme to produce low boar taint pigs. PMID- 18162106 TI - Bounds on direct effects in the presence of confounded intermediate variables. AB - This article considers the problem of estimating the average controlled direct effect (ACDE) of a treatment on an outcome, in the presence of unmeasured confounders between an intermediate variable and the outcome. Such confounders render the direct effect unidentifiable even in cases where the total effect is unconfounded (hence identifiable). Kaufman et al. (2005, Statistics in Medicine 24, 1683-1702) applied a linear programming software to find the minimum and maximum possible values of the ACDE for specific numerical data. In this article, we apply the symbolic Balke-Pearl (1997, Journal of the American Statistical Association 92, 1171-1176) linear programming method to derive closed-form formulas for the upper and lower bounds on the ACDE under various assumptions of monotonicity. These universal bounds enable clinical experimenters to assess the direct effect of treatment from observed data with minimum computational effort, and they further shed light on the sign of the direct effect and the accuracy of the assessments. PMID- 18162107 TI - Robust covariate-adjusted log-rank statistics and corresponding sample size formula for recurrent events data. AB - Recurrent events data are frequently encountered in clinical trials. This article develops robust covariate-adjusted log-rank statistics applied to recurrent events data with arbitrary numbers of events under independent censoring and the corresponding sample size formula. The proposed log-rank tests are robust with respect to different data-generating processes and are adjusted for predictive covariates. It reduces to the Kong and Slud (1997, Biometrika 84, 847-862) setting in the case of a single event. The sample size formula is derived based on the asymptotic normality of the covariate-adjusted log-rank statistics under certain local alternatives and a working model for baseline covariates in the recurrent event data context. When the effect size is small and the baseline covariates do not contain significant information about event times, it reduces to the same form as that of Schoenfeld (1983, Biometrics 39, 499-503) for cases of a single event or independent event times within a subject. We carry out simulations to study the control of type I error and the comparison of powers between several methods in finite samples. The proposed sample size formula is illustrated using data from an rhDNase study. PMID- 18162108 TI - Estimating cumulative treatment effects in the presence of nonproportional hazards. AB - Often in medical studies of time to an event, the treatment effect is not constant over time. In the context of Cox regression modeling, the most frequent solution is to apply a model that assumes the treatment effect is either piecewise constant or varies smoothly over time, i.e., the Cox nonproportional hazards model. This approach has at least two major limitations. First, it is generally difficult to assess whether the parametric form chosen for the treatment effect is correct. Second, in the presence of nonproportional hazards, investigators are usually more interested in the cumulative than the instantaneous treatment effect (e.g., determining if and when the survival functions cross). Therefore, we propose an estimator for the aggregate treatment effect in the presence of nonproportional hazards. Our estimator is based on the treatment-specific baseline cumulative hazards estimated under a stratified Cox model. No functional form for the nonproportionality need be assumed. Asymptotic properties of the proposed estimators are derived, and the finite-sample properties are assessed in simulation studies. Pointwise and simultaneous confidence bands of the estimator can be computed. The proposed method is applied to data from a national organ failure registry. PMID- 18162109 TI - Variable selection in penalized model-based clustering via regularization on grouped parameters. AB - Penalized model-based clustering has been proposed for high-dimensional but small sample-sized data, such as arising from genomic studies; in particular, it can be used for variable selection. A new regularization scheme is proposed to group together multiple parameters of the same variable across clusters, which is shown both analytically and numerically to be more effective than the conventional L(1) penalty for variable selection. In addition, we develop a strategy to combine this grouping scheme with grouping structured variables. Simulation studies and applications to microarray gene expression data for cancer subtype discovery demonstrate the advantage of the new proposal over several existing approaches. PMID- 18162110 TI - Analysis of longitudinal data in the presence of informative observational times and a dependent terminal event, with application to medical cost data. AB - In longitudinal observational studies, repeated measures are often taken at informative observation times. Also, there may exist a dependent terminal event such as death that stops the follow-up. For example, patients in poorer health are more likely to seek medical treatment and their medical cost for each visit tends to be higher. They are also subject to a higher mortality rate. In this article, we propose a random effects model of repeated measures in the presence of both informative observation times and a dependent terminal event. Three submodels are used, respectively, for (1) the intensity of recurrent observation times, (2) the amount of repeated measure at each observation time, and (3) the hazard of death. Correlated random effects are incorporated to join the three submodels. The estimation can be conveniently accomplished by Gaussian quadrature techniques, e.g., SAS Proc NLMIXED. An analysis of the cost-accrual process of chronic heart failure patients from the clinical data repository at the University of Virginia Health System is presented to illustrate the proposed method. PMID- 18162111 TI - Exploiting gene-environment independence for analysis of case-control studies: an empirical Bayes-type shrinkage estimator to trade-off between bias and efficiency. AB - Standard prospective logistic regression analysis of case-control data often leads to very imprecise estimates of gene-environment interactions due to small numbers of cases or controls in cells of crossing genotype and exposure. In contrast, under the assumption of gene-environment independence, modern "retrospective" methods, including the "case-only" approach, can estimate the interaction parameters much more precisely, but they can be seriously biased when the underlying assumption of gene-environment independence is violated. In this article, we propose a novel empirical Bayes-type shrinkage estimator to analyze case-control data that can relax the gene-environment independence assumption in a data-adaptive fashion. In the special case, involving a binary gene and a binary exposure, the method leads to an estimator of the interaction log odds ratio parameter in a simple closed form that corresponds to an weighted average of the standard case-only and case-control estimators. We also describe a general approach for deriving the new shrinkage estimator and its variance within the retrospective maximum-likelihood framework developed by Chatterjee and Carroll (2005, Biometrika92, 399-418). Both simulated and real data examples suggest that the proposed estimator strikes a balance between bias and efficiency depending on the true nature of the gene-environment association and the sample size for a given study. PMID- 18162112 TI - A joint model for longitudinal measurements and survival data in the presence of multiple failure types. AB - In this article we study a joint model for longitudinal measurements and competing risks survival data. Our joint model provides a flexible approach to handle possible nonignorable missing data in the longitudinal measurements due to dropout. It is also an extension of previous joint models with a single failure type, offering a possible way to model informatively censored events as a competing risk. Our model consists of a linear mixed effects submodel for the longitudinal outcome and a proportional cause-specific hazards frailty submodel (Prentice et al., 1978, Biometrics 34, 541-554) for the competing risks survival data, linked together by some latent random effects. We propose to obtain the maximum likelihood estimates of the parameters by an expectation maximization (EM) algorithm and estimate their standard errors using a profile likelihood method. The developed method works well in our simulation studies and is applied to a clinical trial for the scleroderma lung disease. PMID- 18162113 TI - Bayesian optimal design for phase II screening trials. AB - Most phase II screening designs available in the literature consider one treatment at a time. Each study is considered in isolation. We propose a more systematic decision-making approach to the phase II screening process. The sequential design allows for more efficiency and greater learning about treatments. The approach incorporates a Bayesian hierarchical model that allows combining information across several related studies in a formal way and improves estimation in small data sets by borrowing strength from other treatments. The design incorporates a utility function that includes sampling costs and possible future payoff. Computer simulations show that this method has high probability of discarding treatments with low success rates and moving treatments with high success rates to phase III trial. PMID- 18162114 TI - Tests and diagnostic plots for detecting lack-of-fit for circular-linear regression models. AB - Regression diagnostics and lack-of-fit tests mainly focus on linear-linear regression models. When the design points are distributed on the circumference of a circle, difficulties arise as there is no natural starting point or origin. Most classical lack-of-fit tests require an arbitrarily chosen origin, but different choices may result in different conclusions. We propose a graphical diagnostic tool and a closely related lack-of-fit test, which does not require a natural starting point. The method is based on regional residuals which are defined on arcs of the circle. The graphical method formally locates and visualizes subsets of poorly fitting observations on the circle. A data example from the food technology is used to point out the before-mentioned problems with conventional lack-of-fit tests and to illustrate the strength of the methodology based on regional residuals in detecting and localizing departures from the no effect hypothesis. A small simulation study shows a good performance of the regional residual test in case of both global and local deviations from the null model. Finally, the ideas are extended to the case of more than one predictor variable. PMID- 18162115 TI - Bayesian nonparametric meta-analysis using Polya tree mixture models. AB - Summary. A common goal in meta-analysis is estimation of a single effect measure using data from several studies that are each designed to address the same scientific inquiry. Because studies are typically conducted in geographically disperse locations, recent developments in the statistical analysis of meta analytic data involve the use of random effects models that account for study-to study variability attributable to differences in environments, demographics, genetics, and other sources that lead to heterogeneity in populations. Stemming from asymptotic theory, study-specific summary statistics are modeled according to normal distributions with means representing latent true effect measures. A parametric approach subsequently models these latent measures using a normal distribution, which is strictly a convenient modeling assumption absent of theoretical justification. To eliminate the influence of overly restrictive parametric models on inferences, we consider a broader class of random effects distributions. We develop a novel hierarchical Bayesian nonparametric Polya tree mixture (PTM) model. We present methodology for testing the PTM versus a normal random effects model. These methods provide researchers a straightforward approach for conducting a sensitivity analysis of the normality assumption for random effects. An application involving meta-analysis of epidemiologic studies designed to characterize the association between alcohol consumption and breast cancer is presented, which together with results from simulated data highlight the performance of PTMs in the presence of nonnormality of effect measures in the source population. PMID- 18162116 TI - Evaluation of 280,000 cases in Dutch midwifery practices: a descriptive study. AB - OBJECTIVE: To assess the nature and outcome of intrapartum referrals from primary to secondary care within the Dutch obstetric system. DESIGN: Descriptive study. SETTING: Dutch midwifery database (LVR1), covering 95% of all midwifery care and 80% of all Dutch pregnancies (2001-03). POPULATION: Low-risk women (280,097) under exclusive care of a primary level midwife at the start of labour either with intention to deliver at home or with a personal preference to deliver in hospital under care of a primary level midwife. METHODS: Women were classified into three categories (no referral, urgent referral and referral without urgency) and were related to maternal characteristics and to neonatal outcomes. MAIN OUTCOME MEASURES: Distribution of referral categories, main reasons for urgent referral, Apgar score at 5 minutes, perinatal death within 24 hours and referral to a paediatrician within 24 hours. RESULTS: In our study, 68.1% of the women completed childbirth under exclusive care of a midwife, 3.6% were referred on an urgency basis and 28.3% were referred without urgency. Of all referrals, 11.2% were on an urgency basis. The main reasons for urgent referrals were fetal distress and postpartum haemorrhage. The nonurgent referrals predominantly took place during the first stage of labour (73.6% of all referrals). Women who had planned a home delivery were referred less frequently than women who had planned a hospital delivery: 29.3 and 37.2%, respectively (P < 0.001). On average, the mean Apgar score at 5 minutes was high (9.72%) and the peripartum neonatal mortality was low (0.05%) in the total study group. No maternal deaths occurred. Adverse neonatal outcomes occurred most frequently in the urgent referral group, followed by the group of referrals without urgency and the nonreferred group. CONCLUSIONS: Risk selection is a crucial element of the Dutch obstetric system and continues into the postpartum period. The system results in a relatively small percentage of intrapartum urgent referrals and in overall satisfactory neonatal outcomes in deliveries led by primary level midwives. PMID- 18162117 TI - Molecular determination of the breakpoints of a 161 556 bp deletion at chromosome 13q34 that presented as severe factor VII deficiency in a neonate. PMID- 18162118 TI - Nodular pulmonary lesions in children after autologous stem cell transplantation: a source of misinterpretation. AB - In children with malignant disorders, autologous haematopoietic stem cell transplantation (HSCT) represents a therapeutic option, but several possible complications, such as life-threatening pulmonary disease, make appropriate diagnostic procedures essential. We describe two cases with bronchiolitis obliterans with organizing pneumonia after HSCT, with a brief review of important differential diagnoses. PMID- 18162119 TI - Usefulness of the eosin-5'-maleimide cytometric method as a first-line screening test for the diagnosis of hereditary spherocytosis: comparison with ektacytometry and protein electrophoresis. PMID- 18162120 TI - Safety and efficacy of cyclophosphamide, adriamycin, vincristine, prednisone and rituximab in patients with human immunodeficiency virus-associated diffuse large B-cell lymphoma: results of a phase II trial. AB - Immunochemotherapy with cyclophosphamide, adriamycin, vincristine, prednisone and rituximab (R-CHOP) is the standard treatment in non-immunosuppressed patients with diffuse large B-cell lymphoma (DLBCL), but its adequacy has not been definitively established in patients with human immunodeficiency virus (HIV) related lymphoma. This phase II trial aimed to evaluate the safety and efficacy of six cycles of R-CHOP in patients with HIV-related DLBCL and to determine whether response to highly active antiretroviral therapy (HAART) had prognostic impact. Patients were eligible if they had performance status <3 and absence of active opportunistic infections. Eighty-one patients were enrolled, 57 in stages III or IV, International Prognostic Index (IPI) 0 or 1 (n = 26), 2 (n = 19), 3 (n = 20) and 4 or 5 (n = 16), and median CD4 lymphocyte count of 0.158 x 10(9)/l. The main adverse events were neutropenia (48% of cycles) and infections (10% of cycles), which were fatal in seven patients. Complete response was achieved in 55 (69%) patients, with an estimated 3-year disease-free survival of 77% and 3-year overall survival of 56%. IPI score and virological response to HAART were the prognostic parameters for response and survival. In HIV-related DLBCL R-CHOP is feasible, safe and effective. The prognosis depends on lymphoma-related parameters and on the response to HAART. PMID- 18162121 TI - Perspectives in treatment of AL amyloidosis. AB - Light chain (AL) amyloidosis is the most frequently diagnosed form of systemic amyloid in the western world. The historically poor prognosis of AL amyloidosis appears to be improving with currently reported median survival of c. 40 months compared to 13 months in the early 1990s when low-dose oral melphalan was the mainstay of treatment. Autologous stem cell transplantation (ASCT) achieves the highest rates of complete clonal response but is confounded by substantial treatment-related mortality in AL amyloidosis unless it is restricted to highly selected patients. Newer chemotherapy regimens appear to have a balance of better safety and respectable efficacy with overall outcomes nearly similar to ASCT, but which may be used more widely. There are few data comparing durability, depth of clonal response, rate of organ response and overall survival following ASCT or chemotherapy, but a recent small, randomized trial did not suggest superiority of ASCT to oral melphalan and dexamethasone. There is a compelling need for further and larger randomized trials in this context. At the same time, various new specific anti-amyloid drugs have shown (in early phase studies or animal models) some very promising results. This review attempts to highlight the challenges, controversies and progress in AL amyloidosis. PMID- 18162122 TI - Flexural cutaneous eruption due to palifermin. PMID- 18162123 TI - Tumour necrosis factor-induced gene expression in human marrow stroma: clues to the pathophysiology of MDS? AB - Aberrant regulation of the tumour necrosis factor alpha gene (TNF) and stroma derived signals are involved in the pathophysiology of myelodysplasia. Therefore, KG1a, a myeloid leukaemia cell line, was exposed to Tnf in the absence or presence of either HS-5 or HS-27a cells, two human stroma cell lines. While KG1a cells were resistant to Tnf-induced apoptosis in the absence of stroma cells, Tnf promoted apoptosis of KG1a cells in co-culture experiments with stroma cells. To investigate the Tnf-induced signals from the stroma cells, we examined expression changes in HS-5 and HS-27a cells after Tnf exposure. DNA microarray studies found both discordant and concordant Tnf-induced expression responses in the two stroma cell lines. Tnf promoted an increased mRNA expression of pro-inflammatory cytokines [e.g. interleukin (IL)6, IL8 and IL32]. At the same time, Tnf decreased the mRNA expression of anti-apoptotic genes (e.g. BCL2L1) and increased the mRNA expression of pro-apoptotic genes (e.g. BID). Overall, the results suggested that Tnf induced a complex set of pro-inflammatory and pro-apoptotic signals in stroma cells that promote apoptosis in malignant myeloid clones. Additional studies will be required to determine which of these signals are critical for the induction of apoptosis in the malignant clones. Those insights, in turn, may point the way to novel therapeutic approaches. PMID- 18162124 TI - A phase II clinical trial of intensive chemotherapy followed by consolidative stem cell transplant: long-term follow-up in newly diagnosed mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is associated with high relapse rates and poor survival when treated with conventional chemotherapy, with or without rituximab. We report the long-term follow-up of a phase II clinical trial using a new intensive multiagent chemotherapeutic regimen [cyclophosphamide, teniposide, doxorubicin and prednisone (CTAP) alternating with vincristine and high-dose methotrexate and cytarabine (VMAC)] in newly diagnosed MCL. Following 4-6 cycles of CTAP/VMAC induction, patients aged < or =65 years proceeded to consolidative autologous haematopoietic stem cell transplantation (auto-HSCT), while patients < or =55 years who had a HLA-identical sibling received allogeneic-HSCT (busulfan/cyclophosphamide conditioning for both). Twenty-five untreated MCL patients enrolled on the protocol between 1997 and 2002. Among evaluable patients, overall response rate (ORR) was 74% following induction chemotherapy. Seventeen patients received HSCT (autologous-13/allogeneic-4). On intent-to-treat analysis, ORR for patients who received consolidative HSCT was 100% (complete remission 76%). Therapy was well-tolerated with 4% treatment-related mortality (including HSCT). The 5-year event-free-survival (EFS) and overall survival (OS) for all patients was 35% and 50% respectively. Furthermore, at 66-months median follow-up, the 5-year EFS and OS for patients who received consolidative auto HSCT was 54% and 75% respectively. Patients who received auto-HSCT had improved outcomes compared to no auto-HSCT (EFS P = 0.001; OS P = 0.0002). CTAP/VMAC induction followed by consolidative auto-HSCT for newly diagnosed MCL is associated with high ORR and durable survival. PMID- 18162125 TI - Telomere length shortening in Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis (LCH) is a clonal, proliferative disorder of phenotypically immature CD1a(+) Langerhans cells (LC). The aetiology of LCH is unknown and data supporting an immune dysregulatory disorder as well as a clonal neoplasm have been reported. Telomere shortening has been associated with cancers and premalignant lesions as well as promoting chromosomal instability. To determine whether LCH LC have altered telomere lengths, we used dual detection of CD1a expression by immunofluorescence and telomere length by fluorescence in situ hybridization of LCH LC and lymphocytes in local, multisystem and systemic LCH and compared these with telomere lengths of LC and lymphocytes in reactive lymph nodes. LCH LC showed significantly shorter telomere lengths than LC from reactive lymph nodes or unaffected skin. Lymphocyte telomere lengths showed similar profiles among the different samples. These data show a significant telomere shortening in LCH LC in all stages of disease involvement compared with LC from reactive lymph nodes, suggesting that LCH may share mechanisms of telomere shortening and survival with clonal preneoplastic disorders and cancer, although an initiating infectious or immune event is still possible. PMID- 18162126 TI - Frequency of the von Willebrand factor Tyr1584Cys polymorphism in arterial thrombosis. PMID- 18162127 TI - Prevalence of enteropathogenic viruses and molecular characterization of group A rotavirus among children with diarrhea in Dar es Salaam Tanzania. AB - BACKGROUND: Different groups of viruses have been shown to be responsible for acute diarrhea among children during their first few years of life. Epidemiological knowledge of viral agents is critical for the development of effective preventive measures, including vaccines. METHODS: In this study we determined the prevalence of the four major enteropathogenic viruses - rotavirus, norovirus, adenovirus and astrovirus - was determined in 270 stool samples collected from children aged 0 - 60 months who were admitted with diarrhea in four hospitals in Dar es Salaam, Tanzania, using commercially available ELISA kits. In addition, the molecular epidemiology of group A rotavirus was investigated using reverse transcriptase multiplex polymerase chain reaction (RT PCR). RESULTS: At least one viral agent was detected in 87/270 (32.2%) of the children. The prevalence of rotavirus, norovirus, adenovirus and astrovirus was 18.1%, 13.7%, 2.6% and 0.4%, respectively. In most cases (62.1%) of viruses were detected in children aged 7-12 months. The G and P types (VP7 and VP4 genotypes respectively) were further investigated in 49 rotavirus ELISA positive samples. G9 was the predominant G type (81.6%), followed by G1 (10.2%) and G3 (0.2%). P[8] was the predominant P type (83.7%), followed by P[6] (0.4%) and P[4] (0.2%). The following G and P types were not detected in this study population; G2, G4, G8 G10, P[9], P[10] and P[11]. The dominating G/P combination was G9P[8], accounting for 39 (90.7%) of the 43 fully characterized strains. Three (6.1%) of the 49 rotavirus strains could not be typed. CONCLUSION: Nearly one third of children with diarrhea admitted to hospitals in Dar es Salaam had one of the four viral agents. The predominance of rotavirus serotype G9 may have implication for rotavirus vaccination in Tanzania. PMID- 18162128 TI - Expression of genes for bone morphogenetic proteins BMP-2, BMP-4 and BMP-6 in various parts of the human skeleton. AB - BACKGROUND: Differences in duration of bone healing in various parts of the human skeleton are common experience for orthopaedic surgeons. The reason for these differences is not obvious and not clear. METHODS: In this paper we decided to measure by the use of real-time RT-PCR technique the level of expression of genes for some isoforms of bone morphogenetic proteins (BMPs), whose role is proven in bone formation, bone induction and bone turnover. Seven bone samples recovered from various parts of skeletons from six cadavers of young healthy men who died in traffic accidents were collected. Activity of genes for BMP-2, -4 and -6 was measured by the use of fluorescent SYBR Green I. RESULTS: It was found that expression of m-RNA for BMP-2 and BMP-4 is higher in trabecular bone in epiphyses of long bones, cranial flat bones and corpus mandibulae then in the compact bone of diaphyses of long bones. In all samples examined the expression of m-RNA for BMP-4 was higher than for BMP-2. CONCLUSION: It was shown that m-RNA for BMP-6 is not expressed in the collected samples at all. It is postulated that differences in the level of activation of genes for BMPs is one of the important factors which determine the differences in duration of bone healing of various parts of the human skeleton. PMID- 18162129 TI - Background frequencies for residue variability estimates: BLOSUM revisited. AB - BACKGROUND: Shannon entropy applied to columns of multiple sequence alignments as a score of residue conservation has proven one of the most fruitful ideas in bioinformatics. This straightforward and intuitively appealing measure clearly shows the regions of a protein under increased evolutionary pressure, highlighting their functional importance. The inability of the column entropy to differentiate between residue types, however, limits its resolution power. RESULTS: In this work we suggest generalizing Shannon's expression to a function with similar mathematical properties, that, at the same time, includes observed propensities of residue types to mutate to each other. To do that, we revisit the original construction of BLOSUM matrices, and re-interpret them as mutation probability matrices. These probabilities are then used as background frequencies in the revised residue conservation measure. CONCLUSION: We show that joint entropy with BLOSUM-proportional probabilities as a reference distribution enables detection of protein functional sites comparable in quality to a time costly maximum-likelihood evolution simulation method (rate4site), and offers greater resolution than the Shannon entropy alone, in particular in the cases when the available sequences are of narrow evolutionary scope. PMID- 18162130 TI - Association between long-term neuro-toxicities in testicular cancer survivors and polymorphisms in glutathione-s-transferase-P1 and -M1, a retrospective cross sectional study. AB - BACKGROUND: To assess the impact of polymorphisms in Glutathione S-transferase (GST) -P1, -M1, and -T1 on self-reported chemotherapy-induced long-term toxicities in testicular cancer survivors (TCSs). METHODS: A total of 238 TCSs, who had received cisplatin-based chemotherapy at median twelve years earlier, had participated in a long-term follow-up survey which assessed the prevalence of self-reported paresthesias in fingers/toes, Raynaud-like phenomena in fingers/toes, tinnitus, and hearing impairment. From all TCSs lymphocyte-derived DNA was analyzed for the functional A-->G polymorphism at bp 304 in GSTP1, and deletions in GST-M1 and GST-T1. Evaluation of associations between GST polymorphisms and self-reported toxicities included adjustment for prior treatment. RESULTS: All six evaluated toxicities were significantly associated with the cumulative dose of cisplatin and/or bleomycin. Compared to TCSs with either GSTP1-AG or GSTP1-AA, the 37 TCSs with the genotype GSTP1-GG, were significantly less bothered by paresthesias in fingers and toes (p = 0.039, OR 0.46 [0.22-0.96] and p = 0.023, OR 0.42 [0.20-0.88], respectively), and tinnitus (p = 0.008, OR 0.33 [0.14-0.74]). Furthermore, absence of functional GSTM1 protected against hearing impairment (p = 0.025, OR 1.81 [1.08-3.03]). CONCLUSION: In TCSs long-term self-reported chemotherapy-induced toxicities are associated with functional polymorphisms in GSTP1 and GSTM1. Hypothetically, absence of GST-M1 leaves more glutathione as substrate for the co-expressed GST P1. Also intracellular inactivation of pro-apoptotic mediators represents a possible explanation of our findings. Genotyping of these GSTs might be a welcomed step towards a more individualized treatment of patients with metastatic testicular cancer. PMID- 18162131 TI - Pancreatectomy for non-pancreatic malignancies results in improved survival after R0 resection. AB - BACKGROUND: Pancreatectomy has a high morbidity but remains the only chance of cure for pancreatic cancer. Its efficacy for non-pancreatic malignancies is less clear. We reviewed our experience with pancreatectomy for non-pancreatic malignancies to determine outcomes and identify predictors of survival. PATIENTS AND METHODS: The records of patients who underwent pancreatectomy for non pancreatic malignancies between 1990 and 2005 were reviewed. Survival curves were constructed using the Kaplan-Meier method and compared using log-rank analysis. Cox proportional hazards was used to identify predictors of survival. RESULTS: 29 patients (18 M/11 F) with a mean age of 59.9 years (range 29-86) underwent pancreatectomy for non-pancreatic malignancies. 19 (66%) primary malignancies were GI in origin. Most operations were undertaken with curative intent (76%), whereas the remainder was for symptom palliation. Pancreatectomy was completed for metastatic disease in 7 patients (24%) or en bloc to achieve negative margins in 22 patients (76%). Complete (i.e., R0) resection was achieved in 17 (59%). Perioperative mortality was 3%. Median follow-up was 15 months (range 7-172). Median overall survival was 12 months with 1-year survival of 48%. Significant predictors of improved survival by univariate analysis were R0 resection, non-GI primary, and pancreatic metastasectomy (vs. en bloc resection). Only R0 resection was predictive of long-term survival by multivariate analysis (median 21 months vs. 6). CONCLUSION: Pancreatic resection for non-pancreatic malignancies can be completed with minimal mortality. However, incomplete resection results in poor overall survival. Pancreatectomy for non-pancreatic malignancies should only be undertaken if complete resection is possible. PMID- 18162132 TI - Should right ventricle dilatation during exercise have clinical implications in patients with chronic thromboembolic pulmonary hypertension? Case report. AB - We describe the case of a 30-year-old female patient with chronic thromboembolic pulmonary hypertension that has an excellent functional capacity under treatment with sildenafil. She did an exercise stress echocardiography that revealed marked right ventricular dilatation during exercise. This information was used for clinical decision and the authors discuss the potential utility of this echocardiographyc sign. PMID- 18162133 TI - DSS and DHS: longitudinal and cross-sectional viewpoints on child and adolescent mortality in Ethiopia. AB - BACKGROUND: In countries where routine vital registration data are scarce, Demographic Surveillance Sites (DSS: locally defined populations under longitudinal surveillance for vital events and other characteristics) and Demographic and Health Surveys (DHS: periodic national cluster samples responding to cross-sectional surveys) have become standard approaches for gathering at least some data. This paper aims to compare DSS and DHS approaches, seeing how they complement each other in the specific instance of child and adolescent mortality in Ethiopia. METHODS: Data from the Butajira DSS 1987-2004 and the Ethiopia DHS rounds for 2000 and 2005 formed the basis of comparative analyses of mortality rates among those aged under 20 years, using Poisson regression models for adjusted rate ratios. RESULTS: Patterns of mortality over time were broadly comparable using DSS and DHS approaches. DSS data were more susceptible to local epidemic variations, while DHS data tended to smooth out local variation, and be more subject to recall bias. CONCLUSION: Both DSS and DHS approaches to mortality surveillance gave similar overall results, but both showed method-dependent advantages and disadvantages. In many settings, this kind of joint-source data analysis could offer significant added value to results. PMID- 18162134 TI - A large-scale proteomic analysis of human embryonic stem cells. AB - BACKGROUND: Much of our current knowledge of the molecular expression profile of human embryonic stem cells (hESCs) is based on transcriptional approaches. These analyses are only partly predictive of protein expression however, and do not shed light on post-translational regulation, leaving a large gap in our knowledge of the biology of pluripotent stem cells. RESULTS: Here we describe the use of two large-scale western blot assays to identify over 600 proteins expressed in undifferentiated hESCs, and highlight over 40 examples of multiple gel mobility variants, which are suspected protein isoforms and/or post-translational modifications. Twenty-two phosphorylation events in cell signaling molecules, as well as potential new markers of undifferentiated hESCs were also identified. We confirmed the expression of a subset of the identified proteins by immunofluorescence and correlated the expression of transcript and protein for key molecules in active signaling pathways in hESCs. These analyses also indicated that hESCs exhibit several features of polarized epithelia, including expression of tight junction proteins. CONCLUSION: Our approach complements proteomic and transcriptional analysis to provide unique information on human pluripotent stem cells, and is a framework for the continued analyses of self renewal. PMID- 18162135 TI - Infra-specific folk taxonomy in sorghum (Sorghum bicolor (L.) Moench) in Ethiopia: folk nomenclature, classification, and criteria. AB - BACKGROUND: Sorghum is one of the main staple food crops for the poorest and most food insecure people of the world. As Ethiopia is the centre of origin and diversity for sorghum, the crop has been cultivated for many thousands of years. Hence, indigenous knowledge based sorghum classification and naming has a long tradition. METHODS: In order to assess folk taxonomy, various research methods were employed, including, focus group interviews with 360 farmers, direct on-farm participatory monitoring with 120 farmers, key informant interviews with 60 farmers and development agents and semi-structured interviews with 250 farmers. In addition, diversity fairs were conducted with over 1200 farmers. Assessment of folk taxonomy consistency was assessed by 30 farmers' evaluation of 44 folk species. RESULTS: Farmers have been growing sorghum for at least 500 years (20 generations). Sorghum is named as Mishinga in the region. Farmers used twenty five morphological, sixty biotic and abiotic and twelve use-related traits in folk taxonomy of sorghum. Farmers classified their gene-pool by hierarchical classifications into parts that represented distinguishable groups of accessions. Folk taxonomy trees were generated in the highland, intermediate and lowland sorghum ecologies. Over 78 folk species have been identified. The folk species were named after morphological, use-related and breeding methodology used. Relative distribution of folk species over the region, folk taxonomy consistency, and comparison of folk and formal taxonomy are described. CONCLUSION: New folk taxonomy descriptors have been identified and suggested to be used as formal taxonomy descriptors. It is concluded that integrated folk-formal taxonomy has to be used for enhanced collection, characterisation and utilization of on farm genetic resources. PMID- 18162136 TI - Expression of junctional proteins in choroid plexus epithelial cell lines: a comparative study. AB - BACKGROUND: There is an increasing interest in using choroid plexus (CP) epithelial cell lines to study the properties of the blood-cerebrospinal fluid barrier (BCSFB). Currently, there are three major CP-derived cell lines available. Z310 and TR-CSFB3, two immortalized cell lines carrying the simian virus 40 large T-antigen gene, were derived from rat CP epithelium, whereas the CPC-2 cell line was derived from human CP carcinoma. Although these cell lines have previously been used in various functional studies, the expression of adherens junction (AJ) and tight junction (TJ) proteins in these epithelial cells has not been systematically studied. Accordingly, in the present study, we sought to characterize the expression of these junctional proteins in these three cell lines. METHODS: The cells were grown in six-well cell culture plates. Reverse transcriptase polymerase chain reaction, Western blotting, and immunocytochemistry were used to characterize the expression of AJ and TJ proteins in the CP cell lines. RESULTS: Z310 and TR-CSFB3 cells expressed a TJ protein, occludin, and its cytosolic binding partner, zonula occludens 1, as well as an AJ protein, E-cadherin, and beta-catenin, a cytoplasmic protein that interacts with E-cadherin. However, the expression of occludin and E-cadherin in TR-CSFB3 cells at both the mRNA and protein level was weaker than that found in Z301 cells. The immunocytochemical analysis also demonstrated that the staining pattern for these junctional proteins in TR-CSFB3 cells was discontinuous and the staining intensity was weaker than that observed in Z310 cells. The message for claudin 1 and claudin 2 was expressed at low levels in TR-CSFB3 cells and these cells were weakly immunopositive for claudin 1. In comparison, the message for these TJ proteins could not be detected in Z310 cells. CPC-2 cells expressed occludin, which was localized to areas of cell-cell contact, but the staining pattern for this TJ protein was found to be variable and irregular. Although CPC 2 cells expressed mRNA for claudin 1, claudin 2, and claudin 11, only claudin 1 was expressed at the protein level and it was localized to the nuclei rather than to areas of cell-cell contact. An AJ protein, E-cadherin, was also found to be mislocalized in CPC-2 cells, even though its cytosolic binding partner, beta catenin, was restricted to areas of cell-cell contact, as in normal CP. CONCLUSION: The three CP cell lines analyzed in this study vary considerably with regard to the expression of AJ and TJ proteins, which is likely reflected by different barrier properties of these in vitro models of BCSFB. PMID- 18162137 TI - Withdrawal of inhaled corticosteroids in people with COPD in primary care: a randomised controlled trial. AB - BACKGROUND: Guidelines recommend inhaled corticosteroids (ICS) for patients with severe chronic obstructive pulmonary disease (COPD). Most COPD patients are managed in primary care and receive ICS long-term and irrespective of severity. The effect of withdrawing ICS from COPD patients in primary care is unknown. METHODS: In a pragmatic randomised, double-blind, placebo-controlled trial in 31 practices, 260 COPD patients stopped their usual ICS (median duration of use 8 years) and were allocated to 500 mcg fluticasone propionate twice daily (n = 128), or placebo (n = 132). Follow-up assessments took place at three monthly intervals for a year at the patients' practice. Our primary outcome was COPD exacerbation frequency. Secondary outcomes were time to first COPD exacerbation, reported symptoms, peak expiratory flow rate and reliever inhaler use, and lung function and health related quality of life. RESULTS: In patients randomised to placebo, COPD exacerbation risk over one year was RR: 1.11 (CI: 0.91-1.36). Patients taking placebo were more likely to return to their usual ICS following exacerbation, placebo: 61/128 (48%); fluticasone: 34/132 (26%), OR: 2.35 (CI: 1.38-4.05). Exacerbation risk whilst taking randomised treatment was significantly raised in the placebo group 1.48 (CI: 1.17-1.86). Patients taking placebo exacerbated earlier (median time to first exacerbation: placebo (days): 44 (CI: 29-59); fluticasone: 63 (CI: 53-74), log rank 3.81, P = 0.05) and reported increased wheeze. In a post-hoc analysis, patients with mild COPD taking placebo had increased exacerbation risk RR: 1.94 (CI: 1.20-3.14). CONCLUSION: Withdrawal of long-term ICS in COPD patients in primary care increases risk of exacerbation shortens time to exacerbation and causes symptom deterioration. Patients with mild COPD may be at increased risk of exacerbation after withdrawal. TRIAL REGISTRATION: ClinicalTrials.gov NCT00440687. PMID- 18162138 TI - Recent breast cancer trends among Asian/Pacific Islander, Hispanic, and African American women in the US: changes by tumor subtype. AB - BACKGROUND: Recently, unprecedented drops in breast cancer incidence have been reported for populations of mostly White European descent. Incidence patterns in non-White racial/ethnic groups are less described. Therefore, we examined population-based breast cancer incidence trends separately for US Asian/Pacific Islander, Hispanic, African-American, and non-Hispanic White women by etiologically relevant tumor subtype characteristics, including hormone receptor status, histology, size, and in situ behavior. METHODS: We obtained breast cancer data from 13 Surveillance, Epidemiology, and End Results (SEER) cancer registries to calculate age-adjusted incidence rates and trends, stratified by race/ethnicity and tumor subtype for the period 1992-2004. Detailed analyses were limited to women 50 years old or older. Joinpoint regression was used to assess incidence trends by annual quarter of diagnosis. RESULTS: Between 2001 and 2004, incidence rates of invasive breast cancer in women 50 years old or older declined appreciably among Asians/Pacific Islanders (-8.5%) and Hispanics (-2.9%) and were stable in African-Americans (+0.5%), reductions substantially lower than those observed among non-Hispanic Whites (-14.3%). In Asian/Pacific Islander women, perceptible but statistically nonsignificant decreases were observed for hormone receptor-positive, lobular, and small tumors only. Rates of hormone receptor negative tumors increased among African-Americans (26.1%) and Hispanics (26.9%) during 2001-2004. Incidence trends in most groups, except African-American women, peaked between 1999 and mid-2002. Rates of in situ cancer remained stable in all groups. CONCLUSION: Recently reported reductions in breast cancer incidence varied considerably by race/ethnicity. These patterns are consistent with documented racial/ethnic differences in the prevalence and discontinuation of hormone therapy (HT) after July 2002 but do not correspond as well to patterns of mammography use in these groups. The data presented in this analysis provide further evidence that population-level HT use is a major influence on population level rates of particular breast cancer subtypes, especially receptor-positive tumors. PMID- 18162139 TI - Effects of high-fat diet and/or body weight on mammary tumor leptin and apoptosis signaling pathways in MMTV-TGF-alpha mice. AB - INTRODUCTION: Obesity is a risk factor for postmenopausal breast cancer and is associated with shortened mammary tumor (MT) latency in MMTV-TGF-alpha mice with dietary-induced obesity. One link between obesity and breast cancer is the adipokine, leptin. Here, the focus is on diet-induced obesity and MT and mammary fat pad (MFP) leptin and apoptotic signaling proteins. METHODS: MMTV-TGF-alpha mice were fed low-fat or high-fat diets from 10 to 85 weeks of age. High-Fat mice were divided into Obesity-Prone and Obesity-Resistant groups based on final body weights. Mice were followed to assess MT development and obtain serum, MFP, and MT. RESULTS: Incidence of palpable MTs was significantly different: Obesity-Prone > Obesity-Resistant > Low-Fat. Serum leptin was significantly higher in Obesity Prone compared with Obesity-Resistant and Low-Fat mice. Low-Fat mice had higher MFP and MT ObRb (leptin receptor) protein and Jak2 (Janus kinase 2) protein and mRNA levels in comparison with High-Fat mice regardless of body weight. Leptin (mRNA) and pSTAT3 (phosphorylated signal transducer and activator of transcription 3) (mRNA and protein) also were higher in MTs from Low-Fat versus High-Fat mice. Expression of MT and MFP pro-apoptotic proteins was higher in Low Fat versus High-Fat mice. CONCLUSION: These results confirm a connection between body weight and MT development and between body weight and serum leptin levels. However, diet impacts MT and MFP leptin and apoptosis signaling proteins independently of body weight. PMID- 18162140 TI - Two or more enteropathogens are associated with diarrhoea in Mexican children. AB - BACKGROUND: Diarrhoeal diseases constitute a major public health problem, particularly in the developing world, where the rate of mortality and morbidity is very high. The purpose of this study was to conduct a 2 years and 3 months study in order to determine the prevalence of five enteropathogen diarrheogenic agents in Mexico City. METHODS: Faecal samples were obtained from 300 Mexican children diagnosed as positive for diarrhoea, aged > 2 to < 12 years old, and from 80 children matched for age but with no symptoms of the disease (control group). Two multiplex PCR were used to detect Escherichia coli, Salmonella spp., and Shigella spp. In addition, the two protozoan parasites Entamoeba histolytica/Entamoeba dispar and Giardia intestinalis were detected by conventional methods. RESULTS: All diarrhoeal samples were positive for one or more enteropathogens. The most common enteropathogens in diarrhoeal samples were E. histolytica/E. dispar (70.3%), Salmonella (ohio 28.3%; typhimurium 16.3%; infantis 8%; anatum 0.6%; Newport 0.3%), G. intestinalis (33%), E. coli (ETEC 13.3%; EPEC 9.3%; VTEC 8.6%; EIEC 1%) and Shigella spp. (flexneri 1.6%, sonnei 1%). Infections by two (24%) three (16%) and four (12%) pathogens were observed. CONCLUSION: This study revealed that 52% of the patients were infected by more than one enteropathogen, notably E. histolitica/E. dispar and Salmonella ohio. These results are useful for clinicians to improve the empiric treatment used in such cases. PMID- 18162141 TI - Fixed-links models for investigating experimental effects combined with processing strategies in repeated measures designs: a cognitive task as example. AB - Fixed-links models enable the investigation of experimental effects in combination with processing strategies in the framework of repeated measures designs. The consideration of different processing strategies is reasonable whenever the task requirements can be met in different ways. Effects due to specific processing strategies become obvious in comparing the models representing these processing strategies and also by investigating the variances of the corresponding latent variables. The usefulness of such models is demonstrated by the reaction time data of a cognitive task including three treatment levels. Two different processing strategies were considered: analytic and holistic. The results indicated that the participants applied both processing strategies. The analytic strategy was primarily applied for performing with respect to the second and third treatment levels and the holistic strategy with respect to the first treatment level. PMID- 18162142 TI - [Objectively understand and evaluate computer assisted orthopedic surgery and steadily promote the research on its clinical application]. PMID- 18162143 TI - [Computer assisted imaging guided total knee arthroplasty: preliminary clinical study of 46 cases]. AB - OBJECTIVE: To discuss the approach and curative effect of total knee arthroplasty by intraoperative digital 3D bone mode. METHODS: 28 patients with bone arthritis, 7 with rheumatoid arthritis, 7 with ankylosing spondylitis, totally 46 patients with 54 knees affected, 29 males and 17 females, aged 48 (31 - 76), underwent total knee arthroplasty by digital 3D bone mode, including osteotomy, adjustment of soft tissue, and rotational positioning of prostheses. The operative time, hemorrhage and drainage 48 h after operation, and complication were observed. The pre-postoperative knee function was evaluated by HISS scoring system. The pre postoperative range of motion of knee joint, and angle of KNA, variance of angle in exterior and interior stress and separation of joint space in tensile force, and postoperative rotational angle of tibia-femur prostheses were measured. X-ray tangential projection photography of patella was used to evaluate the apposition of patellofemoral joint. The postoperative curative effect was observed for 3 months. RESULTS: No complication as injury of nerves and blood vessel, bone fracture, fat embolism and so on was observed. The operative time was 80 +/- 15 min per knee. The hemorrhage and drainage in 48 h was 550 +/- 60 ml per knee. The Hand Injury Severity Score (HISS) 2 weeks after the operation was (92 +/- 4), significantly higher than that before operation [(46 +/- 6), P < 0.01]. There were statistically significant differences in the KNA angle, variance of angle under exterior stress, interior stress, and separation of joint space in tensile force pre- and postoperatively (all P < 0.01). The postoperative rotational angle of tibia-femur prostheses was (4.2 +/- 0.4) degrees. The X-ray tangential projection photograph of patella showed excellent apposition of patellofemoral joint, and no dislocation and unsteadiness. The postoperative range of motion of knee joint was 120 degrees +/- 9 degrees 3 months after operation. No joint laxity was found. CONCLUSION: Computer assisted imaging guidance actually and dynamically observe the changes of line of force of the lower limbs, knee joint rotational apposition and ligament balance, provides geometrical and morphological data and rebuild the knee joint bio-mechanically and anatomically, thus elevating the reproducibility of total knee arthroplasty and ensuring the safety of the system. PMID- 18162144 TI - [Treatment of complex calcaneal fractures under computer navigation: report of 130 feet]. AB - OBJECTIVE: To investigate the effects of computer navigation in the treatment of intra-articular calcaneal fractures. METHODS: 130 feet in 110 patients with intra articular calcaneal fractures, 57 calcanei with fracture of Sander's type II, 45 of type III, and 28 cases of type IV, were treated with internal fixation under computer navigation, and were followed up for 16.3 months (6 - 24 months). RESULTS: According to the Maryland Foot Score system, excellent result was noted in 63 feet, good result in 57 feet, and fair result in 10 feet, with the excellent and good rates being 92.31% together. CONCLUSION: Using computer navigation to treat intra-articular calcaneal fractures is one of the best ways for treatment of calcaneal fractures. PMID- 18162146 TI - [Comparison of 3D C-arm fluoroscopy-based and CT-based navigation systems in the lumbar pedicle puncture: in vitro experiment on a cadaveric lumbar trunk specimen]. AB - OBJECTIVE: To compare the accuracy and operating features of 3D C-arm fluoroscopy based and CT-based navigation systems in the lumbar pedicle punctures. METHODS: A specimen of cadaveric lumbar trunk underwent lumbar pedicle punctures at the levels of L3, L4, and L5 under the guidance of the 3D C-arm fluoroscopy-based and CT-based navigation systems. During the procedure C-arm fluoroscopy was used to monitor the accuracy of the puncture. Generally, in comparison with the 3D C-arm fluoroscopy-based navigation system, the best operation route and protocol could be drawn up pre-operatively, matched registration needed to be renewed for each vertebra, and the images thus obtained were of high quality in CT-based navigation. RESULTS: Both navigation systems had excellent accuracy in the guidance of lumbar pedicle punctures, and had different operating features. CONCLUSION: Both navigation systems had its special advantages. The operating process of the 3D C-arm fluoroscopy-based navigation system was more convenient and rapid, and suitable for percutaneous vertebral puncture. CT based navigation system had clearer pictures, especially for the osteoporotic vertebral bodies, and it had less requirements for the equipments. PMID- 18162148 TI - [Cerebral vascular accidents after cardiac catheterization in patients with anamnesis of stroke]. AB - OBJECTIVE: To investigate the risk factors of relapse of cerebral vascular accident (CVA) after cardiac catheterization (CC) in patients with anamnesis stroke. METHODS: The clinical data of 892 patients with anamnesis stroke who received cardiac catheterization (CC) from Jan. 2002 to Oct. 2006, 555 males and 337 females, aged 61 +/- 10, were analyzed retrospectively. RESULTS: 101 of the 892 patients (11.32%) suffered from CVA during the procedure of CC or within 24 hours after the operation, including 33 cases of transient ischemic attack (TIA), 64 cases of cerebral infarction (CI) and 4 cases of cerebral hemorrhage (CH). Logistic regression analysis showed that the risk factors of relapse of CVA after CC included male gender (OR = 0.308, 95% CI = 0.141 - 0.674), alcohol consumption (OR = 0.319, 95% CI = 0.128 - 0.797), hyperlipidemia (OR = 3.676, 95% CI = 1.94 - 6.950), peripheral vascular diseases (OR = 7.419, 95% CI = 2.477 - 22.219), and number of cardiac vascular diseases (OR = 1.809, 95% CI = 1.284 - 2.548). The patients undergoing percutaneous transluminal coronary angioplasty (PTCA) and stenting were much more liable to CVA compared with the other patients. CONCLUSION: The risk factors of relapse of CVA after CC include male gender, alcohol consumption, hyperlipidemia, peripheral vascular diseases, number of cardiac vascular diseases, and the procedures of PTCA and stenting. PMID- 18162149 TI - [Impact of pulmonary hypertension on early hemodynamics after orthotopic heart transplantation]. AB - OBJECTIVE: To determine the influence of normal pulmonary artery pressure, and mild to moderate and severe pulmonary hypertension on the early hemodynamics, morbidity, and mortality after orthotopic heart transplantations. METHODS: Sixty seven heart disease patients, 54 males and 13 females, aged (46.4 +/- 14.6), including ischemic heart diseases (n = 16), myocardiopathy (n = 43), and other heart diseases (n = 8), underwent orthotopic heart transplantation. Before and after transplantation routine right heart catheterization was conducted. According to the preoperative pulmonary arterial pressure the patients were divided into 3 groups: Group I (n = 15) without pulmonary hypertension (PH) with the pulmonary vascular resistance (PVR) < or = 2.5 Wood's units; Group II (n = 42) with mild to moderate PH with the PVR between 2.5 and 5.0 Wood's units; and Group III (n = 10) with severe PH with the PVR > or = 5.0 Wood's units. Heart rate (HR), mean artery pressure (MAP), central venous pressure (CVP), mean pulmonary artery pressure (MPAP), pulmonary artery wedge pressure (PAWP), PVR, cardiac output index (CI), and mixed venous oxygen saturation (S(V)O(2)) were measured preoperatively, immediately and 12, 24, and 48 hours postoperatively. 1, 3, and 7 days, and 1 and 3 months post-operatively echocardiography was conducted to measure the left ventricular end-diastolic diameter (LVEDD), left ventricular ejection fraction (EF), mitral valvular regurgitation (MVR), and tricuspid valvular regurgitation (TVR). Postoperative complications and mortality were recorded. RESULTS: The 30-day mortality was zero in all 3 groups. The cardiovascular support used for weaning CPB and postoperative period included dopamine, ephedrine and isoproterenol. In addition, nitroglycerin, NO and iloprost were administered for pulmonary artery vasodilation if the pulmonary artery pressure was higher than 45 mm Hg. The EF value of Group III was significantly lower compared with group (P < 0.05). Before the heart transplantation, 52 patients (86%) had mild to severe PH, of which 10 patients (27%) had severe PH. The patients of Group III had longer CPB time and tracheal intubation time in comparison with the patients of Group I (P < 0.05). Postoperatively 6 patients, 1 in Group I, 2 in Group II, and 3 in Group III, had acute right ventricular failure and 3 patients, 1 in Group II and 2 in Group III, had renal failure. Basiliximab, cyclosporine A, mycophenolate mofetil, and methylprednisolone were administered for immunosuppressive treatment perioperatively. CONCLUSION: The patients with severe PH pre-operatively have significantly lower pulmonary resistance and pressure, and have more complications, such as acute right heart failure, post-operatively. Active treatment with cardiac stimulant and diuretics is helpful. Smooth anesthetic induction and maintenance, peri-operative stable hemodynamic managements, especially the protection of right ventricle function for the first 2 weeks after the transplantation, myocardial protection of donor heart, and immunosuppressive regimen all play important roles for successive heart transplantation. PMID- 18162150 TI - [Stress myocardial perfusion single photon emission computed tomography imaging in the detection of coronary artery disease in woman]. AB - OBJECTIVE: To assess the diagnostic efficacy of stress myocardial perfusion imaging (MPI) in female coronary artery disease (CAD) patients. METHODS: Two hundred and fifty-nine consecutive female patients, aged 60 +/- 8, underwent stress myocardial perfusion single photon emission computed tomography (SPECT) imaging and coronary angiography with an interval of < 60 days. Among them, 227 patients underwent exercise MPI, injected intravenously with (99m)Tc methoxyisobutyl isonitrile (MIBI) 740 - 925 MBq when the exercise end point was reached, and 32 patients underwent pharmacologic stress MPI, injected intravenously with persantine or adenosine and then (99m)Tc-MIBI. RESULTS: Among the 227 patients undergoing exercise MPI 79 had significant coronary artery stenosis with the overall sensitivity and specificity for detecting coronary artery disease of 63% and 97% respectively. According to the exercise heart rate, the 227 patients were divided into two groups: group 1 (n = 137) the patients of which achieved adequate exercise end points, and group 2 (n = 90) the patients of which only reached submaximal exercise. The sensitivity of exercise MPI for detecting CAD was 86% in the group 1 and 38% in the group 2. Among the 32 patients who underwent pharmacologic stress MPI 13 had significant CAD with the sensitivity and specificity of 85% and 84% respectively. CONCLUSION: Stress MPI is an efficient protocol for the detection of CAD in women, and pharmacologic stress MPI is more suitable for the women with decreased exercise capacity and advanced age. PMID- 18162151 TI - [Diagnosis and treatment of intralobar pulmonary sequestration]. AB - OBJECTIVE: To summarize the diagnosis and surgical treatment of intralobar pulmonary sequestration (PS). METHODS: The clinical data of 7 cases of intralobar PS, 5 males and 2 females, aged 15 - 38, was collected and analyzed. Macroscopic and microscopic pathological findings were recorded. The expression of protein p53 and carcinoembryonic antigen (CEA) was evaluated immunohistochemically in 6 samples obtained from lobectomy. RESULTS: All 7 patients were admitted with major features of intermittent lung infection. Diagnosis was confirmed in all 7 cases before operation by contrast enhanced helical CT or MRI. All patients were treated with surgical excision, of which lobectomy was performed in 6 cases and wedge resection in 1 case. No surgical death was reported. All the aberrant systemic arteries arose from the thoracic aorta. The histological pictures showed polycystic lesion in sequestrated area with fibrosis formation and chronic inflammatory cell infiltration in the surrounding pulmonary stroma. Hyperplasia of epithelium occurred in some parts of the cystic lesions. Positive protein p53 staining and diffuse CEA expression were detected in all the 6 cases, showing stronger protein p53 staining in whose superficial layer of hyperplastic epithelium than in the basal layer. The normal bronchial epithelium was not stained with p53 or CEA. CONCLUSION: The diagnosis of intralobar PS can be confirmed by enhanced contrast helical CT with 3-dimensional reconstruction, a non-invasive method. Surgical intervention, such as lobectomy, can be applied after complete control of pulmonary infection. Aberrant accumulation of p53 protein and CEA expression in the cystic epithelium inside PS tissues show a relationship with chronic inflammation. PMID- 18162152 TI - [Incidence and pathological features of incidental prostate cancer and clinical significance thereof]. AB - OBJECTIVE: To investigate the morbidity and pathological features of incidental prostate cancer and their clinical significance. METHODS: 1483 prostate specimens obtained during operation, including transurethral resection of prostate (TURP) and total resection of the prostate, for the diagnoses of benign prostatic hypertrophy (BPH) or bladder cancer between January 1999 and August 2005 underwent pathological examination and 34beta12 and p63 immunohistochemical staining so as to detect incidental prostate cancer. The volume of incidental prostate cancer was calculated by the image analysis system. The clinical data were analyzed retrospectively. Comparison between the clinical and pathological feature of incidental prostate cancer was made. RESULTS: 53 cases of incidental prostate cancer, with the Gleason scores from 2 (1+1) to 9 (4+5) and the volumes from 0.18 to 1440.00 mm(3) were detected. The morbidity of incidental prostate cancer was 3.6%. The volume of 47 cases (88.7%) were less than 0.5 cm(3) as the threshold of insignificant cancer, and the volumes of 6 cases (11.3%) were more than 0.5 cm(3). All incidental prostate cancers of clinical significance were detected in the TURP samples. Among the incidental prostate cancers found in the TURP samples 20% were clinically significant cancers. The clinically significant incidental cancers were located in the central or transitional zone with the Gleason scores of 3 + 4 (2 cases), 4 + 2 (1 case), or 4 + 5 (3 cases). These clinically significant cancers were of diffuse distribution, and their preoperative clinical features were negative in palpation/image examination, elevation of serum PSA, and negative in puncture examination. CONCLUSION: Nowadays, the morbidity of incidental prostate cancer is lower than that of 1980s'. Among the incidental cancers 11.3% were of clinical significance. That the preoperative clinical examination cannot find these clinical significant cancers is partially caused by the pathological features of these tumors. PMID- 18162153 TI - [The effects of different ventilatory modes and spontaneous breathing after recruitment maneuver in acute respiratory distress syndrome. Experiment with dogs]. AB - OBJECTIVE: To investigate the role of different ventilatory modes and spontaneous breathing (SB) in prevention of derecruitment in acute respiratory distress syndrome (ARDS). METHODS: Eighteen dog ARDS models were established by injection of oleic acid into the central vein and ventilated with volume controlled ventilation (VCV,) with the positive end expiratory pressure (PEEP) of 16 cm H(2)O, tidal volume (V(T)) of 10 ml/kg, and respiratory rate (RR) of 30 bpm, and the steady state in this mode was defined as baseline (0 min). After the baseline state was reached, all dogs accepted recruitment maneuver (RM) in pressure controlled ventilation (PCV) mode at the level of 15 cm H(2)O and PEEP of 35 cm H(2)O for 1 min, and then assigned into three groups and ventilated for 4 hs: VCV group (VCV with V(T) of 15 ml/kg), PCV group in which the peak inspiratory airway pressure was adjusted for keeping 15 ml/kg of V(T), and SB grouping which PCV with SB activity and inspiratory airway pressure were adjusted for keeping 15 ml/kg of V(T). PEEP of 10 cm H(2)O, RR of 20 bpm, and inspiratory/expiratory ratio of 1/2 were kept for all animals. Four hours later the dogs were killed and their lungs were taken out. RESULTS: (1) The lower inflection points were identified as 16.1 +/- 1.9, 17.4 +/- 1.5, and 17.1 +/- 1.6 cm H(2)O for the VCV, PCV, and SB group respectively. (2) After RM, the PaO(2) levels of the PCV and SB groups were kept higher than baseline, but the PaO(2) level was as same as that in the baseline state in the VCV group. Compared with the VCV group, higher PaO(2) the PaO(2) level 30 min after RM in the SB group was found (P < 0.05). The intrapulmonary shunt 5 min, 30 min and 1 hr after RM in the SB group were all lower than those in the VCP group, however, not significantly (P = 0.077, 0.058, 0.076). (3) Compared with baseline levels, the plateau pressure and mean airway pressure in all animals were decreased significantly after RM. The plateau pressure in the SB group was significantly lower than that in the VCV group (P < 0.05). CONCLUSION: PCV with spontaneous breathing activity can offset the effect of antiderecruitment for lower PEEP; it would be more safe and effective than VCV. PMID- 18162154 TI - [Impact of RNA interference targeting hypoxia-inducible factor-1alpha on chemosensitivity in esophageal squamous cell carcinoma cells under hypoxia]. AB - OBJECTIVE: To investigate the impact of RNA interference (RNAi) targeting hypoxia inducible factor 1alpha (HIF-1 alpha) on chemosensitivity of esophageal squamous cell carcinoma cells under hypoxia. METHODS: Human esophageal squamous cell carcinoma cells of the line EC9706 were cultured and divided into 3 groups: untransfected group, added with cobalt chloride (CoCl(2)), a chemical hypoxia inducer, for 8 h so as to establish a hypoxia model; control siRNA transfected group, transfected with control siRNA, and 30 h after the transfection exposed to CoCl(2) for 8 h; and HIF-1 alpha siRNA-transfected group, transfected with HIF-1 alpha siRNA, and 30 h later exposed to CoCl(2) for 8 h. Western blotting was used to detect the protein expression of HIF-1 alpha. Another EC9706 were cultured and divided into 3 groups to be treated as mentioned above, and then exposed to cisplantin or platixal under normoxic or hypoxic condition. 24 hours later 3-(4, 5-carboxymethoxypheny1)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay was used to detect the inhibition rates of the cells. Further another EC9706 cells were cultured and then divided into 5 groups: cultured under normoxic condition, cultured under hypoxic condition for 8 h, transfected with control siRNA for 30 h and then under hypoxic condition for 8 h, transfected with HIF-1 alpha siRNA for 30 h and then under hypoxic condition for 8 h. The cell cycle was measured by flow cytometry. RESULTS: The HIF-1alpha protein expression of the HIF 1alpha siRNA group was significantly lower than those of the untransfected and control siRNA transfected groups. The inhibition rates of the EC9706 cells of the groups treated by cisplatin of different concentrations under normoxic condition were all significantly higher than the corresponding levels under hypoxic condition (all P < 0.01). The inhibition rates of the EC9706 cells of the groups treated by platixal of different concentrations under normoxic condition were all significantly higher than the corresponding levels under hypoxic condition (all P < 0.05) Under hypoxic condition, the inhibition rates of the HIF-1alpha siRNA transfected EC9706 cells treated by cisplatin and platixal of different concentrations were all significantly higher than those of the control siRNA transfected and untransfected EC9706 cells (all P < 0.05). Flow cytometry showed that under hypoxic condition the proportion of cells in G(1)-phase of the EC9706 cells was significantly higher, and the proportion of S-phase cells was significantly lower than those of the normoxic group (both P < 0.05), and under the same hypoxic condition the proportion of the EC9706 cells in G(1)-phase was significantly lower, and the proportion the EC9706 cells in S-phase was significantly higher than those of the normoxic group (all P < 0.05). CONCLUSION: The cell cycle arrest induced by HIF-1alpha may be the mechanism of the resistance to anticancer drugs of the esophageal squamous cell carcinoma cells under hypoxic condition. Blocking HIF-1alpha in esophageal squamous cell carcinoma cells may reverse the multidrug resistance of the tumor cells, so it may offer an avenue for gene therapy. PMID- 18162155 TI - [Effects of antisense 6 gene on colon cancer cells]. AB - OBJECTIVE: To investigate the effect of antisense integrin beta6 gene on the growth of colon cancer cells. METHODS: Expressing vector of antisense alphavbeta6 was constructed. Human colon cancer cells of the line HT29 were cultured and divided into 3 groups: Group A, remaining wild type; Group B, transfected with antisense integrin beta6 gene; and Group C, transfected with blank vector. RT-PCR was used to detect the integrin beta6 mRNA expression of in the HT29 cells. The integrinbeta6 protein expression on the surface of the cells was detected by immunohistochemistry and flow cytometry. The binding between the cells and fibronectin was examined. (3)H-labeled thymidine (T) was added into the culture fluid of the cells, and then the radiation amount was detected every 6 days so as to determine the capacity to proliferation of the cells in vitro. Thirty female nude mice were divided into 3 groups to be injected subcutaneously with suspension of HT29 cells of Groups A, B, and C as mentioned above. Six weeks later the size of tumors was measured and part of the tumor nodules were resected 5 weeks after the inoculation to undergo pathological examination. RESULTS: Compared with Groups A and C, no corresponding band at 141 bp was found in Group B by RT-PCR. Flow cytometry showed that the expression level of beta6 protein had was (0.30 +/- 0.051, 30%), significantly lower than those of Groups A and C [(0.80 +/- 0.038, 80%) and (0.85 +/- 0.045, 85%), both P < 0.01]. The binding between the HT29 cells and fibronectin of Group B was significantly degraded after the further addition of anti-beta1 and anti-alphav in comparison of Groups A and C (both P < 0.01). The accumulation values of (3)H-labeled T of Group B 2, 4, and 6 days after addition were all significantly lower than those of Groups A and C (all P < 0.01). The tumors in 9 of the 10 mice injected with the HT29 cells of Group B disappeared and the tumor in the only one mice in Group B was only less than 1 mm(3), significantly smaller then those in Groups A and C (15 mm(3) on average, all P < 0.01). CONCLUSION: Antisense beta6 gene significantly inhibits the mRNA and protein expression of the beta6 gene, and then inhibits the growth and proliferation of colon cancer cells, thus proving that integrin beta6 plays an important role in the regulation of colon cancer cells. PMID- 18162156 TI - [Concentration of endogenous agmatine in normal and injured spinal cord: experiment with rats]. AB - OBJECTIVE: To detect the concentration of endogenous agmatine in normal and injured rats' spinal cord. METHODS: Forty-two SD rats were randomized to sham operation group and 1 hour, 4 hours, 8 hours, 24 hours, 48 hours, 72 hours after operation groups. The modified Allen's model of spinal cord injury was established. The endogenous agmatine was detected both in normal and injured rats' spinal cord by high-performance liquid chromatography (HPLC) with fluorescence detection and OPA derivatization. RESULTS: The endogenous agmatine level in naive rats' spinal cord were (0.643 +/- 0.111) microg/g wet weight. After spinal cord injury, the concentration had a transient drop followed by significantly increase, and then decreased by degrees. CONCLUSION: Up to now, little has been known about the endogenous agmatine levels in mammalian tissues. The results reported by different authors varied up to hundred times. According to our results and other articles, the endogenous agmatine levels in mammals' central nervous system arranged from 0.2 microg/g to 1.1 microg/g wet weight. Although there was a significant increase after spinal cord injury, the peak concentration was extremely lower than what it needed to take its neuroprotective effect in vitro. PMID- 18162157 TI - [Impact of sirolimus on collagen synthesis in vascular smooth muscle cells: experiment with rat]. AB - OBJECTIVE: To evaluate effects of sirolimus on collagen synthesis and collagen type I and type III mRNA expression in vascular smooth muscle cells (VSMCs). METHODS: Rat VSMCs from thoracic aorta were cultured and added with sirolimus of the concentrations of 0, 0.1, 1.0, 10, and 100 ng/ml respectively for 24 h, then co-cultured with L-(3)H-proline for 12 h. L-(3)H-proline incorporation was monitored by liquid scintillation counting. RT-PCR was used to detect the mRNA expression of collagen type I and collagen type III. Logarithmic phase VSMC were treated with sirolimus by 0 ng/ml, 0.1 ng/ml, 1 ng/ml, 10 ng/ml and 100 ng/ml respectively for 24 h prior to 12 h exposure to L-(3)H-proline. RESULTS: The proline incorporation values were 1369 +/- 186, 1211 +/- 157, 1037 +/- 139, 910 +/- 144, and 741 +/- 160 cpm respectively of the 0, 0.1, 1, 10, and 100 ng/ml sirolimus groups. Paired t-test analysis showed that the proline incorporation value of the group with higher sirolimus concentration was significantly lower than that of the group with lower sirolimus concentration (F = 18.936, P < 0.001), except between the 0.1 ng/ml group and 0 ng/ml group (control group) and between the 1 ng/ml group and 10 ng/ml group (P = 0.057, P = 0.12). RT-PCR showed that the collagen I levels were 112 +/- 23, 140 +/- 23, 152 +/- 12, 166 +/- 22, and 179 +/- 18 respectively in the groups of sirolimus of the concentrations of 100, 10, 1, 0.1, and 0 ng/ml respectively, the higher the concentration of sirolimus the lower the collagen I level (all P < 0.05), however, there was no significant difference in the collagen I level between the 0.1 mg/ml group and the control group (P = 0.236). There was no significant difference in the collagen III level among the groups of sirolimus of different concentrations (F = 2.409, P = 0.070). CONCLUSION: Sirolimus inhibits the collagen synthesis in the VSMCs and attenuates the mRNA expression of type I collagen concentration dependently. PMID- 18162158 TI - [Expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule in cardiac allografts and significance thereof: experiment with rats]. AB - OBJECTIVE: To investigate the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in allograft with chronic rejection (CR) after heart transplantation. METHODS: Seventy-two SD rats receiving transplantation of the heats of 71 Wistar rats were divided into 4 equal groups: Group A, undergoing heterotopic cervical heart transplantation (transplanted with donor's heart subcutaneously at the neck) and receiving none intervention, and with the transplanted hearts taken out 3 d after transplantation to the end of CR; Group B, injected intravenously with the splenocytes (SPCs) of the donors on day 0, injected with cyclophomide (CP) on d2, transplanted with the donor hearts on d15, with the transplanted hearts taken out 15 - 120 days after transplantation; Group C, transplanted with the donor's heart and injected intraperitoneally with cyclosporine A (Cs A) 10 mg/kg every other day for 8 - 10 times, and with the transplanted hearts taken out 60 d after transplantation to the end of CR; and Group D, injected intravenously with the SPCs of the donors on day 0, injected with CP on d2, transplanted with the donor hearts on d15, with the transplanted hearts taken out 150 - 420 days after transplantation. Immunohistochemical method was used to detect the expression of ICAM-1 and VCAM-1 in the allografts. RESULTS: The cardiac allograft survival time of Group B was the longest. Less ICAM-1 and VCAM-1 expression and lymphocyte infiltration were observed in SPC and CP-pretreated group. Reversely, protein expression levels of ICAM-1 and VCAM-1 were both high and significant lymphocyte infiltration was seen in Groups B and D. There was no significant difference in the expression of ICAM-1 and VCAM-1 between Groups A and C (P > 0.05). CONCLUSION: The expression levels of ICAM-1 and VCAM-1 are associated with the occurrence and development of rejection and have positive correlation with the severity of graft rejection. Determination of ICAM-1 and VCAM-1 can predict the function of allograft and provide evidence for early diagnosis and prevention of CR. PMID- 18162161 TI - Association of MAPT haplotype-tagging SNPs with sporadic Parkinson's disease. AB - Mutations in the tau gene (MAPT) have been found in families with frontotemporal dementia with parkinsonism linked to chromosome 17. In addition, the MAPT H1 clade specific sub-haplotype, H1c, has been strongly associated with the tauopathies, progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) and, to a lesser extent, with Alzheimer's disease (AD). In Parkinson's disease (PD), there have been several reports of association with the MAPT H1 clade. Although weak to inconclusive, this association is supported by meta analyses of the various studies. To further investigate this baffling role of MAPT in PD, six haplotype-tagging SNPs were genotyped in a large cohort of sporadic PD cases; 324 pathologically confirmed and 248 clinically diagnosed, and 660 controls. In the single-locus association analysis, the H1-clade was associated with an increased risk of PD (p=0.032). In the haplotype-analysis, the sole H2-derived haplotype was under-represented in all of the PD cases compared to controls (p=0.03). There was no significant difference in the distribution of any of the common haplotypes derived from the H1-clade background. Our study supports the hypothesis that genetic variability in the MAPT gene confers susceptibility to PD. However, the effect is not strong, and the H1c haplotype is not involved, suggesting a mechanism that is distinct to that involved in the associated tauopathies and may be explained by the H1/H2 inversion. PMID- 18162162 TI - Analysis of steroidal estrogens as pyridine-3-sulfonyl derivatives by liquid chromatography electrospray tandem mass spectrometry. AB - Sulfonyl chlorides substituted with functional groups having high proton affinity can serve as derivatization reagents to enhance the sensitivity for steroidal estrogens in liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). The most commonly used reagent for derivatization of estrogens for LC-ESI-MS/MS is dansyl chloride. In this study, we compared dansyl chloride, 1,2-dimethylimidazole-4-sulfonyl (DMIS) chloride, pyridine-3-sulfonyl (PS) chloride, and 4-(1H-pyrazol-1-yl)benzenesulfonyl (PBS) chloride for derivatization of 17beta-estradiol (E2) prior to LC-ESI-MS/MS. The product ion spectra of the dansyl and DMIS derivatives were dominated by ions representing derivatization reagent moieties. In contrast, the product ion spectrum of the PS derivative of E2 and, to a lesser extent, the PBS derivative, showed analyte specific fragment ions. Derivatization with PS chloride was therefore chosen for further investigation. The product ion spectrum of the PS derivative of E2 showed intense ions at m/z 272, assigned to the radical E2 cation, and at m/z 350, attributed to the loss of SO(2) from the [M+H](+) ion. Third-stage mass spectrometry of the PS derivative of E2 with isolation and collisional activation of the m/z 272 ion resulted in steroid C and D ring cleavages analogous to those observed in electron ionization mass spectrometry. The product ion spectra of the PS derivatives of estrone, 17alpha-ethinylestradiol, equilin, and equilenin showed similar estrogen-specific ions. Using derivatization with PS chloride, we developed an LC-ESI-MS/MS method with multiple reaction monitoring of primary and confirmatory precursor-to-product ion transitions for the determination of E2 in serum. PMID- 18162163 TI - A fluorescence resonance energy transfer activation sensor for Arf6. AB - The involvement of the small GTPase Arf6 in Rac activation, cell migration, and cancer invasiveness suggests that it is activated in a spatially and temporally regulated manner. Small GTPase activation has been imaged in cells using probes in which the GTPase and a fragment of a downstream effector protein are fused to fluorescent reporter proteins that constitute a fluorescence resonance energy transfer (FRET) donor/acceptor pair. Unlike other Ras family GTPases, the N terminus of Arf6 is critical for membrane targeting and, thus, cannot be modified by fusion to a fluorescent protein. We found that the previously described C terminal green fluorescent protein (GFP) derivative also shows diminished membrane targeting. Therefore, we inserted a fluorescent protein into an inert loop within the Arf6 sequence. This fusion showed normal membrane targeting, nucleotide-dependent interaction with the downstream effector GGA3, and normal regulation by a GTPase-activating protein (GAP) and a guanine nucleotide exchange factor (GEF). Using the recently developed CyPET/YPET fluorescent proteins as a FRET pair, we found that Arf6-CyPET underwent efficient energy transfer when bound to YPET-GGA3 effector domain in intact cells. The addition of platelet derived growth factor (PDGF) to fibroblasts triggered a rapid and transient increase in FRET, indicative of Arf6 activation. These reagents should be useful for investigations of Arf6 activation and function. PMID- 18162164 TI - Facilitated in vivo synthesis of ribonucleic acid and protein via T7 RNA polymerase. AB - Ribozyme and small interfering RNA (siRNA) now are widely used to suppress target genes bearing homologous sequences. In this study, commonly used cell lines (e.g., HEK, HeLa, H1299) were stably transfected with gene encoding T7 RNA polymerase. The cytoplasm-restricted transcription activity of T7 RNA polymerase confers a continuous and robust transcription from T7 promoter-containing oligonucleotide (ODN) template for siRNA or ribozyme and leads to 70 to 80% inhibition of the tested target genes. ODN template offers the advantages of being more stable and economical than synthetic or in vitro-transcribed siRNA or ribozyme. Compared with the use of siRNA/ribozyme-expressing plasmids, our system does not require procedures with preparations of recombinant plasmids and enrichment of transfected cells and can be applied to synthesize protein in which different levels of translation could be modulated via variations in the presence of polyA tail or internal ribosome entry site (IRES) in the T7-transcribed RNAs. The results of our current study provide a rapid and efficient system for the assay of in vivo synthesis and expression of RNAs and proteins. PMID- 18162165 TI - Isotope-coded, iodoacetamide-based reagent to determine individual cysteine pK(a) values by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Cysteine reactivity in enzymes is imparted to a large extent by the stabilization of the deprotonated form of the reduced cysteine (i.e., the thiolate) within the active site. Although this is likely to be an important chemical attribute of many thiol-based enzymes, including cysteine-dependent peroxidases (peroxiredoxins) and proteases, only relatively few pK(a) values have been determined experimentally. Presented here is a new technique for determining the pK(a) value of cysteine residues through quantitative mass spectrometry following chemical modification with an iodoacetamide-based reagent over a range of pH buffers. This isotope-coded reagent, N-phenyl iodoacetamide (iodoacetanilide), is readily prepared in deuterated (d(5)) and protiated (d(0)) versions and is more reactive toward free cysteine than is iodoacetamide. Using this approach, the pK(a) values for the two cysteine residues in Escherichia coli thioredoxin were determined to be 6.5 and greater than 10.0, in good agreement with previous reports using chemical modification approaches. This technique allows the pK(a) of specific cysteine residues to be determined in a clear, fast, and simple manner and, because cysteine residues on separate tryptic peptides are measured separately, is not complicated by the presence of multiple cysteines within the protein of interest. PMID- 18162166 TI - Determination of the structure of lipid vesicle-bound angiotensin II and angiotensin I. AB - A mass spectrometry (MS)-based strategy was developed to determine the structure of lipid vesicle-bound angiotensin II (AII) and angiotensin I (AI). It involves hydrogen-deuterium exchange (HDX), chemical modifications (e.g., nitration of tyrosine, acetylation of free amino group), and ladder sequencing. HDX is also combined with tandem mass spectrometry (MS/MS) to provide structural details at individual amino acid residues. It was observed that a major portion of both of these peptide hormones interacts with the phospholipid head groups on the surface of the vesicles and that Tyr residue is embedded in the vesicles. Both peptides have a U-shaped structure in the lipid environment. PMID- 18162167 TI - Novel chromatographic resolution of chiral diacylglycerols and analysis of the stereoselective hydrolysis of triacylglycerols by lipases. AB - In the present study, we propose a general and accessible method for the resolution of enantiomeric 1,2-sn- and 2,3-sn-diacylglycerols based on derivatization by isocyanates, which can be easily used routinely by biochemists to evaluate the stereopreferences of lipases in a time course of triacylglycerol (TAG) hydrolysis. Diacylglycerol (DAG) enantiomers were transformed into carbamates using achiral and commercially available reagents. Excellent separation and resolution factors were obtained for diacylglycerols present in lipolysis reaction mixtures. This analytical method was then applied to investigate the stereoselectivity of three model lipases (porcine pancreatic lipase, PPL; lipase from Rhizomucor miehei, MML; and recombinant dog gastric lipase, rDGL) in the time course of hydrolysis of prochiral triolein as a substrate. From the measurements of the diglyceride enantiomeric excess it was confirmed that PPL was not stereospecific (position sn-1 vs sn-3 of triolein), whereas MML and rDGL preferentially hydrolyzed the ester bond at position sn-1 and sn-3, respectively. The enantiomeric excess of DAGs was not constant with time, decreasing with the course of hydrolysis. This was due to the fact that DAGs can be products of the stereospecific hydrolysis of TAGs and substrates for stereospecific hydrolysis into monoacylglycerols. PMID- 18162168 TI - A novel sensitive immunoassay method based on the Invader technique. AB - A novel and sensitive immunoassay method has been developed in which the conventional sandwich immunoassay and the highly sensitive DNA detection method, the Invader method, are combined. The signal amplification function of the latter method has been successfully used to enhance the sensitivity of the sandwich immunoassay. The new assay method may be called the Immuno-Invader assay. The assay format involves three important steps: (1) a target antigen is captured and flagged with a biotin-conjugated detection antibody by the sandwich method, (2) streptavidin and a biotin-conjugated oligonucleotide are added to form a complex with the detection antibody, and (3) the oligonucleotide in the complex is detected using the Invader method. The method was applied to the assay of human tumor necrosis factor-alpha (hTNF-alpha). Detection limits obtained were 0.1 pg/ml hTNF-alpha when a luminescent europium chelate was used with a time resolved measurement mode, and 0.8 pg/ml when fluorescein was used with a normal prompt fluorescence measurement mode. On the other hand, the detection limit of a commercially available hTNF-alpha enzyme-linked immunosorbent assay that uses horseradish peroxidase was 3.5 pg/ml. These results demonstrate the feasibility and potential of the new assay method for highly sensitive immunoassay. PMID- 18162169 TI - Near-infrared fluorescence detection permits accurate imaging of loading controls for Western blot analysis. AB - Housekeeping proteins are typically chosen as internal loading controls for Western blot analysis because of their high, relatively constant expression. It was previously reported that antibodies against beta-actin did not reliably identify differences in sample loading, and extended antibody incubations caused a failure to discriminate differences in target protein levels. Here, beta-actin and GAPDH were evaluated as loading controls using near-infrared fluorescence. A load-dependent response in signal intensity was observed over a 250-fold range of sample concentrations, with R(2) values as high as 0.9939. Longer antibody incubations continued to detect differences in protein level and load-dependent responses became more linear. PMID- 18162170 TI - The dysfunction of ATPases due to impaired mitochondrial respiration in phosgene induced pulmonary edema. AB - Phosgene is a toxic gas that is widely used in modern industry, and its inhalation can cause severe pulmonary edema. There is no effective clinical treatment because the mechanism of phosgene-induced pulmonary edema still remains unclear. Many studies have demonstrated that the Na(+)/K(+)-ATPase plays a critical role in clearing pulmonary edema and the inhibition of Na(+)/K(+)-ATPase protein expression has been found in many other pulmonary edema models. In the present study, after the mice were exposed to phosgene, there was serious pulmonary edema, indicating the dysfunction of the ATPases in mice. However, in vitro enzyme study showed that there were increases in the activities of the Na(+)/K(+)-ATPase and Ca(2+)-ATPase. Further investigation showed that the ATP content and mitochondrial respiratory control ratio (RCR) in the lungs decreased significantly. The oxidative stress product, malondialdehyde (MDA), increased while the antioxidants (GSH, SOD, and TAC) decreased significantly. These results indicate that mitochondrial respiration is the target of phosgene. The dysfunction of ATPases due to impaired mitochondrial respiration may be a new mechanism of phosgene-induced pulmonary edema. PMID- 18162171 TI - Interaction of cardiac troponin with cardiotonic drugs: a structural perspective. AB - Over the 40 years since its discovery, many studies have focused on understanding the role of troponin as a myofilament based molecular switch in regulating the Ca(2+)-dependent activation of striated muscle contraction. Recently, studies have explored the role of cardiac troponin as a target for cardiotonic agents. These drugs are clinically useful for treating heart failure, a condition in which the heart is no longer able to pump enough blood to other organs. These agents act via a mechanism that modulates the Ca(2+)-sensitivity of troponin; such a mode of action is therapeutically desirable because intracellular Ca(2+) concentration is not perturbed, preserving the regulation of other Ca(2+)-based signaling pathways. This review describes molecular details of the interaction of cardiac troponin with a variety of cardiotonic drugs. We present recent structural work that has identified the docking sites of several cardiotonic drugs in the troponin C-troponin I interface and discuss their relevance in the design of troponin based drugs for the treatment of heart disease. PMID- 18162172 TI - Mapping of phosphorylation-dependent anti-tau monoclonal antibodies in immunoblots using human tau-constructs synthesized by native chemical ligation. AB - In Alzheimer's disease (AD) neurofibrillary tangles (NFT) are formed by hyperphosphorylated microtubule-associated tau protein. It is still a matter of controversy which phosphorylation sites are AD-specific and how these might be linked to the cause or progress of the disease. Whereas most research projects in this field rely on phosphorylation-dependent tau-specific monoclonal antibodies (mAbs), the phosphorylation patterns recognized by these mAbs are often not characterized in detail. Therefore, we synthesized unphosphorylated, two monophosphorylated (pThr231, pSer235), and the bisphosphorylated (pThr231+pSer235) tau226-240 peptides. The phosphopeptides were ligated via an N terminal cysteine to the thioester-activated C-terminus of human aldo/keto reductase AKR1A1. After purification by preparative gel electrophoresis, the ligation products were analyzed by Western blotting and probed with phosphorylation-dependent anti-tau mAbs HPT-101, HPT-103, HPT-104, and HPT-110. The obtained specificities were very similar to the data obtained by ELISA, showing that ELISA-based epitope mapping studies are also valid for immunoblot analyses. PMID- 18162173 TI - Hes1 stimulates transcriptional activity of Runx2 by increasing protein stabilization during osteoblast differentiation. AB - Runt-related transcription factor 2 (Runx2) is a key transcription factor for osteogenic gene expression and osteoblast differentiation. In order to maintain bone homeostasis, the transcriptional activity of Runx2 is tightly modulated by many intra- and extra-cellular factors. Here, we reveal the mechanism by which Hairy Enhancer of Split1 (Hes1) regulates the transcriptional activity of Runx2, and elucidate the potential role of Hes1 during osteoblast differentiation. Coexpression of Hes1 with Runx2 promoted an increase in Runx2 protein levels by increasing the half-life of Runx2; Hes1 thereby augmented the formation of a Runx2-DNA complex at Runx2 target sites. During osteoblast differentiation, the retroviral overexpression of Hes1 accelerated osteogenesis and stimulated the expression of osteogenic marker genes, including osteopontin and type 1 collagen. Taken together, these results suggest that Hes1 augments the protein level and transcriptional activity of Runx2, resulting in the stimulation of osteoblast differentiation. PMID- 18162174 TI - Gpx1 is a stationary phase-specific thioredoxin peroxidase in fission yeast. AB - The genome sequence of Schizosaccharomyces pombe reveals only one gene for a putative glutathione peroxidase (gpx1(+)). The Gpx1 protein has a peroxidase activity but preferred thioredoxin to glutathione as an electron donor when examined in vitro and in vivo, and therefore is a thioredoxin peroxidase. Besides H(2)O(2), it can reduce alkyl and phospholipid hydroperoxides. Expression of the gpx1 gene was elevated at the stationary phase, and we found that it supported long-term survival of S. pombe. The mutant also exhibited some defect in the activity of aconitase, an oxidation-labile Fe-S enzyme in mitochondria. Activity of sulfite reductase, a labile Fe-S enzyme in the cytosol, was also dramatically lowered in the mutant in the stationary phase. The Gpx1 protein, without any obvious targeting sequence, was localized in mitochondria as well as in the cytosol. Therefore, Gpx1 must serve to ensure optimal mitochondrial function and cytosolic environment, especially in the stationary phase. PMID- 18162175 TI - Recombinant CBD-HepII polypeptide of fibronectin inhibits alphavbeta3 signaling and hematogenous metastasis of tumor. AB - The interaction of integrin alphavbeta3 and its ligands are crucial for tumor metastasis. Recombinant CBD-HepII polypeptide of fibronectin, designated as CH50, suppressed the binding of tumor cells to ECM molecules, and abolished the promoting effect of soluble fibronectin and fibrinogen on tumor cell adhesion to ECM molecules. The underlying mechanisms involve the blockade and downregulation of alphavbeta3 and its co-receptor syndecan 1 by CH50. The activation of FAK, upregulation of cdc2, the production and activation of MMP-2 and MMP-9 by ECM molecules-stimulated tumor cells were inhibited by CH50. CH50 reduced the tumor cell arrest during blood flow, and also inhibited the invasive ability of tumor cells. The in vivo expressed CH50 suppressed the lung metastasis of circulating tumor cells, and prolonged the survival of mice after tumor cell inoculation. These findings suggest a prospective utility of CH50 in the gene therapy for prevention of tumor metastasis. PMID- 18162176 TI - The Ixodes scapularis salivary protein, salp15, prevents the association of HIV-1 gp120 and CD4. AB - Ixodes scapularis salivary protein, Salp15, inhibits CD4(+) T cell activation by binding to the most-extracellular domains of the CD4 molecule, potentially overlapping with the gp120-binding region. We now show that Salp15 inhibits the interaction of gp120 and CD4. Furthermore, Salp15 prevents syncytia formation between HL2/3 (a stable HeLa cell line expressing the envelope protein) and CD4 expressing cells. Salp15 prevented gp120-CD4 interaction at least partially through its direct interaction with the envelope glycoprotein. A phage display library screen provided the interacting residues in the C1 domain of gp120. These results provide a potential basis to define exposed gp120 epitopes for the generation of neutralizing vaccines. PMID- 18162177 TI - Sustained up-regulation of semaphorin 3A, Neuropilin1, and doublecortin expression in ischemic mouse brain during long-term recovery. AB - Strategies to provide neuroprotection and to promote regenerative axonal outgrowth in the injured brain are thwarted by the plethora of axon growth inhibitors and the ligand promiscuity of some of their receptors. Especially, new neurons derived from ischemia-stimulated neurogenesis must integrate this multitude of inhibitory molecular cues, generated as a result of cortical damage, into a functional response. More often than not the response is one of growth cone collapse, axonal retraction and neuronal death. Therefore, characterization of the expression of inhibitory molecules in long-term surviving ischemic brains following stroke is important for designing selective therapeutics. Here, we describe a long-term recovery mouse model for cerebral ischemia in which a brief transient occlusion of the middle cerebral artery (30min) was followed by up to 30 days of long-term reperfusion. Significantly decreased grip strength motor function and increased expression of one of the major repulsive guidance cues, Semaphorin 3A (Sema3A) and its receptor Neuropilin1 (NRP1) occurred in brains of these mice. Interestingly, increased Doublecortin (DCX) expression occurred only in the lateral ventricular wall zone, but not in the dentate gyrus granule cell layer on the ischemic side of the brain. Importantly, no DCX positive cells were detected in the infarct core region after 30d ischemic recovery. Collectively, these studies demonstrated the sustained elevation of Sema3A/NRP1 expression in the ischemic territory, which may contribute to the inhibitory microenvironment responsible for preventing new neurons from entering the infarct area. This model will be of use as a platform for testing anti-inhibitory therapies to stroke. PMID- 18162178 TI - The unique functions of cardiac troponin I in the control of cardiac muscle contraction and relaxation. AB - We review development of evidence and current perceptions of the multiple and significant functions of cardiac troponin I in regulation and modulation of cardiac function. Our emphasis is on the unique structure function relations of the cardiac isoform of troponin I, especially regions containing sites of phosphorylation. The data indicate that modifications of specific regions cardiac troponin I by phosphorylations either promote or reduce cardiac contractility. Thus, a homeostatic balance in these phosphorylations is an important aspect of control of cardiac function. A new concept is the idea that the homeostatic mechanisms may involve modifications of intra-molecular interactions in cardiac troponin I. PMID- 18162179 TI - Covalent modification of stathmin by CCNU determined by FTMS analysis of modified proteins and tryptic peptides. AB - Chemical modification of proteins is often carried out to generate protein-small molecule conjugates for various applications. The high resolution and mass accuracy of a Fourier transform mass spectrometer is particularly useful for assessing the extent or sites of covalent modifications. As protein-small molecule reactions often produce products with variable numbers of the compound incorporated at different sites, a direct mass analysis of the reaction products at times yields mass spectra hard to interpret. Chromatographic separation at protein level could reduce the complexity of a sample, thus allowing more accurate mass spectrometric analysis. In this report, we demonstrate the utility of reversed-phase protein chromatography and FT-ICR mass spectrometry in analyzing CCNU (lomustine, 1-(2-chloroethyl)-3-cyclohexyl-1-nitroso-urea, MW: 233.7Da) modification of stathmin. With this combined approach, we determined the stoichiometry as well as sites of CCNU incorporation into the protein, demonstrating differential reactivity of several lysyl residues to CCNU alkylation. PMID- 18162180 TI - Unique agonist-bound cannabinoid CB1 receptor conformations indicate agonist specificity in signaling. AB - Cannabinoid drugs differ in their rank order of potency to produce analgesia versus other central nervous system effects. We propose that these differences are due to unique agonist-bound cannabinoid CB1 receptor conformations that exhibit different affinities for individual subsets of intracellular signal transduction pathways. In order to test this hypothesis, we have used plasmon waveguide resonance (PWR) spectroscopy, a sensitive method that can provide direct information about ligand-protein and protein-protein interactions, and can detect conformational changes in lipid-embedded proteins. A recombinant epitope tagged human cannabinoid CB1 receptor was expressed in insect Sf9 cells, solubilized and purified using two-step affinity chromatography. The purified receptor was incorporated into a lipid bilayer on the surface of the PWR resonator. PWR spectroscopy demonstrated that cannabinoid agonists exhibit high affinity (KD=0.2+/-0.03 nM and 2+/-0.4 nM for CP 55,940 and WIN 55,212-2, respectively) for the purified epitope tagged hCB(1) receptor. Interestingly however, these structurally different cannabinoid agonists shifted the PWR spectra in opposite directions, indicating that CP 55,940 and WIN 55,212-2 binding leads to different hCB1 receptor conformations. Furthermore, PWR experiments also indicated that these CP 55,940-and WIN 55,212-bound hCB1 receptor conformations exhibit slightly different affinities to an inhibitory G protein heterotrimer, Gi1 (KD=27+/-8 nM and KD=10.7+/-4.7 nM, respectively), whereas they strikingly differ in their ability to activate this G protein type. PMID- 18162181 TI - Novel pharmacological activity of loperamide and CP-339,818 on human HCN channels characterized with an automated electrophysiology assay. AB - Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels underlie the pacemaker currents in neurons (I(h)) and cardiac (I(f)) cells. As such, the identification and characterization of novel blockers of HCN channels is important to enable the dissection of their function in vivo. Using a new IonWorks HT electrophysiology assay with human HCN1 and HCN4 expressed stably in cell lines, four HCN channel blockers are characterized. Two blockers known for their activity at opioid/Ca(2+) channels and K(+) channels, loperamide and CP 339,818 (respectively), are described to block HCN1 more potently than HCN4. The known HCN blocker ZD7288 was also found to be more selective for HCN1 over HCN4, while the HCN blocker DK-AH269 was equipotent on HCN4 and HCN1. Partial replacement of the intracellular Cl(-) with gluconate reduced the potency on both channels, but to varying degrees. For both HCN1 and HCN4, ZD7288 was most sensitive in lower Cl(-) solutions, while the potency of loperamide was not affected by the differing solutions. The block of HCN1 for all compounds was voltage-dependent, being relieved at more negative potentials. The voltage dependent, Cl(-) dependent, HCN1 preferring compounds described here elaborate on the current known pharmacology of HCN channels and may help provide novel tools and chemical starting points for the investigation of HCN channel function in natively expressing systems. PMID- 18162182 TI - Characterization of Rab21-positive tubular endosomes induced by PI3K inhibitors. AB - We found that wortmannin, a potent phosphoinositide 3-kinase (PI3K) inhibitor, markedly induced the formation of Rab21-positive tubular compartments in A431 cells. By time-lapse fluorescence microscopy of live cells co-expressing fluorescent protein-fused Rab21 and other marker proteins, it was shown that the Rab21-positive tubules in wortmannin-treated cells were derived from Rab5 positive early endosomes, but not from late endosomes, recycling endosomes, lysosomes or the trans-Golgi network. The formation of Rab21-positive tubules was very dynamic and required microtubules. Rab21-positive tubules were also formed by the treatment of cells with 3-methyladenine (3-MA), which inhibits class III PI3K rather than class I PI3K. Furthermore, the loss of PI(3)P correlated with the tubulation of Rab21-positive endosomes in cells co-expressing fluorescent protein-fused Rab21 and a tandem FYVE domain. These results suggest that the lowering of PI(3)P as a result of class III PI3K inhibition may be an important cue for the morphological change of Rab21-positive early endosomes from vesicular to tubular form. PMID- 18162183 TI - Unilateral vs. bilateral STN DBS effects on working memory and motor function in Parkinson disease. AB - Bilateral subthalamic nucleus deep brain stimulation (STN DBS) can reduce working memory while improving motor function in Parkinson disease (PD), but findings are variable. One possible explanation for this variability is that the effects of bilateral STN DBS on working memory function depend in part on functional or disease asymmetry. The goal of this study was to determine the relative contributions of unilateral DBS to the effects seen with bilateral DBS. Motor (Unified Parkinson Disease Rating Scale Part III, UPDRS) and working memory function (Spatial Delayed Response, SDR) were measured in 49 PD patients with bilateral STN DBS while stimulators were Both-off, Left-on, Right-on and Both-on in a randomized, double-blind manner. Patients were off PD medications overnight. Effects of unilateral DBS were compared to effects of bilateral STN DBS. Mean UPDRS and SDR responses to Left-on vs. Right-on conditions did not differ (p>.20). However, improvement in contralateral UPDRS was greater and SDR performance was more impaired by unilateral DBS in the more affected side of the brain than in the less affected side of the brain (p=.008). The effect of unilateral DBS on the more affected side on contralateral UPDRS and SDR responses was equivalent to that of bilateral DBS. These results suggest that motor and working memory function respond to unilateral STN DBS differentially depending on the asymmetry of motor symptoms. PMID- 18162184 TI - Subthreshold oscillations facilitate neuropathic spike discharge by overcoming membrane accommodation. AB - We have used computer simulation to better understand how the prolonged epoques of repetitive discharge that underlie chronic neuropathic pain are generated. When subjected to step depolarization the cell soma of most primary afferents produces a single spike, or a brief spike burst, and then falls silent. Slow ramp depolarization typical of physiological stimuli does not evoke any spikes, due to the pronounced membrane accommodation of these neurons. Prior work in live DRG neurons suggests that prolonged neuropathic discharge occurs in neurons that generate subthreshold membrane potential oscillations; the rising (depolarizing) phase of the oscillation sinusoid triggers discharge that can last indefinitely. The specific contribution of oscillations to prolonged electrogenesis is not fully understood, however, as they typically add no more than approximately 5 mV to overall cell depolarization. We constructed a computer simulation of a dorsal root ganglion neuron that generates subthreshold oscillations and prolonged electrogenesis. We found that the slope of the rising phase of the oscillation sinusoid, in addition to its amplitude, is an important factor in the ability of oscillations to generate spike trains. The relatively steep slope of oscillation sinusoids facilitates electrogenesis by transiently overcoming the membrane accommodation associated with physiological slow ramp depolarization. PMID- 18162185 TI - Heparanase expression in B16 melanoma cells and peripheral blood neutrophils before and after extravasation detected by novel anti-mouse heparanase monoclonal antibodies. AB - Degradation of extracellular matrix is associated with extravasation of metastatic tumor cells and inflammatory cells. Heparanase, the heparan sulfate specific endo-beta-glucuronidase, is a key enzyme for the matrix degradation, yet its involvement in extravasation and invasion during pathological processes was not fully clarified in vivo. In the present study, we examined heparanase expression in mouse experimental models, lung metastasis of melanoma and skin infiltration of neutrophils. Sixteen novel monoclonal antibodies specific for mouse heparanase were established by enzyme-linked immunosorbent assay with a recombinant mouse proheparanase, immunocytochemical staining of B16F10 melanoma cells cultured in vitro, and immunoprecipitation of the lysate of heparanase transfectant cells. Heparanase expression in metastatic nodules of B16F10 melanoma cells and in neutrophils localized in the inflamed skin was immunohistochemically detected using a monoclonal antibody RIO-1 that recognized the C-terminus of mouse heparanase. Homogeneous and strong heparanase staining was observed in 46% of the lung micrometastases of B16F10 melanoma cells. The staining was intensely positive on the invasive front of larger established metastasis nodules, but it was weak or heterogeneous inside the nodules. Heparanase expression in skin-infiltrating neutrophils was examined after inducing local inflammation with croton oil. The monoclonal antibody stained a significant portion of neutrophils inside and along the blood vessels, whereas it did not stain dermal neutrophils located distant from the vasculatures. The present study strongly suggests that both melanoma cells and neutrophils transiently express heparanase before and during the invasive process in vivo. PMID- 18162186 TI - Activation of natriuretic peptide receptor-C attenuates the enhanced oxidative stress in vascular smooth muscle cells from spontaneously hypertensive rats: implication of Gialpha protein. AB - We have recently shown that vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) exhibit enhanced expression of Gialpha proteins, which was attributed to the enhanced oxidative stress. Since C-ANP(4 23) that specifically interacts with natriuretic peptide C (NPR-C) receptor has been shown to decrease the expression of Gialpha protein in VSMC, the present study was undertaken to examine if C-ANP(4-23) can also decrease the enhanced expression of Gialpha protein in VSMC from SHR and whether it is attributed to its ability to attenuate the enhanced oxidative stress. Aortic VSMC from 12-week old SHR and their age-matched Wistar-Kyoto (WKY) rats were used for the present studies. VSMC from SHR exhibited enhanced expression of Gialpha-2 and Gialpha-3 proteins, different subunits of NADPH oxidase such as Nox(4) and p(47phox) proteins but not of p(22phox), enhanced production of superoxide anion as well as NADPH oxidase activity as compared to age-matched WKY rats. Treatment of VSMC from SHR with C-ANP(4-23) decreased towards control levels the enhanced expression of Gialpha proteins, enhanced superoxide anion production and enhanced NADPH oxidase activity as well as the enhanced expression of Nox(4) and p(47phox). However, C-ANP(4-23)-induced attenuation of the enhanced level of O(2)(-) and NADPH oxidase activity occurs at 4 h before the decrease in the enhanced expression of p(47phox) that occurs at 16 h of C-ANP(4-23) treatment. The decreased expression of NADPH oxidase in SHR was also associated with further decrease in O(2)(-) and NADPH oxidase activity. Furthermore, treatment of VSMC from SHR with pertussis toxin (PT) decreased the enhanced levels of superoxide anion as well as NADPH oxidase activity; however, the enhanced levels of different subunits of NADPH oxidase were not attenuated by PT treatment. These results suggest that C-ANP(4-23) decreases the enhanced oxidative stress in SHR by attenuating the enhanced expression of Gialpha proteins and also the enhanced levels of NADPH oxidase. PMID- 18162187 TI - Patterns of habitual activity across weekdays and weekend days in 9-11-year-old children. AB - OBJECTIVE: To characterize the pattern of activity in boys and girls across weekdays and weekend days. METHODS: Physical activity was recorded every 2 s by uniaxial accelerometry in 84 children, aged 9-11 years, for up to four weekdays and two weekend days. Activity bouts (>or= 4 s and >or= 5 min) greater than light (>or= LIGHT), moderate (>or= MOD) and vigorous (>or= VIG) intensity were recorded. The study took place in the South-West of England in 2007. RESULTS: The mean duration of activity bouts decreased as intensity increased from 11.0+/-1.3 s for >or= LIGHT activity to 6.1+/-1.0 s for >or= VIG activity. The frequency, duration and intensity of bouts were greater in boys than girls, and the frequency and duration of bouts were greater on weekdays than weekend days. Girls accumulated more activity sporadically than boys, whereas boys accumulated more activity in >or= 5-min bouts. CONCLUSION: Sex differences and weekday/weekend differences in activity were largely due to the intensity of the most frequent bouts of activity and frequency of the most intense bouts. Information regarding the pattern of children's habitual activity can be used to inform activity interventions and assess the aspects of the activity pattern that are related to health. PMID- 18162188 TI - Advance directives in bipolar disorder, a cognitive behavioural conceptualization. AB - Mental Health Advance Directives (MHADs) are potentially useful for bipolar patients due to the episodic characteristic of their disease. An interest for the development of a Cognitive Behavioural Therapy (CBT) approach in the creation process of MHADs arises because of a lack of efficiency of the non-collaborative processes, the potential impact of psychopathology and the awareness of the illness in MHADs' content, the link between the patient's directives interest and the case manager's interest, and the lower interest reported by the potentially high MHADs beneficiary. The CBT intervention in the MHADs creation process that is proposed in this article is based on: the self-determination model for adherence, the cognitive representation of illness model, and the concordance model. The principles of the intervention were adapted from Motivational interviewing's (MI), emphasising personal choice and responsibility, and focusing on the patient's concerns about the treatment through Socratic dialogue. During the course of 2004, 20 advance directives written by patients (17 with bipolar disorder, 3 with schizoaffective disorder) have been collected following the described intervention. Therefore, this intervention seems to be relevant for patients with bipolar disorder. PMID- 18162189 TI - Examining age effects on prototypic melancholic symptoms as a strategy for refining definition of melancholia. AB - BACKGROUND: Melancholic depression appears to have a later age of onset than the non-melancholic disorders, and its phenotypic picture also appears to change with age. The latter phenomenon allows clarification of key symptoms of melancholia by examining for age effects on putative melancholic symptoms, thus enabling identification and refinement of the melancholic sub-type. METHODS: We studied 158 patients receiving a diagnosis of unipolar depression (65 melancholic: 93 non melancholic), dichotomised by age and with a higher representation of those with melancholia in the older age band. The severity of individual DSM-IV-TR melancholic candidate symptom constructs were quantified across age groups and diagnostic sub-type. RESULTS: Symptom constructs identified as most clearly associated with age effects in those with melancholia were anhedonia, non reactivity, diurnal mood variation and, to a lesser degree, psychomotor slowing. When melancholic and non-melancholic patients were compared, non-reactivity, psychomotor slowing and diurnal mood variation were the most differentiating in the older age group. CONCLUSIONS: The capacity of certain symptoms to mark the changing phenotypic expression of melancholia with age may not only assist refined definition of melancholia but inform about underlying causes and, of key importance, explain the suggested differential impact of narrow-action and broad action antidepressant on those with melancholia across differing age groups. PMID- 18162190 TI - High levels of the proNGF peptides LIP1 and LIP2 in the serum and synovial fluid of rheumatoid arthritis patients: evidence for two new cytokines. AB - The proNGF peptides LIP1 and LIP2 display multiple biological and physiological properties several of which share common features with the nerve growth factor (NGF). The objective of this study was firstly to demonstrate the presence of these peptides in the human sera and secondly to provide evidence for their involvement in inflammatory diseases. Their levels measured by specific enzyme linked immunosorbent assays (ELISA) were found to be more than 10-fold higher in sera of patients with rheumatoid arthritis (RA), as compared to healthy controls. High levels of LIP1 and LIP2 were also detected in the synovial fluid (SF) of RA patients. These results provide first evidence for a cytokine-like role of the LIP1 and LIP2 peptides. PMID- 18162191 TI - Fecal collection, ambient preservation, and DNA extraction for PCR amplification of bacterial and human markers from human feces. AB - Feces contain intestinal bacteria and exfoliated epithelial cells that may provide useful information concerning gastrointestinal tract health. Intestinal bacteria that synthesize or metabolize potential carcinogens and produce anti tumorigenic products may have relevance to colorectal cancer, the second most common cause of cancer deaths in the USA. To facilitate epidemiological studies relating bacterial and epithelial cell DNA and RNA markers, preservative/extraction methods suitable for self-collection and shipping of fecal samples at room temperature were tested. Purification and PCR amplification of fecal DNA were compared after preservation of stool samples in RNAlater (R) or Paxgene (P), or after drying over silica gel (S) or on Whatman FTA cards (W). Comparisons were made to samples frozen in liquid nitrogen (N2). DNA purification methods included Whatman (accompanying FTA cards), Mo-Bio Fecal (MB), Qiagen Stool (QS), and others. Extraction methods were compared for amount of DNA extracted, DNA amplifiable in a real-time SYBR-Green quantitative PCR format, and the presence of PCR inhibitors. DNA can be extracted after room temperature storage for five days from W, R, S and P, and from N2 frozen samples. High amounts of total DNA and PCR-amplifiable Bacteroides spp. DNA (34%+/-9% of total DNA) with relatively little PCR inhibition were especially obtained with QS extraction applied to R preserved samples (method QS-R). DNA for human reduced folate carrier (SLC19A1) genomic sequence was also detected in 90% of the QS-R extracts. Thus, fecal DNA is well preserved by methods suitable for self collection that may be useful in future molecular epidemiological studies of intestinal bacteria and human cancer markers. PMID- 18162192 TI - Effect of chitin on the antagonistic activity of Cryptococcus laurentii against Penicillium expansum in pear fruit. AB - This study was designed to evaluate the impact of chitin on the antagonistic activity of Cryptococcus laurentii against the postharvest blue mold rot caused by Penicillium expansum in pear fruit. The results showed that the antagonistic activity of C. laurentii obtained from the culture media of nutrient yeast dextrose broth (NYDB) amended with chitin at 0.5-1.0% was improved greatly compared with the case that without chitin. The addition of chitin to NYDB did not influence the growth of C. laurentii, however, its population was found to increase rapidly thereafter in pear fruit wounds compared to that harvested from NYDB without chitin. Moreover, the cell-free filtrate of the chitin-supplement culture media in which the yeast was incubated for 24 h emerged a direct antifungal activity against P. expansum in pear fruit wounds, with the associated high level of chitinase activity. These results suggested that the use of chitin may be an effective method to induce the antagonistic activity of C. laurentii. To our knowledge, this is the first report regarding the chitin could enhance the efficacy of postharvest biocontrol yeasts. PMID- 18162193 TI - Effect of aw and CO2 level on Aspergillus flavus growth and aflatoxin production in high moisture maize post-harvest. AB - The potential for using modified atmospheres of 25-75% CO2 (balanced with N2) and water activity (aw, 0.95, 0.92) to control Aspergillus flavus development and aflatoxin B1 production has been evaluated (a) on synthetic medium and (b) on maize grain during storage for up to 21 days at 25 degrees C. On agar medium up to 75% CO2 at both 0.95 and 0.92 aw significant inhibition of growth was obtained (P<0.05). In stored grain inoculated with spores of A. flavus there was significantly higher populations of the species at 0.95 aw than 0.92 aw. Up to 75% CO2 resulted in an inhibition of the populations of A. flavus isolated from the grain. Contrasting aflatoxin B1 production was obtained on agar and in stored maize grain. On agar, greatest amounts were produced at 0.92 aw, while more was produced at 0.95 aw on maize grain. Overall, the efficacy of controlled atmospheres x aw showed that treatment with 25% CO2 could be sufficient to efficiently reduce A. flavus development but at least 50% CO2 was required to obtain a significant reduction of aflatoxin synthesis. PMID- 18162194 TI - Bioreducible poly(amido amine)s with oligoamine side chains: synthesis, characterization, and structural effects on gene delivery. AB - A group of bioreducible poly(amido amine)s containing multiple disulfide linkages in main chain and oligoamines in side chain (SS-PAOAs) were prepared by Michael type polyaddition of N-tert-butyloxycarbonyl (N-Boc) protected oligoamine to the disulfide-containing cystaminebisacrylamide, followed by deprotection of the Boc protective groups. These linear polymers show strong DNA condensation capability at low N/P ratios. The chemical structure of oligoamine side chains (i.e. amine type and amino spacer length) in the SS-PAOAs has a distinct effect on their buffer capacity, transfection efficiency and toxicity profile. The SS-PAOAs containing secondary amino functions in the side chain show high buffer capacities and are able to transfect COS-7 cells in vitro at low N/P ratios, with transfection efficiencies similar or even higher than those of 25 kDa branched pEI, along with very low cytotoxicity as determined by XTT assay. Increase of the alkyl spacer from ethylene to propylene between the amino units in side chains results in significant lower transfection and increased toxicity. This study presents detailed factors influencing the relationship between structure and gene delivery properties and may provide helpful insights for the further development of safe and efficient non-viral vectors. PMID- 18162195 TI - o-Diphenol oxidase activity of molluscan hemocyanins. AB - Diphenoloxidase activities of two molluscan hemocyanins, isolated from the marine snails Rapana venosa and garden snails Helix vulgaris were studied using o diphenol and L-Dopa as substrates. The dimers of H. vulgaris Hc show both, diphenol (K(m)=2.86 mM and K(cat)=4.48) and L-Dopa activity due to a more open active sites of the enzyme and better access of the substrates. The K(m) value of molluscan H. vulgaris Hc is very close to those of Helix pomatia and Sepia officinalis Hcs, but several times higher compared to those of Rapana and Octopus Hcs. Also HvH has a very high enzyme activity compared with other molluscan Hcs. Kinetic measurements with native RvH and both structural subunits, RvH1 and RvH2, show that RvH and only one structural subunit, RvH2, exhibited only o-diphenol activity, but no L-Dopa oxidizing activity. PMID- 18162196 TI - Up-regulation of PPARgamma in myocardial infarction. AB - BACKGROUND: Peroxisome proliferator activated receptors (PPARs) are key regulators for cardiac energy metabolism after myocardial injury. We hypothesized, that PPARs are regulated in myocardial infarction (MI) and their activity is modulated by angiotensin receptor blockers (ARBs). METHODS: Following induction of MI, male rats were treated with placebo or the ARB irbesartan for three weeks. PPARalpha, beta/delta and gamma protein expression and gene expression of PPAR target genes and glucose transporters were measured. PPARgamma protein expression was analyzed by immunofluorescence. RESULTS: MI decreased LVP and dp/dtmax and increased LVEDP, this effect was counteracted by irbesartan. PPARalpha and PPARbeta/delta protein expression was not altered in MI and was not affected by irbesartan. PPARgamma protein content was increased in the infarcted area and localized to cardiac myocytes and fibroblasts. In parallel, expression of CTGF was increased 10-fold in the infarcted zone. PPAR target genes (CD36, MCAD, ACO and GLUT4) were significantly decreased in infarcted tissue, and this was unaffected by irbesartan. However, CD36 and ACO in the non-infarcted areas were up-regulated by irbesartan. CONCLUSION: Endogenous up-regulation of PPARgamma in MI is insufficient to counteract the decrease in metabolic genes, but parallels an increase in the profibrotic mediator CTGF. Irbesartan increases fatty acid oxidating enzymes after MI independent of PPARgamma regulation. PMID- 18162197 TI - Muscle receptors close to the myotendinous junction play a role in eliciting exercise pressor reflex during contraction. AB - Although a muscle mechanosensitive reflex contributes to regulation of the cardiovascular responses during exercise, the precise location of muscle mechanoreceptors responding to contraction has not been identified yet. We have recently reported that mechanosensitive receptors located at or close to the myotendinous junction play a role in eliciting the cardiovascular responses to passive stretch of skeletal muscle. The mechanoreceptors located at or near the myotendinous junction are hypothesized to respond to static contraction as well. To test this hypothesis, we had two interventions for the reflex cardiovascular responses to static contraction of the triceps surae muscle with the same tension development in decerebrate or pentobarbital-anesthetized rats; cutting the Achilles tendon and local injection of lidocaine into the myotendinous junction. The cardiovascular responses were evoked by static contraction regardless of the achillotomy, suggesting that mechanoreceptors terminating in the more distal part of the cut Achilles tendon did not contribute to the reflex cardiovascular responses. Lidocaine (volume, 0.04-0.1 ml) injected into the myotendinous junction blunted the reflex cardiovascular responses, indicating that muscle afferent fibers terminating at or passing through the myotendinous junction contribute to the exercise pressor reflex. The achillotomy did not affect the cardiovascular responses to passive stretch with the same tension as static contraction, but the localized injection of lidocaine similarly blunted the responses to passive stretch as contraction. We conclude that the mechanosensitive receptors eliciting the reflex cardiovascular responses may at least partly locate close to the myotendinous junction, to monitor tension development during muscular activity. PMID- 18162198 TI - Obesity in the parturient: an increasing burden. PMID- 18162199 TI - Comparison of epidural analgesia with combined spinal-epidural analgesia for labor: a retrospective study of 6497 cases. AB - BACKGROUND: Combined spinal-epidural analgesia provides rapid-onset analgesia with minimal motor block, but it is a more invasive technique than epidural analgesia and the risk of complications may be increased. This study compared the safety and effect on delivery of combined spinal-epidural and epidural analgesia in labor. METHODS: A retrospective observational study was performed. Data were collected from 6497 women who received regional analgesia in our tertiary hospital in 2005. The incidence of complications during labor and the day after delivery was compared. The effect on labor outcome with both techniques was also assessed. RESULTS: 1964 received combined spinal-epidural (30.2%) and 4533 epidural analgesia (69.8%). Quality of analgesia was better in the combined spinal-epidural group. Labor outcome was similar in the two groups. Pruritus, paresthesia and back pain were more frequent in the combined spinal-epidural group. No differences were observed in the incidence of accidental dural puncture or post dural puncture headache. CONCLUSIONS: We found that epidural and combined spinal-epidural analgesia were comparable in terms of safety, and had a similar effect on delivery type. PMID- 18162200 TI - Assessment of knowledge regarding cardiopulmonary resuscitation of pregnant women. AB - INTRODUCTION: The 2000-2002 triennial UK Report on Confidential Enquiries into Maternal Deaths concluded that over 50% of maternal deaths involved substandard care and that many could have been prevented. Catastrophic events leading to cardio-respiratory arrest may necessitate the resuscitation of pregnant women in various hospital locations. This study was designed to evaluate knowledge about resuscitation of parturients among anesthesiologists, obstetricians and emergency physicians. METHODS: A 12-question survey was distributed anonymously to residents and faculty in the anesthesia (ANES), obstetrics (OB), and emergency medicine (EM) departments at Stanford University Medical Center/Lucile Packard Children's Hospital, Stanford, California. Questions were designed to elicit knowledge deficiencies in four critical areas: need for left uterine displacement (LUD), advanced cardiac life support algorithms (ACLS), physiologic changes of pregnancy (PHYS), and the recommendation to perform cesarean delivery in parturients (>20 weeks gestation) after 4-5 min of unsuccessful resuscitation for cardiac arrest (5CD). RESULTS: In total, 74/75 physicians (43% ANES, 37% OB, and 20% EM) completed the test. ANES scored highest in overall test scores, and in knowledge of PHYS (P<0.05). Scores for LUD and 5CD were similar among groups, but 25-40% of these questions were answered incorrectly. In the ACLS category, the EM group scored highest (93%). CONCLUSION: We conclude that knowledge of important basic concepts, including the need for LUD and the potential benefit of early cesarean delivery during cardiac arrest, is inadequate among all three specialties. All three departments should provide ACLS physician training with emphasis on the special considerations for parturients. PMID- 18162201 TI - Cell salvage in obstetrics. AB - The safety of cell salvage in obstetrics has been questioned because of the presumed risk of precipitating amniotic fluid embolism and, to a lesser extent, maternal alloimmunisation. For these reasons, experience in this field is limited and has lagged far behind that in other surgical specialties. There has, however, been renewed interest in its use over recent years, mainly as a result of problems associated with allogeneic blood transfusion. Our aim was to review the medical literature to ascertain the principles of cell salvage, the ability of the process to remove contaminants, and its safety profile in the obstetric setting. The search engines PubMed and Google Scholar were used and relevant articles and websites hand searched for further references. Existing cell salvage systems differ in their ability to clear contaminants and all require the addition of a leucocyte depletion filter. Although large prospective trials of cell salvage with autotransfusion in obstetrics are lacking, to date, no single serious complication leading to poor maternal outcome has been directly attributed to its use. Cell salvage in obstetrics has been endorsed by several bodies based on current evidence. Current evidence supports the use of cell salvage in obstetrics, which is likely to become increasingly commonplace, but more data are required concerning its clinical use. PMID- 18162202 TI - Transfusion-related acute lung injury (TRALI) in an obstetric patient. AB - Transfusion-related lung injury (TRALI) is the leading cause of mortality following transfusion of blood products. Despite increasing awareness, the condition often remains unrecognised and therefore underreported. A 28-year-old with moderate preeclampsia had a post-partum haemorrhage following emergency caesarean section. Shortly after receiving three units of packed red cells she went into respiratory failure, which progressed to cardiac arrest. She was successfully resuscitated and made a slow but full recovery. Investigation through the National Blood Service confirmed the diagnosis of TRALI. TRALI is an increasingly common life-threatening complication of blood transfusion and should be included in the differential diagnosis of collapse in an obstetric patient who has recently received a blood product transfusion. PMID- 18162203 TI - Selective arterial embolisation: a first line measure for obstetric haemorrhage? AB - Selective arterial embolisation is increasingly used to arrest intractable postpartum haemorrhage. We report a case of postpartum haemorrhage following a placenta praevia, which had a successful outcome with selective arterial embolisation as the first treatment option. This technique should be more widely available, and in many instances should be considered before and in lieu of any surgical intervention. PMID- 18162204 TI - Management of a parturient with thrombocytopenia-absent-radius syndrome undergoing urgent caesarean section. AB - Thrombocytopenia-absent-radius syndrome is a rare congenital condition characterised by a low platelet count and limb abnormalities. There may be airway difficulties and cardiac disease is frequently associated. We present a case of successful general anaesthesia for urgent caesarean section. The major anaesthetic difficulties encountered were severe thrombocytopenia with a platelet count 30x10(9)/L, which precluded regional anaesthesia, and extensive limb abnormalities resulting in difficulty with vascular access and cardiovascular monitoring. Platelet transfusion was required but airway difficulties were not encountered. PMID- 18162205 TI - Trigeminal nerve palsy and Horner's syndrome following epidural analgesia for labour: not a subdural block. PMID- 18162206 TI - Hyponatraemia complicating vaginal delivery. PMID- 18162207 TI - Epistaxis in pregnancy--not to be sniffed at! PMID- 18162208 TI - Post caesarean delivery pain management: do not dismiss pethidine. PMID- 18162211 TI - The 2008 education supplement on hematopoietic stem cell transplantation. PMID- 18162210 TI - The visual fields: an interdisciplinary history I. The evolution of knowledge. AB - A subjective study of a hemianopic field defect was reported to the London Royal Society in 1824. The German ophthalmologist Albrecht von Graefe devised a means of mapping field defects, which evolved into quantitative perimetry as an exact method of localizing lesions in the visual pathways. Knowledge of these pathways increased during the nineteenth century; final identification of the visual cortex in the occipital lobe was achieved by Japanese and British ophthalmologists and neurologists on the basis of wartime studies of field defects due to cerebral missile wounds. PMID- 18162212 TI - The graft-versus-leukemia effect--breaking the black box open. PMID- 18162213 TI - GVL against murine chronic phase and blast phase CML. PMID- 18162214 TI - Engineering GVL through T cell gene transfer. PMID- 18162215 TI - Amyloidosis: 2008 BMT Tandem Meetings (February 13-17, San Diego). AB - AL amyloidosis is a rare disease in which immunoglobulin light chains are deposited as aggregated interstitial fibrils that can compromise organ function and lead to death. The risks that patients with amyloidosis face include late diagnosis, misdiagnosis of amyloid type, untimely and ineffective therapy, and toxicities of therapy. The goals of treatment are (1) reduction or elimination of the amyloid-forming protein, usually a free immunoglobulin light chain measured by the serum-free light chain assay; (2) support of the patient pending hematologic response and improvement; and (3) stabilization of organ function. Whenever possible, patients should be treated on clinical trials. PMID- 18162216 TI - Cancer stem cells in hematopoietic malignancies. AB - Most hematopoietic malignancies are comprised of cells that are functionally heterogeneous, with only a subset being responsible for tumor maintenance. These cancer stem cells are so named because they possess qualities reminiscent of normal tissue stem cells including self-renewal, prolonged survival, and the ability to give rise to cells with more differentiated characteristics. Effort is now focused on identifying cancer stem cells in various hematopoietic malignancies, and defining the cells of origin such that the stepwise accumulation of genetic/epigenetic events necessary for cancer stem cell development can be delineated. A detailed understanding of these processes could lead to development of therapeutics that more effectively treat hematopoietic malignancies and potentially other cancers. PMID- 18162217 TI - Memory T cells. PMID- 18162218 TI - The contribution of endothelial activation and injury to end-organ toxicity following allogeneic hematopoietic stem cell transplantation. AB - Over the last 25 years, allogeneic hematopoietic stem cell transplantation (HSCT) has been used increasingly as a curative treatment option for patients with hematologic and neoplastic diseases. Despite major advances in transplant immunology and improvements in supportive and critical care medicine, HSCT is still plagued by several life-threatening complications. As such, the establishment of effective therapeutic options for these complications will be crucial as increasing numbers of high-risk transplants are performed each year. This brief review will discuss the contribution of vascular endothelial cell activation and injury to inflammation and end-organ toxicity that occurs following allogeneic HSCT, and will highlight translational research efforts that have paved the way to the development of novel strategies to treat and prevent disease. Finally, we will discuss in detail the clinical manifestations and challenges encompassed by the syndrome of thrombotic microangiopathy following HSCT. PMID- 18162219 TI - Immunotherapy for pediatric cancer. AB - Improvements in adult cancer survivorship can be achieved from behavioral changes and adopting screening programs. Yet, these approaches cannot be readily applied to lower the morbidity and mortality from childhood cancers. Rather, pediatric oncologists must rely on procedures and therapies to treat, rather than prevent malignancies. The systematic application of chemotherapy, radiation therapy, and surgery has led to remarkable advances in survival but these improvements have come at a cost. Children routinely receive chemotherapy agents that were designed decades ago, and these drugs have predictable side effects that result in the loss of potential for long-term survivors. The advent of targeted applications of immune-based therapies offers children with cancer a new class of oncolytic therapies that may be used to treat disease refractory to conventional approaches and lessen the toxicity of current treatment regimens without compromising remission. This review explores how 3 components of the immune system--T cells, natural killer (NK) cells, and antibodies--can be used for therapy of pediatric malignancies. PMID- 18162220 TI - Alternative allogeneic donor sources for transplantation for childhood diseases: unrelated cord blood and haploidentical family donors. AB - Allogeneic stem cell transplantation has been demonstrated to be curative in a wide variety of pediatric malignant and nonmalignant diseases, and can be traced back over 50 years ago to the original report of Thomas et al. HLA matched sibling donors have been the gold standard for pediatric recipients requiring allogeneic donors for both nonmalignant and malignant conditions. However, only 25% of potential pediatric recipients possesses an HLA-matched sibling donor, and the frequency is even less in those with genetic nonmalignant conditions because of genetically affected other siblings within the family. Therefore, 75% to 90% of potential pediatric recipients require alternative allogeneic donor cells for treatment of their underlying conditions. Potential alternative allogeneic donor sources include unrelated cord blood donors, unrelated adult donors, and haploidentical family donors. In this article we review the experience of both unrelated cord blood donor and haploidentical family donor transplants in selected pediatric malignant and nonmalignant conditions. PMID- 18162221 TI - Vaccination of children following allogeneic stem cell transplantation. PMID- 18162222 TI - Cellular immune reconstitution after haploidentical transplantation in children. AB - Delayed immune reconstitution is 1 of the major contributions to the morbidity and mortality after haploidentical transplantation. Patients with a slow recovery of the innate and especially of the adaptive immune system are at high risk for severe and often lethal infections. The reason for delayed immune reconstitution after haploidentical transplantation include the T cell depletion (TCD) of the graft, the thymic dysfunction induced by pretransplant chemotherapies and by the conditioning regimens, and the occurrence of graft-versus-host disease (GVHD) and its treatment. The detailed analysis, understanding, and manipulation of the reconstitution of the cellular immune system will be of utmost importance to overcome the posttransplant immunodefcient status, and should result in a reduced risk of severe and overwhelming infections and hopefully also to a reduced risk of relapse through better immunological control of residual malignant cells. PMID- 18162223 TI - Immune reconstitution in children after unrelated cord blood transplantation. AB - There is a great need to learn more about the biology of immune recovery after UCBT. Fundamental gaps in knowledge remain regarding the biology and kinetics of developing antigen-specific protective immunity and understanding the impact of recipient age and immunosuppressive agents. However, there is also realistic hope that clinical translation of new immunotherapy strategies could enhance immune competence after UCBT either by having an impact on the thymic-independent early period or by fostering thymic recovery. PMID- 18162224 TI - Hematopoietic stem cell transplantation for severe combined immunodeficiency diseases. AB - Hematopoietic stem cell transplantation (HSCT) is the only curative option for most children with severe combined immunodeficiency disease (SCID). Survival for SCID following HSCT has significantly improved over the past several decades, and ranges from 70% to 95% depending on the clinical condition of the child at the time of transplant, the availability of an HLA-matched sibling donor, and the SCID genotype/phenotype. In this article we will review the types of SCID and discuss the critical HSCT issues that confront us today, including the optimal source of donor cells when an HLA-matched sibling is not available, as well as the pros and cons of using conditioning therapy pretransplant. As SCID children have been followed for several decades, it is becoming apparent that long-term outcome and durable T and B cell immune reconstitution are quite variable depending on the initial treatment and source of donor cells. Finally, the development of methods to improve the early diagnosis of SCID along with designing prospective trials to evaluate the best approaches to curing these diseases with minimal toxicity are critical to improving outcomes for children with SCID. PMID- 18162225 TI - Oral and dental late effects after pediatric stem cell transplantation. PMID- 18162226 TI - Chronic kidney disease after pediatric hematopoietic cell transplant. AB - There are 3 clearly distinct clinical entities that occur after HCT: TMA, idiopathic CKD, and nephrotic syndrome. The potentially independent role of GVHD and chronic inflammation in the development and progression of idiopathic CKD warrants further investigation. CKD after HCT is a relatively common occurrence. As the indications for and number of transplants performed world wide increases, so will the burden of kidney disease. Identifying those patients at risk for the development of CKD will be important for potential intervention and prevention of CKD and progression to end-stage renal disease in this patient population. There are those patients who will develop CKD that is not related to TBI or the conditioning regimen but rather to complications and/or therapy that occur after HCT, specifically aGVHD and cGVHD and prolonged calcinuerin inhibitor use. The burden of management will fall not only to the nephrologists but the oncologist as well to ensure close monitoring of renal function, blood pressure, and urinalyses posttransplant. It may be that our energies have been misdirected in trying to reduce exposure to TBI, and rather we should try to decrease the inflammatory and cytokine effects of GVHD and reduce exposure to calcineurin inhibitors to prevent CKD in this population of patients. PMID- 18162227 TI - Late effects following hematopoietic cell transplantation for children. AB - High-dose chemotherapy or chemoradiotherapy followed by hematopoietic cell transplantation (HCT) is being used for an ever increasing number of children with malignant or nonmalignant disorders, and improved results are leading to an ever increasing number of long-term survivors. Understanding the delayed effects that may occur after HCT is important in determining the evaluations to perform for this unique group of patients. The articles that follow detail what has been observed to date regarding the oral cavity and dental late effects, the kidney after HCT, and the evaluations to perform to determine late effects when patients return to the centers after their HCT. PMID- 18162228 TI - Clinical strategies to enhance posttransplant immune reconstitution. PMID- 18162229 TI - Controversies in lymphoma: the role of hematopoietic cell transplantation for mantle cell lymphoma and peripheral T cell lymphoma. AB - Mantle cell lymphoma (MCL) and peripheral T cell lymphoma (PTCL) are distinct lymphoma subtypes that each comprise about approximately 10% of the non-Hodgkin lymphomas. Although both subtypes are characterized by high remission rates to frontline chemotherapy, the prognosis is generally poor because of inevitable relapse within 1-2 years or less, depending on the specific histology. Patients with MCL who achieve a complete remission with upfront conventional chemotherapy currently have several options for consolidative therapy including maintenance therapy with rituximab, autologous hematopoietic cell transplantation (HCT), and more recently, allogeneic HCT utilizing a reduced intensity conditioning (RIC) regimen. In the autologous HCT setting, the added efficacy of rituximab is under active investigation as a method of in vivo purging during hematopoietic cell mobilization, as part of the conditioning regimen and as post-HCT maintenance therapy. For patients with PTCL, autologous HCT is commonly offered at relapse but there are a few prospective series utilizing autologous HCT as consolidation of CR1 with encouraging results. There is no conclusive evidence regarding the efficacy of allogeneic HCT, but outcomes with RIC regimens appear promising. This review summarizes the current role of HCT for patients with MCL in first remission and for patients with PTCL as consolidation and for relapsed/refractory disease. PMID- 18162230 TI - Diffuse large B cell lymphoma: from gene expression profiling to prediction of outcome. AB - Diffuse large B cell lymphoma (DLBCL) is a subtype of non-Hodgkin lymphoma (NHL), characterized by a markedly heterogeneous clinical course and response to therapy that is not appreciated with standard histopathologic and immunophenotypic evaluations. Recent studies have focused on the use of genome-scale expression profiles that provide a snap fingerprint of the tumor and identifying tumors with similar genetic alterations and clinical features. Gene expression studies have the ability to recognize distinct subgroups of patients based on similar molecular characteristics and markedly different outcomes that were independent of the International Prognostic Index (IPI). Further, DNA microarray studies also allow identification of new prognostic biomarkers in DLBCL. However, new methods for immunohistochemical analysis of tissue microarray and RNA extraction from paraffin-embedded blocks are required to overcome the major pitfall of this technology--the requirement for fresh tissue. Herein, we summarize the progress made in better prediction of prognosis of DLBCL patients as a result of gene expression profiling. PMID- 18162231 TI - Principles and tools for selection of umbilical cord blood and unrelated adult donor grafts. AB - An analysis of NMDP data shows that allele-level matching for HLA A, B, C, and DRB1 is preferred in the selection of adult unrelated donors. If mismatching is unavoidable, mismatches at HLA B or C may be better tolerated than those at A or DRB1. Whether mismatches are at the allele level (ie, within an antigen group) or at the antigen level makes no difference in outcome, except at HLA C where allele mismatches are better tolerated. Matching for HLA DQ and DP should be prioritized below matching at the 4 major loci. These findings are compared and contrasted with previous publications. The impact of HLA matching on major outcomes in umbilical cord blood (UCB) transplantation continues to be refined. Total nucleated cell dose was previously thought to be sole determinant of outcome with partially HLA matched UCB transplantation, but HLA matching, particularly at low total nucleated cell dose, appears to play an important role. Relatively small sample sizes limit the consistency of findings from cord blood studies, but the consensus supports consideration of both total nucleated cell dose and HLA matching in the selection of optimal UCB units. As search considerations for both adult donors and umbilical cord blood units have become more complex, the National Marrow Donor Program has developed software, services and relationships to ease the burden on transplant teams. PMID- 18162232 TI - Human polymorphism and variable outcomes of cancer chemotherapy and transplantation. PMID- 18162233 TI - Mouse models of bone marrow transplantation. AB - Over the last 50 years, mouse models of bone marrow transplantation have provided the critical links between graft-versus-host disease (GVHD) and graft-versus leukemia (GVL) pathophysiology and clinical practice. The initial insight from mouse models that GVHD and GVL were T cell dependent has long been confirmed clinically. More recent translations from mouse models have included the important role of inflammatory cytokines in GVHD. Newly developed concepts relating to the ability of antigen presenting cell (APC) and T cell subsets to mediate GVHD now promise significant clinical advances. The ability to use knockout and transgenic approaches to dissect mechanisms of GVHD and GVL mean that mouse systems will continue as the predominant preclinical platform. The basic transplant approach in these models, coupled with modern "real-time" immunologic imaging of GVHD and GVL is discussed. PMID- 18162234 TI - T cell therapy in allogeneic stem cell transplantation. PMID- 18162235 TI - Chronic graft-versus-host disease: how can we release Prometheus? PMID- 18162236 TI - Human stem cells for tissue repair. PMID- 18162237 TI - Indications for hematopoietic cell transplantation in acute leukemia. AB - Based on available data, all adults with AML under age 60 years with matched siblings should be considered for allogeneic transplantation in first remission, except for those with favorable risk cytogenetics and possibly those whose disease has normal cytogenetics and is FLT3/ITD negative and NPM1 positive. Patients with matched siblings not transplanted in first remission should be followed closely so that transplantation in early first relapse can be considered. Those without matched siblings should receive a MUD transplant in first CR if they have higher risk disease. All others should receive standard chemotherapy and consider a matched unrelated transplant or autologous transplant, should they relapse. RIC allogeneic transplantation using either a matched family member or a MUD can be considered for patients age 60 years or greater with AML in second or subsequent remission, or AML in first remission with intermediate or high risk disease. PMID- 18162239 TI - Vaccines at the turn of the 21st century: a new era for immunization in public health. PMID- 18162238 TI - Graft failure after allogeneic hematopoietic cell transplantation. AB - Graft failure is a significant complication following allogeneic hematopoietic cell transplantation (alloHCT). It may result from rejection caused by recipient T cells, natural killer (NK) cells, or antibodies. It is increased in HLA mismatched grafts, unrelated grafts, T cell-replete transplants, sensitized patients, and in patients treated with reduced-intensity conditioning (RIC). In recipients of unrelated grafts, graft failure is increased in patients receiving major AB0 blood group mismatched transplants (P = .008). Recent data also suggest that donor-specific antibodies to CD34(+)/VEGFR-2(+) cells may be involved in graft failure after alloHCT. Graft failure may be overcome by more intensified conditioning, increased cell dose, or more effective immunosuppression. With more frequent use of RIC, cord blood grafts and other HLA-mismatched transplants, graft failure is an increasing problem after alloHCT. PMID- 18162240 TI - Correlating immunity with protection for HPV infection. AB - BACKGROUND: Cervical cancer is one of the most common types of cancer in women and is largely attributable to persistent infection with high-oncogenic risk human papillomavirus (HPV) types. Most HPV infections resolve spontaneously over 5 years, but, in a small fraction of women, persistent infection with high-risk HPV results in cervical intraepithelial neoplasia (CIN) grade 2/3 and eventually cervical cancer. PERSPECTIVES: Prophylactic vaccines based on HPV virus-like particles (VLPs) exploit the propensity of the immune system to protect against HPV infection and resulting diseases, including cancer. The quadrivalent (HPV types 6, 11, 16, 18) vaccine was 100% effective in preventing HPV 16- and 18 related CIN 2/3 in young, HPV-naive women and possessed similar efficacy against anogenital warts and vaginal and vulvar neoplasias. HPV VLP vaccine-induced protection is mediated by anti-HPV antibodies specific for conformational "neutralizing" determinants on the HPV capsid. CONCLUSIONS: The quadrivalent vaccine is highly immunogenic in human subjects; induces persistent, high-titer, neutralizing antibodies for at least 5 years; and induces immune memory responses. The quadrivalent HPV vaccine induces the greatest immune response in prepubertal children (9 to 12 years). Long-term follow-up studies are planned through the Nordic cancer registries to assess duration of HPV protection. PMID- 18162241 TI - Overview of the clinical development and results of a quadrivalent HPV (types 6, 11, 16, 18) vaccine. AB - BACKGROUND: Human papillomaviruses (HPVs) play an obligatory role in cervical cancer development. Thus, immunization of women using a prophylactic vaccine against the most common high-oncogenic risk types (e.g., HPV 16 and 18) and HPV 6 and 11, which contribute to development of low-grade cervical lesions and cause most anogenital warts, represents a logical primary prevention strategy. PERSPECTIVES: At the time of licensure, Phase II/Phase III studies showed that administration of a quadrivalent HPV (types 6, 11, 16, 18) vaccine to young women (16 to 26 years) naive to the vac- cine HPV types resulted in 100% efficacy against HPV 16- and 18-related precancerous cervical lesions, 100% efficacy against HPV 16- and 18-related high-grade vulvar/vaginal neoplasias, 95% efficacy against HPV 6, 11, 16, or 18-related cervical intraepithelial neoplasia/adenocarcinoma in situ, and 99% efficacy against HPV 6, 11, 16, or 18 related genital lesions. The quadrivalent HPV vaccine is highly immunogenic in adolescent males and females, and long-term follow up of young women did not detect evidence of waning immunity through 5 years. CONCLUSIONS: The quadrivalent vaccine is generally well tolerated. The efficacy and safety of the quadrivalent vaccine is continuing to be investigated in young men and mid-adult women. Nordic cancer registries are providing ongoing long-term pharmacovigilance. PMID- 18162242 TI - Programmatic issues in the implementation of an HPV vaccination program to prevent cervical cancer. AB - BACKGROUND: Cervical cancer remains an important health problem even in countries with effective cervical screening programs. HPV vaccines offer great potential for primary prevention of cervical cancer and other HPV-related diseases. PERSPECTIVES: Eventual implementation of an HPV vaccination program raises several key issues, including universal vs. targeted vaccinations, the age and gender of vaccine recipients, the acceptability of this vaccine to health care providers, adolescents, and parents, and the effect of this vaccine on cervical cancer screening. These issues were explored among symposium attendees during an interactive question-and-answer session using computerized voting pads. CONCLUSIONS: Preventative HPV vaccination programs should ideally be executed universally in both women and men with an emphasis on children and adolescents prior to their first sexual experience. Parent education on HPV disease and vaccine efficacy and safety will be critical to the acceptability of HPV vaccination for their children. HPV vaccination will not eliminate the need for Pap screening. Further research will be needed to develop rational and cost effective cervical surveillance programs for women protected by HPV vaccines. PMID- 18162243 TI - Efficacy of a pentavalent rotavirus vaccine in reducing rotavirus-associated health care utilization across three regions (11 countries). AB - OBJECTIVE: To evaluate the effect of a human-bovine reassortant pentavalent rotavirus vaccine (PRV) on health care encounters in nearly 70 000 subjects randomized in three regions - Europe, the United States, and Latin America/the Caribbean - in the Rotavirus Efficacy and Safety Trial (REST). METHODS: Healthy 6 to 12-week-old infants received 3 doses of PRV or placebo at 4- to 10-week intervals. The exact binomial method for ratios of Poisson counts was used to evaluate the effect of PRV on the rate of rotavirus-related hospitalizations and emergency department (ED) visits involving rotavirus G-types 1-4 occurring > or =14 days after the third dose of vaccine for up to 2 years. RESULTS: In fully vaccinated infants, reductions in rotavirus-associated hospitalizations and ED visits were 94.7% (95% CI: 90.9, 96.9) in Europe, 94.9% (95% CI: 84.0, 98.9) in the United States, and 90.0% (95% CI: 29.4, 99.8) in the Latin American/Caribbean regions. CONCLUSIONS: PRV reduced hospitalizations and ED visits within each region in REST. Results were consistent across regions and across the overall study cohort. PMID- 18162244 TI - Human papillomavirus infection and the development of cervical cancer and related genital neoplasias. AB - BACKGROUND: The human papillomaviruses (HPV) are simple, nonenveloped, double stranded DNA viruses, which are responsible for an enormous global burden of genital disease. HPV is associated with 500,000 new cases of cervical cancer and 250,000 cervical cancer deaths worldwide each year. Oncogenic HPV types 16 and 18 are responsible for a majority of cervical cancers and can also cause low- and high-grade cervical lesions (CIN 1, 2, 3) as well as high-grade vulvar or vaginal intraepithelial neoplasia (VIN or VaIN 2/3). Nononcogenic types HPV 6 and 11 also contribute to the overall burden of HPV disease, giving rise to CIN 1, anogenital warts, cutaneous lesions, and respiratory papillomatosis. PERSPECTIVES: A substantial body of clinical evidence demonstrates the effectiveness of cytological screening in preventing cervical cancer, but these techniques have not eradicated the disease and are not widely available in most developing countries. Furthermore, evaluation and management of HPV-associated cytologic abnormalities is costly, drains health care resources, and increases the risk for adverse pregnancy outcome. CONCLUSIONS: Targeting cervical cancer through universal immunization with a quadrivalent HPV 6, 11, 16, 18 vaccine may herald the beginning of the end of this deadly disease and substantially reduce the overall global burden of HPV-related genital diseases. PMID- 18162245 TI - The integrated phase III safety profile of the pentavalent human-bovine (WC3) reassortant rotavirus vaccine. AB - BACKGROUND: Rotavirus gastroenteritis is a significant cause of morbidity and mortality. OBJECTIVE: To perform an integrated safety analysis of data from the Phase III studies of the pentavalent rotavirus vaccine (PRV). METHODS: Healthy 6- to 12-week-old infants received 3 doses of PRV or placebo at 4- to 10-week intervals in 3 Phase III, blinded, randomized, placebo-controlled trials. Active surveillance for serious adverse events (AE), including intussusception, was performed at 7, 14, and 42 days after each dose. Other AEs occurring within 42 days after each dose were documented on Vaccination Report Cards. Fecal shedding of vaccine-virus strains was evaluated by plaque assay and electropherotyping. RESULTS: Intussusception and other serious AEs were evaluated among 71,799 vaccinated subjects. Within 42 days after any dose, intussusception occurred among 6 PRV and 5 placebo recipients. All AEs were evaluated among 11 722 vaccinated subjects. Within the week following the first dose, the incidences of fever and irritability were similar among PRV and placebo recipients, although diarrhea and vomiting occurred more frequently among PRV recipients versus placebo recipients (10.4% vs. 9.1% and 6.7% vs. 5.4%, respectively). Fecal shedding of vaccine-virus strains occurred in 8.9% of 360 PRV recipients after the first dose. CONCLUSIONS: Across the 3 Phase III clinical trials, PRV was well tolerated, with no increased clinical risk of intussusception. Fecal shedding of vaccine-virus strains occurred infrequently and in low amounts, suggesting the risk of transmission is unlikely. PMID- 18162246 TI - Prevention of herpes zoster and its painful and debilitating complications. AB - BACKGROUND: Reactivation of latent varicella-zoster virus in sensory neurons to cause herpes zoster (shingles) is common in adults 50 years of age and older; half of adults experience an episode by age 85 years. Herpes zoster is attributable to the progressive decline in the VZV-specific cell-mediated immunity that occurs with aging or other conditions that cause immune compromise. Herpes zoster and complications, such as postherpetic neuralgia (PHN), can have a substantial negative impact on quality of life. DISCUSSION: The incidence of herpes zoster and its associated morbidity is increasing worldwide as the population ages. Nevertheless, the severity and impact of this condition, and its painful sequelae, are often unrecognized. Many patients delay seeking medical attention, complicating both diagnosis and treatment. Prevention appears to be the best option. A new herpes zoster vaccine significantly reduced the burden of illness (61.1%), the incidence of PHN (66.5%), and the incidence of herpes zoster (51.3%) (p < 0.001). Vaccine tolerability was good, with minor local injection site reactions the most common adverse event. CONCLUSIONS: Herpes zoster and PHN represent a substantial burden in terms of suffering and associated costs. Immunization of older adults is a good option to prevent herpes zoster and PHN. PMID- 18162247 TI - A new challenge for Europe: introducing a pediatric quadrivalent vaccine for measles, mumps, rubella, and varicella. AB - BACKGROUND: Varicella is often considered to be a benign disease of childhood. In fact, varicella is associated with serious complications and mortality even among healthy individuals. DISCUSSION: Although the course of varicella can be uncomplicated, it can also be associated with serious complications such as pneumonia, fluid and electrolyte disturbances, skin and soft tissue infections and central nervous system disturbances. Worldwide studies have confirmed the high frequency of disease as well as the resultant morbidity, mortality and medical resource use. A quadrivalent vaccine is now available in certain countries to protect against measles, mumps, rubella and varicella (MMRV). Countries that have initiated routine vaccination pro- grams have reported substantial reductions in morbidity and mortality as well as improved health outcomes. The MMRV vaccine facilitates coverage against all four diseases, and would be expected to improve compliance as well as coverage of varicella. CONCLUSIONS: Universal vaccination programs with MMRV should be considered as a way to reduce the medical and economic impact of varicella. The MMRV vaccine provides a means to achieve universal coverage. PMID- 18162248 TI - Detection of respiratory viruses using non-molecular based methods. AB - The detection of respiratory viruses from clinical samples is important for patient management, promoting the rational use of antivirals and antibiotics, implementing infection control measures and for epidemiology studies. Respiratory viruses can be identified using a variety of techniques including direct antigen testing (non-immunofluorescent and immunofluorescent methods), conventional and rapid cell culture. This article presents an overview of each method. PMID- 18162249 TI - Nucleic acid amplification tests for detection of respiratory viruses. AB - Nucleic acid amplification tests (NATs) are increasingly being used for diagnosis of respiratory virus infections. The most familiar formats use DNA or RNA target amplification methods for enhanced sensitivity above culture and antigen-based procedures. Although gel and plate-hybridisation methods are still utilised for analysis of amplified products, detection using "real-time" methods which do not require handling of amplified products are favoured in many laboratories. Assays based on nucleic acid amplification and detection can be designed against a broad range of respiratory viruses and have been particularly useful for detection of recently identified viruses such as human metapneumovirus and coronaviruses NL63 and HKU1. However, the wide range of potential pathogens which can cause similar respiratory symptomology and disease makes application of individual diagnostic assays based on detection of DNA and RNA both complex and expensive. One way to resolve this potential problem is to undertake multiplexed nucleic acid amplification reactions with analysis of amplified products by suspension microarray. The Respiratory Virus Panel (RVP) from Luminex Molecular Diagnostics is one example of such an approach which could be made available to diagnostic and public health laboratories for broad spectrum respiratory virus detection. PMID- 18162250 TI - Why diagnose respiratory viral infection? PMID- 18162251 TI - Respiratory virus surveillance and outbreak investigation. AB - Sensitive, rapid detection of respiratory viruses is needed for surveillance and for investigation of epidemiologically linked cases. The utility of rapid antigen based methods for detection of common respiratory viruses and to confirm the cause of outbreaks is well established. However, nucleic acid amplification tests (NATs) offer some benefits above antigen or culture-based procedures, with the main advantages being sensitivity and range of pathogens detectable. It is important to understand how changes in our testing methodology alter respiratory virus detection and information for epidemiological studies. For viruses such as influenza A, influenza B and respiratory syncytial virus, NATs offer enhanced sensitivity above antigen assays but still identify the seasonal peaks important for predicting disease and managing time-sensitive prophylaxis. For other viruses, such as rhinoviruses, coronaviruses, human bocavirus and parainfluenza virus type 4, culture and antigen-based procedures are not available and/or lack sensitivity. Thus such targets would be missed if NATs were not included in testing for surveillance and outbreak investigation. As more respiratory viruses are identified there is a need to expand surveillance and further evaluate new technologies and automation beyond currently-available diagnostics to address detection of a broad range of potential pathogens. PMID- 18162252 TI - Principles of the xTAG respiratory viral panel assay (RVP Assay). PMID- 18162253 TI - The clinical need for the RVP test. PMID- 18162254 TI - xTAG RVP assay: analytical and clinical performance. PMID- 18162255 TI - Issues for reporting results. PMID- 18162256 TI - The pathogens. PMID- 18162257 TI - The value of RVP in children's hospitals. PMID- 18162258 TI - Value of RVP in clinical settings: older adults. PMID- 18162259 TI - Value of RVP in clinical settings: intensive care. PMID- 18162260 TI - Respiratory virus panels for global surveillance of emerging infectious diseases. PMID- 18162261 TI - Respiratory viruses. Introduction. PMID- 18162262 TI - Oligo(3-hydroxybutanoate) conjugates with acetylsalicylic acid and their antitumour activity. AB - In this paper we discuss the anticancer activity of acetylsalicylic acid with oligo(3-hydroxybutanoate) conjugates, their characteristics and in vitro biological evaluation. Acetylsalicylic acid (aspirin) attached via hydrolysable ester bonds to non-toxic well-defined 3-hydroxybutanoic acid oligomers shows novel method of drug modification. The resulting conjugates were more effective than aspirin in growth inhibition of human colon adenocarcinoma cells HT-29 and human colon carcinoma cells HCT 116 in vitro. Treatment of rats with doses as high as 2g of the conjugate (equivalent to 0.6g of pure aspirin) per kg of body weight did not exhibit toxic effects. PMID- 18162263 TI - [Persistent hypogastric pain]. PMID- 18162264 TI - The role of complement in the success of vaccination with conjugated vs. unconjugated polysaccharide antigen. AB - The complement system, a well-characterised arm of the innate immune system, significantly influences the adaptive immune response via direct cell-cell interaction and maintenance of lymphoid organ architecture. Development of vaccines is a major advance in modern health care. In this review, we highlight the importance of the marginal zone in response to both, polysaccharide and conjugated vaccines, and discuss the relevance of complement herein, based on findings obtained from animal models with specific deletions of certain complement components and from vaccination reports of complement-deficient individuals. We conclude that both, intactness of the complement system and maturity of expression of its components, are relatively more important to aid in the immune response to polysaccharide vaccine than to conjugated vaccines. PMID- 18162265 TI - Protective immune response in mice vaccinated with a recombinant adenovirus containing capsid precursor polypeptide P1, nonstructural protein 2A and 3C protease genes (P12A3C) of encephalomyocarditis virus. AB - Encephalomyocarditis virus (EMCV) infection can cause acute myocarditis and sudden death in pre-weaned piglets as well as severe reproductive failure in sows. In this study, two recombinant adenoviruses containing capsid precursor polypeptide P1 alone (Ad-P1) and P1 plus nonstructural protein 2A and 3C protease coding regions (Ad-P12A3C) of EMCV were respectively constructed using replication-defective human adenovirus serotype 5 as vector, and their antibody responses and protective efficacies against a lethal EMCV challenge were evaluated in mice. Both Ad-P1 and Ad-P12A3C were confirmed to be capable of expressing VP1 protein in BHK21 cells by immunoperoxidase monolayer assay (IPMA). The results showed that mice vaccinated once or twice with Ad-P1 and Ad-P12A3C generated specific antibody response against VP1 protein of EMCV. Although Ad-P1 induced higher antibody titers, virus-neutralizing antibody response was considerably less (p<0.05), compared to that of Ad-P12A3C. Upon challenging with a virulent EMCV strain, Ad-P12A3C elicited efficacious protection (100% for both vaccination once and twice) in the vaccinated mice; whereas the mice immunized with Ad-P1 showed a lower protection (12.5% for vaccination once and 75% for twice). Our work suggests that the recombinant adenovirus (Ad-P12A3C) containing the capsid precursor polypeptide coding region (P1) plus nonstructural protein 2A and 3C protease genes have an excellent potential to be used as a vaccine that can provide sufficient protective efficacy against EMCV infection in animals. PMID- 18162266 TI - Combination adjuvants for the induction of potent, long-lasting antibody and T cell responses to influenza vaccine in mice. AB - Influenza is controlled by protective titres of neutralizing antibodies, induced with the help of CD4 T-cells, and by antiviral T-cell effector function. Adjuvants are essential for the efficient vaccination of a naive population against avian influenza. We evaluated a range of adjuvants for their ability to enhance, in naive mice, protective hemagglutination inhibition (HI) titres, which represent the generally accepted correlate of protection, virus-neutralizing titres and T-cell responses to a new generation influenza vaccine produced in cell culture. The selected adjuvants include alum, calcium phosphate (CAP), MF59, the delivery system poly-(lactide co-glycolide) (PLG) and the immune potentiator CpG. MF59 was clearly the most potent single adjuvant and induced significantly enhanced, long-lasting HI and neutralizing titres and T-cell responses in comparison to all alternatives. The combination of alum, MF59, CAP or PLG with CpG generally induced slightly more potent titres. The addition of CpG to MF59 also induced a more potent Th1 cellular immune response, represented by higher IgG2a titres and the induction of a strongly enhanced IFN-gamma response in splenocytes from immunized mice. These observations have significant implications for the development of new and improved flu vaccines against pandemic and inter pandemic influenza virus strains. PMID- 18162267 TI - Concentration and specific activity of fallout 137Cs in extracted and particle size fractions of cultivated soils. AB - Fallout (137)Cs and stable Cs in soils were separated with two extractants (1M CH(3)COONH(4) solution and 0.8M CH(3)COONH(4) in 5% HNO(3) solution after H(2)O(2) oxidization). The residue remaining after removal of the oxidizable organic-bound fraction was separated into the particle-size fractions including clay, silt, fine sand and coarse sand with a sieving and sedimentation method. Then, the concentrations of (137)Cs and stable Cs in the extracted fractions and the particle-size fractions were determined. The (137)Cs contents in the exchangeable and organic-bound fractions in the soil were approximately 10 and 20%, respectively. The (137)Cs content in the strongly bound fraction was about 70%, and the concentration of (137)Cs in the clay was the richest among the particle-size fractions. The specific activity of (137)Cs (concentration ratio of fallout (137)Cs/stable Cs) decreased in the order exchangeable, organic-bound and strongly bound fractions. The data suggest that equilibrium between (137)Cs and stable Cs was not reached among those fractions, even though most of the (137)Cs that had been deposited on the soil was derived from fallout weapons tests that occurred several decades ago. The concentration of (137)Cs among the particle size fractions in each soil was different, whereas the specific activity of (137)Cs in the particle-size fractions had a relatively similar value. PMID- 18162268 TI - Assessment of pollution along the Northern Iberian shelf by the combined use of chemical and biochemical markers in two representative fish species. AB - Muscle concentrations of organochlorinated compounds as well as biliary levels of polycyclic aromatic hydrocarbons (PAHs) and alkylphenols (APEs) were determined in two different fish species, the four-spotted megrim (Lepidorhombus boscii) and the pouting (Trisopterus luscus) collected along the Northern Iberian coast. Additionally, a set of biochemical markers namely, 7-ethoxyresorufin O-deethylase (EROD), UDP-glucuronosyltransferase (UGT) and catalase (CAT) were measured in liver subcellular fractions. Chemical analysis indicated geographical differences in pollutant loads that were further reinforced by biomarker responses. Thus, EROD activity showed a good correlation with the amount of PCBs bioaccumulated in muscle tissue of both fish species. Elevated UGT activity was observed in those individuals highly exposed to APEs and 1-naphthol. The study reinforces the need to select representative sentinel species from different habitats for biomonitoring purposes and provides further support for the use of biomarkers in assessing the health of coastal areas. PMID- 18162269 TI - Airborne PBDEs in specialized occupational settings, houses and outdoor urban areas in Greece. AB - Airborne polybrominated diphenyl ethers (PBDEs) were measured in workplaces, homes and urban outdoor air in Greece. The geometric mean concentrations of total PBDEs (sum of 19 congeners) in offices (205 pg m(-3)), internet cafes/computer rooms (127 pg m(-3)) and computers/electronics shops (85 pg m(-3)) were significantly higher than those in furniture stores (12 pg m(-3)), homes (8 pg m( 3)) and outdoor air (18 pg m(-3)). The daily inhalation intake of PBDEs estimated for the employees of the four occupational settings ranged from 0.2 to 1.4 ng day(-1) and it was significantly lower than the expected dietary intake (approximately 77 ng day(-1)). Although inhalation generally represented a small fraction of the overall daily exposure to PBDEs (approximately 1%), the results from a heavily contaminated office (10,848 pg m(-3) of total PBDEs) indicated that the intake from this route (65 ng day(-1)) may, in some extreme cases, be as important as diet. PMID- 18162270 TI - Environmental mercury contamination of an artisanal zinc smelting area in Weining County, Guizhou, China. AB - To investigate the extent of Hg contamination due to artisanal Zn smelting activities in Weining County, northwestern Guizhou, China, total Hg and methyl mercury (MeHg) concentrations in soil and surface water were determined. Samples of corn plants growing in the study area were also collected for total Hg analysis. A high geometric mean Hg emission factor of 75gHgt(-1)Zn was estimated and significantly elevated total gaseous mercury (TGM) concentrations were found in the atmosphere adjacent to the Zn smelting sites, ranging from 30 to 3814ngm( 3). Total Hg and MeHg concentrations in topsoil samples ranged from 62 to 355microgkg(-1) and from 0.20 to 1.1microgkg(-1), respectively. Total Hg Concentrations in corn plant tissues increased in the order of grains or =9 cm in diameter and postmenopausal women were independent predictive factors of patients with development of ovarian cancer. CONCLUSIONS: Some endometriosis lesions may predispose to clear cell and endometrioid ovarian cancers. Advancing age and the size of endometriomas were independent predictors of development of ovarian cancer among women with ovarian endometrioma. PMID- 18162284 TI - Producer responsibility for e-waste management: key issues for consideration - learning from the Swiss experience. AB - E-waste, a relatively recent addition to the waste stream in the form of discarded electronic and electric equipment, is getting increasing attention from policy makers as the quantity being generated is rising rapidly. One of the most promising policy options to address this issue is to extend the producers responsibility for their products beyond the point of sale, until end-of-product life. This paper briefly introduces the concept of extended producer responsibility (EPR) and its applicability in the area of the end-of-life management of electronic and electrical equipment (EEE). It then examines the decade-long experience of Switzerland in using EPR to manage its e-waste, elaborating on the experience of the Swiss system in overcoming specific issues, and finally wrapping up with a synopsis of the lessons for policy makers. We consider each issue as an enquiry of questions confronting a policy maker and the choices that may present themselves. The five issues discussed are: (i) the challenges in getting an EPR based system started; (ii) securing financing to ensure a self-sustaining and smooth functioning system; (iii) organising a logistics network for the take back and collection of the e-waste; (iv) ensuring compliance of the various actors involved; and finally (v) reducing the threat of monopolistic practices. PMID- 18162285 TI - Urinary markers in bladder cancer. AB - OBJECTIVES: Many markers for the detection of bladder cancers have been tested. Almost all urinary markers reported are better than cytology with regard to sensitivity, but they score lower in specificity. The purpose of this review is to highlight the most important urinary biomarkers studied and reported recently. METHODS: Literature on bladder cancer markers has been reviewed regularly in the last few years. In the current review we have tried to summarise the most recent literature of urinary markers. RESULTS: The results of this review show that the first-generation urinary markers did not add much to urinary cytology. The current generation of markers is promising but larger clinical trails are needed. The future of marker development is bright with new techniques emerging, but the perfect marker is still to be found. CONCLUSION: Currently, no single marker can yet guide us in surveillance and lower the frequency of urethrocystoscopy. PMID- 18162286 TI - Gap junctions in the ovary: expression, localization and function. AB - Gap junctions that allow the direct communication between cytoplasmic compartments of neighboring cells are present in a variety of tissues and organs and play pivotal roles in a wide range of physiological processes. In the ovary, gap junctions consist mainly of connexin (Cx) 43 and Cx37, and their indispensable role in regulating folliculogenesis and oogenesis is well established. The ovarian Cx43 is regulated by gonadotropins at the transcriptional, translational and post-translational levels whereas the regulation of the ovarian Cx37 is yet unknown. In addition to their involvement in normal ovarian functions, gap junction proteins, particularly Cx43, seem to act as cancer suppressors. A summary of our present knowledge regarding gap junctional communication (GJC) and the ovarian gap junction proteins in normally developing ovaries and under pathological conditions is presented in this review. PMID- 18162287 TI - Characterization of recombinant human growth differentiation factor-9 signaling in ovarian granulosa cells. AB - Growth differentiation factor-9 (GDF9) is an oocyte secreted paracrine factor essential for mammalian ovarian folliculogenesis. Like other members of the transforming growth factor-beta (TGFbeta) superfamily, GDF9 is synthesized as a prepropeptide which needs processing by furin-like proteases to result in an active mature protein. We have previously characterized a preparation of unpurified recombinant mouse GDF9 which is bioactive as produced by human embryonic kidney 293T (HEK-293T) cells. However, we find that unpurified recombinant human GDF9 (hGDF9) produced by HEK-293T cells is not bioactive. Purified recombinant hGDF9 is bioactive and here we report the characterization of this protein. We find that the purified untagged mature region of hGDF9 is active in transcriptional reporter assays specific for Smad3/4 in human granulosa luteal (hGL) cells. We also demonstrate the use of a BMP (Smad1/5) responsive (BRE-luciferase) adenovirus in primary cultures of hGL cells to detect BMP responses. Using this adenovirus we find that purified human GDF9 does not activate the Smad1/5 pathway. Purified hGDF9 mature region activated the Smad3 pathway also in the FSH responsive human granulosa tumor cell line KGN. Primary cultures of rat granulosa cells responded to purified hGDF9 with an increase in DNA synthesis as measured by [3H]-thymidine uptake. Here we also report that the inclusion of a C-terminal affinity purification tag destroys GDF9 bioactivity. This study is the first characterization of purified biologically active human GDF9 and as such is of importance for studies on human fertility, and efforts aimed at treating infertility conditions. PMID- 18162288 TI - Vancomycin-resistant enterococcal meningitis treated with intrathecal streptomycin. AB - Enterococcal meningitis is a rare complication of neurosurgical procedures. We present a patient who developed vancomycin-resistant enterococcal ventriculitis - meningitis after a brain tumor resection and ventriculoperitoneal shunt placement, treated successfully with intrathecal streptomycin through bilateral cerebrospinal fluid drainage catheters in addition to systemic antibiotics. This is the first report of such treatment for this resistant organism. PMID- 18162289 TI - Kinetin riboside preferentially induces apoptosis by modulating Bcl-2 family proteins and caspase-3 in cancer cells. AB - Here, we demonstrate that kinetin riboside (KR), a cytokinin analog, induces apoptosis in HeLa and mouse melanoma B16F-10 cells. KR disrupted the mitochondrial membrane potential and induced the release of cytochrome c and activation of caspase-3. Bad were upregulated while Bcl-2 was down-regulated under KR exposure. A tumor growth in mice was dramatically suppressed by KR. In contrast, human skin fibroblast CCL-116 and bovine primary fibroblast cells show resistances to KR and no significant changes in Bad, Bcl-X(L,) and cleaved PARP were observed. Our data suggest that KR selectively induces apoptosis in cancer cells through the classical mitochondria dependent apoptosis pathway. PMID- 18162290 TI - Proto-oncogene ACTR/AIB1 promotes cancer cell invasion by up-regulating specific matrix metalloproteinase expression. AB - Overexpression of ACTR/AIB1 is frequently found in different cancers with distant metastasis. To address its possible involvement in tumor metastasis, we performed invasion assays to examine the effect of ACTR alteration on the invasiveness of breast cancer cells (MDA-MB-231 or T-47D) and found that high levels of ACTR are required for their strong invasiveness. Molecular analysis indicates that ACTR functions as a coactivator of AP-1 to up-regulate the expression of matrix metalloproteinases such as MMP-7 and MMP-10 and reduce cell adhesion to specific extracellular matrix proteins. These novel findings provide a mechanistic link between ACTR and MMPs, and suggest that ACTR may also play an important role in cancer progression by facilitating tumor invasion. PMID- 18162291 TI - Laser enhanced photo-catalytic removal of phenol from water using p-type NiO semiconductor catalyst. AB - Phenol is a carcinogen to humans and is a serious health concern, even at low concentrations. The efficient removal of phenol inside waste water is highly desirable. Laser induced photo-catalysis has been applied for removal of phenol from waste water using p-type NiO semiconductor catalyst for the first time. The characteristics of phenol removal using laser induced photo-catalytic process were investigated. The dependence of phenol removal on laser energy (100-250 mJ) and laser irradiation time (0-60 min) was studied. The rate of phenol removal process was characterized using UV absorption spectrometry, Raman spectroscopy and gas chromatography. In addition, the role of pH changes during the removal process was also investigated. Laser induced photo-catalytic process is highly efficient in removal of phenol in water over a short period of time (in minutes) as compared to conventional methods using lamps (hours). The phenol removal process pursued the first order kinetics with decay constant=0.0125 min(-1). PMID- 18162292 TI - Prediction for vented explosions in chambers with multiple obstacles. AB - The predictive ability between existing models on explosion venting, such as the NFPA, Molkov and Yao equations, was examined against experimental data of peak pressures obtained in various chambers with internal obstacles. The NFPA equation yielded the highest overpressures in most cases. The Molkov and Yao equations obtained much better agreement with experiments. However, the statistical diagnosis of the data showed an underprediction of the pressures. This is undesirable for designing calculations where some margin of safety is preferable. A new empirical model derived for characterising chambers with internal obstacles correlated well with the data. In addition the new equation was further validated against a dataset published from the literature and also gave a good correlation. PMID- 18162293 TI - Investigation of the effect of different electrodes and their connections on the removal efficiency of 4-nitrophenol from aqueous solution by electrocoagulation. AB - This study investigates the influence of variables on the removal efficiency of solution containing 4-NP (4-nitrophenol) by D. C. electrocoagulation (EC). The efficiency of different electrode connections and materials (steel 310, Fe, Al, graphite and steel 304) for 4-NP removal is compared. Current density, time of electrolysis, interelectrode distance, supporting electrolyte concentration and stirring rate of the solution were the variables that mostly influenced the 4-NP removal. Initially, a simple electrochemical cell was prepared with an anode and a cathode. Then the effect of each variable was studied separately using aqueous 4-NP in a batch mode. For a solution of 20 mg/L 4-NP+300 mg/L NaCl with chemical oxygen demand (COD) of approximately 40 mg O2/L, almost up to 99% 4-NP and 65% COD were removed, when the pH was about 9, time of electrolysis was approximately 10 min, current density was 100 A m(-2), interelctrode distance was 15 mm and stirring rate was 400 rpm. In the second series of experiments, the efficiency of EC cells with monopolar electrodes in series and parallel connections and an EC cell with bipolar electrodes was compared with that of a simple electrochemical cell. The best results obtained when steel 310 and Fe are used as anodes and employing Al and graphite as anodes would not be satisfactory. Also findings show that the types of sacrificial electrodes are not very significant in the removal of 4-NP. In the real wastewater obtained from Tabriz petrochemical plant 52% removal could be achieved after 10 min with using steel 310 as anode and steel 304 as cathode. PMID- 18162294 TI - Two-step method for preparation of NaA-X zeolite blend from fly ash for removal of cesium ions. AB - Pure zeolites can be synthesized from silica extracts obtained from fly ash by alkaline leaching. The extraction potential of industrial by-product fly ash was investigated under repeated fusion process conditions. The amount of extracted silica was 131.43 g/kg ash while the amount extracted alumina was limited to 41.72 g/kg ash. The results of zeolite synthesis from the Si-bearing extracts demonstrated that pure zeolites with high cation exchange capacity (4.624 meq/g) can be produced. The sorption potential of synthesized A-X zeolite blend for the removal of cesium ions has been investigated. The influences of pH, contact time and temperature have been reported. Thermodynamic parameters such as changes in Gibbs free energy (DeltaG degrees), enthalpy (DeltaH degrees) and entropy (DeltaS degrees) were calculated. A comparison of kinetic models applied to the sorption data was evaluated for pseudo first-order, pseudo second-order and homogeneous particle diffusion models. The results showed that both the pseudo second-order and the homogeneous particle diffusion models were found to best correlate the experimental rate data. PMID- 18162295 TI - Degradation of alachlor and pyrimethanil by combined photo-Fenton and biological oxidation. AB - Biodegradability of aqueous solutions of the herbicide alachlor and the fungicide pyrimethanil, partly treated by photo-Fenton, and the effect of photoreaction intermediates on growth and DOC removal kinetics of the bacteria Pseudomonas putida CECT 324 are demonstrated. Toxicity of 30-120 mg L(-1) alachlor and pyrimethanil has been assayed in P. putida. The biodegradability of photocatalytic intermediates found at different photo-treatment times was evaluated for each pesticide. At a selected time during batch-mode phototreatment, larger-scale biodegradation kinetics were analysed in a 12 L bubble column bioreactor. Both alachlor and pyrimethanil are non-toxic for P. putida CECT 324 at the test concentrations, but they are not biodegradable. A approximately 100 min photo-Fenton pre-treatment was enough to enhance biodegradability, the biological oxidation response being dependent on the pesticide tested. The different alachlor and pyrimethanil respiration and carbon uptake rates in pre-treated solutions are related to change in the growth kinetics of P. putida. Reproducible results have shown that P. putida could be a suitable microorganism for determining photo-Fenton pre-treatment time. PMID- 18162296 TI - Effect of Pb toxicity on leaf growth, physiology and ultrastructure in the two ecotypes of Elsholtzia argyi. AB - Hydroponics experiments were conducted to study the effects of Pb on mined ecotype (ME) and non-mined ecotype (NME) of Elsholtzia argyi from Pb/Cu mining and the non-contaminated agricultural areas, respectively. The results showed that at 200 microM Pb treatment, although concentrations of Pb in leaves and stem of the ME were 2.6 and 4.5 times higher than those of the NME, these plants exhibited higher tolerance to excessive levels of Pb in the growth medium. In both the ecotypes, Pb caused inhibition of leaf growth and photosynthesis, and induced the membrane damage which was more obvious in the NME. Pb treatment decreased the activities of superoxide dismutase (SOD) and guaiacol peroxidase (G POD) while activity of catalase (CAT) and levels of total soluble proteins (TSP), ascorbic acid (AsA) and glutathione (GSH) were significantly induced after Pb exposure, however, increase was sharp in the ME plants. Leaf ultrastructural analysis of the spongy mesophyll cells revealed that the excessive Pb concentrations caused adverse effects on chloroplast ultrastructure of both ecotypes whereby internal damage was more severe in NME. The higher tolerance to Pb displayed by ME is mainly attributed to maintenance of its leaf growth and physiology, induction of GSH and integrity of cell organelles especially chloroplast ultrastructure. PMID- 18162297 TI - Adsorption of Cr(VI) from aqueous solutions by spent activated clay. AB - Adsorption of Cr(VI) onto spent activated clay (SAC), a waste produced from an edible oil refinery company, was investigated for its beneficial use in wastewater treatment. After pressure steam treatment, SAC was used as an adsorbent. The adsorption kinetic data were analyzed and fitted well in a pseudo first-order equation and the rate of removal was found to speed up with decreasing pH and increasing temperature. Activation energy for the adsorption process was found to be 4.01-5.47 kcal/K mol. The Langmuir adsorption isotherm was used to fit the equilibrium data and the effect of pH, temperature and ionic strength were studied. The maximum adsorption capacities for Cr(VI) ranged from 0.743 to 1.422 mg/g for temperature between 4 and 40 degrees C under a condition of pH 2.0. The studies conducted show the process of Cr(VI) removal to be spontaneous at high temperature and endothermic in nature. From the waste utilization and environment point of view, the work carried out is important and useful. Results obtained can serve as baseline data for designing a treatment process using this low-cost adsorbent for the treatment of wastewater rich in Cr(VI). PMID- 18162298 TI - Removal of cadmium (II) from aqueous solutions by adsorption on agricultural waste biomass. AB - This paper reports the feasibility of using various agricultural residues viz. sugarcane bagasse (SCB), maize corncob (MCC) and Jatropha oil cake (JOC) for the removal of Cd(II) from aqueous solution under different experimental conditions. Effect of various process parameters, viz., initial metal ion concentration, pH, and adsorbent dose has been studied for the removal of cadmium. Batch experiments were carried out at various pH (2-7), adsorbent dose (250-2000 mg), Cd(II) concentration (5-500 mg l(-1)) for a contact time of 60 min. The maximum cadmium removal capacity was shown by JOC (99.5%). The applicability of Langmuir and Freundlich isotherm suggests the formation of monolayer of Cd(II) ions onto the outer surface of the adsorbents. Maximum metal removal was observed at pH 6.0 with a contact time of 60 min at stirring speed of 250 rpm with an adsorbent dose of 20 g l(-1) of the test solution. The maximum adsorption of cadmium (II) metal ions was observed at pH 6 for all the adsorbents viz; 99.5%, 99% and 85% for JOC, MCC, and SCB, respectively. Order of Cd(II) removal by various biosorbents was JOC>MCC>SCB. JOC may be an alternative biosorbent for the removal of Cd(II) ions from the aqueous solution. FT-IR spectra of the adsorbents (before use and after exhaustion) were recorded to explore number and position of the functional groups available for the binding of Cd(II) ions on to studied adsorbents. These results can be helpful in designing a batch mode system for the removal of cadmium from dilute wastewaters. PMID- 18162299 TI - Removal of rhodamine B from aqueous solution by adsorption onto sodium montmorillonite. AB - The adsorption of rhodamine B dye was carried out using sodium montmorillonite clay. The effect of parameters such as pH, adsorbent dosage and initial dye concentration was studied. The Langmuir and Freundlich isotherm models were applied and the Langmuir model was found to best fit the equilibrium isotherm data. Langmuir adsorption capacity was found to be 42.19 mg/g. Kinetic data followed pseudo-second-order kinetics. Maximum color removal was observed at pH 7.0. The DeltaG degrees value was found to be negative, the adsorption process was feasible. The results indicate that sodium montmorillonite clay can be used for the removal of basic dyes from aqueous solutions. PMID- 18162300 TI - Adsorptive, thermodynamic and kinetic performances of Al/Ti and Al/Zr-pillared clays from the Brazilian Amazon region for zinc cation removal. AB - Smectite clay samples from the Amazon region, Brazil, were pillarized by intercalating the species obtained from the chemical reactions: (i) AlCl3.6H2O/NaOH, (ii) titanium ethoxide in hydrochloric acid and (iii) direct use of ZrOCl2.8H2O solution. The natural matrices and the pillaring solutions were maintained under vigorous stirring at 298 K for 3 h and then subjected to calcination at temperatures of 723 and 873 K. Natural and pillared matrices were characterized by XRD, FTIR, TG-DTG and nitrogen adsorption-desorption isotherms. The resulting materials were used for zinc adsorption from aqueous solution at room temperature. The Langmuir, Freundlich and Temkin adsorption isotherm models have been applied to fit the experimental data and the Freundlich model is limited for higher concentrations. The pillaring process increases the thermal stability, the basal spacing of the natural clay sample (A1) from 1.55 to 2.06 nm and the surface area from 44.30 to 223.73 m2 g(-1). Kinetic studies demonstrated an equilibrium time of 180 min for zinc adsorption on the pillared matrices. Pseudo-first-order, Lagergren pseudo-second-order and Elovich equations demonstrated a better agreement with second-order kinetics was obtained with K2=4.17-10.43 x 10(-3)g mg(-1)min(-1) for the A1 sample. PMID- 18162301 TI - Decolorization of Acid Orange 7 solution by gas-liquid gliding arc discharge plasma. AB - The decolorization of 180 microM aqueous solutions of Acid Orange 7 (AO7) by means of a non-thermal plasma technique (i.e., the gas-liquid gliding arc discharge, which is generated between at least two metal electrodes with AC high voltage) was investigated in this paper. The effects of the plasma treatment time and the type of feeding gas, including air, oxygen, nitrogen and argon of the dye removal were determined. It is found that the voltage cycles of the gas-liquid gliding arc discharge are characterized by a moderate increase in the tension which is represented by a peak followed by an abrupt decrease and a current peak in the half period (10 ms); the concentration of AO7 solution decreases exponentially to reach 58.9, 77.4, 89.1, 95.1 and 99% in 25, 50, 75, 100 and 125 min, respectively, and the ln(Ct/C0) varies linearly with the treatment time t, indicating that decolorization reaction follow first pseudo-order kinetics with a constant rate of 0.03327 min(-1) when air was used as feeding gas; the decolorization rate during the plasma treatment is the greatest for oxygen as the feeding gas, in turn followed by air and argon, and was the least when using nitrogen. The variations of pH and conductivity and the formations of hydrogen peroxide and ozone are measured. PMID- 18162302 TI - Suppressive effects of magnesium oxide materials on cadmium uptake and accumulation into rice grains I: Characteristics of magnesium oxide materials for cadmium sorption. AB - The objective of this study is to assess the applicability of a commercial magnesium oxide (MgO) and a composite material containing MgO and natural minerals ('MgO-SH-A') as the soil amendments for suppression of cadmium (Cd) uptake and accumulation into rice grains. Firstly, the mineralogical and physicochemical properties, soil neutralizing capacities and Cd sorption characteristics of these materials were investigated. Both materials were strongly alkaline and possessed large surface areas. The X-ray diffraction pattern of MgO-SH-A indicated the presence of MgO and a magnesium-silicate mineral (antigorite) as the main components. MgO-SH-A showed a milder soil neutralizing capacity as compared to commercial MgO. The sorptions of Cd on commercial MgO and MgO-SH-A both fitted Langmuir isotherm. The maximum Cd sorption capacity of commercial MgO (46.8 mmol g(-1) DW) was higher than that of MgO-SH-A (5.87 mmol g(-1) DW), although the latter material showed higher affinity to Cd as compared to the former one. The dominant reaction involved in the Cd sorptions was suggested to be precipitation of Cd(OH)2 on the material surface. About 40% of Cd sorbed on MgO-SH-A was resistant to desorption by 0.1 M HCl, implying that this portion was strongly retained on the material surface. PMID- 18162303 TI - Extraction of extracellular polymeric substances from aerobic granule with compact interior structure. AB - Extracellular polymeric substances (EPS) were extracted from aerobic granules of compact interior structure using seven extraction methods. Ultrasound followed by the chemical reagents formamide and NaOH outperformed other methods in extracting EPS from aerobic granules of compact interior. The collected EPS revealed no contamination by intracellular substances and consisted mainly of proteins, polysaccharides, humic substances and lipids. The quantity of extracted proteins exhibited a weak correlation with quantity of extracted carbohydrates but no correlation with quantity of extracted humic substances. The total polysaccharides/total proteins (PN/PS) ratios for sludge flocs were approximately 0.9 regardless of extraction method. Protein content was significantly enriched in the granules, producing a PN/PS ratio of 3.4-6.2. This experimental result correlated with observations using excitation-emission matrix (EEM) and confocal laser scanning microscope technique. However, detailed study disproved the use of EEM results as a quantitative index of extracted EPS from sludge flocs or from granules. PMID- 18162304 TI - Defluoridation chemistry of synthetic hydroxyapatite at nano scale: equilibrium and kinetic studies. AB - This study describes the advantages of nano-hydroxyapatite (n-HAp), a cost effective sorbent for fluoride removal. n-HAp possesses a maximum defluoridation capacity [DC] of 1845 mg F(-)/kg which is comparable with that of activated alumina, a defluoridation agent commonly used in the indigenous defluoridation technology. A new mechanism of fluoride removal by n-HAp was proposed in which it is established that this material removes fluoride by both ion-exchange and adsorption process. The n-HAp and fluoride-sorbed n-HAp were characterized using XRD, FTIR and TEM studies. The fluoride sorption was reasonably explained with Langmuir, Freundlich and Redlich-Peterson isotherms. Thermodynamic parameters such as DeltaG degrees , DeltaH degrees , DeltaS degrees and Ea were calculated in order to understand the nature of sorption process. The sorption process was found to be controlled by pseudo-second-order and pore diffusion models. Field studies were carried out with the fluoride containing water sample collected from a nearby fluoride endemic area in order to test the suitability of n-HAp material as a defluoridating agent at field condition. PMID- 18162305 TI - Continuous biosorption of Pb/Cu and Pb/Cd in fixed-bed column using algae Gelidium and granulated agar extraction algal waste. AB - Continuous metal ions biosorption from Pb/Cu and Pb/Cd solutions onto seaweed Gelidium sesquipedale and a composite material prepared from an industrial algal waste was performed in a packed bed column. A binary Langmuir equation describes well the equilibrium data and indicates a good adsorption capacity. In the sorption process, Cd and Cu break through the column faster than Pb due to its lower affinity for the biosorbent. An overshoot in the outlet Cd concentration was observed and explained by competitive adsorption between Pb and Cd, whereby the higher Pb affinity for the biosorbent displaces bound Cd ions. A small overshoot happens for Cu adsorption in the presence of Pb ions. Desorption using 0.1 M HNO3 as eluant, was 100% effective. A mass transfer model for the adsorption and desorption processes, considering an external and intraparticle film resistance, adequately simulates the column performance. A binary Langmuir equation was used to describe equilibrium for the saturation process and a mass action law for the desorption process. Elution process is defined as an ion exchange mechanism, between protons and metal ions. PMID- 18162306 TI - Modelling the dispersion of flashing jets using CFD. AB - Risk assessments related to industrial environments where gas is kept in liquid form under high pressure rely on the results from predictive tools. Computational Fluid Dynamics (CFD) is one such predictive tool and it is currently used for a range of applications. One of the most challenging application areas is the simulation of multiphase flows resulting from a breach or leakage in a pressurised pipeline or a vessel containing liquefied gas. The present paper deals with the modelling of the post-flashing scenario of a jet emanating from a circular orifice. In addition to being based on the equations governing fluid flow, the models used are those related to turbulence, droplet transport, evaporation, break-up and coalescence. Some of these models are semi-empirical and based on the data from applications other than flashing. However, these are the only models that are currently available in commercial codes and that would be used by consulting engineers for the type of modelling discussed above, namely the dispersion of a flashing release. A method for calculating inlet boundary conditions after flashing is also presented and issues related to such calculations are discussed. The results from a number of CFD based studies are compared with available experimental results. The results show that whilst a number of features of the experimental results can be reproduced by the CFD model, there are also a number of important shortcomings. The shortcomings are highlighted and discussed. Finally, an optimum approach to modelling of this type is suggested and methods to overcome modelling difficulties are proposed. PMID- 18162307 TI - Cements obtained from rice hull: encapsulation of heavy metals. AB - Rice hull is an agricultural residue containing organic materials (carbohydrates, cellulose, etc.) and about 10% of silica, as inorganic component. Usually rice hull is burned at the fields generating small silica particles that may cause respiratory and environmental damage. On the other hand, beta-Ca2SiO4 is the second most important component of commercial portland cement. The utilization of beta-Ca2SiO4 brings many environmental benefits, such as reduction of carbon dioxide emissions and the saving of raw materials and energy. This work describes the synthesis of beta-Ca2SiO4 and related cements prepared from rice hull and the insertion of heavy metals (copper and cadmium). The insertion of these metals, chemically bounded to dicalcium silicate may represent a useful method to permanent encapsulation of hazardous chemicals. Silica was obtained from burning rice hull at 600 degrees C. The synthesis was carried out by a combination of sonochemical and solid-state chemical reactions and is completed at 800 degrees C. It was found that copper and cadmium can be inserted into beta-Ca2SiO4 structure in proportions up to 10%. PMID- 18162308 TI - Effect of chronic acamprosate treatment on voluntary alcohol intake and beta endorphin plasma levels in rats selectively bred for high alcohol preference. AB - Our previous studies have shown that repeated acamprosate administration to ethanol-naive Warsaw high preferring (WHP) rats resulted in increased plasma beta endorphin levels and at least partially prevents increases in levels of this peptide after a single administration of ethanol compared with untreated control rats. The objective of the present study, which included 45 WHP rats, was to continue the past research and investigate the effect of 10-day acamprosate treatment (200 mg/kg p.o.) on alcohol intake using a free-choice procedure and on changes in plasma beta-endorphin levels while alcohol is available, and 10 days after alcohol withdrawal. Voluntary alcohol consumption increases plasma levels of beta-endorphin from 440+/-25 pg/ml to 711+/-57 pg/ml (p=0.0002). After a 10 day of alcohol withdrawal, the levels of this peptide were significantly reduced compared with levels in rats with free access to ethanol (711+/-57 pg/ml vs. 294+/-38 pg/ml, p=0.000001) and in control naive rats (440+/-25pg/ml vs. 294+/ 38pg/ml, p=0.044). Chronic treatment with acamprosate increased plasma beta endorphin levels both in WHP rats with free access to ethanol (440+/-25 pg/ml vs. 616+/-49 pg/ml, p=0.008) and in rats after ethanol withdrawal (440+/-25 pg/ml vs. 620+/-56 pg/ml, p=0.007). In the group with free access to ethanol, there was a significant reduction in mean ethanol intake, from 6.75+/-0.20 g/kg body weight/day to 4.68+/-0.25 g/kg/day. Our results indicate that chronic acamprosate treatment may have beneficial effects, as it increases the beta-endorphin concentration thereby compensating for beta-endorphin deficiency during ethanol withdrawal. As the endogenous opioid system has an important role in the development of craving for alcohol, restoring the alcohol-induced deficits in beta-endorphin levels may be an important factor to prevent craving and maintaining abstinence. We suppose that the anti-craving mechanism of acamprosate that has been reported to abolish excessive glutamate release during alcohol withdrawal may be accompanied by compensation for the beta-endorphin deficiency. PMID- 18162309 TI - Regulation of sympathetic neuron differentiation by endogenous nerve growth factor and neurotrophin-3. AB - Nerve growth factor (NGF) and neurotrophin-3 (NT3) play distinctive roles in sympathetic axon growth and target field innervation and are required for sympathetic neuron survival in vivo. To ascertain if these neurotrophins selectively regulate the expression of genes that determine the functional characteristics of differentiated sympathetic neurons, we measured the mRNA levels for several such genes in the superior cervical ganglion of NGF(-/-), NT3( /-) and wild type mouse embryos at a stage before excessive neuronal loss occurs in the absence of these neurotrophins. Despite the extensively documented ability of NGF to regulate the noradrenergic phenotype of sympathetic neurons, we found that tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DbetaH) mRNA levels were normal in NGF(-/-) embryos, but significantly reduced in NT3(-/-) embryos. In contrast, the beta2 nicotinic acetylcholine receptor and PACAP receptor 1 mRNA levels were normal in NT3(-/-) embryos, but significantly reduced in NGF(-/-) embryos. Studies of mice lacking neurotrophin receptors suggested that the effects of NGF on gene expression require TrkA whereas those of NT3 require TrkA and p75(NTR). These findings demonstrate that endogenous NGF and NT3 have distinctive and separate effects on gene expression in early sympathetic neurons and that these selective effects on gene expression require a different combination of neurotrophin receptors. PMID- 18162310 TI - The effect of stimulus intensity on the right ear advantage in dichotic listening. AB - The dichotic listening test is non-invasive behavioural technique to study brain lateralization and it has been shown, that its results can be systematically modulated by varying stimulation properties (bottom-up effects) or attentional instructions (top-down effects) of the testing procedure. The goal of the present study was to further investigate the bottom-up modulation, by examining the effect of differences in the right or left ear stimulus intensity on the ear advantage. For this purpose, interaural intensity difference were gradually varied in steps of 3 dB from -21 dB in favour of the left ear to +21 dB in favour of the right ear, also including a no difference baseline condition. Thirty-three right-handed adult participants with normal hearing acuity were tested. The dichotic listening paradigm was based on consonant-vowel stimuli pairs. Only pairs with the same voicing (voice or non-voiced) of the consonant sound were used. The results showed: (a) a significant right ear advantage (REA) for interaural intensity differences from 21 to -3 dB, (b) no ear advantage (NEA) for the -6 dB difference, and (c) a significant left ear advantage (LEA) for differences form -9 to -21 dB. It is concluded that the right ear advantage in dichotic listening to CV syllables withstands an interaural intensity difference of -9 dB before yielding to a significant left ear advantage. This finding could have implications for theories of auditory laterality and hemispheric asymmetry for phonological processing. PMID- 18162311 TI - Stoichiometric analysis of the TM2 6' phenylalanine mutation on desensitization in alpha1beta2 and alpha1beta2gamma2 GABA A receptors. AB - The presence of phenylalanine (F) at the 6' position of transmembrane domain 2 (TM2) in the alpha4 subunit of alpha4beta2 nicotinic receptors enhances desensitization. As the GABA A receptor affords the ability to study the influence of as few as one and as many as five Fs at this position, we have used it to investigate potential subunit- and stoichiometry-dependent effects of the TM2 6'F mutation on desensitization. Whereas the presence of one F at this position decreased extent of desensitization, desensitization was increased in all configurations that included two or more Fs at the TM2 6' position; desensitization was particularly rapid with 3 or 4 F residues present. Our results demonstrate the ability of F residues at the TM2 6' position to modulate desensitization is likely conserved in the cys-loop family of ligand-gated ion channels. Moreover, our findings demonstrate both stoichiometric- and subunit dependent effects of the ability of this mutation to regulate desensitization in GABA A receptors. PMID- 18162312 TI - A dysbindin risk haplotype associated with less severe manic-type symptoms in psychosis. AB - Delineating relationships between susceptibility genes and clinical symptoms may be an important step in understanding the genetics of psychosis. Recent data suggests that the gene dysbindin (DTNBP1) may confer susceptibility across psychotic disorders and may particularly be associated with negative symptoms, i.e. affective flattening, alogia and avolition. We have previously published evidence of association with a dysbindin risk haplotype derived from alleles C-A T at SNPs P1655 (rs2619539), P1635 (rs3213207) and SNP66961 (rs2619538) in two independent schizophrenia (SZ) case-control samples. The C-A-T haplotype impacts at the level of gene function and phenotype: the haplotype indexes lower cortical expression of the dysbindin gene in post-mortem SZ brain samples and haplotype carriers show greater deficits in spatial working memory and early visual processing than non-carrier SZ patients. The aim of this study was to establish if the C-A-T dysbindin risk haplotype is associated with a specific clinical symptom profile. We investigated the relationship between the haplotype and PANSS derived symptom factors in 262 individuals with schizophrenia/schizoaffective disorder using principal components analysis (PCA) and analysis of variance (ANOVA). Dysbindin risk carriers scored significantly less than non-carriers on the 'hostility/excitability' factor (F 1,196=8.468; p=.004), with a trend for higher negative symptom scores. This suggests that risk variation at the dysbindin gene may contribute to a more prototypical SZ presentation with less severe excitement/manic symptoms and more negative symptoms. PMID- 18162313 TI - Primary somatosensory evoked magnetic fields elicited by sacral surface electrical stimulation. AB - To explore the brain response to sacral surface therapeutic electrical stimulation (SSTES) for the treatment of refractory urinary incontinence and frequent micturition, evoked magnetic fields were measured in six healthy males. Electrical stimuli were applied between bilateral surface electrodes over the second through fourth posterior sacral foramens with intensity just below the pain threshold. Somatosensory evoked magnetic fields (SEFs) for the bilateral median (MN) and posterior tibial nerves (PTN) were also measured for the comparison. Sources of the early SEF peaks were superimposed on individual magnetic resonance images. The first peak latency for sacral stimuli, M30, occurred at 30.2+/-0.8 ms (mean+/-standard deviation, N=6), with shorter latency than those for PTN stimulus (39.3+/-1.4 ms, N=12) and longer latency than those for MN stimulus (21.0+/-0.9 ms, N=12). The second peak latency for sacral stimuli, M50, occurred at 47.2+/-2.9 ms (N=6). Both M30 and M50 peaks showed a single dipole pattern over the vertex in the isofield maps. The equivalent current dipoles of M30 and M50 were both estimated near the medial end of the central sulcus with approximately posterior current direction. These results suggest that the sacral M30 and M50 are responses from the primary somatosensory cortex. The relatively long time lag between the onset and peak of M30 suggests that SSTES directly affects both the cauda equina and cutaneous nerve of the sacral surface. PMID- 18162314 TI - Neuroprotective effects of rotigotine in the acute MPTP-lesioned mouse model of Parkinson's disease. AB - Dopamine agonists used to manage Parkinsonian motor symptoms have been suggested to be neuroprotective. The study was designed to assess the neuroprotective potential of the D(3)/D(2)/D(1) dopamine receptor agonist rotigotine in the acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned (MPTP) mouse model of Parkinson's disease by measuring mesencephalic degenerating neurons using FluoroJade staining and the remaining dopaminergic nerve endings in the striatum using dopamine transporter binding. Continuous administration of rotigotine at a dose of 3mg/kg significantly attenuated MPTP-induced acute cell degeneration in the FluoroJade-staining paradigm. Rotigotine (0.3-3mg/kg) partially protected dopamine nerve endings from MPTP-induced degeneration in a dose-dependent manner. These data suggest that rotigotine, at the doses employed, significantly protected dopamine neurons from degeneration in an acute mouse model of MPTP intoxication. PMID- 18162315 TI - Activation of Nrf2-ARE pathway in brain after traumatic brain injury. AB - Secondary brain injury plays a pivotal role in the outcome of patients suffering from traumatic brain injury (TBI). The mechanisms underlying secondary brain injury are complex and interrelated. Previous studies focused on one of these mechanisms have been proved to be ineffective in clinical practice. Therefore, a target, which can interrupt multi-mechanisms underlying TBI, is desirable. Nrf2 ARE pathway has been proved to be the key regulator in reducing oxidative stress, inflammatory damage and accumulation of toxic metabolites, which are all involved in TBI. However, whether Nrf2-ARE pathway is activated after TBI has not been studied. In the present study, the nuclear Nrf2 protein level was detected by Western blot, and the mRNA levels of heme oxygenase-1 (HO-1) and NAD(P)H: quinone oxidoreductase-1 (NQO1), two Nrf2-regulated gene products, were determined using reverse-transcriptase polymerase chain reaction (RT-PCR) 24h after TBI. Furthermore, we also localized the expression of Nrf2 and HO-1 using immunohistochemical study. After TBI, the nuclear Nrf2 protein level was significantly increased, and the mRNA levels of both HO-1 and NQO1 were also up regulated. Moreover, both Nrf2 and HO-1 were localized in the same types of cells. According to these results, it could be postulated that Nrf2-ARE pathway was activated in brain after TBI. PMID- 18162316 TI - Impaired regulation function in cardiovascular neurons of nucleus tractus solitarii in streptozotocin-induced diabetic rats. AB - This study characterizes neural firing activity of the nucleus tractus solitarii (NTS) and baroreflex sensitivity (BRS) in streptozotocin (STZ)-induced diabetic rats relative to control rats by implantation of multi-wire electrode into rat NTS for direct monitoring of barosensitive NTS neurons before and after baroreflex system challenge by phenylephrine (PE) injection. NTS firing data is correlated with arterial pressure for both control and diabetic rats. In control rats, NTS firing rate and systolic arterial pressure correlate significantly with both pre-PE (baseline) and post-PE (p<0.01). In STZ-induced diabetic rats, positive correlation is observed only after PE injection (p<0.05). Although NTS firing rate was not significantly different between control and diabetic rats (p=0.085) in the baseline condition, it was significantly reduced in STZ-induced diabetic rats (p=0.042) with adjustment for BRS. After PE injection, NTS firing rate is significantly lower in diabetic rats relative to control rats (p<0.01). With adjustment for BRS, multivariate analysis shows that diabetes is independently associated with NTS firing rate after PE injection (p=0.034). Prior physiological and immunofluorescent studies found differing NTS data for control and diabetic rat only after PE challenge, but our data show diabetes-induced barosensitive NTS impairment in the baseline condition for STZ-induced diabetic rats. This latter finding suggests greater sensitivity of multi-wire electrode study of NTS relative to earlier methods. PMID- 18162317 TI - Binding of 5'-O-(2-thiodiphosphate) to rat brain membranes is prevented by diadenosine tetraphosphate and correlates with ecto-nucleotide pyrophosphatase phosphodiesterase 1 (NPP1) activity. AB - The distribution of binding sites for [(35)S]5'-O-(2-thiodiphosphate) ([(35)S]ADPbetaS), a radioligand of P2Y(1,12,13) receptors, and of ecto nucleotide pyrophosphatase phosphodiesterase activity were analyzed in the rat forebrain. Binding sites for the radilogand are widespreadly distributed in the rat forebrain, showing the highest density in hypothalamus. K(d) values were in the range 1-2 nM. Diadenosine tetraphosphate (Ap(4)A) and diethenoadenosine tetraphosphate, epsilon-(Ap(4)A), displaced the radioligand, indicating dinucleotide binding to ADPbetaS-recognizing P2Y receptors. Activity ecto nucleotide pyrophosphatase phosphodiesterase 1 (NPP1), able to hydrolyze Ap(4)A and other diadenosine polyphosphates, is also widely distributed through the rat forebrain, with the highest activity in hypothalamus. These results suggests that Ap(4)A signalling mediated by P2Y(1,12,13) receptors and enzymatically regulated by NPP1 activity may be particularly important in hypothalamus and add new support for neurotransmitter/neuromodulatory functions of diadenosine polyphosphates in brain. PMID- 18162318 TI - Histamine depolarizes neurons in the dorsal vagal complex. AB - We sought to determine whether histamine has effects on single neurons in the dorsal vagal complex of the brainstem since previous studies have suggested a role for histamine receptors in this region. Using whole-cell patch clamp recordings from neurons within the nucleus of the tractus solitarius (NTS) and the dorsal vagal nucleus (DVN), histamine (20 microM) depolarized a small proportion of neurons in these regions accompanied by a decrease in input resistance. Although few neurons were depolarized (21% of NTS neurons and 15% of DVN neurons), those that were affected showed robust depolarizations of 13 mV. These depolarizations were antagonized by the histamine H1 receptor antagonist triprolidine (2 microM) and were subject to a level of desensitization. Neither histamine nor the H3 receptor agonist imetit caused any change in the amplitudes of excitatory or inhibitory postsynaptic potentials elicited in NTS neurons by stimulation of the solitary tract. These data indicate that histamine has a restricted but profound effect on neurons in the dorsal vagal complex. PMID- 18162319 TI - Peripheral administration of CDP-choline and its cholinergic metabolites increases serum insulin: muscarinic and nicotinic acetylcholine receptors are both involved in their actions. AB - The present study was designed to test the effects of CDP-choline and its metabolites on serum insulin concentrations in rats and to investigate the involvements of cholinergic and adrenergic receptors in the effect. Intraperitoneal (i.p.) administration of CDP-choline (200-600 micromol/kg) increased serum insulin in a dose- and time-related manner. Equivalent doses (200 600 micromol/kg; i.p.) of phosphocholine or choline also increased serum insulin dose-dependently. Serum-free choline concentrations increased several-fold following i.p. administration of CDP-choline, phosphocholine or choline itself. In contrast, equivalent doses of cytidine monophosphate and cytidine failed to alter serum insulin concentrations. The increases in serum insulin induced by i.p. 600 micromol/kg of CDP-choline, phosphocholine or choline were abolished by pretreatment with the ganglionic nicotinic acetylcholine receptor antagonist hexamethonium (15 mg/kg; i.p.), or by the muscarinic receptor antagonist atropine methylnitrate (2 mg/kg; i.p.). Pretreatment with prazosin (0.5 mg/kg; i.p.), an alpha(1)-adrenoceptor antagonist, or yohimbine (5 mg/kg, i.p.), an alpha(2) adrenoceptor antagonist, enhanced slightly the increases in serum insulin in response to 600 micromol/kg of CDP-choline, phosphocholine and choline. Serum insulin also increased following central administration of choline; the effect was blocked by intracerebroventricularly injected atropine, mecamylamine or hemicholinium-3 (HC-3). It is concluded that CDP-choline or its cholinergic metabolites phosphocholine and choline increases circulating insulin concentrations by increasing muscarinic and nicotinic cholinergic neurotransmission in the insulin secreting beta-cells. PMID- 18162320 TI - Dissociating love-related attention from task-related attention: an event-related potential oddball study. AB - The present event-related potential (ERP) study was conducted to investigate the P3 component in response to love-related stimuli while controlling for task related factors, and to dissociate the influences of both love-related and task related attention on the P3 amplitude. In an oddball paradigm, photographs of beloved and friends served as target and distractor stimuli. Love-related and task-related attention were separated by varying the target and distractor status of the beloved and friends full factorially. As expected, the P3 amplitude was larger for beloved compared to friends and for targets compared to distractors. Moreover, task-related and love-related attention were unconfounded. These results are in line with findings that the P3 is modulated by both emotion- and task-related factors, supporting the view that the P3 amplitude reflects attention. Furthermore, this study validates the notion that romantic love is accompanied by increased attention for stimuli associated with the beloved, and also shows that this form of attention is different from task-related attention. PMID- 18162321 TI - Environmental enrichment-mediated functional improvement after experimental traumatic brain injury is contingent on task-specific neurobehavioral experience. AB - Environmental enrichment (EE) is superior to standard (STD) housing in promoting functional recovery after traumatic brain injury (TBI). However, whether the EE mediated benefits after TBI are dependent on exposure to enrichment during neurobehavioral training has not been elucidated. To address this issue, isoflurane-anesthetized adult male rats received either a cortical impact or sham injury and were then randomly assigned to early EE, delayed EE, continuous EE or no EE (i.e., STD conditions). Continuous EE or no EE was initiated immediately after surgery and continued for the duration of the study. Early EE began directly after surgery, continued for 1 week, and was then followed by STD living (2 rats per cage) for the remainder of the study, while delayed EE commenced 1 week after early STD housing. Functional outcome was assessed with established motor and cognitive tests on post-injury days 1-5 and 14-18, respectively. CA(1)/CA(3) neurons were quantified at 3 weeks. CA(3) cell loss was significantly attenuated in the TBI+continuous EE group versus the TBI+no EE group. Beam walking was facilitated in the TBI groups that received either early or continuous EE versus those receiving delayed or no EE. Cognitive training was enhanced in the TBI groups that received continuous or delayed EE versus the early EE or no EE groups. These data suggest that EE-mediated functional improvement after TBI is contingent on task-specific neurobehavioral experience. PMID- 18162322 TI - McArdle disease: another systemic low-inflammation disorder? AB - McArdle disease is caused by inherited deficit of human muscle glycogen phosphorylase with subsequent blockade in muscle glycogenolysis. Patients usually experience severe exercise intolerance and 'chronic' skeletal muscle damage. We determined circulating levels of 27 cytokines in a group of 31 adult McArdle patients (15 male 16 female; mean (+/-S.E.M.) age: 39+/-3 years) and 29 healthy sedentary controls (14 male, 15 female) before and after an acute exercise bout involving no muscle damage (cycling). Patients had an ongoing state of muscle breakdown even when following a sedentary lifestyle (serum creatine kinase activity at baseline of 2590+/-461 Ul(-1) vs. 97+/-5 Ul(-1) in controls). Under resting conditions, neutrophil count (+20%) and circulating levels of several cytokines were significantly higher (P0.1). In patients, 24h urinary-free cortisol levels (reflecting glucocorticoid replacement therapy) and testosterone levels were not associated with memory performance. These findings suggest that early steroid imbalances affect memory for negative material in children with CAH. Such memory impairments may result from abnormal brain organization and function following hormonal dysfunction during critical periods of development. PMID- 18162330 TI - Multi-centre observational study of buprenorphine use in 32 Italian drug addiction centres. AB - AIM: To examine how buprenorphine is currently being used across Italy, and to identify simultaneously best practice protocols to guide physicians in optimising the safety and efficacy of this treatment option. DESIGN: Retrospective, observational, multi-centre study. PARTICIPANTS: A total of 979 opioid-dependent patients were included from 32 centres involving the initiation of 1122 treatments. FINDINGS: During the study period 33.4% of patients relapsed during the induction phase. Lower induction doses resulted in markedly higher relapse rates (51.2% of those who received 2 mg versus 20.6% of those who received 10mg of buprenorphine relapsed). Over 89% of patients who received 16 mg of buprenorphine during the induction phase successfully went on to maintenance treatment. The percentage of drug-positive urines also decreased over time on buprenorphine treatment (cocaine-positive urines decreased from 25.8% at study entrance to 0% at 24 months). Psychosocial support in addition to buprenorphine pharmacotherapy further decreased the risk of relapse and was associated with lower levels of heroin craving. Retention in treatment was increased by less-than daily dosing of buprenorphine. CONCLUSIONS: Higher induction doses of buprenorphine significantly decreased relapse rates and increased the percentage of patients achieving maintenance treatment. Psychosocial support and/or less than-daily dosing also appeared to promote positive treatment outcomes. PMID- 18162331 TI - Predictors of dropout from group therapy among patients with bipolar and substance use disorders. AB - OBJECTIVE: Bipolar and substance use disorders frequently co-occur. Integrated treatment for these disorders has been shown to be effective at reducing substance use, but no study has examined attrition from dual diagnosis group therapy. The current study identified baseline demographic and clinical characteristics that predict treatment dropout among patients with co-occurring bipolar and substance use disorders. METHOD: Using binary and multivariate analyses, baseline data were analyzed as part of a randomized controlled trial of integrated group therapy for bipolar and substance use disorders. RESULTS: Cigarette smoking, recent mood episode, and lack of a college education were strong predictors of dropout after controlling for demographic and substance use variables. CONCLUSIONS: Given the strength of smoking as a predictor of dropout as well as the high rate of smoking among this population, a greater focus on the relationship between smoking and bipolar disorder is warranted. PMID- 18162332 TI - Prevalence of border disease virus in Spanish lambs. AB - The prevalence of border disease virus (BDV) viraemia in Spanish lambs was determined from 2089 sera randomly collected at two slaughterhouses in 2001 and 2003, as well as in 126 sera obtained in 2004 from a fattening unit with an acute disease problem. BDV was detected with an indirect peroxidase monolayer assay (IPMA), and for the fattening unit sera also by an antigen ELISA. A subset of sera was additionally tested for BDV antibodies. The BDV prevalence in the slaughterhouse sera was 0.24%, whereas 7.1% of randomly selected and 38.6% of sera from clinically affected lambs in the fattening unit were virus positive. Pestivirus antibodies were found in 17.6% of the slaughterhouse sera and 28.6% of those from randomly selected lambs in the fattening unit. In total, 33 virus isolates and 3 antigen positive samples were identified. Genetic typing of 5'-UTR sequences classified all 36 pestiviruses as of BDV type 4. This shows that from a low BDV prevalence in apparently healthy lambs in the entire sheep population, clinical problems associated with BDV can develop when viraemic sheep are brought into intense rearing units. PMID- 18162333 TI - Recovery of COCs from ovaries with high follicle numbers enhances in vitro embryo yield in sheep. AB - The aim of the present study was to investigate the correlation between the number of ovarian follicles and in vitro embryo development and quality in sheep. Sarda ewe ovaries were classified according to the number of follicles: or=8 (High). IVM, IVF and IVC were performed under standard conditions. Cleavage rate and blastocyst development were assessed 48 h after fertilization and on Days 6, 7 and 8 of culture, respectively. Expanded blastocysts were vitrified; blastocoel re-expansion and hatching rates were assessed at 8, 16 and 72 h post-thawing and hatched blastocysts were analyzed with the TUNEL assay. In a subset of thawed blastocysts the incorporation of amino acids was evaluated. The proportion of ovaries varied significantly among the three groups (ANOVA F=12.20, P=0), and more ovaries (59%) were assigned to the Low group than to the Intermediate (28%; ANOVA F=8.19, P=0.009) and High group (13%; ANOVA F=18.63, P=0), (High vs. Intermediate F=6.31, P=0.020). The three groups statistically differed in the proportion of total blastocysts (chi(2)(2)=22.616, P=0.00), of blastocysts produced on Days 6 (chi(2)(2)=6.829, P=0.033) and 7 (chi(2)(2)=6.810, P=0.033), while no difference was found in the proportion of blastocysts obtained on Day 8 (chi(2)(2)=3.874, P=0.144) of culture after fertilization. A higher proportion of total blastocysts was obtained from the High (44%) compared with the other two groups (Low: 28%, chi(2)(2)=22.629, P=0; Intermediate: 33%, chi(2)(2)=7.266, P=0.007), while the Low and Intermediate groups did not statistically differ either in the total blastocyst output (chi(2)(2)=3.384, P=0.066), nor in the number of blastocysts produced on Days 6 (Low: 7%, Intermediate: 9%; chi(2)(2)=0.874, P=0.35), 7 (Low: 14%, Intermediate: 16%, chi(2)(2)=1.256, P=0.26) and 8 (Low: 6%, Intermediate: 7% chi(2)(2)=0.554, P=0.45) of culture. The High group produced a significantly higher percentage of embryos on Days 6 (High: 13%, Low: 7%; chi(2)(2)=6.840, P=0.009) and 7 (High: 21%, Low: 14%; chi(2)(2)=6.806, P=0.009) of culture post insemination than the Low group. The three categories did not differ in the blastocoel re-expansion (chi(2)(2)=0.095, P=0.95) and hatching rates (chi(2)(2)=0.754, P=0.68) after 72 h post-warming, in the total number of cells per blastocyst (ANOVA F=1.12, P=0.337) and in the (F=0.46, P=0.639) incorporation of amino acids. The number of TUNEL-positive cells per embryo was higher (ANOVA F=4.32, P=0.022) in the Low group compared to the other groups. In conclusion, high ovarian follicle number enhances in vitro embryo output in sheep, but has no effect on blastocyst quality. PMID- 18162334 TI - Intrafollicular effect of IGF1 on development of follicle dominance in mares. AB - The effect of an injection of a supraphysiologic dose of rhIGF1 into the second largest ovarian follicle (F2) at the expected beginning of deviation (F1, > or =20 mm; Day 0) on development of dominance by F2 was studied in mares (n=16; controls, n=8). F1 became dominant (> or =28 mm) in 8 of 8 and 15 of 16 follicles in the controls and treated groups, respectively. The incidence of dominance (P<0.001) and ovulation (P<0.02) for F2 was greater for the IGF1 group (13 of 16 and 10 of 16) than for the controls (1 of 8 and 1 of 8). There were day effects but no group effects or group-by-day interactions for systemic FSH, LH, estradiol, or ir-inhibin during the 4 days after treatment. In another experiment, treatment of every follicle, excluding F1, when it reached > or =20mm after the expected beginning of deviation resulted in dominance by 8 of 12 follicles treated with rhIGF1 on Days 1-3 (n=8 mares). Results demonstrated that the IGF1 system plays a pivotal intrafollicular role in the deviation mechanism without altering systemic concentrations of the gonadotropins and ovarian follicular hormones. PMID- 18162335 TI - Hyaluronic acid delays boar sperm capacitation after 3 days of storage at 15 degrees C. AB - The present study was undertaken to determine the effects of the addition of hyaluronic acid (HA), ranged from 12.5 to 200 microg/ml, on boar sperm capacitation status during a storage time (up to 3 days) at 15 degrees C in Beltsville thawing solution (BTS). The raw extender was the negative control whereas different concentrations of caffeine (CAF), ranged from 0.25 to 8mM, served as positive controls. Sperm viability, motility, morphology, and osmotic resistance were also determined before and after assessing the treatments. Samples were obtained from 28 healthy and post-pubertal Pietrain boars and sperm parameters were tested immediately after the addition of treatments and after 1, 2 and 3 days of refrigeration at 15 degrees C. Sperm capacitation status was determined by chlortetracycline (CTC) staining and sperm viability by means of a multiple fluorochrome-staining test. Sperm motility and morphology were assessed using phase-contrast microscopy accompanied by a computer assisted sperm analysis system (CASA). Whereas HA delayed sperm capacitation, CAF increased the frequency of capacitated spermatozoa after 2 days of cooling. Moreover, HA did not modify other sperm parameters, such as sperm velocity, whereas CAF increased progressive motility during the first 2 days of cooling and then decreased. It can be concluded that the addition of HA at 50 and 100 microg/ml to the BTS extender may delay sperm capacitation after 3 days of cooling. PMID- 18162336 TI - Testicular and spermatozoan parameters in the pukeko (Porphyrio porphyrio melanotus). AB - The pukeko (Porphyrio porphyrio melanotus) is widespread in New Zealand, and is the closest living relative to the endangered takahe (Porphyrio mantelli), which has a relatively high rate of infertility. In this study, sperm collected from a number of pukeko was analysed in order to model the reproductive physiology of the male takahe. In addition, testicular parameters were measured. To ascertain the best method of sperm collection five techniques for harvesting sperm were used on two occasions during the breeding season. All five techniques resulted in the successful recovery of sperm. However, the float-out technique produced the best quality samples. Following collection, the morphometry of unstained sperm was assessed. Our findings suggest that pukeko sperm is non-motile in the male reproductive tract. We found the mean sperm head length in the pukeko is 16.9mum, but sperm head length varied significantly between birds. Testicular weight and length was significantly correlated with bird weight (P<0.05). Within each bird, testes weights were asymmetric. However, testes length was significantly correlated (P<0.05). There was a significant difference (P<0.05) in testes length between birds. The methodologies presented for obtaining and analysing pukeko sperm morphometry can be used to assist opportunistic studies of the reproductive biology of other New Zealand native birds. PMID- 18162337 TI - Analysis of actinomycin D treated cattle oocytes and their use for somatic cell nuclear transfer. AB - The present work aimed to evaluate the transcription and replication inhibitor, actinomycin D, for oocyte chemical enucleation. Cattle oocytes matured in vitro were treated with actinomycin D according to the following treatments: T1, control; T2=1.0 microg/ml for 16 h; T3=1.0 microg/ml for 14 h; T4=2.5 microg/ml for 14 h; T5=5.0 microg/ml for 14 h. The oocytes were denuded and activated during 24-26 h of maturation. Oocytes were fixed to determine the maturation status and for chromosome morphology evaluation. Furthermore, oocytes treated with actinomycin D were used for somatic cell nuclear transfer (SCNT). Parthenogenetic and SCNT embryos were fixed to evaluate the percentage of apoptotic nuclei by the TUNEL assay. The maturation (T1=90.4%; T2=82.3%; T3=79.1%; T4=83.4%; T5=74.7%), cleavage (T1=68.9%; T2=46.0%; T3=49.7%; T4=33.4%; T5=29.3%) and blastocyst rate at D8 (T1=41.1%; T2=1.8%; T3=1.3%; T4=0.9%; T5=0.0%) after actinomycin D treatment were significantly different. There was a significant chromosome uncoiling when treated with greater concentrations (2.5 and 5.0 microg/ml). After SCNT, the cleavage rate (61.3%) was similar to the actinomycin D-treated control group (61.3%) and less than the non-treated control (70.2%), although the blastocyst rate was greater in the SCNT group (11.8%) comparing with the treated control (3.6%) and less than the untreated control (38.0%). Treated parthenogenetic embryos had more apoptotic cells than the parthenogenetic controls (24.2% compared with 4.8%). However, the SCNT group using treated cytoplasts was similar from the SCNT control (9.3 compared with 13.0%). Actinomycin D treatment was efficient in blocking embryonic development. Moreover, it was possible to obtain reconstructed embryos that possess an apoptotic cell index indistinguishable from controls. PMID- 18162338 TI - Endometrial polyps in pre-and postmenopausal women: factors associated with malignancy. PMID- 18162339 TI - Postmenopausal complete hydatidiform mole: a case report. AB - BACKGROUND: Gestational trophoblastic neoplasia (GTN) is a disease with a course of trophoblastic proliferation, and histologically classified as partial hydatidiform mole, complete mole, invasive and metastatic mole, choriocarcinoma and placental site trophoblastic tumor. Occurrence of GTN in postmenopausal women is rare. CASE: We report the case of a 56-year-old postmenopausal woman with a complete mole. The patient was admitted to gynecology outpatient clinic with abdominal pain, nausea and vomiting for about 1 month. Ultrasound examination revealed enlargement of the uterus with endometrial thickness containing hypo/hyper echogeneous and cystic areas. Serum beta-HCG was tested against the possibility of GTN because of the appearance in sonography and was found >5000. The patient underwent total abdominal hysterectomy and bilateral salpingo oophorectomy. The serum level of beta-HCG has decreased from initially observed 188,000-0 U/ml in 4th week. The resected uterus contained an endometrial, cystic, grapelike tumor. Microscopic examination demonstrated hydropic degeneration of all the chorionic villi with trophoblastic cell proliferation consistent with a complete hydatidiform mole. CONCLUSION: To our knowledge, our case is the fourth description in the world literature of a benign complete hydatidiform mole in a postmenopausal woman. Although benign gestational trophoblastic disease generally occurs in women of reproductive age and is extremely rare in postmenopausal women, when evaluating patients who are in postmenopausal period the diagnosis of hydatidiform mole must always be considered. PMID- 18162340 TI - Formation mechanism of colloidal nanoparticles obtained from probucol/PVP/SDS ternary ground mixture. AB - The purpose of this study was to investigate the formation mechanism of colloidal nanoparticles after dispersion of probucol/polyvinylpyrrolidone (PVP)/sodium dodecyl sulphate (SDS) ternary ground mixture (GM) into water. Probucol, PVP and SDS were mixed at a weight ratio of 1:3:1 and ground for 30 min with a vibrational rod mill. The morphology and physicochemical properties were investigated through high resolution scanning electron microscopy (SEM), environmental SEM, dynamic light scattering, (13)C NMR and zeta potential measurements. SEM images confirmed the presence of 20 nm size primary particles in the GM powder of probucol/PVP K17/SDS. Spherical nanoparticles with a size of around 100 nm, formed after dispersion of the GM into water, suggested an agglomeration of the primary particles. A further agglomeration of around 160 nm was observed with the stability experiment. Zeta potential and particle size measurements using latex beads revealed that PVPK 17/SDS complex was adsorbed on the probucol particle surface forming a layered structure. A similar agglomeration behavior was observed using the GM of probucol/PVP K12/SDS, though the molecular state of the PVPK 12/SDS complex at the particle surface was different from that of the PVPK 17/SDS complex. (13)C NMR results suggested that intermolecular interactions between PVP K12 and SDS did not reach the same level as the interactions between PVP K17 and SDS. This study proposed a formation mechanism of colloidal nanoparticles. PMID- 18162341 TI - How to achieve sustained and complete protein release from PLGA-based microparticles? AB - One of the most challenging tasks in the delivery of therapeutic proteins from PLGA-based microparticles is the sustained and complete release of the protein in its native form. The mechanisms responsible for incomplete protein release from these devices are numerous and complex; the beneficial effect of different formulations has often been evaluated in vitro. Strategies employed for overcoming protein destabilization during the release step are reviewed in this paper. Proteins have been protected in the deleterious environment by adding stabilizers to the formulation, or by modifying the protein or the polymer. Alternatively, some strategies have aimed at avoiding the formation of the destabilizing environment. As experimental conditions may influence the results from in vitro release studies, we initially report precautions to avoid adverse effects. PMID- 18162342 TI - The development of microthermal analysis and photothermal microspectroscopy as novel approaches to drug-excipient compatibility studies. AB - The use of microthermal analysis as a novel means of assessing chemical incompatibility between drugs and excipients is assessed using magnesium stearate and acetylsalicylic acid as a model system. Localised thermomechanical analysis (L-TMA), localised differential thermal analysis (L-DTA), nanosampling, thermally assisted particle manipulation (TAPM) and photothermal microspectrometry (PTMS) are developed as a means of allowing extremely small quantities of drug and excipient to be heated in close proximity to each other. Differential scanning calorimetry (DSC), hot stage microscopy (HSM) and temperature controlled attenuated total internal reflection (ATR) FTIR were used as supportive techniques. L-TMA and macroscopic TMA of magnesium stearate indicated that the endothermic DSC peak normally associated with melting does not correspond to significant liquefaction. An optimised method for detecting the interaction at a particulate level of scrutiny was developed whereby the drug is placed on the excipient surface via TAPM and the construct heated, allowing the interaction to be detected in both the L-TMA and L-DTA signal. PTMS allowed spectra to be obtained on nanogram-sized samples and also allowed the interaction to be detected. The study has therefore demonstrated the potential for using TAPM with PTMS for studying interactions at an individual particle level. PMID- 18162343 TI - An ionically crosslinked hydrogel containing vancomycin coating on a porous scaffold for drug delivery and cell culture. AB - The aim of this study was to prepare and characterize a scaffold with an ionically crosslinked hydrogel coating layer containing a water-soluble drug, vancomycin, via a novel drug loading method for sustained drug delivery and surface modification. The poly(D,L-lactide acid) (PDLLA)/biphasic calcium phosphate (BCP) scaffold with a highly inter-connected porous structure was fabricated by a particle-leaching/thermally induced phase separation (TIPS) method. The pre-vacuumized scaffold was immersed into an alginate/vancomycin solution. Following impregnation by the solution, the scaffold was removed and immersed in a CaCl(2) solution for 30 min to allow gelation of the alginate solution. In this way, the drug was not exposed to organic solvents or detrimental temperature conditions and it could avoid loss of drug during the leaching process. The water contact angles of the scaffold surface decreased after being coated with the hydrogel. The in vitro drug release profile showed sustained release properties which were influenced by the alginate concentration and the dissolution medium. A standardized bacterial assay showed that the drug was still active after association with the scaffold by this gentle method of drug loading. The in vitro osteoblast culture experiments confirmed the biocompatibility of the scaffold for attachment and proliferation of osteoblasts. PMID- 18162344 TI - Empirical kinetic model of propafenone release from Hot Air Coating microparticles. AB - Lipid microparticles, containing 30% and 50% (w/w) propafenone hydrochloride as the active molecule and cetearyl alcohol and Pluronic F68 as excipients, were prepared by Hot Air Coating (HAC). The aim of the work was to identify the kinetics and the mechanism of the drug release process from these microparticulate systems. The application of the Weibull model to the release data from each single fraction of microparticles suggests that a diffusive mechanism governs drug release from microparticles. Thus, we proposed and applied a release kinetic model to the experimental data that takes into account the diffusion as the predominantly mechanism of drug release. The model proposed is a modified version of the exponential equation in which the product of the apparent release rate constant K, specific for each drug/excipient mixture, and the area to-volume ratio of particles was used. The K values of single fractions of HAC microparticles (coded K(fr)) are very similar to those of the mixtures of particles obtained from the process (coded K(pool)). Using the K(pool) constants, the release behaviour of ensembles of different size microparticles of well-known composition was predicted. The strength of the model was proved by the good fitting of the experimental release data versus those predicted (R(2)> or =0.997). PMID- 18162345 TI - Ligand based dendritic systems for tumor targeting. AB - Medications that can selectively target tumors at the same time avoid access of the drug to nontarget areas, employ utilization of homing devices termed as ligands, that can bind to specific epitopes expressed on the surface of the necrotic mass of cells. Molecular signatures for transferrin, Epidermal Growth Factor, Sialic Lewis and folic acid are expressed on the surface of these cells. Dendrimers are nanosized, non-immunogenic, and hyper-branched vehicles that can be efficiently tailored for spatial distribution of bioactives, thereby reducing untoward cytotoxicity on normal cells. These nanoparticulate drug delivery vehicles provide a unique platform that has precisely placed functional groups so that multiple copies of ligands can be attached to it and facilitate targeting to the tumor surface or neo-vascularizing vessels proliferating around these cells. The article reviews the scope of ligand based dendritic system as a prospective for delivery of anti-cancer drugs, via active targeting with interception of minimal side effects. PMID- 18162346 TI - Enhanced immune response induced by BSA loaded in hydroxyethylstarch microparticles. AB - Microparticles and nanoparticles represent promising carriers for the in vivo delivery of peptides, proteins or deoxyribonucleic acid (DNA). In this study, new hydroxyethylstarch (HES) microparticles were obtained by interfacial cross linking with terephtaloyl chloride. These microparticles exhibit the characteristics required to improve antigen release and presentation to antigen presentating cells compared to free antigens. The adjuvant activity of HES microparticles as vaccine carrier was investigated in mice using bovine serum albumin (BSA) as model antigen. We showed HES microparticles were phagocyted by peritoneal mononuclear cells. The immunization with BSA-microparticles induced antibody synthesis that was predominantly immunoglobulin G1 (IgG1). Aluminium hydroxide remained more efficient to induce IgG synthesis. The analysis of the cytokine profile from spleen cells revealed that BSA-microparticles induced the secretion of both interferon-gamma (IFN-gamma) and interleukin-4 (IL-4). However, the immune responses induced by BSA-microparticles were qualitatively and quantitatively affected by the route of injection. Taken together, these results demonstrate that HES microparticles induce a mixed T helper 1/T helper 2 response against BSA and may be a suitable delivery and presentation system in the field of vaccine development. PMID- 18162347 TI - Dynamic changes in level influence spatial coding in the lateral superior olive. AB - It is well established that the responses of binaural auditory neurons can adapt and change dramatically depending on the nature of a preceding sound. Examples of how the effects of ensuing stimuli play a functional role in auditory processing include motion sensitivity and precedence-like effects. To date, these types of effects have been documented at the level of the midbrain and above. Little is known about sensitivity to ensuing stimuli below in the superior olivary nuclei where binaural response properties are first established. Here we report on single cell responses in the gerbil lateral superior olive, the initial site where sensitivity to interaural level differences is established. In contrast to our expectations we found a robust sensitivity to ensuing stimuli. The majority of the cells we tested (86%), showed substantial suppression and/or enhancement to a designated target stimulus, depending on the nature of a preceding stimulus. Hence, sensitivity to ensuing stimuli is already established at the first synaptic station of binaural processing. PMID- 18162348 TI - Combined effect of fluid and pressure on middle ear function. AB - In our previous studies, the effects of effusion and pressure on sound transmission were investigated separately. The aim of this study is to investigate the combined effect of fluid and pressure on middle ear function. An otitis media with effusion model was created by injecting saline solution and air pressure simultaneously into the middle ear of human temporal bones. Tympanic membrane displacement in response to 90 dB SPL sound input was measured by a laser vibrometer and the compliance of the middle ear was measured by a tympanometer. The movement of the tympanic membrane at the umbo was reduced up to 17 dB by the combination of fluid and pressure in the middle ear over the auditory frequency range. The fluid and pressure effects on the umbo movement in the fluid-pressure combination are not additive. The combined effect of fluid and pressure on the umbo movement is different compared with that of only fluid or pressure change in the middle ear. Negative pressure in fluid-pressure combination had more effect on middle ear function than positive pressure. Tympanometry can detect the middle ear pressure of the fluid-pressure combination. This study provides quantitative information for analysis of the combined effect of fluid and pressure on tympanic membrane movement. PMID- 18162349 TI - Anti-diarrhoeal activity of the aqueous extract of Mezoneuron benthamianum Baill (Caesalpiniaceae). AB - The effect of the aqueous extract of Mezoneuron benthamianum (MB) on experimentally induced diarrhoea, intestinal propulsive movement (IPM) and intestinal fluid accumulation (enteropooling) were investigated in rats and mice. The extract (400, 800 and 1600 mg/kg, orally) produced a significant (p<0.05) and dose-dependent reduction in propulsion in the castor oil-induced intestinal transit in mice. The mean peristaltic index (%) for these doses of extract, control, (distilled water, 10 ml/kg, p.o.) and morphine, (10 mg/kg, s.c.) were 73.48, 69.34, 57.27, 89.93 and 31.56, respectively. The effect of the extract at the highest dose was significantly (p<0.05) lower than that of the standard drug. This effect was antagonised by yohimbine (1mg/kg, s.c.). In a dose-dependent manner, the extract delayed the onset of diarrhoea, produced a significant decrease in the frequency of defaecation, severity of diarrhoea and protected the mice treated with castor oil. Total diarrhoea scores were 12.0+/-0.63, 10.3+/ 2.06, 8.5+/-2.15, 7.1+/-0.91 and 5.8+/-0.79 for control, extract (400, 800 and 1600 mg/kg) and morphine, respectively. The extract significantly decreased the volume (ml) of intestinal fluid secretion induced by castor oil (1.75+/-0.02 to 0.93+/-0.04) compared with 1.90+/-0.05 for control. The inhibitory effect on fluid accumulation by the extract was also attenuated by yohimbine (1.0 mg/kg). Preliminary phytochemical screening revealed the presence of flavonoids, tannins, cardiac glycosides, anthraquinones and saponins. Administration of the extract up to 2 g/kg (orally) did not produce any toxic effect in the acute toxicity studies in mice. The LD(50) of the extract when given intraperitoneally was 1021.31 mg/kg. The results obtained show that MB possesses anti-diarrhoeal activity due to its inhibitory effects on gastrointestinal propulsion and intestinal fluid accumulation. The antagonistic actions of yohimbine in the experiments suggest a role for the a(2)-adrenergic receptor system. PMID- 18162350 TI - Shengmai San reduces hepatic lipids and lipid peroxidation in rats fed on a high cholesterol diet. AB - Shengmai San (SMS), which is comprised of the medicinal herbs of Panax ginseng C.A. Meyer, Schisandra chinensis Baill., and Ophiopogon japonicus Ker-Gawl (2:1:2)., is a traditional Chinese medicine being used for treating coronary heart disease. The aim of this study was to investigate the effects of SMS on the plasma and liver lipids, lipid peroxidation and antioxidant systems in liver and heart of cholesterol-fed rats. Rats were fed on a high-cholesterol (0.5%) diet (control group), high-cholesterol diet containing 2% SMS (2% SMS group) and 4% SMS (4% SMS group) for four weeks. The oxidative stress marker (thiobarbituric acid reactive substances, TBARS) and antioxidant defense systems including glutathione (GSH), glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST) and superoxide dismutase (SOD) activities in rat liver and heart were evaluated. Results showed that rats fed with SMS-containing diet had reduced the H(2)O(2)-induced erythrocytes susceptibility to hemolysis, and 4% SMS feeding rats had higher plasma GSH concentration compared to the animals fed with the control diet. However, SMS had no effect on plasma lipids (total cholesterol, triglyceride and high-density lipoprotein cholesterol) and TBARS concentration. On the other hand, rats fed with the 4% SMS diet reduced the hepatic cholesterol and triglyceride contents. Fecal bile acid excretion was significantly increased in rats fed with the SMS-containing diet. Higher hepatic GSH and lower TBARS concentrations were observed in rats fed with the 4% SMS diet compared with the rats fed with the control diet. No significant difference in activities of GSH Px, GST and SOD was found in liver and heart after the SMS treatment. Results from this study indicate that the SMS may reduce hepatic lipids and lipid peroxidation in rats. PMID- 18162351 TI - Multi-detector row CT angiography in the assessment of coronary in-stent restenosis: a systematic review. AB - PURPOSE: The aim of this study was to perform a systematic review of the diagnostic accuracy of multi-detector row computed tomography angiography (MDCT) for detection of coronary in-stent restenosis in patients treated with coronary stenting when compared to invasive catheter angiography. MATERIALS AND METHODS: A search of PUBMED and MEDLINE databases for English literature was performed. Only studies with at least 10 patients comparing 16- or more detector rows MDCT angiography with invasive catheter angiography in the detection of coronary in stent restenosis (more than 50% stenosis) were included for analysis. Sensitivity and specificity estimates pooled across studies were tested using a fixed effects model. RESULTS: 15 studies met selection criteria for inclusion in the analysis. There were eight studies performed with 16-detector row CT scanners, and five studies with 64-detector row scanners and one study with a 40-detector scanner. The remaining study was performed with a mixture of 16-and 64-detector row scanners. Prevalence of in-stent restenosis following coronary stenting was 18% (95% CI: 13, 24%). Pooled estimates of the sensitivity and specificity of overall MDCT angiography for the detection of coronary in-stent restenosis was 85% (95% CI: 78, 90%) and 97% (95% CI: 95, 98%), respectively. No significant difference was found between 16- and 64-detector row scanners regarding the sensitivity and specificity of MDCT for assessment of in-stent restenosis (p>0.05). CONCLUSION: The results showed that MDCT angiography (with 16 or more detector rows) has moderate sensitivity and high specificity for the detection of coronary in-stent restenosis when compared to invasive catheter angiography. A high specificity value of MDCT may be most valuable as a non-invasive technique of excluding coronary stent restenosis or occlusion. The main factors affecting visualization are stent diameters and stent materials. PMID- 18162352 TI - High b-value diffusion-weighted MR imaging of normal brain at 3T. AB - INTRODUCTION: The purpose of this study was to determine the normative apparent diffusion coefficient (ADC) values at 3T using high b-value (3000 s/mm(2)) diffusion-weighted images (DWI) and compare the signal characteristics of the high b value with standard b-value (1000 s/mm(2)) DWI. METHODS: Institutional review board approval was obtained for this prospective study which included 20 volunteers (10 M, 10 F, mean age: 38.7+/-14.9) without any known clinical disease or radiological findings. All brain examinations were performed with 3T MR by using similar parameters of b1000 and b3000 DWI sequences. DWI and ADC maps were obtained. Signal intensity, noise, signal to noise ratio (SNR), contrast to noise (CNR), contrast ratio (CR), and ADC values of bilateral posterior limb of internal capsule, frontal white matter, parietal gray matter, pons, thalamus, splenium of corpus callosum were measured on b1000 and b3000 DW images. RESULTS: In all anatomic locations, MR signal intensity, SNR and ADC values of b3000 images were significantly lower than MR signal intensity, SNR and ADC values of b1000 images (p<0.001). The CNR and CR values at the posterior limb of internal capsule and pons were significantly increased on b3000 images (p<0.001) and decreased in the other regions measured. CONCLUSION: The ADC values calculated from standard b-value DWI were significantly higher than those calculated from high b-value DWI. These results agree with the previous studies. In the regions where CNR values increase with high b value, b3000 DWI images may provide additional clinical information. PMID- 18162353 TI - Artifacts in 3-T MRI: physical background and reduction strategies. AB - Magnetic resonance imaging (MRI) at a field-strength of 3 T has become more and more frequently used in recent years. In an increasing number of radiological sites, 3-T MRI now starts to play the same role for clinical imaging that was occupied by 1.5-T systems in the past. Because of physical limitations related to the higher field strength and because of protocols transferred from 1.5-T MRI that are not yet fully optimized for 3 T, radiologists and technicians working at these systems are relatively often confronted with image artifacts related to 3-T MRI. The purpose of this review article is to present the most relevant artifacts that arise in 3-T MRI, to provide some physical background on the formation of artifacts, and to suggest strategies to reduce or avoid these artifacts. The discussed artifacts are classified and ordered according to the physical mechanism or property of the MRI system responsible for their occurrence: artifacts caused by B0 inhomogeneity and susceptibility effects, B1 inhomogeneity and wavelength effects, chemical-shift effects, blood flow and magnetohydrodynamics, and artifacts related to SNR. PMID- 18162354 TI - Clinical advantages of 3.0 T MRI over 1.5 T. AB - Since approval by the FDA in 2000, human MR imaging (MRI) at 3.0 T has been increasingly used in clinical practice. In spite of the potential technical challenges, a number of clinical advantages of 3.0 T MRI over 1.5 T have been identified in the recent years. This article reviews the benefits and the current knowledge of 3.0 T whole-body MRI from an evidence-based perspective and summarizes its clinical applications. PMID- 18162355 TI - Isolation and characterization of process related impurities and degradation products of bicalutamide and development of RP-HPLC method for impurity profile study. AB - A reversed-phase high-performance liquid chromatographic method was developed for determination of process impurities and degradation products of bicalutamide in bulk drug and pharmaceutical formulations. The separation was accomplished on a Symmetry C(18) (4.6 mm x 250 mm; particle size 5 microm) column under isocratic mode. The mobile phase was 0.01 M KH(2)PO(4) (pH 3.0):acetonitrile (50:50 v/v) and a PDA detector set at 215 nm was used for detection. Forced degradation of bicalutamide was carried out under thermal, photo, acidic, alkaline and peroxide conditions. The unknown process impurities and alkaline degradation products were isolated and characterized by ESI-MS/MS, (1)H NMR and FT-IR spectral data. Under alkaline conditions bicalutamide was degraded in to an acid and an amine. The kinetics of degradation was studied. The proposed method was validated and successfully applied to the analysis of commercial formulations. Thus, the developed method can be used for process development as well as quality assurance of bicalutamide in bulk drug and pharmaceutical formulations. PMID- 18162356 TI - Determination of moisture content of lyophilized allergen vaccines by NIR spectroscopy. AB - Moisture content is an important parameter for lyophilized vaccines. Currently, Karl Fischer titration is widely used for moisture determination in routine analysis. However, this method is time-consuming, sample destructive and requires environment polluting reagents, as well as the results rely on the random samplings. In this study, near infrared spectroscopy was used as a fast, non invasive and non-destructive method to determine the moisture content in lyophilized allergy vaccines. Five different vaccine products were investigated, which contained water in the range of 0.17-1.51% (w/w, KF). Different data pre treatments, wavelength selection and partial least squares regression were applied to construct calibration models. Multi-products model and product specific models were obtained, which show the possibility of NIR as a rapid method to discriminate whether moisture content fit into the specifications of a pharmaceutical company. PMID- 18162357 TI - Patterns of concordance and non-concordance with clinician recommendations and parents' explanatory models in children with asthma. AB - OBJECTIVE: Many children with asthma do not take medications as prescribed. We studied parents of children with asthma to define patterns of non-concordance between families' use of asthma controller medications and clinicians' recommendations, examine parents' explanatory models (EMs) of asthma, and describe relationships between patterns of non-concordance and EM. METHODS: Qualitative study using semi-structured interviews with parents of children with persistent asthma. Grounded theory analysis identified recurrent themes and relationships between reported medication use, EMs, and other factors. RESULTS: Twelve of the 37 parents reported non-concordance with providers' recommendations. Three types of non-concordance were identified: unintentional- parents believed they were following recommendations; unplanned--parents reported intending to give controller medications but could not; and intentional--parents stated giving medication was the wrong course of action. Analysis revealed two EMs of asthma: chronic--parents believed their child always has asthma; and intermittent--parents believed asthma was a problem their child sometimes developed. CONCLUSIONS: Concordance or non-concordance with recommended use of medications were related to EM's and family context and took on three different patterns associated with medication underuse. PRACTICE IMPLICATIONS: Efforts to reduce medication underuse in children with asthma may be optimized by identifying different types of non-concordance and tailoring interventions accordingly. PMID- 18162358 TI - Lymphatic mapping in patients with primary or recurrent breast cancer following previous axillary surgery. AB - AIMS: To evaluate the feasibility of lymphatic mapping in breast cancer patients after previous axillary surgery and to identify parameters associated with mapping failure. METHODS: Lymphatic mapping using peritumoural injection of blue dye and a radiocolloid was attempted in 30 patients with primary (n=7) or recurrent (n=23) breast cancer and a history of previous axillary lymph node dissection or sentinel node biopsy. RESULTS: Lymphatic mapping identified a mean number of 1.6 (range 1-3) lymph nodes in 19 of 30 patients (identification rate 63%). The lymph nodes were removed from the ipsilateral axilla (n=13), the internal mammary chain (n=2), both the internal mammary nodes and the axilla (n=2), the interpectoral space (n=1) and the contralateral axilla (n=1). Four of 19 patients revealed a positive lymph node. Fifteen of 19 patients had a negative lymph node. Axillary lymph node dissection was done in 13 of 15 patients but found no positive nodes (false negative rate=0). A negative lymphoscintigram (p<0.001) and a number of more than 10 lymph nodes removed at the time of initial surgery (p=0.02) were significantly associated with a mapping failure. CONCLUSION: Lymphatic mapping following prior axillary surgery was accurate but associated with a low identification rate. The lymphatic drainage pattern was unpredictable and the use of a radionuclide was necessary for a successful mapping procedure. PMID- 18162359 TI - Screen-detected breast lesions with an indeterminate (B3) core needle biopsy should be excised. AB - BACKGROUND: Screen-detected breast lesions in the National Health Service Breast Screening Programme (NHSBSP) are assessed by core needle biopsy (CB) or fine needle aspiration cytology (FNAC). Most core biopsies are diagnostic and representative, but a small proportion is indeterminate (coded "B3" in the NHSBSP). We studied the surgical outcome of screen-detected breast lesions with indeterminate (B3) CB. METHODS: We retrieved and analysed the data on women who were recalled for assessment of a screen-detected abnormality in whom the initial CB was reported as B3 over a six-year period from a prospectively collected database in one breast screening centre. The main outcome measure was final histology following surgical excision. RESULTS: Among 4080 CB performed, 220 (5.4%) were B3. Mammographically 127 lesions were microcalcifications and 88 were soft tissue lesions. On surgical excision (n=199, 90%), 67 (34%) were malignant. In patients with malignancy, clinical examination, US and concurrent FNAC were either suspicious or definitive of malignancy only in 2%, 4% and 7%, respectively. CONCLUSION: A third of screen-detected breast lesions with indeterminate CB are malignant on excision. Clinical examination, US, and FNAC may identify some of these carcinomas pre-operatively but most malignancies would not be picked up. Thus, these lesions should undergo surgical excision. PMID- 18162360 TI - [Kidney allograft harvesting on a donor patient under cardiopulmonary bypass]. AB - We report the case of a kidney allograft harvesting on a donor patient under cardiopulmonary bypass (CPB) after a cardiac arrest. The two kidneys were successfully transplanted with immediate graft function. CPB may provide an option to expand the number of harvested grafts. PMID- 18162361 TI - Rosuvastatin prevents endothelial cell death and reduces atherosclerotic lesion formation in ApoE-deficient mice. AB - Accumulating evidence suggests that statins have beneficial effects which are independent of their lipid-lowering actions, on vascular cells. Here, we investigated whether the HMG-CoA reductase inhibitor rosuvastatin can inhibit atherosclerotic lesion development with favorable effects on endothelial cells in ApoE-deficient mice. Rosuvastatin rapidly phosphorylated Akt and endothelial nitric oxide synthase (eNOS) in human endothelial cells. Endothelial cell death induced by serum starvation was significantly inhibited by rosuvastatin (percent cell death; 45.9+/-2.4% vs. 37.3+/-1.1%, p<0.05). Eight-week-old ApoE-deficient mice were orally administered vehicle or rosuvastatin at a dose of 20mg/kg/day for 24 weeks. There was no significant difference in cholesterol profile. Rosuvastatin preserved endothelial lining at the aortic root (CD31-positive luminal side; 63.8+/-2.8% vs. 81.7+/-3.9%, p<0.05). En face Sudan IV staining of aorta revealed that rosuvastatin significantly decreased the atherosclerotic area (21.9+/-2.9% vs. 11.9+/-1.9%, p<0.05). Lipid deposition at the atherosclerotic area was also suppressed by rosuvastatin with more stabilized morphologic features as determined by oil red O staining (3.4+/-0.4% vs. 1.7+/-0.4%, p<0.05). Our findings indicate that rosuvastatin protects endothelial cells from death with phosphorylation of Akt and eNOS. These effects may contribute, at least in part, to the anti-atherosclerotic effects of rosuvastatin. PMID- 18162362 TI - Influenza A virus derived from persistently virus-infected cells shows attenuated cytotoxicity in cultured cells but virulent pathogenicity in mice. AB - The IVpi-43 strain of influenza A virus, a progeny virus derived from persistently virus-infected Madin-Darby canine kidney (MDCK) cells, showed a more attenuated nature in cytopathology in cultured cells than the parental wild-type influenza virus (IVwt) that was used for establishment of the virus carrier culture. Upon infection of MDCK cells, growth of the IVpi-43 virus was restrained with an impaired synthesis of virus structural proteins in the cells. Apoptosis induced by IVpi-43 virus was confined at a low level. The IVpi-43 virus was able to easily cause persistent infection in fresh MDCK cells. In contrast to the in vitro phenotype, the IVpi-43 virus proved highly virulent in mice, with massive and broadly disseminated virus multiplication in the lungs. It was suggested that impaired activity of the neuraminidase molecule of the IVpi-43 virus was responsible for the delayed and faint appearance of apoptosis in the IVpi-43 virus-infected respiratory cells, which made it possible for the virus to replicate for a longer period and to spread to a broader area of the lungs and that abundant numbers of the virus-infected lung cells were killed within a short period by the subsequently established virus-specific immune responses, leading to unrecoverable serious pneumonia. PMID- 18162363 TI - Retinoic acid inhibits the infectivity and growth of Chlamydia pneumoniae in epithelial and endothelial cells through different receptors. AB - Chlamydia pneumoniae is a human respiratory pathogen that has also been associated with cardiovascular disease. C. pneumoniae infection accelerates atherosclerotic plaque development in hyperlipidemic animals and promotes oxidation of low density lipoprotein in vitro. All-trans-retinoic acid (ATRA), an antioxidant, has been shown to inhibit C. pneumoniae infectivity for endothelial cells by preventing binding of the organism to the M6P/IGF2 receptor on the cell surface. This current study investigates whether ATRA similarly affects C. pneumoniae infectivity of epithelial cells, which are the primary site of infection in the respiratory tract, and the effects on intracellular growth in both endothelial and epithelial cells. Because ATRA binds to both the nuclear retinoid acid receptor (RAR) and the M6P/IGF2 receptor, 4-[(E)-2-(5,6,7,8 tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1-propenyl]benzoic acid (TTNPB), an ATRA analog, which binds to the RAR but not the M6P/IGF2 receptor was used to differentiate the receptor mediating the effects of ATRA. The results of this study showed two separate effects of ATRA. The first effect is through interaction with the M6P/IGF2 receptor on the cell surface preventing attachment of the organism (inhibition by ATRA but not TTNPB) in endothelial cells and the second is through the nuclear receptor (inhibition by both ATRA and TTNPB) which inhibits growth in both epithelial and endothelial cells. PMID- 18162364 TI - The posttraumatic cognitions inventory-Chinese revised: validation and refinement with a traumatized college sample in Taiwan. AB - The posttraumatic cognitions inventory (PTCI) is a 33-item self-report measure assessing dysfunctional beliefs following trauma. This study reported the factor structure and psychometric properties of the Chinese version of the PTCI (PTCI C), as well as its relationship with PTSD symptoms. Study participants consisted of 240 traumatized college students in Taiwan. Confirmatory factor analyses indicated adequate replication of the original three-factor structure of the PTCI after eliminating four cross-loaded items. The 29-item PTCI-C was thus developed and its psychometric data were evaluated. This measure displays good internal consistency, test-retest stability, concurrent validity, and discriminative validity. The study findings are discussed in terms of intercultural differences in attribution style and the possible contribution of negative cognitions to PTSD development. PMID- 18162365 TI - Trait anxiety, disgust sensitivity, and the hierarchic structure of fears. AB - This paper describes an evaluation of Taylor's (1998) hierarchic model of fears and its relationship to trait anxiety and disgust sensitivity (DS). In Study 1 (N=420), a confirmatory factor analysis supported a hierarchic structure of fears. Next, an analysis using structural equation modeling indicated that trait anxiety is associated with claustrophobic and social fears, whereas DS is associated with all four fear subtypes examined (claustrophobic, social, blood injection-injury and animal). However, trait anxiety and DS did not account for all variance shared by fear subtypes. The addition of a generalized "fear factor" accounted for significant residual shared variance between the four fear subtypes, beyond that accounted for by trait anxiety and DS. Study 2 (N=213) generally replicated these results. Findings suggest that the hierarchic structural model of fears would benefit from inclusion of trait anxiety and DS as higher-order contributors to fearfulness. PMID- 18162367 TI - Pre-filtering improves reliability of Affymetrix GeneChips results when used to analyze gene expression in complex tissues. AB - Affymetrix GeneChip represents a very reliable and standardized technology for genome-wide gene expression screening. However, in experiments carried out on complex biological samples (e.g. brain tissues composed of several diverse cell types), significant noise can arise due to important transcripts being expressed in a relatively small number of cells. This noise results in many observations coming from unreliable hybridization reactions. Here we propose a method for pre filtering Affymetrix data according to measures of hybridization reliability. We used our pre-filtering method on a microarray dataset obtained from the brains of rats chronically treated with a psychostimulant drug. Our pre-filter protocol facilitates selection of biologically relevant candidate genes, which could be validated by real-time PCR with a rate of 98%. PMID- 18162366 TI - Treatment of experimental autoimmune encephalomyelitis with alpha lipoic acid and associative conditioning. AB - We have initiated studies to evaluate the suitability of performing therapeutic conditioning trials in experimental autoimmune encephalomyelitis (EAE) mice treated with alpha lipoic acid (ALA). EAE was induced in SJL mice by active immunization with myelin antigen. Once daily subcutaneous injection of ALA served as the unconditional stimulus (US) administered with the conditional stimulus (CS) saccharin-flavored drinking water under a regimen of restricted water access. In the first study, we found that water restriction and saccharin administration were compatible with disease development and effective ALA treatment of EAE mice. In the second study, mice were conditioned to once daily administration of ALA paired with administration of saccharin-flavored water (US+CS) on days 7-16. Test trials spanned experimental days 17-32 in groups receiving either saccharin-flavored water (CS, in the experimental group) versus unflavored water (CSo, in the control group) and compared several measures of EAE severity using multivariate ANOVA (MANOVA). Reduced disease severity in the experimental group (US+CS:CS) compared to the control group (US+CS:CSo) suggested that conditioning had occurred. These results demonstrate an approach for conducting therapeutic conditioning trials in EAE mice and suggest considerations for future investigations. PMID- 18162368 TI - The physical status of children with autism in China. AB - The height, weight and BMI of children with autism was investigated and analyzed to find the physical status of children with autism in China. Three hundred and eighty boys and 49 girls diagnosed with autistic disorder participated. Their parents were interviewed with a questionnaire about general information, and children were evaluated with Childhood Autism Rating Scale, and height and weight were measured. Children with autism had high level height, weight and BMI; the rate of height >or=P(75) was less in 6-11 years old group than that in 2-5 years old group in boys and all children. The prevalence of at-risk-for or being overweight was 31.8% and 17.0% in 2-5 years old group, were 37.9% and 21.8% in 6 11 years old group. At-risk-for-overweight/overweight of children with autism had no relationship with their core symptoms, the older age was the only predictor for lower height and at-risk-for-overweight. Prevalence of at-risk-for-overweight and overweight in children with autism was high. Children's height level decreased, and being at-risk-for-overweight increased with age. PMID- 18162369 TI - Pathogenesis of PR3-ANCA associated vasculitis. AB - Wegener's Granulomatosis (WG) is closely associated with antineutrophil cytoplasmic autoantibodies (ANCA), particularly those directed to proteinase 3 (PR3). ANCA directed to myeloperoxidase (MPO) are associated with microscopic polyangiitis (MPA) and the Churg Strauss syndrome. PR3-ANCA associated vasculitis differs from MPO-ANCA associated vasculitis particularly in the occurrence of granulomatous inflammation and more widespread disease in the former condition. Studies in experimental animals strongly suggest a pathogenic role for MPO-ANCA, but this is less clear for PR3-ANCA. Here, we review pathogenic pathways in PR3 ANCA associated vasculitis focussing on unique patterns of PR3 expression on the neutrophil membrane in WG, as surface expression of PR3 is a prerequisite for stimulation by PR3-ANCA; (PR3-specific) cellular immunity in WG; and the role of Staphylococcus aureus carriage in this disease. These factors, together, may explain, at least in part, differences in clinical expression between MPO-ANCA and PR3-ANCA associated vasculitis. PMID- 18162370 TI - RDP58 inhibits T cell-mediated bladder inflammation in an autoimmune cystitis model. AB - Interstitial cystitis (IC) is a chronic inflammatory condition of the urinary bladder with a strong autoimmune component. Currently, the major challenge in IC treatment is the development of effective therapies. RDP58 is a novel d-amino acid decapeptide with potent immunosuppressive activity. In this study, we investigated whether RDP58 was effective as an intravesical agent for treating bladder autoimmune inflammation in a transgenic mouse model (URO-OVA mice). URO OVA mice were adoptively transferred with syngeneic activated splenocytes of OT-I mice transgenic for the OVA-specific CD8(+) TCR for cystitis induction and treated intravesically with RDP58 at days 0 and 3. Compared with controls, the RDP58-treated bladders showed markedly reduced histopathology and expressions of mRNAs and proteins of TNF-alpha, NGF and substance P. To determine whether the inhibition of bladder inflammation by RDP58 was due to the interference with effector T cells, we treated the cells with RDP58 in vitro. Cells treated with RDP58 showed reduced production of TNF-alpha and IFN-gamma as well as apoptotic death. Collectively, these results indicate that RDP58 is effective for treating T cell-mediated experimental autoimmune cystitis and may serve as a useful intravesical agent for the treatment of autoimmune-associated bladder inflammation such as IC. PMID- 18162371 TI - The "Annales de l'Institut Pasteur", 1887-2007: a glimpse into history. PMID- 18162372 TI - The International Union of Microbiological Societies, IUMS. PMID- 18162373 TI - Molecular and cellular bases of bacterial virulence: guessing the future episodes of an ongoing saga. PMID- 18162374 TI - Early and advanced molecular biology of amino acid biosynthetic pathways and regulation. PMID- 18162375 TI - Safety of a weekly high dose of liposomal amphotericin B for prophylaxis of invasive fungal infection in immunocompromised patients: PROPHYSOME Study. AB - With its broad spectrum of activity and better tolerability profile than conventional amphotericin B, liposomal amphotericin B (L-AmB) may be the drug of choice for antifungal prophylaxis in haematological patients. An open-label, multicentre, prospective, pilot study was conducted in adult patients receiving chemotherapy for acute leukaemia (AL) or myeloablative allogeneic stem cell transplantation (SCT). Patients received weekly 10mg/kg infusions of L-AmB for 4 weeks for AL and 8 weeks for SCT. The primary objective was safety, with particular attention to infusion-related reactions and nephrotoxicity. Twenty nine adult patients were included: 21 AL (median age 52 years) and 8 SCT (median age 37 years). The most frequent adverse events (AEs) related to study drug were infusion-related reactions, 12 of which (from a total of 76 infusions) led to increased infusion duration for better tolerance. No AE related to the study drug led to discontinuation of prophylactic treatment in AL patients. In SCT patients, eight AEs (in six patients) reported to be related to study treatment led to treatment discontinuation. Enrolment was discontinued in the SCT group as recommended by the independent data review committee in accordance with the 10% limit of AEs (CTC grade 3-4) fixed by the protocol. The appropriate timing of high-dose prophylactic L-AmB remains to be determined in the SCT setting to optimise the safety profile of this regimen. For AL, a 10mg/kg weekly dose appears to be well tolerated during chemotherapy and may represent an important tool towards improving AL patient outcome. PMID- 18162376 TI - Evaluation of acute and chronic bacterial prostatitis and diagnostic management of chronic prostatitis/chronic pelvic pain syndrome with special reference to infection/inflammation. AB - The evaluation of acute and chronic bacterial prostatitis and the diagnostic management of chronic prostatitis/chronic pelvic pain syndrome with special reference to infection and inflammation is well defined. Men with symptoms of acute bacterial prostatitis (NIH I) have to undergo urine analysis and culture of the urine. An initial imaging of the prostate is suggested to exclude prostatic abscess. In chronic bacterial prostatitis (NIH II) the 4- and/or the 2-glass test for white blood cell counts and culture are necessary. Culture of semen has a lower sensitivity but is recommended for evaluation in selected men with problems with infertility. Imaging of the prostate is indicated to exclude abscess formations. In patients with chronic prostatitis/chronic pelvic pain syndrome (NIH III) symptom scoring is mandatory using the NIH-CPSI. The 4- and/or the 2 glass test are suggested to rule out bacterial infections. The routinely done analysis of urine and expressed prostatic secretions for leukocytes is debatable, especially due to the fact that the differentiation between patients with inflammatory and non-inflammatory subgroups of CP/CPPS may not be useful for the daily praxis. Optional investigations include the analysis for leukocytes in the ejaculate. Histopathological and molecular microbiological evaluation of prostatic tissue are investigational tests requiring for evaluation. Routine done serum PSA, routine imaging of the prostate and tests for Chlamydia trachomatis and Ureaplasma are not really proven to provide benefit for the patient. In patients with asymptomatic prostatitis (NIH IV) is no evaluation necessary unless considering antimicrobial therapy for elevated PSA or infertility. PMID- 18162377 TI - Antibiotic resistance of commensal Escherichia coli of food-producing animals from three Vojvodinian farms, Serbia. AB - Commensal bacteria of food-producing animals are considered an important reservoir of antibiotic resistance. The aim of this study was to determine the current prevalence of resistance to 18 different antibiotics in animal commensal Escherichia coli isolated from food-producing animals from three different farms with specific modes of antimicrobial use. A very high prevalence of resistance was found to tetracycline, a moderate level to streptomycin, ampicillin, cefalothin and nalidixic acid and a low of resistance to the other tested antibiotics. Resistance to two or more antibiotics was observed among all swine E. coli, 63.2% of broiler isolates and 37.5% of cattle isolates. The results show that commensals of food-producing animals from Vojvodina region are important reservoirs of resistance to older-generation antibiotics. PMID- 18162378 TI - Epidemiology and clinical outcomes of community-acquired pneumonia in adult patients in Asian countries: a prospective study by the Asian network for surveillance of resistant pathogens. AB - Appropriate antimicrobial treatment of community-acquired pneumonia (CAP) should be based on the distribution of aetiological pathogens, antimicrobial resistance of major pathogens, clinical characteristics and outcomes. We performed a prospective observational study of 955 cases of adult CAP in 14 hospitals in eight Asian countries. Microbiological evaluation to determine etiological pathogens as well as clinical evaluation was performed. Bronchopulmonary disease (29.9%) was the most frequent underlying disease, followed by cardiovascular diseases (19.9%), malignancy (11.7%) and neurological disorder (8.2%). Streptococcus pneumoniae (29.2%) was the most common isolate, followed by Klebsiella pneumoniae (15.4%) and Haemophilus influenzae (15.1%). Serological tests were positive for Mycoplasma pneumoniae (11.0%) and Chlamydia pneumoniae (13.4%). Only 1.1% was positive for Legionella pneumophila by urinary antigen test. Of the pneumococcal isolates, 56.1% were resistant to erythromycin and 52.6% were not susceptible to penicillin. Seventeen percent of CAP had mixed infection, especially S. pneumoniae with C. pneumoniae. The overall mortality rate was 7.3%, and nursing home residence, mechanical ventilation, malignancy, cardiovascular diseases, respiratory rate>30/min and hyponatraemia were significant independent risk factors for mortality by multivariate analysis (P<0.05). The current data provide relevant information about pathogen distribution and antimicrobial resistance of major pathogens of CAP as well as clinical outcomes of illness in Asian countries. PMID- 18162379 TI - pH dependent effect of glycosylation on protein stability. AB - The effect of glycosylation state on the thermal and storage stability of interleukin-2 mutein (IL-2 mutein) was investigated. The thermal stability of IL 2 mutein was studied by DSC and UV. An accelerated storage stability study was conducted at 40 degrees C in the dark and analyzed by UV, SDS-PAGE, and RP-HPLC. The unfolding temperatures (Tu) of both glycosylated and unglycosylated forms of IL-2 mutein are similar (within +/-1 degrees C) at pH 5.5 and 7.5. At pH 4.0, the Tu of glycosylated IL-2 mutein was 4 degrees C lower than that of the unglycosylated form. The precipitation temperature of glycosylated IL-2 mutein is similar to that of the unglycosylated form at pH 5.5 but 4 degrees C higher at pH 7.5. The precipitation temperature is not detectable for both forms at pH 4.0. During storage, both glycosylated and unglycosylated IL-2 mutein form aggregates (soluble and insoluble) and other degradation products. The aggregates are formed by both physical and chemical mechanisms. The major pathway of chemical aggregation appears to be disulfide bond formation/exchange. The glycosylated form is much less stable than the unglycosylated form at pH 4.0 and both forms are most stable at pH 5.5 in terms of thermal stability, precipitation rate and total degradation rate. This study clearly demonstrates that the effect of glycosylation on the stability of a protein is pH-dependent. PMID- 18162380 TI - [Hurler syndrome. Early diagnosis and successful enzyme replacement therapy: a new therapeutic approach. Case report]. AB - Mucopolysaccharidosis type I (MPS I) is a lysosomal storage disorder due to alpha L-iduronidase deficiency. Its severe prognosis has been significantly improved by enzyme replacement therapy using recombinant human alpha-L-iduronidase (laronidase). We report the case of a boy who was diagnosed at 19 months of age with Hurler's disease, the most severe form of MPS I, and received thereafter a treatment by laronidase, resulting in clinical and biological improvement. The aim of this case report is to draw physicians' attention on the presenting signs of Hurler's disease, in order to enable an earlier diagnosis, increasing the treatment's benefits. PMID- 18162381 TI - [Hypercalcemia revealing iatrogenic hypervitaminosis A in a child with autistic troubles]. AB - Hypervitaminosis A is an unusual cause of infant hypercalcemia. The way it occurs can be very surprising, as one can notice from the following case report. CASE REPORTS: A three-year-old boy, presenting important behavioral disorders, was hospitalized because of a deterioration of his general state of health associated with vomiting, cephalgias, fever and cutaneous abnormalities. A 168 mg/L hypercalcemia was found. The only etiology is a deviant consumption of vitamin A within the framework of an "autistic diet": 100000 UI/d during three months, and then 150000 UI/d the three following months. Intoxication was confirmed by the increased vitamin A plasmatic level, and vitamin A/RBP molar ratio and by the presence of plasmatic retinyl palmitate. An emergency treatment by rehydration, biphosphonates and furosemide led to effective calcemia normalization. CONCLUSION: In the case of nonobvious causes of hypercalcemia, a thorough cross examination must look for vitamin A intoxication. Our observation illustrates the danger of certain diets suggested for autistic children. PMID- 18162382 TI - [Development of widely available reference frames of pediatrics in maternity in the Languedoc-Roussillon region]. AB - In France the organization of the care in perinatality is governed by the decrees of October 1998 and was specified by the "plan perinatalite" 2005-2007. This organization allowed an operation in network, declined in a variable way according to areas. In this context, the group of the paediatricians in Languedoc Roussillon took concrete action of harmonization of the reference frames of care of paediatrics in maternity. METHODS: The methodology installation since 2001 lies on the active participation of many paediatricians of the Languedoc Roussillon region, pledge of an appropriation of these reference frames by the professionals. It relies upon a rigorous process of drafting and validation, and on the use of the data processing tool for the drafting and the diffusion via Internet of these reference frames. RESULTS: In June 2006, 60 reference frames were placed at the disposal of the professionals who intervened with the children in maternity (paediatricians, midwifes, nursery nurses). The paediatricians taking part in the drafting of these regional reference frames accounted for 43% of the paediatricians exerting in maternity of Languedoc-Roussillon. The majority (77%) of the establishments of birth of the area were represented, whatever the level (1, 2 or 3). CONCLUSION: We showed that paediatricians could organize themselves to write a great number of reference frames contributing to the improvement of the quality of the care, and harmonization of the practices within the perinatal network. PMID- 18162383 TI - [Utility of interferon gamma assays for diagnosis of tuberculosis in children]. AB - In vitro assays that measure the interferon gamma production by T cells incubated with specific antigen of Mycobacterium tuberculosis may be useful in the diagnosis of tuberculosis in children. Compared to Mantoux test, these tests are easier to perform and have a greater specificity, especially because they do not cross-react with BCG vaccine and with most of environmental Mycobacteria. However, their sensitivity is not well evaluated in children. To date in pediatrics, they can be associated with Mantoux test for the diagnosis of active tuberculosis. In the diagnosis of latent tuberculosis infection, larger studies with commercial kits are required. PMID- 18162384 TI - [HHV-6 infection and acute lymphoblastic leukemia in a child]. AB - We report the case of a child who was infected by HHV-6 and who started an acute lymphoblastic leukemia two months later. This case reminds that an etiologic role have been suggested for many viral infections in some leukemias in childhood, particularly the human herpesvirus 6 (HHV-6). PMID- 18162385 TI - [Congenital transient leukemia: a case report]. AB - Neonates with Down's syndrome have an increased risk for congenital leukaemia, particularly acute megakaryoblastic leukaemia (FAB, M7) which most often resolves spontaneously and is called transient leukaemia. It can be observed in non constitutional trisomy 21 infants then presenting trisomy 21 on blasts cells. OBSERVATION: We report a transient leukaemia with an isolated pericardial effusion in a phenotypically normal neonate. Trisomy 21 was found on blasts cells. Complete remission remains after 32 months. DISCUSSION: Congenital leukaemias, with trisomy 21 on blasts cells have a good prognosis that justifies observation before using chemotherapy. PMID- 18162386 TI - Effect of seabuckthorn leaf extracts on circulating energy fuels, lipid peroxidation and antioxidant parameters in rats during exposure to cold, hypoxia and restraint (C-H-R) stress and post stress recovery. AB - The present study was carried out to study mechanism of adaptogenic activity of seabuckthorn leaf extract, administered orally in rats both in single and five doses at a dose of 100mg/kg body weight 30min prior to C-H-R exposure. The efficacy of the extract was studied on circulating energy fuels, lipid peroxidation and anti-oxidant parameters in rats on attaining the T(rec) 23 degrees C during C-H-R exposure and after recovery (T(rec) 37 degrees C) from C-H R induced hypothermia. Single dose treatment in rats restricted rise in blood malondialdehyde (MDA) levels and decrease in glutathione (GSH) and catalase (CAT) levels. Both single and five doses also restricted the rise in serum free fatty acids (FFA) and lactate dehydrogenase (LDH) levels on attaining T(rec) 23 degrees C during C-H-R exposure, suggesting more efficient utilization of FFA for energy production and better maintained cell membrane permeability. This suggested that the adaptogenic activity of the extract might be due to its anti-oxidative activity, maintained blood glucose levels, better utilization of FFA and improved cell membrane permeability. PMID- 18162387 TI - Positive antibacterial co-action between hop (Humulus lupulus) constituents and selected antibiotics. AB - The research reported here deals with co-action of the hop (Humulus lupulus) derived anti-bacterial compounds, lupulone and xanthohumol, with several antibiotics. Among the antibiotics investigated for their co-action, polymyxin B sulfate, tobramycin and ciprofloxacin had a positive co-action in inhibiting selected test bacteria. The disc/well-diffusion assay and the minimum inhibitory concentration test (MIC) were employed to determine co-action. Both Gram-positive and Gram-negative bacteria were used in the evaluation. There was some co-action against all Gram-positive bacteria tested. Surprisingly, there was some positive co-action even against certain Gram-negative bacteria but not against others. Particularly, there was no co-action against E.coli. An antibacterial cream with lupulone, neomycin and polymyxin B sulfate was prepared and showed co-action. Ideas for other practical applications of this effect are put forth. The mechanism of the synergistic effect is briefly discussed but no attempt was made to prove it experimentally. PMID- 18162388 TI - The Medical Research Council chronic dyspnea score predicts the survival of patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: The Medical Research Council (MRC) chronic dyspnea scale (6-point) is used in different clinical conditions to grade breathlessness on daily activities. We have previously shown that in patients with histologically documented usual interstitial pneumonia/idiopathic pulmonary fibrosis (UIP/IPF), the MRC dyspnea scale is useful in estimating disease severity. The aim of this study was to further investigate the usefulness of the MRC scale in IPF as a marker of survival. METHODS: The records of 25 patients with histologically documented UIP/IPF were retrospectively reviewed. Clinical parameters, pulmonary function tests, and arterial blood gases at the time of diagnosis, as well as survival time were retrieved and recorded for each patient. The impact of the different variables determined at diagnosis on survival was examined using the Kaplan-Meier and uni- and multi-variate Cox-regression analyses. RESULTS: Among the baseline clinical and physiologic parameters determined at the time of IPF diagnosis, the MRC score, the Tiffeneau index, and the total lung capacity were the only significant and independent predictors of survival. In specific, a high MRC score, a high Tiffeneau index, and a low total lung capacity at presentation were associated with shorter survival. CONCLUSION: In accordance with the previous work, our results indicate that the Tiffeneau index and total lung capacity (TLC) are the important determinants of survival in patients with IPF. In addition, we show that the simple MRC chronic dyspnea score estimated at the time of diagnosis is equally predictive of survival and may aid clinicians in assessing the prognosis of new cases of IPF. PMID- 18162389 TI - Increasing the appeal and utilization of services for alcohol and drug problems: what consumers and their social networks prefer. AB - BACKGROUND: A large gap exists in the United States between population need and the utilization of treatment services for substance-related problems. Surveying consumer preferences may provide valuable information for developing more attractive services with greater reach and impact on population health. METHODS: A state-level telephone survey using random digit dialling sampling methods assessed preferences for available professional, mutual help, and lay resources, as well as innovative computerized and self-help resources that enhance anonymity (N=439 households in Alabama). RESULTS: Respondents preferred help that involved personal contact compared to computerized help or self-help, but were indifferent whether personalized help was dispensed by professional or lay providers. Attractive service features included lower cost, insurance coverage, confidentiality, rapid and convenient appointments, and addressing functional problems and risks of substance misuse. Respondents in households with a member who misused substances rated services more negatively, especially if services had been used. CONCLUSION: The findings highlight the utility of viewing substance misusers and their social networks as consumers, and the implications for improving the system of care and for designing and marketing services that are responsive to user preferences are discussed. PMID- 18162390 TI - Vacuum pyrolysis of waste tires with basic additives. AB - Granules of waste tires were pyrolyzed under vacuum (3.5-10 kPa) conditions, and the effects of temperature and basic additives (Na2CO3, NaOH) on the properties of pyrolysis were thoroughly investigated. It was obvious that with or without basic additives, pyrolysis oil yield increased gradually to a maximum and subsequently decreased with a temperature increase from 450 degrees C to 600 degrees C, irrespective of the addition of basic additives to the reactor. The addition of NaOH facilitated pyrolysis dramatically, as a maximal pyrolysis oil yield of about 48 wt% was achieved at 550 degrees C without the addition of basic additives, while a maximal pyrolysis oil yield of about 50 wt% was achieved at 480 degrees C by adding 3 wt% (w/w, powder/waste tire granules) of NaOH powder. The composition analysis of pyrolytic naphtha (i.b.p. (initial boiling point) approximately 205 degrees C) distilled from pyrolysis oil showed that more dl limonene was obtained with basic additives and the maximal content of dl-limonene in pyrolysis oil was 12.39 wt%, which is a valuable and widely-used fine chemical. However, no improvement in pyrolysis was observed with Na2CO3 addition. Pyrolysis gas was mainly composed of H2, CO, CH4, CO2, C2H4 and C2H6. Pyrolytic char had a surface area comparable to commercial carbon black, but its proportion of ash (above 11.5 wt%) was much higher. PMID- 18162391 TI - Effect of granular porous media on the composting of swine manure. AB - This study investigated the feasibility of a bulking agent of granular porous media (GPM) for the composting of swine manure. Two lab-scale composting reactors were operated to evaluate the general performances and maturity parameters using GPM made of wastes from the Portland cement manufacturing processes as an alternative bulking agent. The overall volatile solid (VS) removal was 38.5% (dry basis). During the experiments, moisture content ranged between 41% and 53%, ensuring feasibility of microbial activity in composting. Cured compost showed proper maturity and low phytotoxicity, despite the slight decreases of CO2 production and VS removal at the second batch operation. Various physico-chemical parameters of the cured compost met the regulatory standards reported elsewhere. The pH, carbon-to-nitrogen ratio, ammonia nitrogen and soluble organic carbon (SOC) of the cured compost were significantly correlated to the germination index (GI) using the seeds of Chinese cabbage and lettuce, indicating the progressive biodegradation of phytotoxins as well as organic matter. Consequently, the results obtained in this study demonstrate that GPM could contribute to the environmentally friendly and economical composting of problematic swine manure as a recyclable bulking agent. PMID- 18162392 TI - Integrated microfluidic systems with an immunosensor modified with carbon nanotubes for detection of prostate specific antigen (PSA) in human serum samples. AB - This paper describes the development of an immunosensor coupled to glassy carbon electrode (GCE) modified with multiwall carbon nanotubes (MWCNT) (CNT-GCE) integrated with microfluidic systems for rapid and sensitive quantification of prostate specific antigen (PSA) in human serum samples. Mouse monoclonal (5G6) to PSA antibodies were immobilized on a rotating disk. PSA in the serum sample are allowed to react immunologically with the immobilized anti-tPSA and horseradish peroxidase (HRP) enzyme-labeled second antibodies specific to PSA. HRP, in the presence of hydrogen peroxide (H(2)O(2)) catalyzes the oxidation of 4-tert butylcatechol (4-TBC), whose back electrochemical reduction was detected on CNT GCE at -0.15 V. The electrochemical detection can be done within 1 min and total assay time was 30 min. The calculated detection limits for electrochemical detection and the ELISA procedure are 0.08 and 0.5 microg L(-1), respectively and the intra- and inter-assay coefficients of variation were below 4.5%. The electrochemical immunosensor showed higher sensitivity and lower time consumed than the standard spectrophotometric detection ELISA method, which shows potential for detecting PSA in clinical diagnosis. PMID- 18162393 TI - CGH microarrays and cancer. AB - Genetic alterations are a key feature of cancer cells and typically target biological processes and pathways that contribute to cancer pathogenesis. Array based comparative genomic hybridization (aCGH) has provided a wealth of new information on copy number changes in cancer on a genome-wide level and aCGH data have also been utilized in cancer classification. More importantly, aCGH analyses have allowed highly accurate localization of specific genetic alterations that, for example, are associated with tumor progression, therapy response, or patient outcome. The genes involved in these aberrations are likely to contribute to cancer pathogenesis, and the high-resolution mapping by aCGH greatly facilitates the subsequent identification of these cancer-associated genes. PMID- 18162394 TI - The pathogenesis of cancer related fatigue: could increased activity of pro inflammatory cytokines be the common denominator? AB - Cancer related fatigue (CRF), defined as a persistent subjective sense of tiredness related to cancer or cancer treatment that interferes with daily functioning, is highly prevalent and probably the most underestimated and insufficiently treated complication in cancer patients. Therefore, unravelling the pathogenesis of CRF is of great importance allowing the identification of novel therapeutic targets. It is generally believed that the pathogenesis is multicausal, explaining why most therapeutic interventions directed towards only one potential causative factor are unsuccessful. In this regard, it is of interest that increased activity of pro-inflammatory cytokines might be a common denominator causing CRF. Detailed insight in the central role of increased activity of pro-inflammatory cytokines in CRF will hopefully offer an effective approach in the treatment of CRF by affecting a broad array of proposed causative factors such as anaemia, disturbances of the hypothalamic-pituitary-adrenal axis and altered brain serotonin metabolism. PMID- 18162395 TI - Ergoline derivatives as highly potent and selective antagonists at the somatostatin sst 1 receptor. AB - Non-peptidic compounds containing the octahydro-indolo[4,3-fg]quinoline (ergoline) structural element have been optimized into derivatives with high affinity (pK(d) r sst(1)>9) and selectivity (>1000-fold for h sst(1) over h sst(2)-h sst(5)) for the somatostatin sst(1) receptor. In functional assays, these ergolines act as antagonists at human recombinant sst(1) receptors. Pharmacokinetic studies in rodents reveal good oral bioavailability and brain penetration for some of these compounds. PMID- 18162396 TI - Carbonic anhydrase inhibitors. Interaction of 2-N,N-dimethylamino-1,3,4 thiadiazole-5-methanesulfonamide with 12 mammalian isoforms: kinetic and X-ray crystallographic studies. AB - 2-N,N-Dimethylamino-1,3,4-thiadiazole-5-methanesulfonamide was tested for its interaction with the 12 catalytically active mammalian carbonic anhydrase (CA, EC 4.2.1.1) isozymes, CA I-XIV. The compound is a potent inhibitor of CA IV, VII, IX, XII, and XIII (K(I)s of 0.61-39 nM), a medium potency inhibitor of CA II and VA (K(I)s of 121-438 nM), and a weak inhibitor against the other isoforms (CA III, VB, VI, and XIV), making it a very interesting candidate for situations in which a strong/selective inhibition of certain isozymes is needed. The crystal structure of the hCA II adduct of this sulfonamide revealed interesting interactions between the inhibitor and the enzyme which are quite different from those observed in the adducts of CA II with the structurally related aliphatic derivatives zonisamide, 2-amino-1,3,4-thiadiazolyl-5-difluoromethanesulfonamide, and 2-dimethylamino-5-[sulfonamido-(aminomethyl)]-1,3,4-thiadiazole reported earlier. PMID- 18162397 TI - On the mechanism of demethylation of 5-methylcytosine in DNA. AB - DNA methylation is an important biological process that programmes gene expression in vertebrates. The methylation pattern is generated by a combination of methylation and demethylation reactions catalyzed by DNA methyltransferases and putative demethylases. MBD2 binds methylated DNA and possesses DNA demethylase activity. We use here direct analysis of the reaction mixture by GC MS using a water-tolerant gas chromatographic column to avoid the loss of potential volatile products and identify the leaving residue of the demethylation reaction. We show that the DNA demethylase reaction catalyzed by a recombinant human MBD2 purified from SF9 insect cells releases dideuteroformaldehyde from [Me (2)H(3)]-5-methylcytosine in DNA. A mechanism of the DNA demethylation reaction is proposed based on this observation. PMID- 18162398 TI - Acylguanidine inhibitors of beta-secretase: optimization of the pyrrole ring substituents extending into the S1 and S3 substrate binding pockets. AB - Proteolytic cleavage of amyloid precursor protein by beta-secretase (BACE-1) and gamma-secretase leads to formation of beta-amyloid (A beta) a key component of amyloid plaques, which are considered the hallmark of Alzheimer's disease. Small molecule inhibitors of BACE-1 may reduce levels of A beta and thus have therapeutic potential for treating Alzheimer's disease. We recently reported the identification of a novel small molecule BACE-1 inhibitor N-[2-(2,5-diphenyl pyrrol-1-yl)-acetyl]guanidine (3.a.1). We report here the initial hit-to-lead optimization of this hit and the SAR around the aryl groups occupying the S(1) and S(2') pockets leading to submicromolar BACE-1 inhibitors. PMID- 18162399 TI - SAR-oriented discovery of peroxisome proliferator-activated receptor pan agonist with a 4-adamantylphenyl group as a hydrophobic tail. AB - 3-(4-Alkoxyphenyl)propanoic acid derivatives were prepared as candidate peroxisome proliferator-activated receptor (PPAR) alpha/delta/gamma pan agonists, based on our previous SAR studies directed toward the development of subtype selective PPAR agonists. Those studies indicated that the steric bulkiness of substituents introduced at the distal benzene ring had an important influence on PPAR activity. The finding that a 4-adamantyl derivative exhibited not only PPARalpha/delta activity but also significant PPARgamma activity prompted us to search for structurally novel phenylpropanoic acid derivatives with more potent adipocyte differentiation activity than the well-known PPARgamma agonist, rosiglitazone, as well as well-balanced PPARalpha and PPARdelta agonistic activities. A representative phenylpropanoic acid derivative (12) bearing a 4 adamantylphenyl substituent proved to be a well-balanced PPAR-pan agonist with activities to regulate the expression of genes involved in lipid and glucose homeostasis, and should be useful as a candidate drug for the treatment of altered PPAR function. PMID- 18162400 TI - Fragmentation of axillary fibrofatty tissue during dissection facilitates preservation of the intercostobrachial nerve and the lateral thoracic vein. AB - The preservation of neurovascular elements passing through the axillary fibrofatty tissue (the intercostobrachial nerve and the lateral thoracic vein) could be techniqually demanding if an en bloc axillary dissection is performed in the conventional way. In this paper we describe a surgical technique for more successful preservation of these elements, by which fragmentation of the axillary fibrofatty tissue is planned and performed "in advance". The techniques of axillary sampling biopsies, where lymphatic vessels are always divided, have shown that cutting of the lymph routes does not increase the risk of local regional recurrence. After adopting and applying this technique in a series of 22 consecutive dissections, the nerve was spared in 22 patients (100%) (compared to an earlier series where the nerve was spared in 53 of 65 patients (81.5%)), while the vein was spared in 21 patients (95%) (earlier 22 of 65 patients (34%)). PMID- 18162401 TI - End of life decisions: nurses perceptions, feelings and experiences. AB - Decisions to withdraw treatment are made on a regular basis in intensive care units. While nurses play a central role in patient care, previous studies have found that they are not always involved in withdrawal decisions. AIM: To explore the experiences of ICU nurses caring for patients who have had their treatment withdrawn and to answer two research questions: what role do nurses play and how does this affect them? METHOD: Constructivist grounded theory was used to explore the experiences and feelings of ICU nurses. A purposive sample of eight ICU nurses participated and semi-structured interviews were used to collect data. Framework analysis was used to facilitate systematic analysis. RESULTS: The analysis revealed two major themes (1) the nurse's role: experience counts, not really a nurse's decision, planting the seed, supporting the family and being a patient advocate and (2) perceptions of the withdrawal of treatment: getting the timing right and emotional labour. CONCLUSIONS: Nurses make an important contribution to end of life decisions and care. Guidelines recommend they have input into withdrawal decisions, therefore it is imperative that nurses are supported in this role and their responsibilities to continue to provide care during withdrawal. PMID- 18162402 TI - Characterizing head motion in three planes during combined visual and base of support disturbances in healthy and visually sensitive subjects. AB - Multiplanar environmental motion could generate head instability, particularly if the visual surround moves in planes orthogonal to a physical disturbance. We combined sagittal plane surface translations with visual field disturbances in 12 healthy (29-31 years) and 3 visually sensitive (27-57 years) adults. Center of pressure (COP), peak head angles, and RMS values of head motion were calculated and a three-dimensional model of joint motion was developed to examine gross head motion in three planes. We found that subjects standing quietly in front of a visual scene translating in the sagittal plane produced significantly greater (p<0.003) head motion in yaw than when on a translating platform. However, when the platform was translated in the dark or with a visual scene rotating in roll, head motion orthogonal to the plane of platform motion significantly increased (p<0.02). Visually sensitive subjects having no history of vestibular disorder produced large, delayed compensatory head motion. Orthogonal head motions were significantly greater in visually sensitive than in healthy subjects in the dark (p<0.05) and with a stationary scene (p<0.01). We concluded that motion of the visual field could modify compensatory response kinematics of a freely moving head in planes orthogonal to the direction of a physical perturbation. These results suggest that the mechanisms controlling head orientation in space are distinct from those that control trunk orientation in space. These behaviors would have been missed if only COP data were considered. Data suggest that rehabilitation training can be enhanced by combining visual and mechanical perturbation paradigms. PMID- 18162403 TI - Towards safer reoperations: special aspects in aortic dissection. AB - Surgical treatment of acute aortic dissection has been largely standardized, but some patients develop late complications that require repeat surgery. The 10-year freedom from proximal and distal reoperation is approximately 70-80%, and the risk of proximal reoperation is approximately 10%. Aggressive resection of the aortic root has not been proven to eliminate the need for proximal reoperation, but it appears reasonable for pre-existent root dilatation. The coronary button technique has resulted in greater freedom from death and reoperation and thus appears advisable. The risk of distal reoperations seems generally lower than that of proximal operations (0-4%). Routine extension of replacement into the total arch in the initial operation is associated with increased risk, and it appears not to decrease the probability of reoperation. Aggressive replacement of the total arch may be justified only for connective tissue disease. The choice of thoracic incision is important for aortic reoperations. A median sternotomy provides excellent exposure to the proximal aorta. A posterolateral thoracotomy allows good exposure for distal arch or descending aorta but may be associated with pain-related pulmonary impairment. A bilateral thoracotomy gives easy access for arch and descending aorta but is associated with the highest degree of respiratory impairment. We use a median sternotomy in most instances and choose a bilateral thoracotomy for complex aortic pathology. A posterolateral thoracotomy seems preferable for most distal arch and/or descending aortic replacements. Since femoral arterial cannulation can contribute to an increased incidence of neurological complication, cannulation of the axillary artery appears advisable. We prefer implantation of a Dacron graft and to cannulate the graft. If the aorta is adjacent to the sternum, the patient is placed on cardiopulmonary bypass (CPB) before repeat sternotomy. The patient is cooled to a nasopharyngeal temperature of 28-30 degrees C and at the time of sternotomy CPB is temporarily interrupted. We have not yet had to use transapical venting. Using these approaches we have been able to maintain a procedure-related mortality of 4%, which is not different from primary operations on an aneurysmatic aorta. PMID- 18162404 TI - Kinetic assisted venous drainage for orthotopic heart transplantation in patients under mechanical circulatory support: a double-edged sword. AB - BACKGROUND: Heart transplantation in patients supported with ventricular assist devices (VADs) entails a high risk of injury at resternotomy. Prior femorofemoral bypass is the preferred approach in these patients, but poor venous drainage may restrict arterial flow rate. PATIENTS AND METHODS: We compared bypass parameters, transfusion requirements and postoperative outcome in 33 consecutive patients (40.4+/-12.2 years old, 28 men) assisted with the Thoratec paracorporeal VAD (mean duration, 3.0+/-2.96 months) undergoing transplantation using either gravity siphon drainage (GSD, n=16) or kinetic assisted venous drainage (KAVD, n=17). RESULTS: Cannulation technique, perfusion pressure, temperature and duration were similar between groups. There were no significant differences in arterial re-infusion flow rates (GSD, 3.6+/-0.7 vs KAVD, 3.8+/-0.6l/min, p=0.5). KAVD patients had a lower mean S(v)O(2) and a higher desaturation index than GSD patients (69.5+/-4.6 vs 76.1+/-5.4mmHg, p=0.004; and 0.63+/-0.23 vs 0.25+/-0.63, p=0.0001, respectively). Perioperative requirements in fresh frozen plasma and platelet transfusions were significantly higher in KAVD patients. However, there were no differences in postoperative patient outcome. CONCLUSION: Perceived benefits on venous return associated with KAVD do not necessarily translate into improved arterial re-infusion flow rates and should be weighed against the hazards of increased venous air aspiration and blood product requirements. PMID- 18162405 TI - Long-term outcomes after entry closure and aneurysmal wall plication for type B aortic dissection. AB - OBJECTIVE: To examine the long-term outcomes after entry closure and aneurysmal wall plication for type B chronic dissecting aortic aneurysm. This procedure uses no artificial graft and preserves all intercostal arteries. METHODS: We reviewed the records of 40 consecutive patients who underwent this procedure between September 1983 and December 2002. The mean age at operation was 60+/-12 years (range, 38-79 years). The mean follow-up period was 9.8+/-5.1 years (range, 4-23 years). Follow-up was completed in 38 patients (95%). The latest computed tomography scans (n=22) were obtained 9.5+/-5.1 years (range, 3-18 years) after surgery. RESULTS: There were no operative deaths and 14 late deaths, none of which were related to the aneurysm. No paraplegia or paraparesis occurred. The survival rate was 92+/-4% at 5 years and 64+/-9% at 10 years; 24 patients are still alive. Follow-up computed tomography revealed that the mean diameter of the plicated descending aorta was 31+/-5mm (range, 22-39 mm) except in four patients. One of the four patients required reoperation for recurrent aneurysm of the plicated aorta 3 years postoperatively. In the remaining three patients, the plicated aorta has become enlarged; however, these patients have not yet undergone reoperation. Reoperation for residual dissecting aneurysm was performed in another three patients whose plicated aorta was normal. Freedom from reoperation for residual dissecting aneurysm was 78+/-5% at 10 years. CONCLUSIONS: This procedure produces excellent short-term outcomes and low long term morbidity. It could be the procedure of choice in selected patients to prevent paraplegia, although graft replacement is currently the standard treatment for chronic aortic dissecting aneurysm. PMID- 18162406 TI - The role of external polyamines on photosynthetic responses, lipid peroxidation, protein and chlorophyll a content under the UV-A (352nm) stress in Physcia semipinnata. AB - Physcia semipinnata was exposed to UV-A (352nm) and visible light (210, 800 and 2000mW/cm2) for 30min, 1, 2, 24, and 48h to seek the alterations in the PSII photosynthetic quantum yield, in response to radiation. Chlorophyll a fluorescence did not influence exposure to light, 210, 800 and 2000mW/cm2. Significant alterations of the photosynthetic quantum yield ratio occurred in response to increase in UV-A exposure time. The photosynthetic quantum yield ratio decreased in P. semipinnata following exposure to UV-A for 24 and 48h. The thalli of P. semipinnata treated with 1mM polyamine were not influenced during the exposure to UV-A for 24 and 48h. It was also found that exogenously spd added samples had higher chla content than spm and put added samples. In this study, we showed that lipid peroxidation levels between UV-A-treated samples and exogenously polyamine treated samples that were previously exposed to UV-A for 24 and 48h were significantly decreased. This result is the first record to indicate that external polyamines might have some protective role on photosystem II and membrane against UV-A stress. PMID- 18162408 TI - Molecular cloning and functional characterisation of a cathepsin L-like proteinase from the fish kinetoplastid parasite Trypanosoma carassii. AB - Trypanosoma carassii is a fish kinetoplastid parasite that belongs to the family Trypanosomatida. In the present study we cloned a cathepsin L-like proteinase from T. carassii. The nucleotide sequence of 1371bp translated into a preproprotein of 456 amino acids. The preproprotein contained the oxyanion hole (Gln), the active triad formed by Cys, His and Asn and the conserved ERFNIN-like, GNFD and GCNGG motifs, characteristic for cathepsin L proteinases. Phylogenetic analysis showed that the T. carassii cysteine proteinase clustered with other cathepsin L-like proteinases from the Trypanosomatida. We produced a recombinant T. carassii cysteine proteinase in Escherichia coli and demonstrated that it has cathepsin L activity. Immunization of common carp (Cyprinus carpio L.) with the recombinant protein induced a very high increase in proteinase-specific antibodies but only slightly lowered parasitaemia. Our findings suggest that the T. carassii cysteine proteinase is highly conserved within the Trypanosomatida with respect to structure and activity but is not a major protective antigen in carp. PMID- 18162407 TI - Foreign body reaction to biomaterials. AB - The foreign body reaction composed of macrophages and foreign body giant cells is the end-stage response of the inflammatory and wound healing responses following implantation of a medical device, prosthesis, or biomaterial. A brief, focused overview of events leading to the foreign body reaction is presented. The major focus of this review is on factors that modulate the interaction of macrophages and foreign body giant cells on synthetic surfaces where the chemical, physical, and morphological characteristics of the synthetic surface are considered to play a role in modulating cellular events. These events in the foreign body reaction include protein adsorption, monocyte/macrophage adhesion, macrophage fusion to form foreign body giant cells, consequences of the foreign body response on biomaterials, and cross-talk between macrophages/foreign body giant cells and inflammatory/wound healing cells. Biomaterial surface properties play an important role in modulating the foreign body reaction in the first two to four weeks following implantation of a medical device, even though the foreign body reaction at the tissue/material interface is present for the in vivo lifetime of the medical device. An understanding of the foreign body reaction is important as the foreign body reaction may impact the biocompatibility (safety) of the medical device, prosthesis, or implanted biomaterial and may significantly impact short- and long-term tissue responses with tissue-engineered constructs containing proteins, cells, and other biological components for use in tissue engineering and regenerative medicine. Our perspective has been on the inflammatory and wound healing response to implanted materials, devices, and tissue-engineered constructs. The incorporation of biological components of allogeneic or xenogeneic origin as well as stem cells into tissue-engineered or regenerative approaches opens up a myriad of other challenges. An in depth understanding of how the immune system interacts with these cells and how biomaterials or tissue engineered constructs influence these interactions may prove pivotal to the safety, biocompatibility, and function of the device or system under consideration. PMID- 18162410 TI - Residual subluxation of the elbow after dislocation or fracture-dislocation: treatment with active elbow exercises and avoidance of varus stress. AB - During a 6-year period, 2 surgeons prescribed active elbow exercises and avoidance of shoulder abduction (varus stress) for 23 patients (15 men, 8 women) with a mean age of 43 years (range, 19-74 years) who had slight residual subluxation of a dislocated elbow after either nonoperative (3 simple dislocations and 2 fracture-dislocations) or operative treatment (18 fracture dislocations). A stable elbow was achieved in all 23 patients, although 1 patient, treated late, had a very slight residual posterior subluxation of the radial head on radiographs. After a mean of 24 months (range, 12-45 months), the average flexion arc was 113 degrees , with mean flexion of 132 degrees and mean flexion contracture of 20 degrees . The mean Broberg-Morrey score was 90 points (range, 61-100 points). Ratings were excellent in 9 patients, good in 10, and fair in 4. Mild radiographic signs of arthrosis (type 1 Broberg-Morrey) were present in 2 patients. Slight postreduction subluxation of the elbow, without detrimental contact of the articular surfaces, a type of pseudosubluxation of the elbow sometimes referred to as the "drop sign," can be treated with active exercises and avoidance of varus stress. PMID- 18162411 TI - How should we use the Constant Score?--A commentary. PMID- 18162412 TI - Intermethod agreement and interobserver correlation of radiologic acromiohumeral distance measurements. AB - The acromiohumeral distance (AHD) measured on conventional, true anteroposterior radiographs taken with the patient standing and the arm held in neutral rotation can predict the presence or absence of a large, chronic rotator cuff tears, and narrowing of the acromiohumeral distance can predict irreparability of rotator cuff tears. The relation between AHD measured on conventional radiographs and the integrity of the rotator cuff tendons is well known. Conversely, the correlation between the AHD on conventional radiographs and that measured on magnetic resonance imaging (MRI) or computed tomography (CT) scans is unknown. It has also not been determined which method correlates better with fatty infiltration of the rotator cuff muscles and might thereby be a better prognostic value for the outcome of rotator cuff repair. Conventional radiographs, plus MRI and or CT studies taken at the same time of patients with rotator cuff tears, shoulder instability, primary glenohumeral osteoarthritis, or idiopathic frozen shoulder, were randomly selected, and the AHD was measured both on conventional radiographs and on MRI or CT. The values were correlated with the stage of fatty degeneration of the rotator cuff muscles in the cases of rotator cuff tears. The AHD on conventional radiographs was larger than on MRI and CT, and correlation of the values measured (conventional radiography vs MRI and CT) could be demonstrated. For rotator cuff diseases, a significant but moderate correlation (r = 0.6; P < .05) between conventional and MRI was identified, and the conventional values were approximately 0.6 mm greater than the corresponding MRI values. It is, therefore, possible to use the AHD read on MRI or CT studies for prognostic reasons, provided the values obtained are converted accordingly and remembering that the correlation of the values is only moderate. PMID- 18162409 TI - Molecular phylogenetics of the spider family Micropholcommatidae (Arachnida: Araneae) using nuclear rRNA genes (18S and 28S). AB - The spider family Micropholcommatidae is an enigmatic taxon of uncertain limits and uncertain affinities. Various phylogenetic hypotheses have been proposed for the family, but these hypotheses have never been tested with a robust phylogenetic analysis. The existence of similar Australasian and New World taxa, the possibility of morphological convergence associated with extreme 'smallness', and the apparent paucity of synapomorphic morphological characters, have all clouded generic relationships in this group. We used fragments from two nuclear ribosomal RNA genes (18S and 28S) to test the monophyly and phylogenetic position of the Micropholcommatidae. The analyses incorporated 50 ingroup spider species, including 23 micropholcommatid species and representatives from 14 other spider families. Ribosomal RNA secondary structures were inferred for the V3-V5 region of the 18S rRNA gene, and Domain II of the 28S rRNA gene of Hickmania troglodytes [Higgins, E.T., Petterd, W.F., 1883. Description of a new cave-inhabiting spider, together with notes on mammalian remains from a recently discovered cave in the Chudleigh district. Pap. Proc. R. Soc. Tasman. 1882, 191-192]. These secondary structures were used to guide multiple sequence alignments, and determine the position and nature of indels in different taxa. Secondary structure information was also incorporated into a structurally partitioned rRNA analysis in MrBayes Version 3.1.2, using a doublet model of nucleotide substitution. This structurally partitioned rRNA analysis provided a less resolved but more conservative and informative estimate of phylogeny than an otherwise identical, unpartitioned rDNA analysis. With the exception of the Chilean species Teutoniella cekalovici [Platnick, N.I., Forster, R.R., 1986. On Teutoniella, an American genus of the spider family Micropholcommatidae (Araneae, Palpimanoidea). Am. Mus. Novit. 2854, 1-9], the family Micropholcommatidae was found to be monophyletic with three monophyletic sub-lineages-congruent with the Micropholcommatinae, Textricellinae, and a group of 'taphiassine' species. Teutoniella cekalovici never grouped with the other micropholcommatid taxa, and could not be assigned to any family group with confidence. PMID- 18162413 TI - Determination of in vivo glenohumeral translation using fluoroscopy and shape matching techniques. AB - The purpose of this study was to investigate glenohumeral translation in-vivo during active shoulder abduction in the scapular plane. Three-dimensional (3D) models of 9 shoulders were created from CT scans. Fluoroscopic views aligned to the plane of the scapula were recorded during active arm abduction with neutral rotation. 3D motions were determined using model-based 3D-to-two-dimensional (2D) registration. Humeral translation was referenced to the glenoid center in the superior/inferior direction. The humerus moved an average of 1.7 mm superior with arm abduction, from an inferior location to the glenoid center. The humeral head was centered within 1 mm from the glenoid center above 80 degrees abduction. Variability in glenohumeral translation between shoulders decreased significantly from initial to final arm abduction. Our findings agree with some authors' observations of inferior-to-central translation of the humerus and behavior as a congruent ball and socket. We believe this information will help improve the understanding of shoulder function. PMID- 18162414 TI - Behavior of host and graft cells in the early remodeling process of rotator cuff defects in a transgenic animal model. AB - Autologous tissue graft is one of the treatment options for a large rotator cuff defect. To develop appropriate strategies for enhanced solid graft integration at the bone-tendon interface and tendon-tendon interface, clarifying the fate of the graft and host cells that contribute to repair and remodeling is necessary. We have developed a new grafting model using green fluorescent protein-transgenic rats and wild-type rats to simulate autologous transplantation for examining the behavior of the host and graft cells in the remodeling process after tendon grafting. We found that the host cells commenced proliferation in the graft at 1 day after grafting. The host cells infiltrated into the graft from the subacromial synovium, proximal tendon, and bone-tendon insertion. The number of graft-derived cells decreased with time. Our result clearly demonstrated that host cells, rather than graft cells, were essential for rotator cuff remodeling after tendon grafting for rotator cuff defect. PMID- 18162415 TI - Relative contributions of the infraspinatus and deltoid during external rotation in patients with symptomatic subacromial impingement. AB - A principal cause of subacromial impingement (SAI) is failure of the rotator cuff to center the humeral head in the glenoid during shoulder motion, counteracting the effect of the deltoid. As rehabilitation of the rotator cuff endeavors to restore balance between these muscle groups, the purpose of this companion study was to evaluate, in the symptomatic shoulders of patients with SAI, (1) the conditions of resisted isometric external rotation (ER) that optimized the contribution of the infraspinatus and (2) the load of ER at which adduction was most effective at reducing the deltoid contribution and then to compare this with the relative contribution of the infraspinatus and the posterior and middle deltoid in asymptomatic shoulders. In 14 subjects (18 shoulders) with SAI, surface electromyographic activity of the infraspinatus and the posterior and middle deltoid and pectoralis major was recorded at low, medium, and high loads of resisted isometric ER, with and without adduction. These data were normalized to find each muscle's relative contribution to the task and were compared with normalized data from subjects with healthy shoulders. In subjects with SAI, low loads of isometric ER (10%-40% maximum voluntary isometric contraction) optimized the relative contribution of the infraspinatus. Adduction with isometric ER at 10% maximum voluntary isometric contraction reduced the middle deltoid involvement. Higher loads preferentially activated the middle deltoid over the infraspinatus and may have effected unwanted humeral head superior translation, counteracting the presumed benefits of rotator cuff ER exercises. An individualized loading regimen and the use of surface electromyography may have significant implications during rotator cuff rehabilitation. PMID- 18162416 TI - Gender issues in epilepsy--the science of why it is special. PMID- 18162417 TI - Sexual problems in males with epilepsy--an interdisciplinary challenge! AB - Sexual function can be altered in patients with different types of epileptic disorder, especially those with temporal lobe epilepsy. The awareness of sexual function disturbances, giving an enormous impact on someone's quality of life, should lead to therapeutic measures. The incidence, evaluation and therapeutic options are demonstrated and seen through the urologist's eyes. PMID- 18162418 TI - Joint dependent concentrations of bone alkaline phosphatase in serum and synovial fluids of horses with osteochondral injury: an analytical and clinical validation. AB - OBJECTIVES: Validate use of a commercially available immunoassay for measurement of bone alkaline phosphatase (BAP) in equine serum and synovial fluid (SF), and investigate the effects of osteochondral (OC) injury in horses on BAP concentrations in serum and SF. METHODS: SF was collected from 37 joints of 34 Thoroughbred (TB) racehorses undergoing arthroscopic surgery for the removal of OC fragments from either the carpal joints (n=18) or the metacarpo /metatarsophalangeal (MP) joints (n=19). SF was also obtained from 52 joints of 16 normal TB horses, collected bilaterally from carpal joints of 10 horses (n=40), and MP joints of six horses (n=12). Blood was obtained from all 50 horses. A commercially available immunoassay was validated and subsequently used to determine equine serum and SF BAP concentrations. Correlations to radiographic and arthroscopic scores were assessed. RESULTS: BAP concentrations were significantly lower in serum from horses with OC injury in their carpal or MP joints than in serum from normal horses. SF BAP concentrations in normal and OC injured carpal joints were significantly higher than MP joints. BAP concentrations were significantly higher in SF from OC injured carpal joints than normal. BAP concentrations were affected by joint sampled, with age having a significant interaction. Concentrations of BAP in the serum (<30U/L), SF (>22U/L) and a ratio of SF to serum > or = 0.5 were predictive of OC injury. Radiographic and arthroscopic scores significantly correlated with serum BAP concentrations, and SF:serum BAP correlated with arthroscopic scores. CONCLUSIONS: Determination of serum and SF BAP concentrations may be beneficial in the investigation of early joint injury. Joint and injury dependent differences in BAP concentrations allowed the estimation of predictive value for identifying OC injury. PMID- 18162419 TI - Differential expression of two winter wheat alpha-tubulin genes during cold acclimation. AB - Overwintering crops, such as winter wheat, display significant increase in freezing tolerance during a period of cold acclimation (CA). To gain better understanding of molecular mechanisms of CA, it is important to unravel functions and regulations of CA-associated genes. Differential screening of a cDNA library constructed from cold-acclimated crown tissue of winter wheat identified an alpha tubulin cDNA clone named wca18g11 that showed elevated expression upon cold acclimation. Nucleotide sequence analysis showed that the clone encoded a group 3 alpha-tubulin. Reverse transcription real-time PCR analysis of the expression of both wca18g11 clone and its closest paralogs of the wheat tubulin A-2 homeologous group during the course of cold acclimation revealed that both genes were differentially regulated with distinct expression patterns. The involvement of the two alpha-tubulin genes in cold acclimation and signal transduction is discussed. PMID- 18162420 TI - Plant tubulin intronics. AB - Introns of plant tubulin genes are useful molecular tools to study IME (Intron Mediated Enhancement of gene expression) and to define plant genetic and evolutionary relationships through ILP (Intron Length Polymorphism). Here we show that the intron present within the 5'UTR sequence of some rice beta-tubulin genes can sustain IME in rice transgenic plants and that degenerated oligonucleotide mixtures designed to amplify the first and the second intron present within the coding sequence of plant beta-tubulin genes can successfully detect ILPs among different bean varieties. PMID- 18162421 TI - The substrate for long-lasting memory: if not protein synthesis, then what? AB - The prevailing textbook view that de novo protein synthesis is required for memory (e.g., [Bear, M. F., Connors, B., & Paradiso, M. 2006. Neuroscience. Lippincott, New York]) is seriously flawed and an alternative hypothesis has been proposed in which post-translational modification (PTM) of proteins already synthesized and already present within the synapse is 'the' substrate for long lasting memory. Protein synthesis serves a replenishment role. The first part of this review discusses how long-lasting memory can be achieved with 'only' PTM of existing synaptic proteins. The second part critically reviews a recent report published in Neuron 2007 that exemplifies the current view of protein synthesis and memory while also illustrating how these results can be understood within this new PTM framework. A necessary yet unexpected conclusion to emerge from consideration of the consequences of a PTM mechanism as the necessary, sufficient and exclusive substrate for long-lasting memory, is that the central Hebbian dogma that cells that 'fire together, wire together' is an unlikely mechanism for long-lasting memory. Thus, a unique feature of the PTM model is that longevity of information storage is achieved not by stability of the synaptic mechanism, but by impermanent pseudoredundant circuits. This is so because PTM is a reversible process and thus any permanent connection, any 'lasting effect' cannot be in the form of stable synapse formation. We have therefore proposed a solution in which network level processes regulate cellular mechanisms, even as such mechanisms regulate the network. Thus, synapses are 'meta-stabilized' by regulated feedback mediated by the circuit in which the synapse is embedded. For example, spontaneous activity is proposed to be a substrate feedback mechanism we term 'cryptic rehearsal' to sustain for some period of time after learning an approximation to the state initially created by input. Additionally, because the duplication of these traces is ongoing, this provides a degenerate code for the engram. Stability is thus achieved, not by stabilizing the synapse, but by implementing a pseudo-redundant yet malleable circuitry so that memory can be protected in the face of small catastrophes in network representation. PMID- 18162422 TI - What do different databases tell about the use of opioids in seven European countries in 2002? AB - OBJECTIVE: The objective of this paper was to analyse opioid consumption in a number European countries using different sources of data. METHODS: Data were extracted from the United Nations' International Narcotics Control Board Report (INCB) 2003 and from the registers of the national health authorities in seven countries where data were available for 2002. The amount of opioid used was calculated as daily defined doses per 1000 inhabitants per day (DDD/1000/day). Danish Register of Medicinal Products Statistics was further explored for characteristics of opioid consumption (age, gender, type of opioids consumed) by patients in primary care. Total opioid consumption and consumption of 11 selected opioids (7 strong and 4 weak) were analysed. The amount of opioids consumed by outpatients was also examined. RESULTS: There were considerable differences in the number of opioids reported and significant discrepancies in the amounts of opioids consumed between the national data and the INCB report. The source of data for the national registers on drug consumption varied (pharmacies or wholesale). The INCB data provide information on opioid import and estimated need rather than on medical consumption. CONCLUSIONS: Caution is required when interpreting the data on opioid consumption between countries because of differences in the collection and reporting of data. Better recording of opioid consumption is needed for meaningful analysis of opioid consumption and its possible effect on pain management in different countries. Data on opioids consumed for cancer-related pain in comparison with chronic non-malignant pain are needed. A uniform method of collection of data on analgesic consumption should be established for all European countries. PMID- 18162423 TI - Interactions between glutamate and capsaicin in inducing muscle pain and sensitization in humans. AB - The aim of the study was to investigate the interaction between glutamate and capsaicin in inducing muscle pain and sensitization in humans. Fifteen male volunteers participated. Glutamate or capsaicin or isotonic saline, in a paired sequence order, was injected randomly into the right or left masseter muscle. Two injections were given in a double-blinded design 25 min apart in 1 session/week over 4 weeks: saline (A1) followed by glutamate (A2), capsaicin (B1) followed by glutamate (B2), saline (C1) followed by capsaicin (C2), and glutamate (D1) followed by capsaicin (D2). The subjects drew the area of perceived pain and scored pain intensity on a 0-10 visual analogue scale (VAS). Pressure pain threshold (PPT) at the injection site, at a site 2-cm away, and on the contralateral side, as well as pressure pain tolerance (PPTol) at the injection site and contralateral site, were also measured before and after injection and subsequently at 5-min intervals. Paired t-test analyses showed that the pain drawing area was significantly smaller in the B2 compared to the A2 condition (P=0.028), and significantly larger in the D2 compared to the C2 condition (P=0.027). It also revealed significantly lower VAS peak pain intensity (P=0.008) and smaller VAS area under the curve (P=0.003) for the B2 compared to the A2 condition, and significantly higher VAS peak pain (P=0.015) and larger VAS area under the curve (P=0.037) for the D2 compared to the C2 condition. There was a significant PPT and PPTol decrease at the injection site after glutamate or capsaicin injection (ANOVA: P<0.028). The percentage decrease in PPT or PPTol (at the injection site) was not significantly different for the B2 compared to the A2 condition (Paired t-test: P>0.682) or for the D2 compared to the C2 condition (P>0.133). Significant PPT changes were also observed at the site 2 cm away, but not on the contralateral side. In conclusion, these findings indicate that intramuscular administrations of glutamate and capsaicin interact and influence pain and sensitization of muscle nociceptors: glutamate causes a sensitization to subsequent administration of capsaicin, whereas capsaicin is associated with a desensitization to subsequent injection of glutamate. These findings support previous animal data. PMID- 18162424 TI - Proximal magnetometry in thin films using betaNMR. AB - Low energy ion implantation of hyperpolarized radioactive magnetic resonance probes allows the NMR study of thin film heterostructures by enabling depth resolved measurements on a nanometer lengthscale. By stopping the probe ions in a layer adjacent to a layer of interest, it is possible to study magnetic fields proximally. Here we show that, in the simplest case of a uniformly magnetized layer, this yields an unperturbed in situ frequency reference. We also discuss demagnetization contributions to measured shifts for this case. With a simple illustrative calculation, we show how a nonuniformly magnetized layer causes a strongly depth-dependent line broadening in an adjacent layer. We then give some experimental examples of resonance line broadening in heterostructures. PMID- 18162425 TI - Monte Carlo simulation of DNMR spectra of coupled spin systems. AB - A new program MC-DNMR is presented for the simulation of dynamic nuclear magnetic resonance spectra. The algorithm is a Monte Carlo type method based on the extension of single spin vector model to coupled spin systems. This extension is explained in detail and the theory is justified by examples. The main advantage of this program is the significantly smaller sizes of matrices than that in programs based on density matrix theory. So spectra of systems can be simulated that was impossible previously. PMID- 18162426 TI - Enhanced citrate production through gene insertion in Aspergillus niger. AB - The effect of inserting genes involved in the reductive branch of the tricarboxylic acid (TCA) cycle on citrate production by Aspergillus niger was evaluated. Several different genes were inserted individually and in combination, i.e. malate dehydrogenase (mdh2) from Saccharomyces cerevisiae, two truncated, cytosolic targeted, fumarases (Fum1s and FumRs) from S. cerevisiae and Rhizopus oryzae, respectively, and the cytosolic soluble fumarate reductase (Frds1) from S. cerevisiae. Overexpression of these genes in their native strain backgrounds has been reported to lead to alterations in the intracellular cytosolic dicarboxylate concentrations. It was found that all the transformant strains had enhanced yield and productivities of citrate compared with the wild-type strain. The transformants also had the ability to produce citrate in trace-manganese contaminated medium, where the wild type was unable to produce. Overexpression of FumRs and Frds1 resulted in the best citrate-producing strain in the presence of trace manganese concentrations. This strain gave a maximum yield of 0.9g citrate per g glucose and a maximum specific productivity of 0.025g citrate per g DW per h. Overexpression of mdh2 alone resulted in an increased citrate production rate only in the initial phase of the fermentations compared with the other transformants and the wild type. PMID- 18162427 TI - Glucosylceramide accumulation is not confined to the lysosome in fibroblasts from patients with Gaucher disease. AB - Gaucher disease (GD) is an inborn error of glycosphingolipid metabolism resulting from a deficiency of the lysosomal enzyme beta-glucosidase leading to the accumulation of glucosylceramide (GC) in lysosomes of affected cells. In order to determine the effect of GC accumulation on intracellular lipid content in fibroblasts from patients with GD, we measured individual species of ceramide, di and trihexosylceramide, sphingomyelin, phosphatidylcholine, phosphatidylinositol and phosphatidylglycerol using electrospray ionisation-tandem mass spectrometry. The different subspecies of each lipid class correlated with each other and were summed to give total lipid concentrations. In addition to GC, we also noted secondary elevations in other lipids, especially in type 2 GD. Sub-cellular fractionation showed that GC was not confined to the lysosome but increased throughout the cell. The sequelae of extra-lysosomal accumulation may have implications in the pathogenic mechanisms of GD by interaction with biochemical and metabolic pathways located outside the lysosome. The elevation of ceramide in confluent type 2 GD fibroblasts redistributed from its primary site of accumulation in the lysosome to the endosomal region at four-weeks post confluence. The accumulation of lipids in the endosome and lysosome suggests both impaired trafficking of lipids and reduced capacity of the lysosome to degrade lipids. PMID- 18162428 TI - Measurement of radiotherapy superficial X-ray dose under eye shields with radiochromic film. AB - Accurate measurement and knowledge of dose delivered under eye shield during superficial X-ray radiotherapy is required for patient peripheral dose assessment. Critical structures can include the cornea, lens and retina. Measurement of dose under eye shields has been historically performed with Thermoluminescent Dosimeters (TLD's) due to their small size and design. Restrictions include the energy dependence and the fact that they only provide a point dose assessment. This note investigates the use of a low energy dependence radiochromic thin film for measurement of dose under eye shields in a phantom and compares results to theoretical calculation of dose. Results have shown a good match between predicted and experimentally measured results at the centre of an eye shield irradiated with 50kVp and 150kVp beams. The added advantage of radiochromic film compared to TLD measurements is the two dimensional dose map which is recorded for the assessment of dose providing not only an assessment at the site of the cornea, lens and retina in a phantom but in other areas as well. Radiochromic film has been found to accurately measure dose under eye shield in phantom treatments. PMID- 18162429 TI - Efficacy of linezolid in the treatment of mediastinitis due to methicillin resistant Staphylococcus aureus: an experimental study. AB - INTRODUCTION: The treatment of postoperative mediastinitis is very important because of its high morbidity, mortality, and increased hospital stay and hospital costs. The aims of our research were to investigate whether linezolid alone can be an effective treatment agent for methicillin-resistant Staphylococcus aureus (MRSA) mediastinitis, and to determine whether linezolid can provide synergistic activity when given in combination with rifampin. METHODS: A partial upper median sternotomy was performed on 70 rats. The animals were divided into seven groups: an uncontaminated control group; an untreated contaminated group; three contaminated groups that received antibiotic therapy with either 25 or 50 mg/kg linezolid twice a day, or rifampin 5 mg/kg twice a day; and two contaminated groups that received a combination therapy consisting of 25 or 50 mg/kg linezolid and rifampin 5 mg/kg twice a day. The antibiotic treatment lasted 7 days. Tissue samples from the upper ends of the sternum and swab specimens of the upper mediastinum were obtained and evaluated microbiologically. RESULTS: The 25-mg/kg dose of linezolid, either alone or combined with rifampin, was not effective in reducing the bacterial counts in mediastinum and sternum. Quantitative bacterial cultures of mediastinum and sternum were significantly lower in the groups receiving 50 mg/kg linezolid alone or in combination with rifampin compared with the control. Adding of rifampin to linezolid therapy did not result in a significant change in bacterial counts versus linezolid alone. CONCLUSION: A high dose of linezolid should be considered as a possible therapeutic agent for the treatment of post-sternotomy infection caused by MRSA. PMID- 18162430 TI - [Dupuytren's contracture. A report of three cases in Black Africans]. AB - Having reported the first case of Dupuytren's disease in West Africa in 1990, the authors report two further cases, which brings the number of Dupuytren's disease reported in West Africa to three. The authors demonstrate by a literature review, that numerous cases had in fact been described in black people since 1974. The factors which are incriminated are always alcoholism, microtrauma and other factors like diabetes. Dupuytren's disease appears to be a condition, which is actually not so rare in black people after all. PMID- 18162432 TI - Who begets whom? Plant cell fate determination by asymmetric cell division. AB - Asymmetric cell division generates cell types with different fates. Recent studies have improved our understanding of the molecular mechanisms involved in asymmetric cell division in Arabidopsis thaliana. Genetic approaches have identified candidate intrinsic factors and signaling components that mediate extrinsic cues. WOX genes appear to be putative intrinsic determinants acting in early embryonic asymmetric divisions. A non-canonical mechanism involving specific SHORT ROOT (SHR)-SCARECROW (SCR) nuclear complexes is implicated in ground tissue asymmetric divisions. Asymmetric stem cell division requires extrinsic organizer signaling, whereas the involvement of intrinsic stem cell segregants is unknown. Finally, new studies on stomatal development have identified several intrinsic acting factors that specify cell fate and an extrinsic signaling cascade that controls the number and plane of asymmetric divisions. PMID- 18162431 TI - Interactive sequences in the molecular chaperone, human alphaB crystallin modulate the fibrillation of amyloidogenic proteins. AB - Multiple interactive domains are involved in the activity of the stress protein, alphaB crystallin that protects against the unfolding, aggregation, and toxicity of amyloidogenic proteins. Six peptides corresponding to the interactive sequences 41STSLSPFYLRPPSFLRAP58, 73DRFSVNLDVKHFS85, 101HGKHEERQDE110, 113FISREFHR120, 131LTITSSLSSDGV142, and 156ERTIPITRE164 in human alphaB crystallin were synthesized and evaluated in Thioflavin T fluorescence assays for their effects on the modulation of fibrillation of four disease-related amyloidogenic proteins: amyloid-beta, alpha-synuclein, transthyretin, and beta2 microglobulin. The 73DRFSVNLDVKHFS85 and 101HGKHEERQDE110 peptides in the conserved alpha crystallin core domain of alphaB crystallin were the most effective fibril inhibitors. 73DRFSVNLDVKHFS85 completely inhibited alpha synuclein fibrillation and reduced the fibrillation of amyloid-beta, transthyretin, and beta2-microglobulin by >50%. 101HGKHEERQDE110 completely inhibited amyloid-beta fibrillation and reduced the fibrillation of alpha synuclein, transthyretin, and beta2-microglobulin by >50%. The peptides FSVN, NLDV, HGKH, and HEER, which are synthetic fragments of 73DRFSVNLDVKHFS85 and 101HGKHEERQDE110, inhibited fibrillation of all four amyloidogenic proteins by >75%. In contrast, the peptides FISREFHR, ERTIPITRE, DRFS, KHFS, and EERQ were the strongest promoters of fibrillation. Molecular modeling of the interactions between transthyretin and beta2-microglobulin and the synthetic bioactive peptides determined that residues Phe-75, Ser-76, Val-77, Asn-78, Leu-79, and Asp 80 in 73DRFSVNLDVKHFS85 and residues His-101, Lys-103, His-104, Glu-105, and Arg 107 in 101HGKHEERQDE110 interact with exposed residues in the beta strands, F and D of transthyretin and beta2-microglobulin, respectively, to modulate fibrillation. This is the first characterization of specific bioactive peptides synthesized on the basis of interactive domains in the small heat shock protein, alphaB crystallin that protect against the fibrillation of amyloidogenic proteins. PMID- 18162433 TI - Do oestrogens induce chromosome specific aneuploidy in vitro, similar to the pattern of aneuploidy seen in breast cancer? AB - The study was concerned with investigating the specific effects of non-DNA reactive oestrogens at low "biologically relevant" doses and the causative role they may play in breast cancer through inducing aneuploidy. A review of previous studies identified a non-random pattern of aneuploidy seen in breast cancers. This information was used to select those chromosomes that undergo copy number changes in breast cancer and chromosomes that appear stable. A panel of centromeric specific probes were selected and centromeric specific fluorescence in situ hybridisation (FISH) was carried out on the human lymphoblastoid cell line, AHH-1, which had been pre-treated with the chemical aneugens 17-beta oestradiol, diethylstilbestrol (DES) and bisphenol-A (BP-A). The results suggest that oestrogens may play a causative role in breast cancer by inducing a specific pattern of aneuploidy similar to that seen in breast carcinomas. 17-beta oestradiol appears to induce changes most similar to those seen in breast tumours, BP-A induces the same pattern but at a lower frequency and DES appears to be less chromosome specific in its act. PMID- 18162434 TI - Theoretical calculations of the molecular properties of maleimide and its dimer. AB - B3LYP theoretical calculations with 6-31++G(d,p) basis set have been performed to study the infrared spectrum of maleimide and its dimer. Our calculations have shown that the dimer formation leads to a binding energy of 44.0 kJ mol(-1) involving two intermolecular hydrogen bonds between the amide hydrogen and a carbonyl group of two neighboring maleimides. This value is essentially due to the electrostatic interaction term. Our calculations have also revealed the vibrational changes, in terms of frequencies and IR intensities, after dimer formation. The most affected modes are associated with the N-H stretching and in plane bending bands. This behavior can be adequately interpreted by the hydrogen atomic charge and N-H charge-flux based on the modified charge-charge flux overlap model for infrared intensities. The B3LYP frequency shifts are in very good agreement with the experimental ones. PMID- 18162435 TI - Delivery and follow-up of a healthy newborn from a mother with clinical rabies. PMID- 18162436 TI - Choanoflagellate lorica construction and assembly: the nudiform condition. I. Savillea species. AB - The lorica of Savillea spp. (Choanoflagellida) comprises a two layered arrangement of siliceous costae, the inner layer consists of helical costae and the outer layer longitudinal. In Savillea micropora, the helical costae, in a left-handed conformation, undergo 1.5 turns and extend from a short distance above the base of the lorica to the anterior opening. In S. parva the helical costae undergo two turns from base to anterior opening. Ratios of the numbers of helical to longitudinal costae vary from 1:1 in S. parva to 1:1-1:4 in S. micropora. Cell division in Savillea is of the nudiform type, whereby a cell divides to produce a 'naked' flagellated juvenile that swims away from the parent lorica, settles on to a surface and produces a complete set of costal strips. The first formed strips are those that will form the longitudinal costae, this is followed by the slightly thicker strips which will form the inner helical costae. Lorica assembly occurs as a single continuous process and is mediated by the forward movement of the collar tentacles and a rotational movement by the cell. The longitudinal and helical costae are thereby moved in this one combined movement to their respective positions. The longitudinal costae must rotate freely during assembly whilst the helical costae are held at their front end by the respective longitudinal costae and their rear end on the surface of the cell sheath. The concluding inference, based on lorica construction, that there must be a rotational as well as the observed forward movement during lorica assembly is of seminal importance to understanding the basic pattern of lorica construction and the mechanism of lorica assembly in all choanoflagellates. PMID- 18162437 TI - Characterization of a modular, cell-surface protein and identification of a new gene family in the diatom Thalassiosira pseudonana. AB - We report the characterization of a cell-surface protein isolated from copper stressed cells of the centric diatom Thalassiosira pseudonana Hasle and Heimdal (CCMP 1335). This protein has an apparent molecular weight of 100kDa and is highly acidic. The 100kDa protein (p100) sequence is comprised almost entirely of a novel domain termed TpRCR for T. pseudonana repetitive cysteine-rich domain, that is repeated 8 times and that contains conserved aromatic, acidic, and potential metal-binding amino acids. The analysis of the T. pseudonana genome suggests that p100 belongs to a large family of modular proteins that consist of a variable number of TpRCR domain repeats. Based on cell surface biotinylation and antibody data, p100 appears to migrate more rapidly with SDS-PAGE when extracted from cells exposed to high levels of copper; however, the discovery of a large family of TpRCR domain-containing proteins leaves open the possibility that the antibody may be cross-reacting with members of this protein family that are responding differently to copper. The response of the gene encoding p100 at the mRNA level during synchronized progression through the normal cell cycle is similar to previously characterized genes in T. pseudonana encoding cell wall proteins called silaffins. PMID- 18162438 TI - Efficiency of real-time polymerase chain reaction assay to detect Vibrio vulnificus in seawater. AB - The growth of Vibrio vulnificus in an enriched culture of seawater during the summer in Japan was monitored by a plating technique used as the culture method and a real-time polymerase chain reaction (PCR) assay as the molecular method. V. vulnificus was detected by the real-time PCR assay in the samples of August and September but not by the culture method. Vibrio parahaemolyticus, however, was detected among all of the samples with both the culture method and real-time PCR assay. In the analysis of the bacterial populations in enrichment culture, it was demonstrated that the growth of V. vulnificus on agar media was inhibited by the rapid growth of V. parahaemolyticus after 4h of incubation and the 100 times larger initial populations of bacteria other than V. vulnificus and V. parahaemolyticus. These findings demonstrate that V. vulnificus detection by culture methods is a failure, and molecular methods are effective and detect V. vulnificus accurately. PMID- 18162439 TI - Contribution to the evaluation of reference values for PFOA and PFOS in plasma of children and adults from Germany. AB - Perfluorinated compounds (PFC) are a large group of chemicals produced for several decades and widely used for many industrial and consumer applications. Human Biomonitoring studies reveal a background exposure of the general population to perfluorooctanoic acid (PFOA) and pefluorooctane sulfonate (PFOS) in many parts of the world. Reference values for PFOS and PFOA in the German population are currently not available. However, the data of three PFC human biomonitoring studies are taken as basis for deriving a preliminary reference value. The first two studies were performed in southern Germany with 105 (sampling period 2003-2004) and 356 adults (sampling period 2005). The third study was performed in North Rhine-Westphalia (sampling period October and November 2006) in connection with the high PFOA contamination of drinking water in the Sauerland region. Non PFOA exposed control groups comprised of 80 children and 153 females from Siegen and 103 men from Brilon. The whole study which could be taken as a basis for PFOS reference considerations comprised of 170 children, 317 females and 204 men. Though the studies are not representative for the German population, they provide at present the best available data basis for deriving reference values. The 95th percentile values of the studies were used and the following preliminary reference values are recommended: PFOA, 10microg/l for all groups; for PFOS 10microg/l for children at school beginner age, 15microg/l for adult females and 25microg/l for adult males. PMID- 18162440 TI - Blood lactate, heart rate and rating of perceived exertion responses of elite surf lifesavers to high-performance competition. AB - A paucity of empirical research exists into surf lifesaving competition from which coaches and athletes may formulate training and recovery strategies. Seventeen (male=9; female=8) high-performance surf lifesavers (21.2+/-3.9 years) contested multiple rounds of team and individual events at a 2-day surf lifesaving competition. Individual events consisted of the multi-discipline ironman (IRON), paddle board (BOARD) and surf swim (SWIM). Blood lactate (BLa), rating of perceived exertion (RPE) and heart rate (HR) were determined following heats, semi-finals and final. IRON HR and RPE following semi-finals (153.0+/ 21.6beatsmin(-1) and 14.4+/-1.5) and final (171.0+/-9.1beatsmin(-1) and 19.1+/ 0.2) were greater than heats (141.8+/-17.2beatsmin(-1) and 12.0+/-1.9; p<0.05) and final BLa (10.5+/-2.8mmolL(-1)) was greater than heats (5.8+/-3.6mmolL(-1); p<0.05). BOARD BLa and HR were greater after the final (9.0+/-2.8mmolL(-1) and 159.0+/-19.9beatsmin(-1)) compared to heats (4.7+/-2.4mmolL(-1) and 133.0+/ 17.1beatsmin(-1); p<0.05). No significant differences were identified for SWIM. RPE-HR relationships were identified for pooled IRON and BOARD results following semi-finals (0.668; p<0.05) and finals (r=0.741; p<0.05). In conclusion, high performance surf lifesavers employ race strategies with all-out maximal exercise limited to semi-finals and finals. PMID- 18162441 TI - Medial temporal fMRI activation reflects memory lateralization and memory performance in patients with epilepsy. AB - Memory difficulties are a frequent cognitive complaint of patients with chronic epilepsy. Previous studies have suggested that the presence of a seizure focus causes reorganization of brain mechanisms underlying memory function. Here we examine whether seizure onset in the left hemisphere and onset in the right hemisphere have different effects on memory lateralization and whether longer duration of epilepsy is associated with increased lateralization of memory functions to the unaffected hemisphere. We hypothesized that hemisphere of onset and duration of epilepsy would influence plasticity of memory mechanisms, similar to the plasticity observed for language mechanisms. Healthy controls (HC, N = 10) and patients with epilepsy (N = 23, 11 with a left- and 12 with a right hemisphere focus) performed a scene-encoding fMRI task at 4 T. Active voxels (relative to scrambled image viewing) were identified for each participant. Memory laterality indices (LIs) were calculated in three regions of interest (ROIs) designed on the basis of HC group data: a functional ROI, an anatomical hippocampal ROI, and an anatomical-medial temporal ROI encompassing hippocampus and parahippocampal gyrus. In healthy controls, LIs were suggestive of slight left lateralization of encoding memory for pictures. Patients with right hemisphere epilepsy showed a nonsignificant increase in degree of left lateralization. In contrast, patients with left hemispheric epilepsy showed right lateralized activation, differing significantly from controls and from patients with right hemispheric epilepsy. Neuropsychological measures of memory (WMS-III Story Recall) across epilepsy patients predicted LIs in the anatomical ROIs: higher scores were associated with more left-lateralized medial temporal fMRI activation. Neither age of onset nor duration of epilepsy was significantly related to LI. These results indicate that focal epilepsy may influence the functional neuroanatomy of memory function. PMID- 18162442 TI - The measurement of ammonia blood levels in patients taking valproic acid: looking for problems where they do not exist? PMID- 18162443 TI - HIV-1 protecting CCR5-Delta32 allele in medieval Poland. AB - CCR5-Delta32 is the mutation in the chemokine receptor CCR5 that gives its homozygous carriers nearly complete protections from HIV-1 infection. Restricted almost exclusively to Europe, the mutation is thought to have originated in the continent and risen in frequency to the present-day value of approximately 10% due to a selective advantage it gave its carriers. The mutation bearing allele was initially calculated to be approximately 1000 years old and pandemic diseases, such as Bubonic Plague or smallpox were postulated to have selected it. However, new reports appear, that question these hypotheses. Data from ancient DNA (aDNA) studies prove the mutation to be much older, as suggested by calculations based on newer genetic maps. In order to investigate if the plagues of the last millennium selected the allele, and add to the discussion on CCR5 Delta32 origin and age, we searched for the mutation in aDNA isolated from individuals whose skeletal remains were collected at archaeological sites in Poland, dated back to 11-14th centuries. The calculated mean frequency of the allele in medieval Poland (5.06% as compared to contemporary 10.26%), implies its longer than previously believed presence in European populations, and suggests that historic pandemics had little effect on its present-day frequency. PMID- 18162444 TI - Hormesis defined. AB - Hormesis is a term used by toxicologists to refer to a biphasic dose-response to an environmental agent characterized by a low dose stimulation or beneficial effect and a high dose inhibitory or toxic effect. In the fields of biology and medicine hormesis is defined as an adaptive response of cells and organisms to a moderate (usually intermittent) stress. Examples include ischemic preconditioning, exercise, dietary energy restriction and exposures to low doses of certain phytochemicals. Recent findings have elucidated the cellular signaling pathways and molecular mechanisms that mediate hormetic responses which typically involve enzymes such as kinases and deacetylases, and transcription factors such as Nrf-2 and NF-kappaB. As a result, cells increase their production of cytoprotective and restorative proteins including growth factors, phase 2 and antioxidant enzymes, and protein chaperones. A better understanding of hormesis mechanisms at the cellular and molecular levels is leading to and to novel approaches for the prevention and treatment of many different diseases. PMID- 18162445 TI - Truncation of MBD4 predisposes to reciprocal chromosomal translocations and alters the response to therapeutic agents in colon cancer cells. AB - We previously identified a novel genomic instability phenotype of multiple reciprocal chromosomal translocations in a MLH1-defective, microsatellite unstable (MSI) colon cancer cell line (HCA7) and, further, showed that it was unlikely to be directly caused by the mismatch repair (MMR) defect in this cell line. To gain insight into the molecular basis to this novel translocation phenotype, we examined coding and splice-site nucleotide repeat tracts in DNA repair genes for mutations by direct sequencing together with RT-PCR expression analysis of the associated transcript. The material was a selected panel of 8 MSI cell lines including HCA7. A strong candidate identified through this approach was MBD4 as it showed a homozygous truncating mutation associated with substantial loss of the transcript in HCA7 not seen in the other lines. In previous published studies, heterozygous MBD4 mutations were observed in up to 89% of sporadic MSI microdissected colon tumor foci. Using MFISH, we show that over-expression of the truncated MBD4 (+MBD4(tru)) in DLD1, a MSH6 defective, MSI human colon carcinoma cell line predisposed these cells to acquire structural chromosomal rearrangements including multiple reciprocal translocations after irradiation, reminiscent of those seen in HCA7. We also show that over-expression of MBD4(tru) in DLD1 alters the colony survival after exposure to cisplatin or etoposide. These data suggest a wide role for MBD4 in DNA damage response and maintaining chromosomal stability. PMID- 18162446 TI - Determination of levocetirizine in human plasma by liquid chromatography electrospray tandem mass spectrometry: application to a bioequivalence study. AB - We describe a liquid chromatography-tandem mass spectrometric method (LC-MS/MS) for levocetirizine quantification (I) in human plasma. Sample preparation was made using a fexofenadine (II) addition as internal standard (IS), liquid-liquid extraction using cold dichloromethane, and dissolving the final extract in acetonitrile. I and II (IS) were injected in a C18 column and the mobile phase composed of acetonitrile:water:formic acid (80.00:19.90:0.10, v/v/v) and monitored using positive electrospray source with tandem mass spectrometry analyses. The selected reaction monitoring (SRM) was set using precursor ion and product ion combinations of m/z 389>201 for I and m/z 502>467 for II. The limit of quantification and the dynamic range achieved were 0.5ng/mL and 0.5 500.0ng/mL. Validation results on linearity, specificity, accuracy, precision and stability, as well as its application to the analysis of plasma samples taken up to 48h after oral administration of 5mg of levocetirizine dichloridrate in healthy volunteers demonstrate its applicability to bioavailability studies. PMID- 18162447 TI - Genetic syndromes and prenatally detected renal anomalies. AB - Renal anomalies are frequently detected on the routine second trimester scan offered to all pregnant women in the UK. These anomalies may be isolated but can also be associated with other congenital anomalies. Many combinations of ultrasound scan findings constitute recognised genetic entities. Knowledge of these conditions is essential for adequate management of the pregnancy and subsequent balanced parental counselling. This short review discusses the common genetic syndromes associated with the renal abnormalities identified on the antenatal ultrasound scan, and also provides an overview of renal symptoms in chromosome imbalances and after teratogenic influences. PMID- 18162448 TI - Reconstruction of cheek skin defects by the 'Yin-Yang' rotation of the Mustarde flap and the temporoparietal scalp. AB - BACKGROUND AND PURPOSES: When the skin's laxity is limited, the preauricular cutaneous defect resulting from the Mustarde cheek rotation flap for the coverage of suborbital skin defects is very difficult to close even with a large undermining of the cervicofacial skin. The rotation of the temporoparietal scalp in an opposite direction compared to the Mustarde flap or the 'Yin-Yang' rotation of these flaps is interesting for two reasons: it allows an easy closure of this preauricular defect with a limited facial undermining and it suspends with efficiency the Mustarde flap at the temporal area avoiding the lower lid's ectropion. MATERIAL AND METHODS: From September 2001 to April 2005, nine patients aged between 35 and 78 years old, with no facial skin laxity, have benefited from this technique to cover suborbital defects secondary to excision of basal cell carcinomas with a mean diameter of 6 cm. The design of the Mustarde flap was classical but the skin undermining stopped at the mandible's lower border. The triangular preauricular defect has served as a geometrical base to design an opposite temporoparietal rotation flap which is undermined under the galea. RESULTS: This technique has allowed an easy primary closure of all these defects. No complications were reported. With a mean follow up of 36 months the aesthetic quality of these reconstructions was satisfying and there was no malposition of the lower lid. CONCLUSION: This is a good technique for the simple coverage of medium-sized suborbital skin defects. PMID- 18162449 TI - Improved fat graft viability by delayed fat flap with ischaemic pretreatment. AB - The aim of this study was to investigate the efficacy of the delayed fat flap on viability and longevity of grafted fat in an animal model. Sixty New Zealand rabbits were used for the long-term study. In each animal, a U-shaped fat flap was raised randomly in the inguinal region on one leg and then resutured in place to delay the flap for ischaemic pretreatment. The contralateral inguinal region was left untreated as a matched control. At 12h after the delaying operation, vascular endothelial growth factor (VEGF) expression in the fat from both groups of 10 rabbits was assessed. At 3 weeks, fat grafts from both the pretreated and control inguinal regions were transferred into a pocket that had been created on each side of the dorsal midline of the rabbit. Ten rabbits were sacrificed at intervals of 1, 3, 6, 9 and 12 months after transplantation for gross and histomorphometric analysis. Results showed that VEGF protein level in the pretreated fat flaps was significantly higher than that in the controls at 12h after the delaying operation. The number of vessels was increased in pretreated groups at 1 month, but no significant difference observed between two groups at 3 months. Histomorphometric analysis showed that the capillary density of the fat grafts was significantly higher in the pretreated group than that in controls at 1 and 3 months, respectively (P<0.01). There was no statistical difference in fat graft size and weight between the pretreated and control groups at 1 and 3 months after transplantation (P>0.05). However, the fat grafts in control groups disappeared after 6 months, and those in the pretreated groups were still maintained up to 12 months. This study demonstrated that the pretreatment of fat tissues with ischaemia insult could be advantageous for their viability and longevity. These results encourage further studies on potential clinical application of this new and simple technique to enable long lasting results. PMID- 18162450 TI - [Haemodialysis and its impact on the quality of life]. AB - The haemodialysis has considerably transformed the evolution of chronic renal failure which usually has a precarious evolution. However, this palliative treatment disturbed in several manners the universe of the patient. Our purposes were to evaluate the quality of life of patient on haemodialysis and to identify the risk factors that can deteriorate it. Our work was a transversal study realized in four centers of haemodialysis during a period of three months (2004). One hundred and sixty-two patients participated. The quality of life was assessed with a specific scale Kidney Disease Quality of Life Short-Form (KDQDL-SF). A standardization of the initial average scores of SF-36 was realized. The global average score, according to KDQOL-SF, was 55,2. The standardization showed a physical component more alterated than the mental one with respective scores 36,8 and 44,7. The logistic regression permitted us to identify that the quality of life's alteration was correlated with an absent autonomy, a diabetic origin of renal insufficiency, a predialytic serum creatine concentration inferior to 800micromol/l and haemodialysis demographic social features. This clinimetric approach not only precises the more deteriorated sectors through the regard of patient, but also reintroduces its point of view about the treatment permitting a personalization of therapies and consequently a better adhesion. PMID- 18162451 TI - Birth of a very low birth weight preterm infant and the intention to breastfeed 'naturally'. AB - An interpretive phenomenological study involving 17 Australian parents was undertaken to explore parents' experiences of breastfeeding very low birth weight (VLBW) preterm infants from birth to 12 months of age. Data were collected from 45 individual interviews held with both mothers and fathers, which were then transcribed verbatim and analysed using thematic analysis. From this study, the analysis identified the following themes: the intention to breastfeed naturally; breast milk as connection; the maternal role of breast milk producer; breastmilk as the object of attention; breastfeeding and parenting the hospitalised baby and the demise of breastfeeding. The discussion presented here presents the theme of the intention to breastfeed 'naturally'. This study found that all of the participant women decided to breastfeed well before the preterm birth, and despite the birth of a VLBW preterm infant continued to expect the breastfeeding experience to be normal regardless of the difference of the postpartum experience. It is without doubt that for these parents the pro-breastfeeding rhetoric is powerfully influential and thus successful in promoting breastfeeding. Furthermore, all participants expected breastfeeding to be 'natural' and satisfying. There is disparity between parents' expectations of breastfeeding 'naturally' and the commonplace reality of long-term breast expression and uncertain at-breast feeding outcomes. How the parents came to make the decision to breastfeed their unborn child -- including the situations and experiences that have influenced their decision making -- and how the preterm birth and the dominant cultures subsequently affected that decision will be discussed. The findings have implications for midwifery education and maternity care professionals who support parents making feeding decisions early in pregnancy and those striving to breastfeed preterm infants. PMID- 18162452 TI - Microglia of teleosts: facing a challenge in neurobiology. AB - This review is concerned with recent literature on teleost fish CNS microglia. It covers not only various aspects of these cells, notably comparing them with mammalian microglia, but also points out the several potentialities neural tissue of teleosts exhibits in neurobiological research. The relationships between neurons and glial cells are considered in fish, aiming at an integrated picture of the complex ways neurons and glia communicate and collaborate in normal and injured neural tissues. In addition, attention has been paid to different teleost models according to their availability, easy maintenance in experimental conditions, possibilities of embryos manipulation and sequenced genome. The recent setting up of successful protocols for fish glia and mixed neuron-glia cultures, together with the molecular facilities offered from genome knowledge, should provide a new boost to studies about microglia and neuron-microglia relationships. PMID- 18162453 TI - Thymic development in surgically bursectomized embryonic chicken: expression of PCNA, CD3, CD4 and CD8 markers. AB - Little information is available on the functional relationship between bursa and thymus during chicken embryogenesis. We, therefore, investigated embryonic thymuses taken at 17 days in ovo from chickens bursectomized at 68-72 hours, with histological, histochemical (PAS, Alcian blue), and immunoreaction (anti cytokeratin B, anti-PCNA/cyclin and anti-CD3, CD4 and CD8 antibodies) methods and compared these data with those from normal and sham-operated chickens of the same age. The bursectomized thymuses distinctly differed from normal and sham-operated thymuses: they were smaller, and the cortical zone was thinner and contained fewer epithelial cells and thymocytes. Only few cortical thymocytes were immunoreactive for PCNA, indicating low proliferative rate. More cortical thymocytes as compared with the normal, expressed CD3 on their cell membrane, whereas the thymocytes at the cortical-medullary border expressing anti- CD4 and anti-CD8 antidodies were less numerous than in normal thymus. The medullary zone contained few epithelial clusters made up of fewer cells than medullary clusters in normal chickens. Some cystic formations were enlarged and contained PAS- or Alcian-blue positive amorphous material. All these data suggest that early bursectomy affects both morphological and functional thymic development. PMID- 18162454 TI - Expression of neurotransmitters and neurotrophins in neurogenic inflammation of the rat retina. AB - Antidromic stimulation of the rat trigeminal ganglion triggers the release of substance P (SP) and calcitonin gene-related peptide (CGRP) from sensory nerve terminals of the capsaicin sensitive C-fibers. These pro-inflammatory neuropeptides produce a marked hyperemia in the anterior segment of the eye, accompanied by increased intraocular pressure, breakdown of the blood-aqueous barrier and myosis. To assess the effects of neurogenic inflammation on the retina, specifically on the immunostaining of neurotransmitters and neurotrophins, as well as on the expression of neurotrophin receptors in the retina. RT-PCR was also accomplished in control and stimulated animals to confirm the immunohistochemical results. In the electrically stimulated eyes, immunostaining for SP, CGRP, VIP and nNOS demonstrated a marked increase in the RPE/POS (Retinal Pigment Epithelium/Photoreceptor Outer Segments), in the inner and outer granular layers and in the ganglion cells in comparison to the control eyes. CGRP and SP were found increased in stimulated animals and this result has been confirmed by RT- PCR. Changes in neurotrophin immunostaining and in receptor expression were also observed after electric stimulation of trigeminal ganglia. Decrease of BDNF and NT4 in the outer and inner layers and in ganglion cells was particularly marked. In stimulated rat retinas immunostaining and RT-PCR showed a NGF expression increase. Neurotrophin receptors remained substantially unchanged. These studies demonstrated, for the first time, that antidromic stimulation of the trigeminal ganglion and subsequent neurogenic inflammation affect immunostaining of retinal cell neurotransmitter/neuropeptides and neurotrophins as well as the expression of neurotrophin receptors. PMID- 18162455 TI - Connexin 43 expression in human and mouse testes with impaired spermatogenesis. AB - Connexin 43 (Cx43) belongs to a family of proteins that form gap junction channels. The aim of this study was to examine the expression of Cx43 in the testis of a patient with Klinefelter's syndrome and of mice with the mosaic mutation and a partial deletion in the long arm of the Y chromosome. These genetic disorders are characterized by the presence of numerous degenerated seminiferous tubules and impaired spermatogenesis. In mouse testes, the expression and presence of Cx43 were detected by means of immunohistochemistry and Western blot analysis, respectively. In testes of Klinefelter's patient only immunoexpression of Cx43 was detected. Regardless of the species Cx43 protein was ubiquitously distributed in testes of reproductively normal males, whereas in those with testicular disorders either a weak intensity of staining or no staining within the seminiferous tubules was observed. Moderate to strong or very strong staining was confined to the interstitial tissue. In an immunoblot analysis of testicular homogenates Cx43 appeared as one major band of approximately 43 kDa. Our study adds three more examples of pathological gonads in which the absence or apparent decrease of Cx43 expression within the seminiferous tubules was found. A positive correlation between severe spermatogenic impairment and loss of Cx43 immunoreactivity observed in this study supports previous data that gap junctions play a crucial role in spermatogenesis. Strong Cx43 expression detected mostly in the interstitial tissue of the Klinefelter's patient may presumably be of importance in sustaining Leydig cell metabolic activity. However, the role of gap junction communication in the control of Leydig cell function seems to be more complex than originally thought. PMID- 18162456 TI - Mean diameter of nucleolar bodies in cultured human leukemic myeloblasts is mainly related to the S and G2 phase of the cell cycle. AB - Mean diameter of nucleolar bodies (nucleoli without the perinucleolar chromatin) per cell was studied in human leukemic myeloblasts represented by K 562 and Kasumi 1 cell lines which originated from chronic and acute myeloid leukaemia. The measurement of mean diameter of nucleolar bodies in specimens stained for RNA was very simple. Such approach eliminated the variability of the perinucleolar chromatin discontinuous shell which might influence the measured nucleolar size as suggested by earlier studies. Ageing of K 562 myeloblasts produced a significant decrease of cells in S+G2 phase of the cell cycle accompanied by a significant reduction of mean diameter of nucleolar bodies (MDNoBs) per cell. In contrast, treatment of Kasumi 1 myeloblasts with histone deacetylase inhibitor - Trichostatin A - produced a large incidence of resistant cells in S+G2 phase which were characterised by a large increase of MDNoBs. Thus, MDNoBs in leukemic myeloblasts might be a helpful tool to estimate the incidence of cells in the S+G2 phase at the single cell level in smear preparations when the number of cells is very small. PMID- 18162457 TI - In vitro cultured progenitors and precursors of cardiac cell lineages from human normal and post-ischemic hearts. AB - The demonstration of the presence of dividing primitive cells in damaged hearts has sparked increased interest about myocardium regenerative processes. We examined the rate and the differentiation of in vitro cultured resident cardiac primitive cells obtained from pathological and normal human hearts in order to evaluate the activation of progenitors and precursors of cardiac cell lineages in post-ischemic human hearts. The precursors and progenitors of cardiomyocyte, smooth muscle and endothelial lineage were identified by immunocytochemistry and the expression of characteristic markers was studied by western blot and RT-PCR. The amount of proteins characteristic for cardiac cells (alpha-SA and MHC, VEGFR 2 and FVIII, SMA for the precursors of cardiomyocytes, endothelial and smooth muscle cells, respectively) inclines toward an increase in both alpha-SA and MHC. The increased levels of FVIII and VEGFR2 are statistically significant, suggesting an important re-activation of neoangiogenesis. At the same time, the augmented expression of mRNA for Nkx 2.5, the trascriptional factor for cardiomyocyte differentiation, confirms the persistence of differentiative processes in terminally injured hearts. Our study would appear to confirm the activation of human heart regeneration potential in pathological conditions and the ability of its primitive cells to maintain their proliferative capability in vitro. The cardiac cell isolation method we used could be useful in the future for studying modifications to the microenvironment that positively influence cardiac primitive cell differentiation or inhibit, or retard, the pathological remodeling and functional degradation of the heart. PMID- 18162458 TI - Effect of a topical treatment in organotypic culture of human breast skin after exposure to gamma-rays. AB - The early radiation of epidermal reactions can lead to healing of the lesion or radiation necrosis. There is no general agreement for either the prevention and/or treatment of radiation skin response, also as little is known about the immediate phases of this phenomenon. We investigated the early effects exerted by Healing and Wound Emulsion (HWE) on human skin response after ionizing radiation. Epidermal morphology, Heat Shock Protein (HSP) 70, and Transforming Growth Factor beta1 (TGF-beta1) gene expression were investigated in organotypic human skin cultures undergoing a double dose of gamma-rays (2 Gy). HSP70 gene expression tended to be induced in the HWE group 6 hours after cream administration and was significantly up-regulated after 48 hours, when epidermal morphological alterations were evident. TGF-beta1 seems not affected in cream treated samples. HWE may stimulate skin to mount an early defensive response against damage induced by gamma rays. PMID- 18162459 TI - Expression and distribution of S-100 protein, CD83 and apoptosis-related proteins (Fas, FasL and Bcl-2) in thyroid tissues of autoimmune thyroid diseases. AB - Previous studies have shown that dendritic cells (DCs) and apoptosis-related proteins play a critical role in the pathogenesis of autoimmune thyroid diseases (ATD). This study was designed to investigate the expression and distribution of S-100 protein, CD83 and apoptosis-related proteins (Fas, FasL and Bcl-2) in the thyroid tissues of ATD and their role in ATD pathogenesis as determined by immunochemical staing techniques and other methods. Pathological tissues of 30 patients with Hashimoto's thyroiditis (HT), 30 patients with Graves' disease (GD) and 30 cases of thyroid follicular adenoma (TFA, as control) were used for this study. A higher expression of S-100 in HT (4.2+/-3.1%) and GD (3.9+/-2.8%) vs TFA (0.95+/-0.64%) (p<0.001). was observed as well as a higher expression of CD83 in HT (22.58+/-13.96% and GD (29.92+/-14.43%) vs TFA (5.19+/-8.08%) (p<0.001). HT thyrocytes adjacent to thyroid infiltrating lymphocytes (TILs) showed greater increases in the levels of Fas and FasL than did the GD thyrocytes while HT TILs exhibited lower expression of Fas and FasL than did the GD TILs. GD thyrocytes expressed increased levels of the antiapoptotic protein Bcl-2 as compared to the low levels detected in HT thyrocytes. An opposite pattern was observed in the TILs in GD (low expression of Bcl-2) and HT (high expression of Bcl-2). The findings suggest that the high expression of DC markers is related to the pathogenesis of HT and GD. Up-regulation of both the number and matured functions of DCs may lead to the presentation of more antigens to lymphocytes which are related to the development of autoimmune thyroid diseases. The regulation of Fas/FasL/Bcl-2 in GD favors apoptosis of infiltrating lymphocytes and thyrocyte survival. The regulation of Fas/FasL/Bcl-2 in HT may promote thyrocyte apoptosis leading to hypothyroidism. PMID- 18162460 TI - Efficiency of two different transfection reagents for use with human NTERA2 cells. AB - The teratocarcinoma cell line NTERA2 is recently used in a wide range of researches (from developmental biology to toxicology, for their ability to be induced to neural differentiation. In order to study the genetic potential of these cells, it is needed to use methods for gene silencing and/or mRNA interference, allowing cell viability and further differentiation. To check these features, we simultaneously tested the transfection efficiency of NTERA2, A549 and HeLa cells with Metafectene PRO (Biontex, Germany) and another optimal transfection reagent currently used in our Laboratory, using as a reporter gene the DsRed2 vector (Clontech, Mountain View, CA). Under our culture conditions for NTERA2 and HeLa cells, Metafectene PRO transfection method was found to possess high throughput performance, that allows low concentration rate and low exposure time to excitation light source, thus reducing both toxicity and phototoxicity. PMID- 18162461 TI - Physical training is associated with changes in nuclear magnetic resonance and morphometrical parameters of the skeletal muscle in senescent mice. AB - The effect of a three-month training period on T2 relaxation time as well as on myofibre size and type was investigated in the lower limbs of senescent mice. After training, T2 (which is a magnetic resonance imaging parameter known to increase during acute exercise) was significantly higher in trained mice (36.37+/ 1.27 vs 37.76+/-2.06 ms, p=0.003, n=8), whereas no change was found in non trained animals (36.35+/-1.02 vs 36.24+/-1.15 ms, p=0.278, n=8). The percentage of muscle limb area evaluated in vivo on magnetic resonance images before and after the experimental period was unchanged in trained mice (69.84+/-2.50 vs 70.29+/-2.29, p=0.896, n=3) and decreased in non-trained animals (72.98+/-1.68 vs 64.62+/-2.34, p=0.006, n=3). Cross-sectional area of fast and slow myofibres, evaluated on paraffin-embedded samples after immunolabelling for skeletal fast fibre myosin, was lower in non-trained than in trained mice in both gastrocnemius and quadriceps muscle, but no change in slow/fast fibre ratio nor in apoptotic rate was found. These data show that training can prevent sarcopenia in senescent mice by affecting muscle status and inducing myofibre hypertrophy in the absence of significant muscle damage. PMID- 18162462 TI - Reinvestigation of the catalytic mechanism of formyl-CoA transferase, a class III CoA-transferase. AB - Formyl-coenzyme A transferase from Oxalobacter formigenes belongs to the Class III coenzyme A transferase family and catalyzes the reversible transfer of a CoA carrier between formyl-CoA and oxalate, forming oxalyl-CoA and formate. Formyl CoA transferase has a unique three-dimensional fold composed of two interlaced subunits locked together like rings of a chain. We here present an intermediate in the reaction, formyl-CoA transferase containing the covalent beta-aspartyl-CoA thioester, adopting different conformations in the two active sites of the dimer, which was identified through crystallographic freeze-trapping experiments with formyl-CoA and oxalyl-CoA in the absence of acceptor carboxylic acid. The formation of the enzyme-CoA thioester was also confirmed by mass spectrometric data. Further structural data include a trapped aspartyl-formyl anhydride protected by a glycine loop closing down over the active site. In a crystal structure of the beta-aspartyl-CoA thioester of an inactive mutant variant, oxalate was found bound to the open conformation of the glycine loop. Together with hydroxylamine trapping experiments and kinetic as well as mutagenesis data, the structures of these formyl-CoA transferase complexes provide new information on the Class III CoA-transferase family and prompt redefinition of the catalytic steps and the modified reaction mechanism of formyl-CoA transferase proposed here. PMID- 18162463 TI - ECRG2 disruption leads to centrosome amplification and spindle checkpoint defects contributing chromosome instability. AB - Cancer cells contain an abnormal number of chromosomes (aneuploidy), which is a prevalent form of genetic instability in human cancers. Abnormal amplification of centrosomes and defects of spindle assembly checkpoint are the major causes of chromosome instability in cancer cells. Here we present biochemical evidence to suggest a role of ECRG2, a novel tumor suppressor gene, in maintaining chromosome stability. ECRG2 localized to centrosomes during interphase and kinetochores during mitosis. Further analysis revealed that ECRG2 participates in centrosome amplification in a p53-dependent manner. Depletion of ECRG2 not only destabilized p53, down-regulated p21, and increased the cyclin E/CDK2 activity, thus initiating centrosome amplification, but also abolished the ability of p53 localize to centrosomes. Overexpression of ECRG2 restored the p53-dependent suppression of centrosome duplication. Furthermore, ECRG2-depleted cells show severely disrupted spindle phenotype but fail to maintain the mitotic arrest due to minimal BUBR1 protein levels. Taken together, our results indicate that ECRG2 is important for ensuring centrosome duplication, spindle assembly checkpoint, and accurate chromosome segregation, and its depletion may contribute to chromosome instability and aneuploidy in human cancers. PMID- 18162464 TI - Both G i and G o heterotrimeric G proteins are required to exert the full effect of norepinephrine on the beta-cell K ATP channel. AB - The effects of norepinephrine (NE), an inhibitor of insulin secretion, were examined on membrane potential and the ATP-sensitive K+ channel (K ATP) in INS 832/13 cells. Membrane potential was monitored under the whole cell current clamp mode. NE hyperpolarized the cell membrane, an effect that was abolished by tolbutamide. The effect of NE on K ATP channels was investigated in parallel using outside-out single channel recording. This revealed that NE enhanced the open activities of the K ATP channels approximately 2-fold without changing the single channel conductance, demonstrating that NE-induced hyperpolarization was mediated by activation of the K ATP channels. The NE effect was abolished in cells preincubated with pertussis toxin, indicating coupling to heterotrimeric G i/G o proteins. To identify the G proteins involved, antisera raised against alpha and beta subunits (anti-G alpha common, anti-G beta, anti-G alpha i1/2/3, and anti-G alpha o) were used. Anti-G alpha common totally blocked the effects of NE on membrane potential and K ATP channels. Individually, anti-G alpha i1/2/3 and anti-G alpha o only partially inhibited the action of NE on K ATP channels. However, the combination of both completely eliminated the action. Antibodies against G beta had no effects. To confirm these results and to further identify the G protein subunits involved, the blocking effects of peptides containing the sequence of 11 amino acids at the C termini of the alpha subunits were used. The data obtained were similar to those derived from the antibody work with the additional information that G alpha i3 and G alpha o1 were not involved. In conclusion, both G i and G o proteins are required for the full effect of norepinephrine to activate the K ATP channel. PMID- 18162465 TI - ATR-Chk2 signaling in p53 activation and DNA damage response during cisplatin induced apoptosis. AB - Cisplatin is one of the most effective anti-cancer drugs; however, the use of cisplatin is limited by its toxicity in normal tissues, particularly injury of the kidneys. The mechanisms underlying the therapeutic effects of cisplatin in cancers and side effects in normal tissues are largely unclear. Recent work has suggested a role for p53 in cisplatin-induced renal cell apoptosis and kidney injury; however, the signaling pathway leading to p53 activation and renal apoptosis is unknown. Here we demonstrate an early DNA damage response during cisplatin treatment of renal cells and tissues. Importantly, in the DNA damage response, we demonstrate a critical role for ATR, but not ATM (ataxia telangiectasia mutated) or DNA-PK (DNA-dependent protein kinase), in cisplatin induced p53 activation and apoptosis. We show that ATR is specifically activated during cisplatin treatment and co-localizes with H2AX, forming nuclear foci at the site of DNA damage. Blockade of ATR with a dominant-negative mutant inhibits cisplatin-induced p53 activation and renal cell apoptosis. Consistently, cisplatin-induced p53 activation and apoptosis are suppressed in ATR-deficient fibroblasts. Downstream of ATR, both Chk1 and Chk2 are phosphorylated during cisplatin treatment in an ATR-dependent manner. Interestingly, following phosphorylation, Chk1 is degraded via the proteosomal pathway, whereas Chk2 is activated. Inhibition of Chk2 by a dominant-negative mutant or gene deficiency attenuates cisplatin-induced p53 activation and apoptosis. In vivo in C57BL/6 mice, ATR and Chk2 are activated in renal tissues following cisplatin treatment. Together, the results suggest an important role for the DNA damage response mediated by ATR-Chk2 in p53 activation and renal cell apoptosis during cisplatin nephrotoxicity. PMID- 18162466 TI - The phosphatase PHLPP controls the cellular levels of protein kinase C. AB - The life cycle of protein kinase C (PKC) is controlled by multiple phosphorylation and dephosphorylation steps. The maturation of PKC requires three ordered phosphorylations, one at the activation loop and two at COOH-terminal sites, the turn motif and the hydrophobic motif, to yield a stable and signaling competent enzyme. Dephosphorylation of the enzyme leads to protein degradation. We have recently discovered a novel family of protein phosphatases named PH domain leucine-rich repeat protein phosphatase (PHLPP) whose members terminate Akt signaling by dephosphorylating the hydrophobic motif on Akt. Here we show that the two PHLPP isoforms, PHLPP1 and PHLPP2, also dephosphorylate the hydrophobic motif on PKC betaII, an event that shunts PKC to the detergent insoluble fraction, effectively terminating its life cycle. Deletion mutagenesis reveals that the PH domain is necessary for the effective dephosphorylation of PKC betaII by PHLPP in cells, whereas the PDZ-binding motif, required for Akt regulation, is dispensable. The phorbol ester-mediated dephosphorylation of the hydrophobic site, but not the turn motif or activation loop, is insensitive to okadaic acid, consistent with PHLPP, a PP2C family member, controlling the hydrophobic site. In addition, knockdown of PHLPP expression reduces the rate of phorbol ester-triggered dephosphorylation of the hydrophobic motif, but not turn motif, of PKC alpha. Last, we show that depletion of PHLPP in colon cancer and normal breast epithelial cells results in an increase in conventional and novel PKC levels. These data reveal that PHLPP controls the cellular levels of PKC by specifically dephosphorylating the hydrophobic motif, thus destabilizing the enzyme and promoting its degradation. PMID- 18162467 TI - Mastermind-like domain-containing 1 (MAMLD1 or CXorf6) transactivates the Hes3 promoter, augments testosterone production, and contains the SF1 target sequence. AB - Although chromosome X open reading frame 6 (CXorf6) has been shown to be a causative gene for hypospadias, its molecular function remains unknown. To clarify this, we first examined CXorf6 protein structure, identifying homology to mastermind-like 2 (MAML2) protein, which functions as a co-activator in canonical Notch signaling. Transactivation analysis for wild-type CXorf6 protein by luciferase assays showed that CXorf6 significantly transactivated the promoter of a noncanonical Notch target gene hairy/enhancer of split 3 (Hes3) without demonstrable DNA-binding capacity. Transactivation analysis was also performed for the previously described three apparently pathologic nonsense mutations, indicating that E124X and Q197X proteins had no transactivation function, whereas R653X protein retained a nearly normal transactivation function. Subcellular localization analysis revealed that wild-type and R653X proteins co-localized with MAML2 protein in nuclear bodies, whereas E124X and Q197X proteins were incapable of localizing to nuclear bodies. Thus, further studies were performed for R653X, revealing the occurrence of nonsense mediated mRNA decay in vivo. Next, transient knockdown of CXorf6 was performed using small interfering RNA, showing reduced testosterone production in mouse Leydig tumor cells. Furthermore, steroidogenic factor 1 (SF1) protein bound to a specific sequence in the upstream of the CXorf6 coding region and exerted a transactivation activity. These results suggest that CXorf6 transactivates the Hes3 promoter, augments testosterone production, and contains the SF1 target sequence, thereby providing the first clue to clarify the biological role of CXorf6. We designate CXorf6 as MAMLD1 (mastermind-like domain-containing 1) based on its characteristic structure. PMID- 18162468 TI - Protein conformation changes of HemAT-Bs upon ligand binding probed by ultraviolet resonance Raman spectroscopy. AB - HemAT from Bacillus subtilis (HemAT-Bs) is a heme-based O2 sensor protein that acts as a signal transducer responsible for aerotaxis. HemAT-Bs discriminates its physiological effector (O2) from other gas molecules (CO and NO), although all of them bind to a heme. To monitor the conformational changes in the protein moiety upon binding of different ligands, we have investigated ultraviolet resonance Raman (UVRR) spectra of the ligand-free and O2-, CO-, and NO-bound forms of full length HemAT-Bs and several mutants (Y70F, H86A, T95A, and Y133F) and found that Tyr70 in the heme distal side and Tyr133 and Trp132 from the G-helix in the heme proximal side undergo environmental changes upon ligand binding. In addition, the UVRR results confirmed our previous model, which suggested that Thr95 forms a hydrogen bond with heme-bound O2, but Tyr70 does not. It is deduced from this study that hydrogen bonds between Thr95 and heme-bound O2 and between His86 and heme 6-propionate communicate the heme structural changes to the protein moiety upon O2 binding but not upon CO and NO binding. Accordingly, the present UVRR results suggest that O2 binding to heme causes displacement of the G-helix, which would be important for transduction of the conformational changes from the sensor domain to the signaling domain. PMID- 18162469 TI - Deciphering the structural role of histidine 83 for heme binding in hemophore HasA. AB - Heme carrier HasA has a unique type of histidine/tyrosine heme iron ligation in which the iron ion is in a thermally driven two spin states equilibrium. We recently suggested that the H-bonding between Tyr75 and the invariantly conserved residue His83 modulates the strength of the iron-Tyr75 bond. To unravel the role of His83, we characterize the iron ligation and the electronic properties of both wild type and H83A mutant by a variety of spectroscopic techniques. Although His83 in wild type modulates the strength of the Tyr-iron bond, its removal causes detachment of the tyrosine ligand, thus giving rise to a series of pH dependent equilibria among species with different axial ligation. The five coordinated species detected at physiological pH may represent a possible intermediate of the heme transfer mechanism to the receptor. PMID- 18162470 TI - Requirements for the interaction of mouse Polkappa with ubiquitin and its biological significance. AB - Polkappa protein is a eukaryotic member of the DinB/Polkappa branch of the Y family DNA polymerases, which are involved in the tolerance of DNA damage by replicative bypass. Despite universal conservation through evolution, the precise role(s) of Polkappa in this process has remained unknown. Here we report that mouse Polkappa can physically interact with ubiquitin by yeast two-hybrid screening, glutathione S-transferase pulldown, and immunoprecipitation methods. The association of Polkappa with ubiquitin requires the ubiquitin-binding motifs located at the C terminus of Polkappa. In addition, Polkappa binds with monoubiquitinated proliferating cell nuclear antigen (PCNA) more robustly than with non-ubiquitinated PCNA. The ubiquitin-binding motifs mediate the enhanced association between monoubiquitinated PCNA and Polkappa. The ubiquitin-binding motifs are also required for Polkappa to form nuclear foci after UV radiation. However, the ubiquitin-binding motifs do not affect Polkappa half-life. Finally, we have examined levels of Polkappa expression following the exposure of mouse cells to benzo[a]pyrene-dihydrodiol epoxide or UVB radiation. PMID- 18162471 TI - S-Phase-specific activation of PKC alpha induces senescence in non-small cell lung cancer cells. AB - Protein kinase C (PKC) has been widely implicated in positive and negative control of cell proliferation. We have recently shown that treatment of non-small cell lung cancer (NSCLC) cells with phorbol 12-myristate 13-acetate (PMA) during G1 phase inhibits the progression into S phase, an effect mediated by PKC delta induced up-regulation of the cell cycle inhibitor p21 Cip1. However, PMA treatment in asynchronously growing NSCLC cells leads to accumulation of cells in G2/M. Studies in post-G1 phases revealed that PMA induced an irreversible G2/M cell cycle arrest in NSCLC cells and conferred morphological and biochemical features of senescence, including elevated SA-beta-Gal activity and reduced telomerase activity. Remarkably, this effect was phase-specific, as it occurred only when PKC was activated in S, but not in G1, phase. Mechanistic analysis revealed a crucial role for the classical PKC alpha isozyme as mediator of the G2/M arrest and senescence, as well as for inducing p21(Cip1) an obligatory event for conferring the senescence phenotype. In addition to the unappreciated role of PKC isozymes, and specifically PKC alpha, in senescence, our data introduce the paradigm that discrete PKCs trigger distinctive responses when activated in different phases of the cell cycle via a common mechanism that involves p21 Cip1 up-regulation. PMID- 18162472 TI - Limited mitochondrial permeabilization is an early manifestation of palmitate induced lipotoxicity in pancreatic beta-cells. AB - Involvement of the mitochondrial permeability transition (MPT) pore in early stages of lipotoxic stress in the pancreatic beta-cell lines MIN6 and INS-1 was the focus of this study. Both long term (indirect) and acute (direct) effects of fatty acid (FA) application on beta-cell susceptibility to Ca(2+)-induced MPT induction were examined using both permeabilized and intact beta-cells. Long term exposure to moderate (i.e. below cytotoxic) levels of the saturated FA palmitate sensitized beta-cell mitochondria to MPT induced by Ca(2+). Long term exposure to palmitate was significantly a more efficient inducer of MPT than the unsaturated FA oleate, although upon acute application both caused similar MPT activation. Application of antioxidants, inhibitors of the ceramide pathway, or modifiers of membrane fluidity did not protect beta-cell mitochondria from FA exposure. However, significant protection was provided by co-application of the unsaturated FA oleate in a phosphatidylinositol 3-kinase-dependent manner. Characterization of MPT pore opening in response to moderate palmitate treatment revealed the opening of a unique form of MPT in beta-cells as it encompassed features of both low and high conductance MPT states. Specifically, this MPT showed solute selectivity, characteristic of a low conductance MPT; however, it affected mitochondrial respiration and membrane potential in a way typical of a high conductance MPT. Activation of the full-size/high conductance form of MPT required application of high levels of FA that reduced growth and initiated apoptosis. These findings suggest that in the beta-cell, MPTs can act as both initiators of cell death and as versatile modulators of cell metabolism, depending on the mode of the MPT pore induced. PMID- 18162473 TI - A comparative synteny map of Burkholderia species links large-scale genome rearrangements to fine-scale nucleotide variation in prokaryotes. AB - Genome rearrangement events, including inversions and translocations, are frequently observed across related microbial species, but the impact of such events on functional diversity is unclear. To clarify this relationship, we compared 4 members of the Gram-negative Burkholderia family (Burkholderia pseudomallei, Burkholderia mallei, Burkholderia thailandensis, and Burkholderia cenocepacia) and identified a core set of 2,590 orthologs present in all 4 species (metagenes). The metagenes were organized into 255 synteny blocks whose relative order has been altered by a predicted minimum of 242 genome rearrangement events. Functionally, metagenes within individual synteny blocks were often related. The molecular divergence of metagenes adjacent to synteny breakpoints (boundary metagenes) was significantly greater compared with metagenes within blocks, suggesting an association between breakpoint locations and local fine-scale nucleotide alterations. This phenomenon, referred to as boundary element associated divergence, was also observed in Pseudomonas and Shigella, suggesting that this is a common phenomenon in prokaryotes. We also observed preferential localization of species-specific genes and insertion sequence element to synteny breakpoints in Burkholderia. Our results suggest that in prokaryotes, genome rearrangements may influence functional diversity through the enhanced divergence of boundary genes and the creation of foci for acquiring and deleting species-specific genes. PMID- 18162474 TI - Letter regarding: "Paraquat: the red herring of Parkinson's disease research". PMID- 18162475 TI - Re: "the general epidemiologist: is there a place in today's epidemiology?". PMID- 18162476 TI - Re: "the general epidemiologist: is there a place in today's epidemiology?". PMID- 18162477 TI - Physical activity and mammographic breast density in the EPIC-Norfolk cohort study. AB - Physical inactivity and high mammographic breast density have both been associated with increased breast cancer risk. However, the association between physical activity and mammographic breast density remains inconsistent. In the European Prospective Investigation of Cancer (EPIC)-Norfolk population-based cohort study (United Kingdom), the authors investigated the cross-sectional association between physical activity level at baseline during 1993-1997 and breast density among 1,394 postmenopausal, cancer-free women. Usual physical activity was assessed by a brief, validated questionnaire. Percentage breast density was determined visually from mammograms by three trained radiologists using the Boyd six-category scale. The association between physical activity level and breast density risk category was examined. No statistically significant association between physical activity and percentage breast density was observed in the unadjusted or adjusted regression models. A suggested increase in breast density for the most active women in the unadjusted regression analysis (odds ratio = 1.13, 95% confidence interval: 0.71, 1.80) was reversed after inclusion of body mass index and reproductive and lifestyle variables (odds ratio = 0.78, 95% confidence interval: 0.45, 1.34). The lack of an association between physical activity and percentage breast density suggests that an association between physical activity and breast cancer risk is unlikely to be mediated through an effect on mammographic breast density. PMID- 18162478 TI - Meta- and pooled analyses of the methylenetetrahydrofolate reductase C677T and A1298C polymorphisms and gastric cancer risk: a huge-GSEC review. AB - Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in the metabolism of folate, whose role in gastric carcinogenesis is controversial. The authors performed a meta-analysis and individual data pooled analysis of case-control studies that examined the association between C677T and A1298C polymorphisms (the former being associated with low folate serum levels) and gastric cancer (meta analyses: 16 studies, 2,727 cases and 4,640 controls for C677T and seven studies, 1,223 cases and 2,015 controls for A1298C; pooled analyses: nine studies, 1,540 cases and 2,577 controls for C677T and five studies, 1,146 cases and 1,549 controls for A1298C). An increased risk was found for MTHFR 677 TT in the meta analysis (odds ratio (OR) = 1.52, 95% confidence interval (CI): 1.31, 1.77) and pooled analysis (OR = 1.49, 95% CI: 1.14, 1.95). No association resulted for MTHFR 1298 CC (meta-OR = 0.94, 95% CI: 0.65, 1.35; pooled OR = 0.90, 95% CI: 0.69, 1.34). Results from the pooled analysis of four studies on C677T stratified according to folate levels showed an increased risk for individuals with low (OR = 2.05, 95% CI: 1.13, 3.72) versus high (OR = 0.95, 95% CI: 0.54, 1.67) folate levels. Overall, these findings support the hypothesis that folate plays a role in gastric carcinogenesis. PMID- 18162479 TI - Corticosteroids increase glutamine utilization in human splanchnic bed. AB - Glutamine is the most abundant amino acid in the body and is extensively taken up in gut and liver in healthy humans. To determine whether glucocorticosteroids alter splanchnic glutamine metabolism, the effect of prednisone was assessed in healthy volunteers using isotope tracer methods. Two groups of healthy adults received 5-h intravenous infusions of l-[1-(14)C]leucine and l [(2)H(5)]glutamine, along with q. 20 min oral sips of tracer doses of l-[1 (13)C]glutamine in the fasting state, either 1) at baseline (control group; n = 6) or 2) after a 6-day course of 0.8 mg.kg(-1).day(-1) prednisone (prednisone group; n = 8). Leucine and glutamine appearance rates (Ra) were determined from plasma [1-(14)C]ketoisocaproate and [(2)H(5)]glutamine, respectively, and leucine and glutamine oxidation from breath (14)CO(2) and (13)CO(2), respectively. Splanchnic glutamine extraction was estimated by the fraction of orally administered [(13)C]glutamine that failed to appear into systemic blood. Prednisone treatment 1) did not affect leucine Ra or leucine oxidation; 2) increased plasma glutamine Ra, mostly owing to enhanced glutamine de novo synthesis (medians +/- interquartiles, 412 +/- 61 vs. 280 +/- 190 mumol.kg(-1).h( 1), P = 0.003); and 3) increased the fraction of orally administered glutamine undergoing extraction in the splanchnic territory (means +/- SE 64 +/- 6 vs. 42 +/- 12%, P < 0.05), without any change in the fraction of glutamine oxidized (means +/- SE, 75 +/- 4 vs. 77 +/- 4%, not significant). We conclude that high dose glucocorticosteroids increase in splanchnic bed the glutamine requirements. The role of such changes in patients receiving chronic corticoid treatment for inflammatory diseases or suffering from severe illness remains to be determined. PMID- 18162480 TI - The cross-bridge cycle and skeletal muscle fatigue. AB - The functional correlates of fatigue observed in both animals and humans during exercise include a decline in peak force (P0), maximal velocity, and peak power. Establishing the extent to which these deleterious functional changes result from direct effects on the myofilaments is facilitated through understanding the molecular mechanisms of the cross-bridge cycle. With actin-myosin binding, the cross-bridge transitions from a weakly bound low-force state to a strongly bound high-force state. Low pH reduces the number of high-force cross bridges in fast fibers, and the force per cross bridge in both fast and slow fibers. The former is thought to involve a direct inhibition of the forward rate constant for transition to the strong cross-bridge state. In contrast, inorganic phosphate (Pi) is thought to reduce P0 by accelerating the reversal of this step. Both H+ and Pi decrease myofibrillar Ca2+ sensitivity. This effect is particularly important as the amplitude of the Ca2+ transient falls with fatigue. The inhibitory effects of low pH and high Pi on P0 are reduced as temperature increases from 10 to 30 degrees C. However, the H+-induced depression of peak power in the slow fiber type, and Pi inhibition of myofibrillar Ca2+ sensitivity in slow and fast fibers, are greater at high compared with low temperature. Thus the depressive effects of H+ and Pi at in vivo temperatures cannot easily be predicted from data collected below 25 degrees C. In vitro, reactive oxygen species reduce myofibrillar Ca2+ sensitivity; however, the importance of this mechanism during in vivo exercise is unknown. PMID- 18162481 TI - Sex-specific influence of aging on exercising leg blood flow. AB - Our previous work suggests that healthy human aging is associated with sex specific differences in leg vascular responses during large muscle mass exercise (2-legged cycling) (Proctor DN, Parker BA. Microcirculation 13: 315-327, 2006). The present study determined whether age x sex interactions in exercising leg hemodynamics persist during small muscle mass exercise that is not limited by cardiac output. Thirty-one young (20-30 yr; 15 men/16 women) and 31 older (60-79 yr; 13 men/18 women) healthy, normally active adults performed graded single-leg knee extensions to maximal exertion. Femoral artery blood velocity and diameter (Doppler ultrasound), heart rate (ECG), and beat-to-beat arterial blood pressure (mean arterial pressure, radial artery tonometry) were measured during each 3-min work rate (4.8 and 8 W/stage for women and men, respectively). The results (means +/- SE) were as follows. Despite reduced resting leg blood flow and vascular conductance, older men exhibited relatively preserved exercising leg hemodynamic responses. Older women, by contrast, exhibited attenuated hyperemic (young: 52 +/ 3 ml.min(-1).W(-1); vs. older: 40 +/- 4 ml.min(-1).W(-1); P = 0.02) and vasodilatory responses (young: 0.56 +/- 0.06 ml.min(-1).mmHg(-1).W(-1) vs. older: 0.37 +/- 0.04 ml.min(-1).mmHg(-1) W(-1); P < 0.01) to exercise compared with young women. Relative (percentage of maximal) work rate comparisons of all groups combined also revealed attenuated vasodilator responses in older women (P < 0.01 for age x sex x work rate interaction). These sex-specific age differences were not abolished by consideration of hemoglobin, quadriceps muscle, muscle recruitment, and mechanical influences on muscle perfusion. Collectively, these findings suggest that local factors contribute to the sex-specific effects of aging on exercising leg hemodynamics in healthy adults. PMID- 18162482 TI - Maintenance of gasping and restoration of eupnea after hypoxia is impaired following blockers of alpha1-adrenergic receptors and serotonin 5-HT2 receptors. AB - In severe hypoxia or ischemia, normal eupneic breathing fails and is replaced by gasping. Gasping serves as part of a process of autoresuscitation by which eupnea is reestablished. Medullary neurons, having a burster, pacemaker discharge, underlie gasping. Conductance through persistent sodium channels is essential for the burster discharge. This conductance is modulated by norepinephrine, acting on alpha 1-adrenergic receptors, and serotonin, acting on 5-HT2 receptors. We hypothesized that blockers of 5-HT2 receptors and alpha 1-adrenergic receptors would alter autoresuscitation. The in situ perfused preparation of the juvenile rat was used. Integrated phrenic discharge was switched from an incrementing pattern, akin to eupnea, to the decrementing pattern comparable to gasping in hypoxic hypercapnia. With a restoration of hyperoxic normocapnia, rhythmic, incrementing phrenic discharge returned within 10 s in most preparations. Following addition of blockers of alpha 1-adrenergic receptors (WB-4101, 0.0625 0.500 microM) and/or blockers of 5-HT2 (ketanserin, 1.25-10 microM) or multiple 5 HT receptors (methysergide, 3.0-10 microM) to the perfusate, incrementing phrenic discharge continued. Fictive gasping was still induced, although it ceased after significantly fewer decrementing bursts than in preparations than received no blockers. Moreover, the time for recovery of rhythmic activity was significantly prolonged. This prolongation was in excess of 100 s in all preparations that received both WB-4101 (above 0.125 microM) and methysergide (above 2.5 microM). We conclude that activation of adrenergic and 5-HT2 receptors is important to sustain gasping and to restore rhythmic respiratory activity after hypoxia induced depression. PMID- 18162483 TI - Respiration during feeding on solid food: alterations in breathing during mastication, pharyngeal bolus aggregation, and swallowing. AB - During feeding, solid food is chewed and propelled to the oropharynx, where the bolus gradually aggregates while the larynx remains open and breathing continues. The aggregated bolus in the valleculae is exposed to respiratory airflow, yet aspiration is rare in healthy individuals. The mechanism for preventing aspiration during bolus aggregation is unclear. One possibility is that alterations in the pattern of respiration during feeding could help prevent inhalation of food from the pharynx. We hypothesized that respiration was inhibited during bolus aggregation in the valleculae. Videofluorography was performed on 10 healthy volunteers eating solid foods with barium. Respiration was monitored concurrently with plethysmography and nasal air pressure. The timing of events during mastication, food transport, pharyngeal bolus aggregation, and swallowing were measured in relation to respiration. Respiratory cycle duration decreased during chewing (P < 0.001) but increased with swallowing (P < 0.001). During 66 recordings of vallecular bolus aggregation, there was inspiration in 8%, expiration in 41%, a pause in breathing in 17%, and multiple phases (including inspiration) in 35%. Out of 98 swallows, 47% started in the expiratory phase and 50% started during a pause in breathing, irrespective of bolus aggregation in the valleculae. Plethysmography was better than nasal manometry for determining the end of active expiration during feeding and swallowing with solid food. The hypothesis is rejected in that respiration was not inhibited during bolus aggregation. These findings suggest that airflow through the pharynx does not have a role in preventing aspiration during bolus aggregation in the oropharynx. PMID- 18162484 TI - Measuring endothelial glycocalyx dimensions in humans: a potential novel tool to monitor vascular vulnerability. AB - The endothelial glycocalyx is increasingly considered as an intravascular compartment that protects the vessel wall against pathogenic insults. The purpose of this study was to translate an established experimental method of estimating capillary glycocalyx dimension into a clinically useful tool and to assess its reproducibility in humans. We first evaluated by intravital microscopy the relation between the distance between the endothelium and erythrocytes, as a measure of glycocalyx thickness, and the transient widening of the erythrocyte column on glycocalyx compression by passing leukocytes in hamster cremaster muscle capillaries. We subsequently assessed sublingual microvascular glycocalyx thickness in 24 healthy men using orthogonal polarization spectral imaging. In parallel, systemic glycocalyx volume (using a previously published tracer dilution technique) as well as cardiovascular risk profiles were assessed. Estimates of microvascular glycocalyx dimension from the transient erythrocyte widening correlated well with the size of the erythrocyte-endothelium gap (r = 0.63). Measurements in humans were reproducible (0.58 +/- 0.16 and 0.53 +/- 0.15 microm, coefficient of variance 15 +/- 5%). In univariate analysis, microvascular glycocalyx thickness significantly correlated with systemic glycocalyx volume (r = 0.45), fasting plasma glucose (r = 0.43), and high-density lipoprotein cholesterol (r = 0.40) and correlated negatively with low-density lipoprotein cholesterol (r = -0.41) as well as body mass index (r = -0.45) (all P < 0.05). In conclusion, the dimension of the endothelial glycocalyx can be measured reproducibly in humans and is related to cardiovascular risk factors. It remains to be tested whether glycocalyx dimension can be used as an early marker of vascular damage and whether therapies aimed at glycocalyx repair can protect the vasculature against pathogenic challenges. PMID- 18162485 TI - Effects of aggression replacement training in young offender institutions. AB - This article reports a study where aggression replacement training (ART), combined with token economy, was compared with relationally oriented treatment at four residential treatment units in a nonrandomized design. In all, 57 adolescents in the ages between 16 and 19 participated. Outcome was measured as weighted indices of sentences and police suspicion reports. The results show no differences between the treatment models. In a separate analysis, the hypotheses were tested that those adolescents would relapse less frequently who admitted their crimes at intake or who could talk about guilt for their criminal acts. These hypotheses were refuted as main effects, but an interaction effect was found that suggests that adolescents with less consciousness of guilt got better results at the ART institutions. Case descriptions suggest that a more individualized approach to treatment, where ART is used for those adolescents who are motivated for it, would give better results. PMID- 18162486 TI - Genetic deletion or pharmacological inhibition of cyclooxygenase-1 attenuate lipopolysaccharide-induced inflammatory response and brain injury. AB - Cyclooxygenase (COX) -1 and -2 metabolize arachidonic acid to prostanoids and reactive oxygen species, major players in the neuroinflammatory process. While most reports have focused on the inducible isoform, COX-2, the contribution of COX-1 to the inflammatory response is unclear. In the present study, the contribution of COX-1 in the neuroinflammatory response to intracerebroventricular lipopolysaccharide (LPS) was investigated using COX-1 deficient (COX-1(-/-)) mice or wild-type (COX-1(+/+)) mice pretreated with SC 560, a selective COX-1 inhibitor. Twenty-four hours after lipopolysaccharide (LPS) injection, COX-1(-/-) mice showed decreased protein oxidation and LPS induced neuronal damage in the hippocampus compared with COX-1(+/+) mice. COX-1( /-) mice showed a significant reduction of microglial activation, proinflammatory mediators, and expression of COX-2, inducible NOS, and NADPH oxidase. The transcriptional down-regulation of cytokines and other inflammatory markers in COX-1(-/-) mice was mediated by a reduced activation of NF-kappaB and signal transducer and activator of transcription 3. Administration of SC-560 prior to LPS injection also attenuated the neuroinflammatory response by decreasing brain levels of prostaglandin (PG)E(2), PGD(2), PGF(2alpha), and thromboxane B(2), as well as the expression of proinflammatory cytokines and chemokine. These findings suggest that COX-1 plays a previously unrecognized role in neuroinflammatory damage. PMID- 18162487 TI - Genetic variability in the SNCA gene influences alpha-synuclein levels in the blood and brain. AB - Genetic variability in the promoter and 3' region of the SNCA gene coding alpha synuclein modulates the risk to develop sporadic Parkinson's disease (PD). Whether this is mediated by regulating alpha-synuclein expression levels remains unknown. Therefore, we analyzed levels of alpha-synuclein in blood and human post mortem brain tissue including the substantia nigra using quantitative real-time reverse transcriptase-polymerase chain reaction and enzyme linked immunosorbent assay in vivo. Single nucleotide polymorphism (SNP) rs356219, a tagging SNP for a disease-associated haplotype in the 3' region of the SNCA gene, has a significant effect on SNCA mRNA levels in the substantia nigra and the cerebellum. Further, the "protective" genotype 259/259 of the PD-associated promoter repeat NACP-Rep1 is associated with lower protein levels in blood than genotypes 261/261, 259/261, and 259/263. In conclusion, we provide evidence that alpha-synuclein levels are influenced by genetic variability in the promoter and 3' region of the SNCA gene in vivo. PMID- 18162488 TI - The gustatory pathway is involved in CD36-mediated orosensory perception of long chain fatty acids in the mouse. AB - The sense of taste informs the body about the quality of ingested foods. Tastant mediated signals are generated by a rise in free intracellular calcium levels ([Ca(2+)]i) in the taste bud cells and then are transferred to the gustatory area of brain via connections between the gustatory nerves (chorda tympani and glossopharyngeal nerves) and the nucleus of solitary tract in the brain stem. We have recently shown that lingual CD36 contributes to fat preference and early digestive secretions in the mouse. We show here that 1) the induction of an increase in [Ca(2+)]i by linoleic acid is CD36-dependent in taste receptor cells, 2) the spontaneous preference for or conversely conditioned aversion to linoleic acid requires intact gustatory nerves, and 3) the activation of gustatory neurons in the nucleus of the solitary tract elicited by a linoleic acid deposition on the tongue in wild-type mice cannot be reproduced in CD36-null animals. We conclude that the CD36-mediated perception of long-chain fatty acids involves the gustatory pathway, suggesting that the mouse may have a "taste" for fatty foods. This system would constitute a potential physiological advantage under conditions of food scarcity by leading the mouse to select and absorb fatty foods. However, it might also lead to a risk of obesity and associated diseases in a context of constantly abundant food. PMID- 18162489 TI - Vibrissa hair bulge houses two populations of skin epithelial stem cells distinct by their keratin profile. AB - Defining the properties of postnatal stem cells is of interest given their relevance for tissue homeostasis and therapeutic applications, such as skin tissue engineering for burn patients. In hair follicles, the bulge region of the outer root sheath houses stem cells. We show that explants from the prominent bulge area, but not the bulb, in rodent vibrissa follicles can produce epidermis in a skin model of tissue engineering. Using morphological criteria and keratin expression, we typified epithelial stem cells of vibrissa bulge. Two types of slow-cycling cells (Bb, Bs1) featuring a high colony-forming capacity occur in the bulge. Bb cells are located in the outermost basal layer, express K5, K15, K17, and K19, and feature a loosely organized keratin network. Bs1 cells localize to the suprabasal layers proximal to Bb cells and express K5/K17, correlating with a network of densely bundled filaments. These prominent bundles are missing in K17-null mice, which lack vibrissa. Atypically, both the Bb and Bs1 keratinocytes lack K14 expression. These findings show heterogeneity within the hair follicle stem cell repository, establish that a subset of slow-cycling cells are suprabasal in location, and point to a special role for K5/K17 filaments in a newly defined subset of stem cells. Our results are discussed in the context of long-term survival of engineered tissues after grafting that requires the presence of stem cells. PMID- 18162490 TI - Inflammatory arthritis and work disability: what is the role of occupational medicine? PMID- 18162492 TI - Safe systems of work are needed for the diagnosis of occupational mental illness. PMID- 18162493 TI - Cardiorespiratory fitness as a predictor of cancer mortality among men with pre diabetes and diabetes. AB - OBJECTIVE: The purpose of this article was to examine the risk of cancer mortality across levels of fitness and to examine the fitness-mortality relation for site-specific cancers in men with pre-diabetes and diabetes. RESEARCH DESIGN AND METHODS: We examined the fitness-mortality relation for all-cause and site specific cancer mortality among 18,858 men with pre-diabetes and 2,805 men with diabetes (aged 46.3 +/- 9.7 years [mean +/- SD]) from the Aerobics Center Longitudinal Study. We identified 719 cancer deaths during 354,558 person-years of risk. The duration of follow-up was 16.4 +/- 7.8 years (range <1-30.0 years). RESULTS: In men with pre-diabetes, moderate (hazard ratio 0.71 [95% CI 0.57 0.88]) and high fitness (0.76 [0.60-0.96]) were associated with lower risks of cancer mortality compared with the low-fit group in a model adjusted for age, examination year, smoking, alcohol use, fasting glucose concentration, previous cancer, and BMI. Similarly, for individuals with diabetes, moderate (0.53 [0.35 0.82]) and high fitness (0.44 [0.26-0.73]) were associated with lower risks of cancer mortality compared with the low-fit group. Among all men, being fit was associated with a lower risk of mortality from gastrointestinal (0.55 [0.39 0.77]), colorectal (0.53 [0.30-0.96]), liver (0.22 [0.07-0.71]), and lung cancer (0.43 [0.30-0.60]). CONCLUSIONS: In men with pre-diabetes and diabetes, higher levels of cardiorespiratory fitness were associated with lower risk of cancer mortality, particularly as a result of cancers of the gastrointestinal tract, compared with those who had low levels of fitness. PMID- 18162494 TI - Comparison of negative pressure wound therapy using vacuum-assisted closure with advanced moist wound therapy in the treatment of diabetic foot ulcers: a multicenter randomized controlled trial. AB - OBJECTIVE: The purpose of this study was to evaluate safety and clinical efficacy of negative pressure wound therapy (NPWT) compared with advanced moist wound therapy (AMWT) to treat foot ulcers in diabetic patients. RESEARCH DESIGN AND METHODS: This multicenter randomized controlled trial enrolled 342 patients with a mean age of 58 years; 79% were male. Complete ulcer closure was defined as skin closure (100% reepithelization) without drainage or dressing requirements. Patients were randomly assigned to either NPWT (vacuum-assisted closure) or AMWT (predominately hydrogels and alginates) and received standard off-loading therapy as needed. The trial evaluated treatment until day 112 or ulcer closure by any means. Patients whose wounds achieved ulcer closure were followed at 3 and 9 months. Each study visit included closure assessment by wound examination and tracings. RESULTS: A greater proportion of foot ulcers achieved complete ulcer closure with NPWT (73 of 169, 43.2%) than with AMWT (48 of 166, 28.9%) within the 112-day active treatment phase (P = 0.007). The Kaplan-Meier median estimate for 100% ulcer closure was 96 days (95% CI 75.0-114.0) for NPWT and not determinable for AMWT (P = 0.001). NPWT patients experienced significantly (P = 0.035) fewer secondary amputations. The proportion of home care therapy days to total therapy days for NPWT was 9,471 of 10,579 (89.5%) and 12,210 of 12,810 (95.3%) for AMWT. In assessing safety, no significant difference between the groups was observed in treatment-related complications such as infection, cellulitis, and osteomyelitis at 6 months. CONCLUSIONS: NPWT appears to be as safe as and more efficacious than AMWT for the treatment of diabetic foot ulcers. PMID- 18162495 TI - Skin replacement therapies for diabetic foot ulcers: systematic review and meta analysis. PMID- 18162496 TI - Maternal insulin therapy increases fetal endothelial progenitor cells during diabetic pregnancy. PMID- 18162497 TI - Plasma concentration of asymmetric dimethylarginine (ADMA) predicts cardiovascular morbidity and mortality in type 1 diabetic patients with diabetic nephropathy. AB - OBJECTIVE: To investigate whether circulating asymmetric dimethylarginine (ADMA) levels are predictive of cardiovascular events, decline in glomerular filtration rate (GFR), end-stage renal disease (ESRD), and all-cause mortality in type 1 diabetic patients. RESEARCH DESIGN AND METHODS: We performed a prospective observational follow-up study including 397 type 1 diabetic patients with overt diabetic nephropathy (243 men aged 42.1 +/- 10.5 years, GFR 76 +/- 34 ml/min per 1.73 m(2)) and a control group of 175 patients with longstanding type 1 diabetes and persistent normoalbuminuria (104 men aged 42.7 +/- 9.7 years, duration of diabetes 27.7 +/- 8.3 years). Patients were followed for a median 11.3 years (range 0.0-12.9) with yearly measurements of GFR ((51)Cr-EDTA plasma clearance) in patients with diabetic nephropathy. Endpoints were fatal and nonfatal cardiovascular disease (CVD), decline in GFR, ESRD, and all-cause mortality. RESULTS: Among patients with diabetic nephropathy, 37 patients (19.4%) with ADMA levels below the median, compared with 79 patients (43.4%) above the median, suffered a major cardiovascular event during the follow-up period (P < 0.001). This effect persisted after adjustment for conventional CVD risk factors including baseline GFR (adjusted hazard ratio [HR] for elevated ADMA 2.05 [95% CI 1.31-3.20], P = 0.002). Furthermore, elevated ADMA levels predicted an increased rate of decline in GFR, development of ESRD, and all-cause mortality (P < 0.001). After adjustment for well-known progression promoters, including baseline GFR, the HR (adjusted) was 1.85 (95% CI 0.99-3.46, P = 0.055) for ESRD comparing upper and lower median ADMA levels. CONCLUSIONS: Plasma ADMA levels predict fatal and nonfatal cardiovascular events in patients with type 1 diabetic nephropathy. Furthermore, increased ADMA levels tend to contribute to increased risk of progressive diabetic kidney disease. PMID- 18162498 TI - Human insulin analog--induced lipoatrophy. AB - OBJECTIVE: To characterize the pathophysiology of recombinant human insulin induced lipoatrophy. RESEARCH DESIGN AND METHODS: We performed immunologic laboratory evaluation and skin testing for different insulin analogs and diluents in patients with type 1 diabetes and severe insulin-induced local lipoatrophy. Subcutaneous adipose tissue biopsies of areas of acute (7 days) and chronic insulin administration were examined. Topical sodium cromolyn was applied twice a day to atrophic areas and prophylactically to new sites of insulin administration. RESULTS: Subcutaneous adipose biopsies showed an elevated population of tryptase-positive, chymase-positive degranulated mast cells. Of five patients treated with topical sodium cromolyn, none had new lipoatrophic sites and four showed improvements in old lesions. CONCLUSIONS: Tryptase positive/chymase-postitive mast cells, known to be sensitive to sodium cromolyn, may contribute to the destructive immune process mediated in response to exogenous insulin. Mast cell stabilizing therapy with topical cromolyn may reverse early and prevent new lipoatrophic lesions. PMID- 18162500 TI - Biomarker discovery: proteome fractionation and separation in biological samples. AB - Proteomics, analogous with genomics, is the analysis of the protein complement present in a cell, organ, or organism at any given time. While the genome provides information about the theoretical status of the cellular proteins, the proteome describes the actual content, which ultimately determines the phenotype. The broad application of proteomic technologies in basic science and clinical medicine has the potential to accelerate our understanding of the molecular mechanisms underlying disease and may facilitate the discovery of new drug targets and diagnostic disease markers. Proteomics is a rapidly developing and changing scientific discipline, and the last 5 yr have seen major advances in the underlying techniques as well as expansion into new applications. Core technologies for the separation of proteins and/or peptides are one- and two dimensional gel electrophoresis and one- and two-dimensional liquid chromatography, and these are coupled almost exclusively with mass spectrometry. Proteomic studies have shown that the most effective analysis of even simple biological samples requires subfractionation and/or enrichment before protein identification by mass spectrometry. Selection of the appropriate technology or combination of technologies to match the biological questions is essential for maximum coverage of the selected subproteome and to ensure both the full interpretation and the downstream utility of the data. In this review, we describe the current technologies for proteome fractionation and separation of biological samples, based on our lab workflow for biomarker discovery and validation. PMID- 18162499 TI - Methods and approaches for the comprehensive characterization and quantification of cellular proteomes using mass spectrometry. AB - Proteomics has been proposed as one of the key technologies in the postgenomic era. So far, however, the comprehensive analysis of cellular proteomes has been a challenge because of the dynamic nature and complexity of the multitude of proteins in cells and tissues. Various approaches have been established for the analyses of proteins in a cell at a given state, and mass spectrometry (MS) has proven to be an efficient and versatile tool. MS-based proteomics approaches have significantly improved beyond the initial identification of proteins to comprehensive characterization and quantification of proteomes and their posttranslational modifications (PTMs). Despite these advances, there is still ongoing development of new technologies to profile and analyze cellular proteomes more completely and efficiently. In this review, we focus on MS-based techniques, describe basic approaches for MS-based profiling of cellular proteomes and analysis methods to identify proteins in complex mixtures, and discuss the different approaches for quantitative proteome analysis. Finally, we briefly discuss novel developments for the analysis of PTMs. Altered levels of PTM, sometimes in the absence of protein expression changes, are often linked to cellular responses and disease states, and the comprehensive analysis of cellular proteome would not be complete without the identification and quantification of the extent of PTMs of proteins. PMID- 18162501 TI - A framework to identify physiological responses in microarray-based gene expression studies: selection and interpretation of biologically relevant genes. AB - In whole genome microarray studies major gene expression changes are easily identified, but it is a challenge to capture small, but biologically important, changes. Pathway-based programs can capture small effects but may have the disadvantage of being restricted to functionally annotated genes. A structured approach toward the identification of major and small changes for interpretation of biological effects is needed. We present a structured approach, a framework, that addresses different considerations in 1) the identification of informative genes in microarray data sets and 2) the interpretation of their biological relevance. The steps of this framework include gene ranking, gene selection, gene grouping, and biological interpretation. Random forests (RF), which takes gene gene interactions into account, is examined to rank and select genes. For human, mouse, and rat whole genome arrays, less than half of the probes on the array are annotated. Consequently, pathway analysis tools ignore half of the information present in the microarray data set. The framework described takes all genes into account. RF is a useful tool to rank genes by taking interactions into account. Applying a permutation approach, we were able to define an objective threshold for gene selection. RF combined with self-organizing maps identified genes with coordinated but small gene expression responses that were not fully annotated but corresponded to the same biological process. The presented approach provides a flexible framework for biological interpretation of microarray data sets. It includes all genes in the data set, takes gene-gene interactions into account, and provides an objective threshold for gene selection. PMID- 18162502 TI - PPARGC1A variation associated with DNA damage, diabetes, and cardiovascular diseases: the Boston Puerto Rican Health Study. AB - OBJECTIVE: Individuals with type 2 diabetes exhibit higher DNA damage and increased risk of cardiovascular disease (CVD). However, mechanisms underlying the association between DNA damage and development of type 2 diabetes and CVD are not understood. We sought to link peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A), a master transcriptional regulator of mitochondrial oxidative phosphorylation and cellular energy metabolism, with DNA damage, type 2 diabetes, and CVD. RESEARCH DESIGN AND METHODS: We measured DNA damage as urinary 8-hydroxydeoxyguanosine (8-OHdG) concentration and examined the relationship between nine PPARGC1A genetic variants, DNA damage, type 2 diabetes, and self-reported CVD in 959 participants of the Boston Puerto Rican Health Study. RESULTS: With respect to urinary 8-OHdG, PPARGC1A variants showed significant association, and PPARGC1A haplotypes exhibited significant association after correction for multiple testing. Two independent PPARGC1A variants associated significantly with type 2 diabetes (odds ratios [ORs] 1.35 and 2.46; P = 0.045 and <0.001). Carriers of minor alleles of two other PPARGC1A variants, both in strong linkage disequilibrium and associated with lower DNA damage, showed lower prevalence of CVD (ORs 0.53 and 0.65; P = 0.030 and 0.175). Moreover, we found that physical activity correlated negatively with DNA damage. CONCLUSIONS: It is plausible that low physical activity combined with risk haplotyes contribute to the high prevalence of type 2 diabetes in this population. We propose that PPARGC1A influences development of type 2 diabetes and CVD via DNA damage. Increasing physical activity, which induces PPARGC1A expression, is a potential strategy to slow DNA damage, thereby decreasing the risk of CVD for individuals with type 2 diabetes. PMID- 18162503 TI - Evidence of interaction between PPARG2 and HNF4A contributing to variation in insulin sensitivity in Mexican Americans. AB - OBJECTIVE: We hypothesized that interaction between PPARG2 Pro12Ala and variants in the promoter region of HNF4A are associated with type 2 diabetes-related quantitative traits in Mexican-American families of a proband with previous gestational diabetes. RESEARCH DESIGN AND METHODS: The BetaGene project genotyped PPARG2 Pro12Ala and nine HNF4A single nucleotide polymorphisms (SNPs) in 473 individuals in 89 families. Members of the proband generation had fasting glucose <126 mg/dl and were phenotyped by oral and intravenous glucose tolerance tests. RESULTS: Neither PPARG2 Pro12Ala nor any of the nine HNF4A SNPs were independently associated with type 2 diabetes-related quantitative traits. However, the interaction between PPARG2 Pro12Ala and HNF4A rs2144908 was significantly associated with both insulin sensitivity (S(I)) (Bonferroni P = 0.0006) and 2-h insulin (Bonferroni P = 0.039). Subjects with at least one PPARG2 Ala allele and homozygous for the HNF4A rs2144908 A allele had 40% higher S(I) compared with individuals with at least one G allele. S(I) did not vary by rs2144908 genotype among PPARG2 Pro/Pro. The interaction result for S(I) was replicated by the Insulin Resistance Atherosclerosis Family Study (P = 0.018) in their San Antonio sample (n = 484) where subjects with at least one PPARG2 Ala allele and homozygous for the HNF4A rs2144908 A allele had a 29% higher S(I) compared with individuals with at least one G allele. However, the interaction was not replicated in their San Luis Valley sample (n = 496; P = 0.401). CONCLUSIONS: Together, these results suggest that variation in PPARG2 and HNF4A may interact to regulate insulin sensitivity in Mexican Americans at risk for type 2 diabetes. PMID- 18162504 TI - Separate impact of obesity and glucose tolerance on the incretin effect in normal subjects and type 2 diabetic patients. AB - OBJECTIVE: To quantitate the separate impact of obesity and hyperglycemia on the incretin effect (i.e., the gain in beta-cell function after oral glucose versus intravenous glucose). RESEARCH DESIGN AND METHODS: Isoglycemic oral (75 g) and intravenous glucose administration was performed in 51 subjects (24 with normal glucose tolerance [NGT], 17 with impaired glucose tolerance [IGT], and 10 with type 2 diabetes) with a wide range of BMI (20-61 kg/m(2)). C-peptide deconvolution was used to reconstruct insulin secretion rates, and beta-cell glucose sensitivity (slope of the insulin secretion/glucose concentration dose response curve) was determined by mathematical modeling. The incretin effect was defined as the oral-to-intravenous ratio of responses. In 8 subjects with NGT and 10 with diabetes, oral glucose appearance was measured by the double-tracer technique. RESULTS: The incretin effect on total insulin secretion and beta-cell glucose sensitivity and the GLP-1 response to oral glucose were significantly reduced in diabetes compared with NGT or IGT (P Golgi) vesicle trafficking, which is rescued by overexpression of a Rab GTPase that regulates ER ->Golgi trafficking. The homologous Rab1 rescues alpha-syn toxicity in dopaminergic neuronal models of PD. Here we investigate this conserved feature of alpha-syn pathobiology. In a cell-free system with purified transport factors alpha-syn inhibited ER-->Golgi trafficking in an alpha-syn dose-dependent manner. Vesicles budded efficiently from the ER, but their docking or fusion to Golgi membranes was inhibited. Thus, the in vivo trafficking problem is due to a direct effect of alpha-syn on the transport machinery. By ultrastructural analysis the earliest in vivo defect was an accumulation of morphologically undocked vesicles, starting near the plasma membrane and growing into massive intracellular vesicular clusters in a dose-dependent manner. By immunofluorescence/immunoelectron microscopy, these clusters were associated both with alpha-syn and with diverse vesicle markers, suggesting that alpha-syn can impair multiple trafficking steps. Other Rabs did not ameliorate alpha-syn toxicity in yeast, but RAB3A, which is highly expressed in neurons and localized to presynaptic termini, and RAB8A, which is localized to post-Golgi vesicles, suppressed toxicity in neuronal models of PD. Thus, alpha-syn causes general defects in vesicle trafficking, to which dopaminergic neurons are especially sensitive. PMID- 18162537 TI - Extracellular matrix production and calcium carbonate precipitation by coral cells in vitro. AB - The evolution of multicellularity in animals required the production of extracellular matrices that serve to spatially organize cells according to function. In corals, three matrices are involved in spatial organization: (i) an organic ECM, which facilitates cell-cell and cell-substrate adhesion; (ii) a skeletal organic matrix (SOM), which facilitates controlled deposition of a calcium carbonate skeleton; and (iii) the calcium carbonate skeleton itself, which provides the structural support for the 3D organization of coral colonies. In this report, we examine the production of these three matrices by using an in vitro culturing system for coral cells. In this system, which significantly facilitates studies of coral cell physiology, we demonstrate in vitro excretion of ECM by primary (nondividing) tissue cultures of both soft (Xenia elongata) and hard (Montipora digitata) corals. There are structural differences between the ECM produced by X. elongata cell cultures and that of M. digitata, and ascorbic acid, a critical cofactor for proline hydroxylation, significantly increased the production of collagen in the ECM of the latter species. We further demonstrate in vitro production of SOM and extracellular mineralized particles in cell cultures of M. digitata. Inductively coupled plasma mass spectrometry analysis of Sr/Ca ratios revealed the particles to be aragonite. De novo calcification was confirmed by following the incorporation of (45)Ca into acid labile macromolecules. Our results demonstrate the ability of isolated, differentiated coral cells to undergo fundamental processes required for multicellular organization. PMID- 18162538 TI - The N-end rule pathway is a sensor of heme. AB - The conjugation of arginine, by arginyl-transferase, to N-terminal aspartate, glutamate or oxidized cysteine is a part of the N-end rule pathway of protein degradation. We report that arginyl-transferase of either the mouse or the yeast Saccharomyces cerevisiae is inhibited by hemin (Fe(3+)-heme). Furthermore, we show that hemin inhibits arginyl-transferase through a redox mechanism that involves the formation of disulfide between the enzyme's Cys-71 and Cys-72 residues. Remarkably, hemin also induces the proteasome-dependent degradation of arginyl-transferase in vivo, thus acting as both a "stoichiometric" and "catalytic" down-regulator of the N-end rule pathway. In addition, hemin was found to interact with the yeast and mouse E3 ubiquitin ligases of the N-end rule pathway. One of substrate-binding sites of the yeast N-end rule's ubiquitin ligase UBR1 targets CUP9, a transcriptional repressor. This site of UBR1 is autoinhibited but can be allosterically activated by peptides that bear destabilizing N-terminal residues and interact with two other substrate-binding sites of UBR1. We show that hemin does not directly occlude the substrate-binding sites of UBR1 but blocks the activation of its CUP9-binding site by dipeptides. The N-end rule pathway, a known sensor of short peptides, nitric oxide, and oxygen, is now a sensor of heme as well. One function of the N-end rule pathway may be to coordinate the activities of small effectors, both reacting to and controlling the redox dynamics of heme, oxygen, nitric oxide, thiols, and other compounds, in part through conditional degradation of specific transcription factors and G protein regulators. PMID- 18162539 TI - Pressure-induced metallization of silane. AB - There is a great interest in electronic transitions in hydrogen-rich materials under extreme conditions. It has been recently suggested that the group IVa hydrides such as methane (CH(4)), silane (SiH(4)), and germane (GeH(4)) become metallic at far lower pressures than pure hydrogen at equivalent densities because the hydrogen is chemically compressed in group IVa hydride compounds. Here we report measurements of Raman and infrared spectra of silane under pressure. We find that SiH(4) undergoes three phase transitions before becoming opaque at 27-30 GPa. The vibrational spectra indicate the material transforms to a polymeric (framework) structure in this higher pressure range. Room-temperature infrared reflectivity data reveal that the material exhibits Drude-like metallic behavior above 60 GPa, indicating the onset of pressure-induced metallization. PMID- 18162540 TI - Binding of rapamycin analogs to calcium channels and FKBP52 contributes to their neuroprotective activities. AB - Rapamycin is an immunosuppressive immunophilin ligand reported as having neurotrophic activity. We show that modification of rapamycin at the mammalian target of rapamycin (mTOR) binding region yields immunophilin ligands, WYE-592 and ILS-920, with potent neurotrophic activities in cortical neuronal cultures, efficacy in a rodent model for ischemic stroke, and significantly reduced immunosuppressive activity. Surprisingly, both compounds showed higher binding selectivity for FKBP52 versus FKBP12, in contrast to previously reported immunophilin ligands. Affinity purification revealed two key binding proteins, the immunophilin FKBP52 and the beta1-subunit of L-type voltage-dependent Ca(2+) channels (CACNB1). Electrophysiological analysis indicated that both compounds can inhibit L-type Ca(2+) channels in rat hippocampal neurons and F-11 dorsal root ganglia (DRG)/neuroblastoma cells. We propose that these immunophilin ligands can protect neurons from Ca(2+)-induced cell death by modulating Ca(2+) channels and promote neurite outgrowth via FKBP52 binding. PMID- 18162541 TI - Modulation of CaV2.1 channels by Ca2+/calmodulin-dependent protein kinase II bound to the C-terminal domain. AB - Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is a key regulator of synaptic responses in the postsynaptic density, but understanding of its mechanisms of action in the presynaptic neuron is incomplete. Here we show that CaMKII constitutively associates with and modulates voltage-gated calcium (Ca(V))2.1 channels that conduct P/Q type Ca(2+) currents and initiate transmitter release. Both exogenous and brain-specific inhibitors of CaMKII accelerate voltage-dependent inactivation, cause a negative shift in the voltage dependence of inactivation, and reduce Ca(2+)-dependent facilitation of Ca(V)2.1 channels. The modulatory effects of CaMKII are reduced by a peptide that prevents binding to Ca(V)2.1 channels but not by a peptide that blocks catalytic activity, suggesting that binding rather than phosphorylation is responsible for modulation. Our results reveal a signaling complex formed by Ca(V)2.1 channels and CaMKII that regulates P/Q-type Ca(2+) current in neurons. We propose an "effector checkpoint" model for the control of Ca(2+) channel fitness for function that depends on association with CaMKII, SNARE proteins, and other effectors of Ca(2+) signals. This regulatory mechanism would be important in presynaptic nerve terminals, where Ca(V)2.1 channels initiate synaptic transmission and CaMKII has noncatalytic effects on presynaptic plasticity. PMID- 18162542 TI - Retinal progenitor cells can produce restricted subsets of horizontal cells. AB - Retinal progenitor cells have been shown to be multipotent throughout development. Similarly, many other structures of the developing central nervous system have been found to contain multipotent progenitor cells. Previous lineage studies did not address whether these multipotent progenitor cells were biased in their production of neuronal subtypes. This question is of interest because subtypes are the basis of distinct types of circuits. Here, lentivirus-mediated gene transfer was used to mark single retinal progenitor cells in vivo, and the different subtypes of horizontal cells (HCs) in each clone were quantified. Clones with two HCs consistently contained a single HC subtype, a pair of either H1 or H3 cells. This suggests that a multipotent progenitor cell produces a mitotic cell fated to make a terminal division that produces two HCs of only one subtype. This bias in production of one HC subtype suggests a previously undescribed mechanism of cell fate determination in at least a subset of retinal cells that involves decisions made by mitotic cells that are inherited in a symmetric manner by both neuronal daughter cells. PMID- 18162543 TI - Synthetic antibodies for specific recognition and crystallization of structured RNA. AB - Antibodies that bind protein antigens are indispensable in biochemical research and modern medicine. However, knowledge of RNA-binding antibodies and their application in the ever-growing RNA field is lacking. Here we have developed a robust approach using a synthetic phage-display library to select specific antigen-binding fragments (Fabs) targeting a large functional RNA. We have solved the crystal structure of the first Fab-RNA complex at 1.95 A. Capability in phasing and crystal contact formation suggests that the Fab provides a potentially valuable crystal chaperone for RNA. The crystal structure reveals that the Fab achieves specific RNA binding on a shallow surface with complementarity-determining region (CDR) sequence diversity, length variability, and main-chain conformational plasticity. The Fab-RNA interface also differs significantly from Fab-protein interfaces in amino acid composition and light chain participation. These findings yield valuable insights for engineering of Fabs as RNA-binding modules and facilitate further development of Fabs as possible therapeutic drugs and biochemical tools to explore RNA biology. PMID- 18162544 TI - Laminopathic mutations interfere with the assembly, localization, and dynamics of nuclear lamins. AB - Lamins are nuclear intermediate filament proteins and the major building blocks of the nuclear lamina. Besides providing nuclear shape and mechanical stability, lamins are required for chromatin organization, transcription regulation, DNA replication, nuclear assembly, nuclear positioning, and apoptosis. Mutations in human lamins cause many different heritable diseases, affecting various tissues and causing early aging. Although many of these mutations result in nuclear deformation, their effects on lamin filament assembly are unknown. Caenorhabditis elegans has a single evolutionarily conserved lamin protein, which can form stable 10-nm-thick filaments in vitro. To gain insight into the molecular basis of lamin filament assembly and the effects of laminopathic mutations on this process, we investigated mutations in conserved residues of the rod and tail domains that are known to cause various laminopathies in human. We show that 8 of 14 mutant lamins present WT-like assembly into filaments or paracrystals, whereas 6 mutants show assembly defects. Correspondingly, expressing these mutants in transgenic animals shows abnormal distribution of Ce-lamin, abnormal nuclear shape or change in lamin mobility. These findings help in understanding the role of individual residues and domains in laminopathy pathology and, eventually, promote the development of therapeutic interventions. PMID- 18162545 TI - Synthetic heterovalent inhibitors targeting recognition E3 components of the N end rule pathway. AB - Multivalent binding allows high selectivity and affinity in a ligand-protein interaction. The N-end rule pathway is a ubiquitin (Ub)-dependent proteolytic system in which specific E3s, called N-recognins, mediate ubiquitylation through the recognition of types 1 and 2, destabilizing N-terminal residues of substrates. We recently identified a set of E3 Ub ligases (named UBR1-UBR7) containing the 70-residue UBR box, and we demonstrated that UBR1, UBR2, UBR4, and UBR5 can bind to destabilizing N-terminal residues. To explore a model of heterovalent interaction to the N-recognin family, we synthesized the small molecule compound RF-C11, which bears two heterovalent ligands designed to target N-recognins, together with control molecules with two homovalent ligands. We demonstrate that heterovalent ligands of RF-C11 selectively and cooperatively bind cognate-binding sites of multiple N-recognins and thereby inhibit both types 1 and 2 N-end rule activities. Furthermore, the efficacy of heterovalent RF-C11 was substantially higher than homovalent inhibitors, which can target either a type 1 or type 2 site, providing the molecular basis of designing multivalent inhibitors for the control of specific intracellular pathways. In addition, RF C11 exhibited higher efficacy and stability, compared with dipeptides bearing destabilizing N-terminal residues, which are known competitive inhibitors of the pathway. We also used the heterovalent compound to study the function of N recognins in cardiac signaling. Using mouse and rat cardiomyocytes, we demonstrate that the N-end rule pathway has a cell-autonomous function in cardiac proliferation and hypertrophy, explaining our earlier results implicating the pathway in cardiac development and proteolysis of multiple cardiovascular regulators. PMID- 18162546 TI - Replication stress induces tumor-like microdeletions in FHIT/FRA3B. AB - Common fragile sites (CFSs) are loci that preferentially exhibit metaphase chromosome gaps and breaks after partial inhibition of DNA synthesis. The fragile site FRA3B, which lies within the FHIT tumor-suppressor gene, is a site of frequent heterozygous and homozygous deletions in many cancer cells and precancerous lesions. The great majority of FHIT and other CFS-associated gene rearrangements in tumors are submicroscopic, intralocus deletions of hundreds of kilobases that often result in inactivation of associated genes. Although CFS instability leads to chromosome gaps and breaks and translocations, there has been no direct evidence showing that CFS instability or replication stress can generate large submicroscopic deletions of the type seen in cancer cells. Here, we have produced FHIT/FRA3B deletions closely resembling those in tumors by exposing human-mouse chromosome 3 somatic hybrid cells to aphidicolin-mediated replication stress. Clonal cell populations were analyzed for deletions by using PCR, array comparative genomic hybridization (aCGH), and FISH. Thirteen percent to 23% of clones exhibited submicroscopic FHIT deletions spanning approximately 200-600 kb within FRA3B. Chromosomes with FRA3B deletions exhibited significantly decreased fragility of this locus, with a 2- to 12-fold reduction in metaphase gaps and breaks compared with controls. Sequence analysis showed no regions of homology at breakpoints and suggests involvement of NHEJ in generating the deletions. Our results demonstrate that replication stress induces a remarkably high frequency of tumor-like microdeletions that reduce fragility at a CFS in cultured cells and suggests that similar conditions during tumor formation lead to intralocus deletion and inactivation of genes at CFSs and perhaps elsewhere in the genome. PMID- 18162547 TI - Costs and benefits of cold acclimation in field-released Drosophila. AB - One way animals can counter the effects of climatic extremes is via physiological acclimation, but acclimating to one extreme might decrease performance under different conditions. Here, we use field releases of Drosophila melanogaster on two continents across a range of temperatures to test for costs and benefits of developmental or adult cold acclimation. Both types of cold acclimation had enormous benefits at low temperatures in the field; in the coldest releases only cold-acclimated flies were able to find a resource. However, this advantage came at a huge cost; flies that had not been cold-acclimated were up to 36 times more likely to find food than the cold-acclimated flies when temperatures were warm. Such costs and strong benefits were not evident in laboratory tests where we found no reduction in heat survival of the cold-acclimated flies. Field release studies, therefore, reveal costs of cold acclimation that standard laboratory assays do not detect. Thus, although physiological acclimation may dramatically improve fitness over a narrow set of thermal conditions, it may have the opposite effect once conditions extend outside this range, an increasingly likely scenario as temperature variability increases under global climate change. PMID- 18162548 TI - The varitint-waddler (Va) deafness mutation in TRPML3 generates constitutive, inward rectifying currents and causes cell degeneration. AB - Varitint-waddler (Va and Va(J)) mice are deaf and have vestibular impairment, with inner ear defects that include the degeneration and loss of sensory hair cells. The semidominant Va mutation results in an alanine-to-proline substitution at residue 419 (A419P) of the presumed ion channel TRPML3. Another allele, Va(J), has the A419P mutation in addition to an I362T mutation. We found that hair cells, marginal cells of stria vascularis, and other cells lining the cochlear and vestibular endolymphatic compartments express TRPML3. When heterologously expressed in LLC-PK1-CL4 epithelial cells, a culture model for hair cells, TRPML3 accumulated in lysosomes and in espin-enlarged microvilli that resemble stereocilia. We also demonstrated that wild-type TRPML3 forms channels that are blocked by Gd(3+), have a conductance of 50-70 pS and, like many other TRP channels, open at very positive potentials and thus rectify outwardly. In addition to this outward current, TRPML3(419P) and (I362T+A419P) generated a constitutive inwardly rectifying current that suggests a sensitivity to hyperpolarizing negative potentials and that depolarized the cells. Cells expressing TRPML3(A419P) or (I362T+A419P), but not wild-type TRPML3, died and were extruded from the epithelium in a manner reminiscent of degenerating hair cells in Va mice. The increased open probability of TRPML3(A419P) and (I362T+A419P) at physiological potentials likely underlies hair cell degeneration and deafness in Va and Va(J) mice. PMID- 18162549 TI - Phase diagram of water in carbon nanotubes. AB - A phase diagram of water in single-walled carbon nanotubes at atmospheric pressure is proposed, which summarizes ice structures and their melting points as a function of the tube diameter up to 1.7 nm. The investigation is based on extensive molecular dynamics simulations over numerous thermodynamic states on the temperature-diameter plane. Spontaneous freezing of water in the simulations and the analysis of ice structures at 0 K suggest that there exist at least nine ice phases in the cylindrical space, including those reported by x-ray diffraction studies and those unreported by simulation or experiment. Each ice has a structure that maximizes the number of hydrogen bonds under the cylindrical confinement. The results show that the melting curve has many local maxima, each corresponding to the highest melting point for each ice form. The global maximum in the melting curve is located at approximately 11 A, where water freezes in a square ice nanotube. PMID- 18162550 TI - Transgenic system for conditional induction and rescue of chronic myocardial hibernation provides insights into genomic programs of hibernation. AB - A key energy-saving adaptation to chronic hypoxia that enables cardiomyocytes to withstand severe ischemic insults is hibernation, i.e., a reversible arrest of contractile function. Whereas hibernating cardiomyocytes represent the critical reserve of dysfunctional cells that can be potentially rescued, a lack of a suitable animal model has hampered insights on this medically important condition. We developed a transgenic mouse system for conditional induction of long-term hibernation and a system to rescue hibernating cardiomyocytes at will. Via myocardium-specific induction (and, in turn, deinduction) of a VEGF sequestering soluble receptor, we show that VEGF is indispensable for adjusting the coronary vasculature to match increased oxygen consumption and exploit this finding to generate a hypoperfused heart. Importantly, ensuing ischemia is tunable to a level at which large cohorts of cardiomyocytes are driven to enter a hibernation mode, without cardiac cell death. Relieving the VEGF blockade even months later resulted in rapid revascularization and full recovery of contractile function. Furthermore, we show that left ventricular remodeling associated with hibernation is also fully reversible. The unique opportunity to uncouple hibernation from other ischemic heart phenotypes (e.g., infarction) was used to determine the genetic program of hibernation; uncovering hypoxia-inducible factor target genes associated with metabolic adjustments and induced expression of several cardioprotective genes. Autophagy, specifically self-digestion of mitochondria, was identified as a key prosurvival mechanism in hibernating cardiomyocytes. This system may lend itself for examining the potential utility of treatments to rescue dysfunctional cardiomyocytes and reverse maladaptive remodeling. PMID- 18162551 TI - Mechanism of activation of the Formin protein Daam1. AB - The Formin proteins are central players in mediating cytoskeletal reorganization and are epistatically positioned in a pathway downstream of Rho activation. These proteins exist in the cytoplasm in an autoinhibited state, which is mediated by intramolecular interactions between the amino-terminal GTPase binding domain (GBD) that encompasses the diaphanous inhibitory domain (DID) and the carboxyl terminal diaphanous autoregulatory domain (DAD). It has been proposed that the binding of Rho within the GBD releases this molecule from autoinhibition by disrupting the DID/DAD interactions. Here we report that Daam1 is not significantly activated by Rho binding but rather by its interaction with Dishevelled (Dvl). Removal of the DAD domain disrupts interactions between Dvl and Daam1, and the binding of Dvl to Daam1 disrupts the interaction between the GBD and DAD that mediates Daam1 autoinhibition. Mutations within or removal of the DAD converts Daam1 into an active protein that can induce Rho activation. We further demonstrate that Dvl synergizes with Daam1 to regulate gastrulation during Xenopus embryogenesis and that expression of activated Daam1 can rescue impaired convergent extension movements resulting from deregulated noncanonical Wnt signaling. Our studies together define the importance of a carboxyl-terminal binding partner, Dvl, that leads to the activation of Daam1. PMID- 18162552 TI - Tendons of myostatin-deficient mice are small, brittle, and hypocellular. AB - Tendons play a significant role in the modulation of forces transmitted between bones and skeletal muscles and consequently protect muscle fibers from contraction-induced, or high-strain, injuries. Myostatin (GDF-8) is a negative regulator of muscle mass. Inhibition of myostatin not only increases the mass and maximum isometric force of muscles, but also increases the susceptibility of muscle fibers to contraction-induced injury. We hypothesized that myostatin would regulate the morphology and mechanical properties of tendons. The expression of myostatin and the myostatin receptors ACVR2B and ACVRB was detectable in tendons. Surprisingly, compared with wild type (MSTN(+/+)) mice, the tendons of myostatin null mice (MSTN(-/-)) were smaller and had a decrease in fibroblast density and a decrease in the expression of type I collagen. Tendons of MSTN(-/-) mice also had a decrease in the expression of two genes that promote tendon fibroblast proliferation: scleraxis and tenomodulin. Treatment of tendon fibroblasts with myostatin activated the p38 MAPK and Smad2/3 signaling cascades, increased cell proliferation, and increased the expression of type I collagen, scleraxis, and tenomodulin. Compared with the tendons of MSTN(+/+) mice, the mechanical properties of tibialis anterior tendons from MSTN(-/-) mice had a greater peak stress, a lower peak strain, and increased stiffness. We conclude that, in addition to the regulation of muscle mass and force, myostatin regulates the structure and function of tendon tissues. PMID- 18162553 TI - Activity-dependent phosphorylation of Ser187 is required for SNAP-25-negative modulation of neuronal voltage-gated calcium channels. AB - Synaptosomal-associated protein of 25 kDa (SNAP-25) is a SNARE protein that regulates neurotransmission by the formation of a complex with syntaxin 1 and synaptobrevin/VAMP2. SNAP-25 also reduces neuronal calcium responses to stimuli, but neither the functional relevance nor the molecular mechanisms of this modulation have been clarified. In this study, we demonstrate that hippocampal slices from Snap25(+/-) mice display a significantly larger facilitation and that higher calcium peaks are reached after depolarization by Snap25(-/-) and Snap25(+/-) cultured neurons compared with wild type. We also show that SNAP-25b modulates calcium dynamics by inhibiting voltage-gated calcium channels (VGCCs) and that PKC phosphorylation of SNAP-25 at ser187 is essential for this process, as indicated by the use of phosphomimetic (S187E) or nonphosphorylated (S187A) mutants. Neuronal activity is the trigger that induces the transient phosphorylation of SNAP-25 at ser187. Indeed, enhancement of network activity increases the levels of phosphorylated SNAP-25, whereas network inhibition reduces the extent of protein phosphorylation. A transient peak of SNAP-25 phosphorylation also is detectable in rat hippocampus in vivo after i.p. injection with kainate to induce seizures. These findings demonstrate that differences in the expression levels of SNAP-25 impact on calcium dynamics and neuronal plasticity, and that SNAP-25 phosphorylation, by promoting inhibition of VGCCs, may mediate a negative feedback modulation of neuronal activity during intense activation. PMID- 18162554 TI - Activity of human hippocampal and amygdala neurons during retrieval of declarative memories. AB - Episodic memories allow us to remember not only that we have seen an item before but also where and when we have seen it (context). Sometimes, we can confidently report that we have seen something (familiarity) but cannot recollect where or when it was seen. Thus, the two components of episodic recall, familiarity and recollection, can be behaviorally dissociated. It is not clear, however, whether these two components of memory are represented separately by distinct brain structures or different populations of neurons in a single anatomical structure. Here, we report that the spiking activity of single neurons in the human hippocampus and amygdala [the medial temporal lobe (MTL)] contain information about both components of memory. We analyzed a class of neurons that changed its firing rate to the second presentation of a previously novel stimulus. We found that the neuronal activity evoked by the presentation of a familiar stimulus (during retrieval) distinguishes stimuli that will be successfully recollected from stimuli that will not be recollected. Importantly, the ability to predict whether a stimulus is familiar is not influenced by whether the stimulus will later be recollected. We thus conclude that human MTL neurons contain information about both components of memory. These data support a continuous strength of memory model of MTL function: the stronger the neuronal response, the better the memory. PMID- 18162555 TI - Instructive role of aPKCzeta subcellular localization in the assembly of adherens junctions in neural progenitors. AB - In the neurogenic phase of CNS development, the proliferating progenitors are found medially within the neuroepithelium. The adherens junctions on the apical membrane of proliferating neural progenitors allow for cell-cell adhesion and medial stratification. In contrast, differentiating neuronal precursors delaminate and migrate laterally, establishing the laminar layers. Apical adherens junctions also establish the apical-basal polarity in neural progenitors, which in turn is postulated to lead to asymmetric inheritance of cell fate determinants during neurogenic divisions. The signaling pathways and cellular mechanisms that regulate the assembly and asymmetric localization of adherens junctions in neural progenitors remain elusive. Here we show that atypical PKCzeta/lambda (aPKCzeta/lambda) localizes at the apical membrane of proliferating neural stem cells, but not postmitotic neuronal precursors, in the developing chicken neural tube. This precise subcellular compartmentalization of the kinase activity provides an instructive signal for apical assembly of adherens junctions in a PI3K, Rac/Cdc42 signaling-dependent pathway. Apical aPKCzeta coordinates neural stem cell proliferation and the overall stratification of cell types within the neural tube. PMID- 18162556 TI - Hip, an HP1-interacting protein, is a haplo- and triplo-suppressor of position effect variegation. AB - The Drosophila heterochromatin protein 1 (HP1) regulates epigenetic gene silencing and heterochromatin formation by promoting and maintaining chromatin condensation. Here we report the identification and characterization of an HP1 interacting protein (Hip). Hip interacts with HP1 in vitro and is associated with HP1 in vivo. This interaction is mediated by at least three independent but similar HP1-binding modules of the Hip protein. Hip and HP1 completely colocalize in the pericentric heterochromatin, and both haplo- and triplo-dosage mutations act as dominant suppressors of position effect variegation. These findings identify a player in heterochromatinization and suggest that Hip cooperates with HP1 in chromatin remodeling and gene silencing. PMID- 18162557 TI - The RCK2 domain of the human BKCa channel is a calcium sensor. AB - Large conductance voltage and Ca(2+)-dependent K(+) channels (BK(Ca)) are activated by both membrane depolarization and intracellular Ca(2+). Recent studies on bacterial channels have proposed that a Ca(2+)-induced conformational change within specialized regulators of K(+) conductance (RCK) domains is responsible for channel gating. Each pore-forming alpha subunit of the homotetrameric BK(Ca) channel is expected to contain two intracellular RCK domains. The first RCK domain in BK(Ca) channels (RCK1) has been shown to contain residues critical for Ca(2+) sensitivity, possibly participating in the formation of a Ca(2+)-binding site. The location and structure of the second RCK domain in the BK(Ca) channel (RCK2) is still being examined, and the presence of a high affinity Ca(2+)-binding site within this region is not yet established. Here, we present a structure-based alignment of the C terminus of BK(Ca) and prokaryotic RCK domains that reveal the location of a second RCK domain in human BK(Ca) channels (hSloRCK2). hSloRCK2 includes a high-affinity Ca(2+)-binding site (Ca bowl) and contains similar secondary structural elements as the bacterial RCK domains. Using CD spectroscopy, we provide evidence that hSloRCK2 undergoes a Ca(2+)-induced change in conformation, associated with an alpha-to-beta structural transition. We also show that the Ca bowl is an essential element for the Ca(2+)-induced rearrangement of hSloRCK2. We speculate that the molecular rearrangements of RCK2 likely underlie the Ca(2+)-dependent gating mechanism of BK(Ca) channels. A structural model of the heterodimeric complex of hSloRCK1 and hSloRCK2 domains is discussed. PMID- 18162558 TI - Stepwise dynamics of epitaxially growing single amyloid fibrils. AB - The assembly mechanisms of amyloid fibrils, tissue deposits in a variety of degenerative diseases, is poorly understood. With a simply modified application of the atomic force microscope, we monitored the growth, on mica surface, of individual fibrils of the amyloid beta25-35 peptide with near-subunit spatial and subsecond temporal resolution. Fibril assembly was polarized and discontinuous. Bursts of rapid (up to 300-nm(-1)) growth phases that extended the fibril by approximately 7 nm or its integer multiples were interrupted with pauses. Stepwise dynamics were also observed for amyloid beta1-42 fibrils growing on graphite, suggesting that the discontinuous assembly mechanisms may be a general feature of epitaxial amyloid growth. Amyloid assembly may thus involve fluctuation between a fast-growing and a blocked state in which the fibril is kinetically trapped because of intrinsic structural features. The used scanning force kymography method may be adapted to analyze the assembly dynamics of a wide range of linear biopolymers. PMID- 18162559 TI - Single-molecule tracking of mRNA exiting from RNA polymerase II. AB - Single-pair fluorescence resonance energy transfer was used to track RNA exiting from RNA polymerase II (Pol II) in elongation complexes. Measuring the distance between the RNA 5' end and three known locations within the elongation complex allows us determine its position by means of triangulation. RNA leaves the polymerase active center cleft via the previously proposed exit tunnel and then disengages from the enzyme surface. When the RNA reaches lengths of 26 and 29 nt, its 5' end associates with Pol II at the base of the dock domain. Because the initiation factor TFIIB binds to the dock domain and exit tunnel, exiting RNA may prevent TFIIB reassociation during elongation. RNA further extends toward the linker connecting to the polymerase C-terminal repeat domain (CTD), which binds the 5'-capping enzyme and other RNA processing factors. PMID- 18162560 TI - Aldosterone acts centrally to increase brain renin-angiotensin system activity and oxidative stress in normal rats. AB - Aldosterone acts upon mineralocorticoid receptors in the brain to increase blood pressure and sympathetic nerve activity, but the mechanisms are still poorly understood. We hypothesized that aldosterone increases sympathetic nerve activity by upregulating the renin-angiotensin system (RAS) and oxidative stress in the brain, as it does in peripheral tissues. In Sprague-Dawley rats, aldosterone (Aldo) or vehicle (Veh) was infused for 1 wk via an intracerebroventricular (ICV) cannula, while RU-28318 (selective mineralocorticoid receptor antagonist), Tempol (superoxide dismutase mimetic), losartan [angiotensin II type 1 receptor (AT(1)R) antagonist], or Veh was infused simultaneously via a second ICV cannula. After 1 wk of ICV Aldo, plasma norepinephrine was increased and mean arterial pressure was slightly elevated, but heart rate was unchanged. These effects were ameliorated by ICV infusion of RU-28318, Tempol or losartan. Aldo increased expression of AT(1)R and angiotensin-converting enzyme (ACE) mRNA in hypothalamic tissue. RU-28318 minimized and Tempol prevented the increase in AT(1)R mRNA; RU 28318 prevented the increase in ACE mRNA. Losartan had no effect on AT(1)R or ACE mRNA. Immunohistochemistry revealed Aldo-induced increases in dihydroethidium staining (indicating oxidative stress) and Fra-like activity (indicating neuronal excitation) in neurons of the hypothalamic paraventricular nucleus (PVN). RU 28318 prevented the increases in superoxide and Fra-like activity in PVN; Tempol and losartan minimized these effects. Acute ICV infusions of sarthran (AT(1)R antagonist) or Tempol produced greater sympathoinhibition in Aldo-treated than in Veh-treated rats. Thus aldosterone upregulates key elements of brain RAS and induces oxidative stress in the hypothalamus. Aldosterone may increase sympathetic nerve activity by these mechanisms. PMID- 18162561 TI - The effects of caloric restriction- and exercise-induced weight loss on left ventricular diastolic function. AB - Little is known about the effects of weight loss on diastolic function. Furthermore, it is not known whether both caloric restriction (CR)- and exercise (Ex)-induced weight loss have salutary effects on diastolic function. Therefore, we assessed the effects of yearlong CR (n = 12) and Ex (n = 13) interventions, which induced approximately 12% weight loss, on diastolic function in healthy, nonobese (body mass index = 23.5-29.9 kg/m2) men and women aged 50 to 60 yr. Recordings of Doppler transmitral flow and Doppler tissue imaging were acquired and analyzed by conventional approaches and a validated parameterized diastolic filling (PDF) formalism. Isovolumic relaxation time decreased after weight loss in both groups (P < 0.05). Septal peak early mitral annular velocity (E') increased (P < 0.01) and peak E-wave velocity/E' decreased (P < 0.05) after weight loss in the CR group. Based on the PDF-derived indexes, CR resulted in a decrease in global ventricular stiffness (k) and increases in longitudinal (septal annulus motion) stored elastic strain (chi'o), peak force (k'chi'o), and peak stored strain energy (1/2k'chi'o2). In the Ex group, k was unchanged, although septal chi'o and 1/2k'chi'o2 increased significantly and k'chi'o (P = 0.13) tended to increase. We conclude that weight loss, whether induced by CR or Ex, has salutary effects on diastolic function. PMID- 18162564 TI - Improving practice through research in and about assisted living: implications for a research agenda. PMID- 18162565 TI - Historical evolution of assisted living in the United States, 1979 to the present. AB - PURPOSE: This article provides a historical overview of the emergence of assisted living in the United States over a 25-year period to identify goals and key concepts that underpinned the emerging form of care. DESIGN AND METHODS: The method is historical analysis based on records and my own personal experiences in conceptualizing and implementing assisted living in Oregon and nationwide. RESULTS: I identified four time periods: (a) 1979 to 1985, when a paradigm shift occurred on both the East and West coasts, motivated by distaste for nursing facilities and idealistic values regarding residential environments, service capacity, and consumer-centered care philosophy; (b) 1986 to 1993, when providers, consumers, and state governments became interested and four identifiable types of assisted living (hybrid, hospitality, housing, and health care) appeared, each of which informed the evolution of assisted living; (c) 1994 to 2000, a period of expansion, Wall Street money, dilution of the ideals, and emerging quality concerns; a crisis of confidence and a crossroads for assisted living; (d) 2000 to the present, a time of regrouping, slow-down in growth, and reexamination of earlier efforts to define and set standards for assisted living. IMPLICATIONS: Well-conceptualized and designed research may provide a mechanism to suggest practice, regulatory, and payment models. I recommend that researchers conduct studies from the values premises underlying the assisted living approach. PMID- 18162566 TI - The place of assisted living in long-term care and related service systems. AB - PURPOSE: The purpose of this article is to describe how assisted living (AL) fits with other long-term-care services. DESIGN AND METHODS: We analyzed the evolution of AL, including the populations served, the services offered, and federal and state policies that create various incentives or disincentives for using AL to replace other forms of care such as nursing home care or home care. RESULTS: Provider models that have emerged include independent senior housing with services, freestanding AL, nursing home expansion, and continuing care retirement communities. Some integrated health systems have also built AL into their array of services. Federal and state policy rules for financing and programs also shape AL, and states vary in how deliberately they try to create an array of options with specific roles for AL. Among state policies reviewed are reimbursement and rate-setting policies, admission and discharge criteria, and nurse practice policies that permit or prohibit various nursing tasks to be delegated in AL settings. Recent initiatives to increase flexible home care, such as nursing home transition programs, cash and counseling, and money-follows-the-person initiatives may influence the way AL emerges in a particular state. IMPLICATIONS: There is no single easy answer about the role of AL. To understand the current role and decide how to shape the future of AL, researchers need information systems that track the transitions individuals make during their long-term-care experiences along with information about the case-mix characteristics and service needs of the clientele. PMID- 18162567 TI - Definition and classification of assisted living. AB - PURPOSE: The purpose of this article is to discuss the benefits and limitations of, and considerations in, developing a typology of assisted living (AL). DESIGN AND METHODS: We conducted a review and comparison of nine AL typologies drawn from the literature. RESULTS: Typologies addressed matters related to the structure, process, population, and philosophy of AL to varying degrees. A lack of available data and different sampling frames hindered attempts to quantitatively compare the typologies. IMPLICATIONS: Typologies are potentially useful for consumers, practitioners, policy makers, and researchers. It is advisable to identify state-based typologies and then empirically determine types that have national representation. Stakeholders should consider the tradeoff between sensitivity and specificity and allow any resulting typology to anticipate ongoing evolution in the field of AL. PMID- 18162568 TI - Defining quality in assisted living: comparing apples, oranges, and broccoli. AB - PURPOSE: The purpose of this article is to discuss and describe various measures of quality, quality indicators, and uses of information on quality with specific reference to the role or purpose of assisted living. DESIGN AND METHODS: We reviewed a variety of major studies of assisted living quality. We elaborated models of assisted living based on differing goals or claims made for it by providers and policy makers. We then searched for available quality measures that might indicate whether assisted living was meeting those goals or living up to those claims. RESULTS: Any meaningful concept of "quality" must embrace a variety of dimensions, including quality of care, quality of life, the physical environment, and resident rights. The ability to use a multidimensional concept of quality is complicated by the lack of consensus, confusion, and disagreement among consumers, providers, and regulators about the role of assisted living. This disagreement significantly confounds the task of comparing quality among assisted living settings and between assisted living and other types of long-term care. IMPLICATIONS: We propose ways that researchers may compare quality along dimensions claimed to be intrinsic to assisted living as part of an effort to inform consumer information systems, quality monitoring and assurance systems, and policy-relevant research. Such comparisons would vary, depending on the intended use of the indicators and role defined for assisted living. However, all uses contain structural, process, and outcome quality indicators, including direct feedback from interviews with residents that go beyond satisfaction measures. PMID- 18162569 TI - Dementia and assisted living. AB - PURPOSE: This article presents an overview of what is known about dementia services in assisted living settings and suggests areas for future research. DESIGN AND METHODS: We undertook a search of Medline, the Journals of Gerontology, and The Gerontologist. We then organized publications dealing with the target subject into 10 topic areas and reviewed them. RESULTS: The article describes the demographic characteristics of cognitively impaired residents in assisted living and related residential settings in the United States, the services they receive, and process and structural elements both in specialized dementia units and in integrated assisted living settings. Finally, we review the literature on methodological issues regarding research in this area. IMPLICATIONS: It is important to generate research on processes as well as outcomes, such as dignity, individualized and pleasurable experiences, and freedom from pain and discomfort. We make recommendations for both content areas that would benefit from further research as well as methodological approaches that will yield important information in this field. PMID- 18162570 TI - Physical environments of assisted living: research needs and challenges. AB - PURPOSE: This article aims to review research measures and findings related to physical environments of assisted living (AL) according to multiple conceptual perspectives--ecological, cultural, and Maslovian hierarchy. DESIGN AND METHODS: A literature and research review was undertaken with two foci: performance measures for physical environments, and environmental research findings themselves. RESULTS: The research review identified a variety of environmental studies with a broad scope of topics, including post-occupancy design multimethod approaches, homeyness, evolution of AL, services, quality of life as an outcome, aging in place, regulatory influences, and environmental design principles. Most studies were descriptive; few dealt with outcomes linked to the environment, and those that did often focused on dementia settings. Some large scale studies with environmental components suggested that physical designs would affect a resident's ability to age in place within an AL setting. Overall, the environmental research was sparse and often characterized by small samples, lack of longitudinal data, or lack of depth. IMPLICATIONS: The field needs research studies that show how resident and environmental characteristics interact to generate both quality-of-life and functioning outcomes, and it also needs work on measures to permit such studies. I suggest eight specific studies in targeted areas and recommend full post-occupancy evaluation studies to develop in-depth understanding about how a setting works for its users. Research on AL environments is most likely to be meaningful if it anchors itself in the study of housing rather than hospitals, nursing homes, and other health settings. PMID- 18162571 TI - Families and assisted living. AB - PURPOSE: Despite growing research on assisted living (AL) as a residential care option for older adults, the social ramifications of residents' transitions to AL are relatively unexplored. This article examines family involvement in AL, including family structures of residents, types of involvement from family members living outside the AL facility, and outcomes for these family members. DESIGN AND METHODS: We reviewed current literature utilizing the MEDLINE, PsycINFO, and CINAHL databases to identify AL studies that examined issues pertaining to families or informal care. Following the screening of abstracts, we retrieved 180 reports for further review and selected 62 studies for inclusion. RESULTS: Families visit residents frequently and provide a wide range of instrumental assistance but provide only minimal personal care. Studies of family outcomes indicated relatively high satisfaction but potential care burden as well. IMPLICATIONS: How family care and involvement occurs in AL in relation to formal care provision and whether various types of formal/informal care integration influence family outcomes remains unclear. We suggest a research agenda that attempts to tease out causal relationships for family involvement, differentiate family roles, and implement longitudinal analyses for a range of family outcomes. PMID- 18162572 TI - Improving health care for assisted living residents. AB - PURPOSE: The purpose of this article is to explore how medical care is delivered to older people in assisted living (AL) settings and to suggest ways for improving it. DESIGN AND METHODS: We present a review of the limited research available on health care for older AL residents and on building testable models of better ways to organize primary health care and other health services for AL residents. RESULTS: AL residents are frequently frail older persons who need good chronic care. The predominant care models today do not respond adequately to this challenge. Medical care for AL residents is currently practiced very much like that for persons living in the community. The potential for using the aggregation of patients has not been effectively tapped. We review some managed care models from other elements of long-term care to look for ways that might be adapted. However, the current funding approach emphasizes living settings rather than inherent client characteristics. IMPLICATIONS: A research agenda might include ways to improve communication between AL and medical providers and to get AL staff more actively involved in daily care. Research support might produce the data necessary to entice the Centers for Medicare & Medicaid Services into changing its current reimbursement policies to create a climate better suited to delivering good chronic disease care in AL facilities. PMID- 18162573 TI - Assisted living and special populations: what do we know about differences in use and potential access barriers? AB - PURPOSE: The purpose of this article was to review existing knowledge about assisted living (AL) use by traditionally underserved populations, including people of color, low-income people, and those living in rural communities. DESIGN AND METHODS: We reviewed methodologies and findings of research on AL and residential care up to June 2004. RESULTS: Some studies suggested that low-income people and people of color either are less likely to utilize AL or receive AL in settings with less space and amenities and/or lower scores in selected quality measures. Generalizations from national or multistate studies to the population have methodological limitations. Numerous topics remain unstudied, including the pricing of AL, the experience of choosing an AL facility, the presence of discriminatory admission and retention practices, and the role of resident preferences. Data on access to AL in rural areas are inconclusive. IMPLICATIONS: We offer recommendations for national surveys to better classify supportive housing settings. Furthermore, we suggest quantitative and qualitative studies to illuminate the experience of low-income, racial/ethnic minority, and rural populations in AL settings. PMID- 18162574 TI - Assisted living and residential care in Oregon: two decades of state policy, supply, and Medicaid participation trends. AB - PURPOSE: The study describes Oregon state policy and supply developments for licensed long-term-care settings, particularly apartment-style assisted living facilities and more traditional residential care facilities. DESIGN AND METHODS: Data came from a variety of sources, including state agency administrative records, other secondary data sources, and key informant interviews. Descriptive statistics examined public financing, Medicaid reimbursement, and licensed bed supply trends from 1986 to 2004, as well as Medicaid resident use between 1990 and 2004. RESULTS: Residential care expansion, combined with nursing facility contraction, has transformed Oregon's supply of licensed long-term-care settings in favor of less institutional options. State financing, reimbursement, and licensing policies varied across provider type, with greater public resources supporting growth of assisted living facilities. By 2004, such settings were more likely to be Medicaid providers than residential care facilities and had a higher proportion of Medicaid residents relative to available bed supply. IMPLICATIONS: State financing and reimbursement policies may play a role in stimulating the supply of apartment-style assisted living available to low-income and/or rural service users. Less favorable policy conditions may have unintended consequences for the supply and use of other residential care settings. PMID- 18162575 TI - Assisted living literature through May 2004: taking stock. AB - PURPOSE: This article assesses the state of research on assisted living (AL) from 1989 to May 2004. DESIGN AND METHODS: We undertook keyword searches for AL research and amplified these with searches of Web sites, conference proceedings, and follow-up inquiries. We annotated and coded the resultant items according to categories reflecting the research methods used and the topics studied. We did additional comparisons for 38 studies with quantitative data that permitted summarizing resident characteristics, settings, and entry and move-out patterns. RESULTS: The 411 identified items ranged across a large number of topics. Qualitative studies outnumbered quantitative ones, and longitudinal studies were rare. We found little standardization in the way variables were measured, making cross-study comparisons difficult. As AL research has become more common, some items are directed at studying ways to proceed within AL as opposed to globally commenting on the worth of AL as a service sector. IMPLICATIONS: The research base for AL has grown rapidly but is still underdeveloped. We recommend using more consistent sets of standardized measures in AL studies and reporting analyses based on them. We also recommend fuller reporting of details on sampling, time frames, and measures in AL research. PMID- 18162576 TI - Developing a research agenda for assisted living. AB - PURPOSE: We describe an approach to identifying knowledge gaps, research questions, and methodological issues for assisted living (AL) research. DESIGN AND METHODS: We undertook an inventory of AL literature and research in progress and commissioned background papers critiquing knowledge on selected subtopics. With an advisory committee, we identified a comprehensive list of researchable questions of potential utility to consumers, providers, and/or policy makers, which AL researchers then rated as to their importance. The preliminary work facilitated a structured working conference of AL researchers. RESULTS: The top five priority topics identified as a result of the polling before the conference were consumer preferences, cost and financing, developing an information system for consumer decision making, developing quality measures, and resident outcomes. From conference discussion, conferees added other emphasis areas and refined the original ones. They flagged lack of standardized definitions and measures as barriers to building an empirically based AL literature. Conferees also identified distinctions between research on AL as a whole and research on interventions within AL. IMPLICATIONS: In an emerging area in which the literature cannot yet support rigorous comparisons, meta-analysis, or consensus conferences, the systematic approaches, including assembling researchers who use widely different methods, generated substantial agreement on a research agenda. PMID- 18162577 TI - Relative structural and functional roles of multiple deubiquitylating proteins associated with mammalian 26S proteasome. AB - We determined composition and relative roles of deubiquitylating proteins associated with the 26S proteasome in mammalian cells. Three deubiquitylating activities were associated with the 26S proteasome: two from constituent subunits, Rpn11/S13 and Uch37, and one from a reversibly associated protein, Usp14. RNA interference (RNAi) of Rpn11/S13 inhibited cell growth, decreased cellular proteasome activity via disrupted 26S proteasome assembly, and inhibited cellular protein degradation. In contrast, RNAi of Uch37 or Usp14 had no detectable effect on cell growth, proteasome structure or proteolytic capacity, but accelerated cellular protein degradation. RNAi of both Uch37 and Usp14 also had no effect on proteasome structure or proteolytic capacity, but inhibited cellular protein degradation. Thus, proper proteasomal processing of ubiquitylated substrates requires Rpn11 plus either Uch37 or Usp14. Although the latter proteins feature redundant deubiquitylation functions, they also appear to exert noncatalyic effects on proteasome activity that are similar to but independent of one another. These results reveal unexpected functional relationships among multiple deubiquitylating proteins and suggest a model for mammalian 26S proteasome function whereby their concerted action governs proteasome function by linking deubiquitylation to substrate hydrolysis. PMID- 18162578 TI - The DEAD-box RNA helicase Ded1p affects and accumulates in Saccharomyces cerevisiae P-bodies. AB - Recent results suggest that cytoplasmic mRNAs can form translationally repressed messenger ribonucleoprotein particles (mRNPs) capable of decapping and degradation, or accumulation into cytoplasmic processing bodies (P-bodies), which can function as sites of mRNA storage. The proteins that function in transitions between the translationally repressed mRNPs that accumulate in P-bodies and mRNPs engaged in translation are largely unknown. Herein, we demonstrate that the yeast translation initiation factor Ded1p can localize to P-bodies. Moreover, depletion of Ded1p leads to defects in P-body formation. Overexpression of Ded1p results in increased size and number of P-bodies and inhibition of growth in a manner partially suppressed by loss of Pat1p, Dhh1p, or Lsm1p. Mutations that inactivate the ATPase activity of Ded1p increase the overexpression growth inhibition of Ded1p and prevent Ded1p from localizing in P-bodies. Combined with earlier work showing Ded1p can have a positive effect on translation, these results suggest that Ded1p is a bifunctional protein that can affect both translation initiation and P-body formation. PMID- 18162579 TI - Identification of novel human Cdt1-binding proteins by a proteomics approach: proteolytic regulation by APC/CCdh1. AB - In mammalian cells, Cdt1 activity is strictly controlled by multiple independent mechanisms, implying that it is central to the regulation of DNA replication during the cell cycle. In fact, unscheduled Cdt1 hyperfunction results in rereplication and/or chromosomal damage. Thus, it is important to understand its function and regulations precisely. We sought to comprehensively identify human Cdt1-binding proteins by a combination of Cdt1 affinity chromatography and liquid chromatography and tandem mass spectrometry analysis. Through this approach, we could newly identify 11 proteins, including subunits of anaphase-promoting complex/cyclosome (APC/C), SNF2H and WSTF, topoisomerase I and IIalpha, GRWD1/WDR28, nucleophosmin/nucleoplasmin, and importins. In vivo interactions of Cdt1 with APC/C(Cdh1), SNF2H, topoisomerase I and IIalpha, and GRWD1/WDR28 were confirmed by coimmunoprecipitation assays. A further focus on APC/C(Cdh1) indicated that this ubiquitin ligase controls the levels of Cdt1 during the cell cycle via three destruction boxes in the Cdt1 N-terminus. Notably, elimination of these destruction boxes resulted in induction of strong rereplication and chromosomal damage. Thus, in addition to SCF(Skp2) and cullin4-based ubiquitin ligases, APC/C(Cdh1) is a third ubiquitin ligase that plays a crucial role in proteolytic regulation of Cdt1 in mammalian cells. PMID- 18162580 TI - The same receptor, G protein, and mitogen-activated protein kinase pathway activate different downstream regulators in the alternative white and opaque pheromone responses of Candida albicans. AB - Candida albicans must undergo a switch from white to opaque to mate. Opaque cells then release mating type-specific pheromones that induce mating responses in opaque cells. Uniquely in C. albicans, the same pheromones induce mating incompetent white cells to become cohesive, form an adhesive basal layer of cells on a surface, and then generate a thicker biofilm that, in vitro, facilitates mating between minority opaque cells. Through mutant analysis, it is demonstrated that the pathways regulating the white and opaque cell responses to the same pheromone share the same upstream components, including receptors, heterotrimeric G protein, and mitogen-activated protein kinase cascade, but they use different downstream transcription factors that regulate the expression of genes specific to the alternative responses. This configuration, although common in higher, multicellular systems, is not common in fungi, and it has not been reported in Saccharomyces cerevisiae. The implications in the evolution of multicellularity in higher eukaryotes are discussed. PMID- 18162581 TI - RNA interference in J774 macrophages reveals a role for coronin 1 in mycobacterial trafficking but not in actin-dependent processes. AB - Macrophages are crucial for innate immunity, apoptosis, and tissue remodeling, processes that rely on the capacity of macrophages to internalize and process cargo through phagocytosis. Coronin 1, a member of the WD repeat protein family of coronins specifically expressed in leukocytes, was originally identified as a molecule that is recruited to mycobacterial phagosomes and prevents the delivery of mycobacteria to lysosomes, allowing these to survive within phagosomes. However, a role for coronin 1 in mycobacterial pathogenesis has been disputed in favor for its role in mediating phagocytosis and cell motility. In this study, a role for coronin 1 in actin-mediated cellular processes was addressed using RNA interference in the murine macrophage cell line J774. It is shown that the absence of coronin 1 in J774 macrophages expressing small interfering RNA constructs specific for coronin 1 does not affect phagocytosis, macropinocytosis, cell locomotion, or regulation of NADPH oxidase activity. However, in coronin 1 negative J774 cells, internalized mycobacteria were rapidly transferred to lysosomes and killed. Therefore, these results show that in J774 cells coronin 1 has a specific role in modulating phagosome-lysosome transport upon mycobacterial infection and that it is dispensable for most F-actin-mediated cytoskeletal rearrangements. PMID- 18162582 TI - Isc1p plays a key role in hydrogen peroxide resistance and chronological lifespan through modulation of iron levels and apoptosis. AB - The inositolphosphosphingolipid phospholipase C (Isc1p) of Saccharomyces cerevisiae belongs to the family of neutral sphingomyelinases that generates the bioactive sphingolipid ceramide. In this work the role of Isc1p in oxidative stress resistance and chronological lifespan was investigated. Loss of Isc1p resulted in a higher sensitivity to hydrogen peroxide that was associated with an increase in oxidative stress markers, namely intracellular oxidation, protein carbonylation, and lipid peroxidation. Microarray analysis showed that Isc1p deficiency up-regulated the iron regulon leading to increased levels of iron, which is known to catalyze the production of the highly reactive hydroxyl radicals via the Fenton reaction. In agreement, iron chelation suppressed hydrogen peroxide sensitivity of isc1Delta mutants. Cells lacking Isc1p also displayed a shortened chronological lifespan associated with oxidative stress markers and aging of parental cells was correlated with a decrease in Isc1p activity. The analysis of DNA fragmentation and caspase-like activity showed that Isc1p deficiency increased apoptotic cell death associated with oxidative stress and aging. Furthermore, deletion of Yca1p metacaspase suppressed the oxidative stress sensitivity and premature aging phenotypes of isc1Delta mutants. These results indicate that Isc1p plays an important role in the regulation of cellular redox homeostasis, through modulation of iron levels, and of apoptosis. PMID- 18162583 TI - Caveolin-1 and -2 interact with connexin43 and regulate gap junctional intercellular communication in keratinocytes. AB - Connexin43 (Cx43) has been reported to interact with caveolin (Cav)-1, but the role of this association and whether other members of the caveolin family bind Cx43 had yet to be established. In this study, we show that Cx43 coimmunoprecipitates and colocalizes with Cav-1 and Cav-2 in rat epidermal keratinocytes. The colocalization of Cx43 with Cav-1 was confirmed in keratinocytes from human epidermis in vivo. Our mutation and Far Western analyses revealed that the C-terminal tail of Cx43 is required for its association with Cavs and that the Cx43/Cav-1 interaction is direct. Our results indicate that newly synthesized Cx43 interacts with Cavs in the Golgi apparatus and that the Cx43/Cavs complex also exists at the plasma membrane in lipid rafts. Using overexpression and small interfering RNA approaches, we demonstrated that caveolins regulate gap junctional intercellular communication (GJIC) and that the presence of Cx43 in lipid raft domains may contribute to the mechanism modulating GJIC. Our results suggest that the Cx43/Cavs association occurs during exocytic transport, and they clearly indicate that caveolin regulates GJIC. PMID- 18162584 TI - StARD13(Dlc-2) RhoGap mediates ceramide activation of phosphatidylglycerolphosphate synthase and drug response in Chinese hamster ovary cells. AB - To identify genes involved in etoposide drug response, we used promoter trap mutagenesis to isolate an etoposide-resistant Chinese hamster ovary (CHO) cell line. This resistant CHO-K1 line, named E91, showed cross-resistance to C(2) ceramide (N-acetylsphingosine). The promoter trap retrovirus was found integrated into intron 1-2 of the Dlc-2 (Stard13) RhoGap gene. The E91 cells showed elevated guanosine triphosphate (GTP)-bound RhoA levels compared with the parental line, suggesting that retrovirus integration had inactivated one of the Dlc-2 RhoGap alleles. To test whether E91 cells were impaired in an intracellular ceramide regulated process not directly related to cell killing, we measured mitochondrial phosphatidylglycerolphosphate (PGP) synthase and phospholipase A2 enzyme activities in cells after C(2)-ceramide addition. Parental cells showed elevated enzyme activities after treatment with C(2)-ceramide or tumor necrosis factor alpha, but not the E91 cells. These results suggested that intracellular ceramide signaling was defective in E91 cells due to increased levels of active GTP-bound RhoA. RNA knockdown experiments of the Dlc2 RhoGap resulted in increased GTP bound RhoA and reduced induction of PGP synthase after C(2)-ceramide addition compared with controls. Expression of a dominant-negative RhoA in the E91 cell line allowed induction of PGP synthase by ceramide. The RNA interference knockdown cell line also showed increased etoposide resistance. This study is the first report for the regulation of a phospholipid biosynthetic enzyme through RhoGap expression. PMID- 18162585 TI - Functions of chloroplastic adenylate kinases in Arabidopsis. AB - Adenosine monophosphate kinase (AMK; adenylate kinase) catalyses the reversible formation of ADP by the transfer of one phosphate group from ATP to AMP, thus equilibrating adenylates. The Arabidopsis (Arabidopsis thaliana) genome contains 10 genes with an adenylate/cytidylate kinase signature; seven of these are identified as putative adenylate kinases. Encoded proteins of at least two members of this Arabidopsis adenylate kinase gene family are targeted to plastids. However, when the individual genes are disrupted, the phenotypes of both mutants are strikingly different. Although absence of AMK2 causes only 30% reduction of total adenylate kinase activity in leaves, there is loss of chloroplast integrity leading to small, pale-looking plantlets from embryo to seedling development. In contrast, no phenotype for disruption of the second plastid adenylate kinase was found. From this analysis, we conclude that AMK2 is the major activity for equilibration of adenylates and de novo synthesis of ADP in the plastid stroma. PMID- 18162586 TI - High temperature-induced abscisic acid biosynthesis and its role in the inhibition of gibberellin action in Arabidopsis seeds. AB - Suppression of seed germination at supraoptimal high temperature (thermoinhibiton) during summer is crucial for Arabidopsis (Arabidopsis thaliana) to establish vegetative and reproductive growth in appropriate seasons. Abscisic acid (ABA) and gibberellins (GAs) are well known to be involved in germination control, but it remains unknown how these hormone actions (metabolism and responsiveness) are altered at high temperature. Here, we show that ABA levels in imbibed seeds are elevated at high temperature and that this increase is correlated with up-regulation of the zeaxanthin epoxidase gene ABA1/ZEP and three 9-cis-epoxycarotenoid dioxygenase genes, NCED2, NCED5, and NCED9. Reverse-genetic studies show that NCED9 plays a major and NCED5 and NCED2 play relatively minor roles in high temperature-induced ABA synthesis and germination inhibition. We also show that bioactive GAs stay at low levels at high temperature, presumably through suppression of GA 20-oxidase genes, GA20ox1, GA20ox2, and GA20ox3, and GA 3-oxidase genes, GA3ox1 and GA3ox2. Thermoinhibition-tolerant germination of loss of-function mutants of GA negative regulators, SPINDLY (SPY) and RGL2, suggests that repression of GA signaling is required for thermoinibition. Interestingly, ABA-deficient aba2-2 mutant seeds show significant expression of GA synthesis genes and repression of SPY expression even at high temperature. In addition, the thermoinhibition-resistant germination phenotype of aba2-1 seeds is suppressed by a GA biosynthesis inhibitor, paclobutrazol. We conclude that high temperature stimulates ABA synthesis and represses GA synthesis and signaling through the action of ABA in Arabidopsis seeds. PMID- 18162587 TI - The NADPH-dependent thioredoxin reductase/thioredoxin system in germinating barley seeds: gene expression, protein profiles, and interactions between isoforms of thioredoxin h and thioredoxin reductase. AB - The NADPH-dependent thioredoxin reductase (NTR)/thioredoxin (Trx) system catalyzes disulfide bond reduction in the cytoplasm and mitochondrion. Trx h is suggested to play an important role in seed development, germination, and seedling growth. Plants have multiple isoforms of Trx h and NTR; however, little is known about the roles of the individual isoforms. Trx h isoforms from barley (Hordeum vulgare) seeds (HvTrxh1 and HvTrxh2) were characterized previously. In this study, two NTR isoforms (HvNTR1 and HvNTR2) were identified, enabling comparison of gene expression, protein appearance, and interaction between individual NTR and Trx h isoforms in barley embryo and aleurone layers. Although mRNA encoding both Trx h isoforms is present in embryo and aleurone layers, the corresponding proteins differed in spatiotemporal appearance. HvNTR2, but not HvNTR1, gene expression seems to be regulated by gibberellic acid. Recombinant HvNTR1 and HvNTR2 exhibited virtually the same affinity toward HvTrxh1 and HvTrxh2, whereas HvNTR2 has slightly higher catalytic activity than HvNTR1 with both Trx h isoforms, and HvNTR1 has slightly higher catalytic activity toward HvTrxh1 than HvTrxh2. Notably, both NTRs reduced Trx h at the acidic conditions residing in the starchy endosperm during germination. Interspecies reactions between the barley proteins and Escherichia coli Trx or Arabidopsis thaliana NTR, respectively, occurred with 20- to 90-fold weaker affinity. This first investigation of regulation and interactions between members of the NTR/Trx system in barley seed tissues suggests that different isoforms are differentially regulated but may have overlapping roles, with HvNTR2 and HvTrxh1 being the predominant isoforms in the aleurone layer. PMID- 18162588 TI - Functional coexpression of the mitochondrial alternative oxidase and uncoupling protein underlies thermoregulation in the thermogenic florets of skunk cabbage. AB - Two distinct mitochondrial energy dissipating systems, alternative oxidase (AOX) and uncoupling protein (UCP), have been implicated as crucial components of thermogenesis in plants and animals, respectively. To further clarify the physiological roles of AOX and UCP during homeothermic heat production in the thermogenic skunk cabbage (Symplocarpus renifolius), we identified the thermogenic cells and performed expression and functional analyses of these genes in this organism. Thermographic analysis combined with in situ hybridization revealed that the putative thermogenic cells surround the stamens in the florets of skunk cabbage and coexpress transcripts for SrAOX, encoding Symplocarpus AOX, and SrUCPb, encoding a novel UCP that lacks a fifth transmembrane segment. Mitochondria isolated from the thermogenic florets exhibited substantial linoleic acid (LA)-inducible uncoupling activities. Moreover, our results demonstrate that LA is capable of inhibiting the mitochondrial AOX pathway, whereas the proportion of pyruvate-stimulated AOX capacity was not significantly affected by LA. Intriguingly, the protein expression levels for SrAOX and SrUCPb were unaffected even when the ambient air temperatures increased from 10.3 degrees C to 23.1 degrees C or from 8.3 degrees C to 24.9 degrees C. Thus, our results suggest that functional coexpression of AOX and UCP underlies the molecular basis of heat production, and that posttranslational modifications of these proteins play a crucial role in regulating homeothermic heat production under conditions of natural ambient temperature fluctuations in skunk cabbage. PMID- 18162589 TI - Quantitative conversion of phytate to inorganic phosphorus in soybean seeds expressing a bacterial phytase. AB - Phytic acid (PA) contains the major portion of the phosphorus in the soybean (Glycine max) seed and chelates divalent cations. During germination, both minerals and phosphate are released upon phytase-catalyzed degradation of PA. We generated a soybean line (CAPPA) in which an Escherichia coli periplasmic phytase, the product of the appA gene, was expressed in the cytoplasm of developing cotyledons. CAPPA exhibited high levels of phytase expression, >or=90% reduction in seed PA, and concomitant increases in total free phosphate. These traits were stable, and, although resulted in a trend for reduced emergence and a statistically significant reduction in germination rates, had no effect on the number of seeds per plant or seed weight. Because phytate is not digested by monogastric animals, untreated soymeal does not provide monogastrics with sufficient phosphorus and minerals, and PA in the waste stream leads to phosphorus runoff. The expression of a cytoplasmic phytase in the CAPPA line therefore improves phosphorus availability and surpasses gains achieved by other reported transgenic and mutational strategies by combining in seeds both high phytase expression and significant increases in available phosphorus. Thus, in addition to its value as a high-phosphate meal source, soymeal from CAPPA could be used to convert PA of admixed meals, such as cornmeal, directly to utilizable inorganic phosphorus. PMID- 18162590 TI - Specificity of RCN1-mediated protein phosphatase 2A regulation in meristem organization and stress response in roots. AB - Protein dephosphorylation by the serine/threonine protein phosphatase 2A (PP2A) modulates a broad array of cellular functions. PP2A normally acts as a heterotrimeric holoenzyme complex comprising a catalytic subunit bound by regulatory A and B subunits. Characterization of the regulatory A subunit isoforms (ROOTS CURL IN NAPHTHYLPHTHALAMIC ACID1 [RCN1], PP2AA2, and PP2AA3) of Arabidopsis thaliana PP2A has shown that RCN1 plays a primary role in controlling root and hypocotyl PP2A activity in seedlings. Here we show that hypocotyl and root growth exhibit different requirements for RCN1-mediated regulation of PP2A activity. Roots of rcn1 mutant seedlings exhibit characteristic abnormalities in cell division patterns at the root apical meristem, as well as reduced growth under ionic, osmotic, and oxidative stress conditions. We constructed chimeric A subunit genes and found that restoration of normal root tip development in rcn1 plants requires both regulatory and coding sequences of RCN1, whereas the hypocotyl elongation defect of rcn1 plants can be complemented by either RCN1 or PP2AA3 transgenes. Furthermore, the RCN1 and PP2AA3 proteins exhibit ubiquitous subcellular localization patterns in seedlings and both associate with membrane compartments. Together, these results show that RCN1-containing PP2A has unique functions that cannot be attributed to isoform-specific expression and localization patterns. Postembryonic RCN1 function is required to maintain normal auxin distribution and stem cell function at the root apex. Our data show that RCN1-regulated phosphatase activity plays a unique role in regulating postembryonic root development and stress response. PMID- 18162591 TI - Arabidopsis branched-chain aminotransferase 3 functions in both amino acid and glucosinolate biosynthesis. AB - In Arabidopsis thaliana, transamination steps in the leucine biosynthetic and catabolic pathways and the methionine (Met) chain elongation cycle of aliphatic glucosinolate formation are catalyzed by branched-chain aminotransferases (BCATs) that are encoded by a small gene family of six members. One member of this family, the plastid-located BCAT3, was shown to participate in both amino acid and glucosinolate metabolism. In vitro activity tests with the recombinant protein identified highest activities with the 2-oxo acids of leucine, isoleucine, and valine, but also revealed substantial conversion of intermediates of the Met chain elongation pathway. Metabolite profiling of bcat3-1 single and bcat3-1/bcat4-2 double knockout mutants showed significant alterations in the profiles of both amino acids and glucosinolates. The changes in glucosinolate proportions suggest that BCAT3 most likely catalyzes the terminal steps in the chain elongation process leading to short-chain glucosinolates: the conversion of 5-methylthiopentyl-2-oxo and 6-methylthiohexyl-2-oxo acids to their respective Met derivatives, homomethionine and dihomo-methionine, respectively. The enzyme can also at least partially compensate for the loss of BCAT4, which catalyzes the initial step of Met chain elongation by converting Met to 4-methylthio-2 oxobutanoate. Our results show the interdependence of amino acid and glucosinolate metabolism and demonstrate that a single enzyme plays a role in both processes. PMID- 18162592 TI - PSY3, a new member of the phytoene synthase gene family conserved in the Poaceae and regulator of abiotic stress-induced root carotenogenesis. AB - Abscisic acid (ABA) plays a vital role in mediating abiotic stress responses in plants. De novo ABA biosynthesis involves cleavage of carotenoid precursors by 9 cis-epoxycarotenoid dioxygenase (NCED), which is rate controlling in leaves and roots; however, additional bottlenecks in roots must be overcome, such as biosynthesis of upstream carotenoid precursors. Phytoene synthase (PSY) mediates the first committed step in carotenoid biosynthesis; with PSY3 described here, maize (Zea mays) and other members of the Poaceae have three paralogous genes, in contrast to only one in Arabidopsis thaliana. PSY gene duplication has led to subfunctionalization, with each paralog exhibiting differential gene expression. We showed that PSY3 encodes a functional enzyme for which maize transcript levels are regulated in response to abiotic stresses, drought, salt, and ABA. Drought stressed roots showed elevated PSY3 transcripts and ABA, responses reversed by rehydration. By blocking root carotenoid biosynthesis with the maize y9 mutation, we demonstrated that PSY3 mRNA elevation correlates with carotenoid accumulation and that blocking carotenoid biosynthesis interferes with stress-induced ABA accumulation. In parallel, we observed elevated NCED transcripts and showed that, in contrast to dicots, root zeaxanthin epoxidase transcripts were unchanged. PSY3 was the only paralog for which transcripts were induced in roots and abiotic stress also affected leaf PSY2 transcript levels; PSY1 mRNA was not elevated in any tissues tested. Our results suggest that PSY3 expression influences root carotenogenesis and defines a potential bottleneck upstream of NCED; further examination of PSY3 in the grasses is of value for better understanding root specific stress responses that impact plant yield. PMID- 18162593 TI - Overexpression of AtMYB44 enhances stomatal closure to confer abiotic stress tolerance in transgenic Arabidopsis. AB - AtMYB44 belongs to the R2R3 MYB subgroup 22 transcription factor family in Arabidopsis (Arabidopsis thaliana). Treatment with abscisic acid (ABA) induced AtMYB44 transcript accumulation within 30 min. The gene was also activated under various abiotic stresses, such as dehydration, low temperature, and salinity. In transgenic Arabidopsis carrying an AtMYB44 promoter-driven beta-glucuronidase (GUS) construct, strong GUS activity was observed in the vasculature and leaf epidermal guard cells. Transgenic Arabidopsis overexpressing AtMYB44 is more sensitive to ABA and has a more rapid ABA-induced stomatal closure response than wild-type and atmyb44 knockout plants. Transgenic plants exhibited a reduced rate of water loss, as measured by the fresh-weight loss of detached shoots, and remarkably enhanced tolerance to drought and salt stress compared to wild-type plants. Microarray analysis and northern blots revealed that salt-induced activation of the genes that encode a group of serine/threonine protein phosphatases 2C (PP2Cs), such as ABI1, ABI2, AtPP2CA, HAB1, and HAB2, was diminished in transgenic plants overexpressing AtMYB44. By contrast, the atmyb44 knockout mutant line exhibited enhanced salt-induced expression of PP2C-encoding genes and reduced drought/salt stress tolerance compared to wild-type plants. Therefore, enhanced abiotic stress tolerance of transgenic Arabidopsis overexpressing AtMYB44 was conferred by reduced expression of genes encoding PP2Cs, which have been described as negative regulators of ABA signaling. PMID- 18162594 TI - GeBP and GeBP-like proteins are noncanonical leucine-zipper transcription factors that regulate cytokinin response in Arabidopsis. AB - Understanding the role of transcription factors (TFs) is essential in reconstructing developmental regulatory networks. The plant-specific GeBP TF family of Arabidopsis thaliana (Arabidopsis) comprises 21 members, all of unknown function. A subset of four members, the founding member GeBP and GeBP-like proteins (GPL) 1, 2, and 3, shares a conserved C-terminal domain. Here we report that GeBP/GPL genes represent a newly defined class of leucine-zipper (Leu zipper) TFs and that they play a redundant role in cytokinin hormone pathway regulation. Specifically, we demonstrate using yeast, in vitro, and split-yellow fluorescent protein in planta assays that GeBP/GPL proteins form homo- and heterodimers through a noncanonical Leu-zipper motif located in the C-terminal domain. A triple loss-of-function mutant of the three most closely related genes gebp gpl1 gpl2 shows a reduced sensitivity to exogenous cytokinins in a subset of cytokinin responses such as senescence and growth, whereas root inhibition is not affected. We find that transcript levels of type-A cytokinin response genes, which are involved in the negative feedback regulation of cytokinin signaling, are higher in the triple mutant. Using a GPL version that acts as a constitutive transcriptional activator, we show that the regulation of Arabidopsis response regulators (ARRs) is mediated by at least one additional, as yet unknown, repressor acting genetically downstream in the GeBP/GPL pathway. Our results indicate that GeBP/GPL genes encode a new class of unconventional Leu-zipper TF proteins and suggest that their role in the cytokinin pathway is to antagonize the negative feedback regulation on ARR genes to trigger the cytokinin response. PMID- 18162595 TI - Structural, biochemical, and phylogenetic analyses suggest that indole-3-acetic acid methyltransferase is an evolutionarily ancient member of the SABATH family. AB - The plant SABATH protein family encompasses a group of related small-molecule methyltransferases (MTs) that catalyze the S-adenosyl-L-methionine-dependent methylation of natural chemicals encompassing widely divergent structures. Indole 3-acetic acid (IAA) methyltransferase (IAMT) is a member of the SABATH family that modulates IAA homeostasis in plant tissues through methylation of IAA's free carboxyl group. The crystal structure of Arabidopsis (Arabidopsis thaliana) IAMT (AtIAMT1) was determined and refined to 2.75 A resolution. The overall tertiary and quaternary structures closely resemble the two-domain bilobed monomer and the dimeric arrangement, respectively, previously observed for the related salicylic acid carboxyl methyltransferase from Clarkia breweri (CbSAMT). To further our understanding of the biological function and evolution of SABATHs, especially of IAMT, we analyzed the SABATH gene family in the rice (Oryza sativa) genome. Forty one OsSABATH genes were identified. Expression analysis showed that more than one half of the OsSABATH genes were transcribed in one or multiple organs. The OsSABATH gene most similar to AtIAMT1 is OsSABATH4. Escherichia coli-expressed OsSABATH4 protein displayed the highest level of catalytic activity toward IAA and was therefore named OsIAMT1. OsIAMT1 exhibited kinetic properties similar to AtIAMT1 and poplar IAMT (PtIAMT1). Structural modeling of OsIAMT1 and PtIAMT1 using the experimentally determined structure of AtIAMT1 reported here as a template revealed conserved structural features of IAMTs within the active-site cavity that are divergent from functionally distinct members of the SABATH family, such as CbSAMT. Phylogenetic analysis revealed that IAMTs from Arabidopsis, rice, and poplar (Populus spp.) form a monophyletic group. Thus, structural, biochemical, and phylogenetic evidence supports the hypothesis that IAMT is an evolutionarily ancient member of the SABATH family likely to play a critical role in IAA homeostasis across a wide range of plants. PMID- 18162596 TI - The Arabidopsis kinase-associated protein phosphatase regulates adaptation to Na+ stress. AB - The kinase-associated protein phosphatase (KAPP) is a regulator of the receptor like kinase (RLK) signaling pathway. Loss-of-function mutations rag1-1 (root attenuated growth1-1) and rag1-2, in the locus encoding KAPP, cause NaCl hypersensitivity in Arabidopsis thaliana. The NaCl hypersensitive phenotype exhibited by rag1 seedlings includes reduced shoot and primary root growth, root tip swelling, and increased lateral root formation. The phenotype exhibited by rag1-1 seedlings is associated with a specific response to Na(+) toxicity. The sensitivity to Na(+) is Ca(2+) independent and is not due to altered intracellular K(+)/Na(+). Analysis of the genetic interaction between rag1-1 and salt overly sensitive1 (sos1-14) revealed that KAPP is not a component of the SOS signal transduction pathway, the only Na(+) homeostasis signaling pathway identified so far in plants. All together, these results implicate KAPP as a functional component of the RLK signaling pathway, which also mediates adaptation to Na(+) stress. RLK pathway components, known to be modulated by NaCl at the messenger RNA level, are constitutively down-regulated in rag1-1 mutant plants. The effect of NaCl on their expression is not altered by the rag1-1 mutation. PMID- 18162597 TI - Glutamate receptor subtypes evidenced by differences in desensitization and dependence on the GLR3.3 and GLR3.4 genes. AB - Ionotropic glutamate (Glu) receptors in the central nervous system of animals are tetrameric ion channels that conduct cations across neuronal membranes upon binding Glu or another agonist. Plants possess homologous molecules encoded by GLR genes. Previous studies of Arabidopsis thaliana root cells showed that the amino acids alanine (Ala), asparagine (Asn), cysteine (Cys), Glu, glycine (Gly), and serine trigger transient Ca(2+) influx and membrane depolarization by a mechanism that depends on the GLR3.3 gene. This study of hypocotyl cells demonstrates that these six effective amino acids are not equivalent agonists. Instead, they grouped into hierarchical classes based on their ability to desensitize the response mechanism. Sequential treatment with two different amino acids separated by a washout phase demonstrated that Glu desensitized the depolarization mechanism to Gly, but Gly did not desensitize the mechanism to Glu. All 36 possible pairs of agonists were tested to characterize the desensitization hierarchy. The results could be explained by a model in which one class of channels contained a subunit that was activated and therefore desensitized only by Glu, while a second class could be activated and desensitized by Ala, Cys, Glu, or Gly. A third class could be activated and desensitized by any of the six effective amino acids. Analysis of knockout mutants indicated that GLR3.3 was a required component of all three classes of channels, while the related GLR3.4 molecule specifically affected only two of the classes. The resulting model is an important step toward understanding the biological roles of these enigmatic ion channels. PMID- 18162598 TI - Silicon uptake in diatoms revisited: a model for saturable and nonsaturable uptake kinetics and the role of silicon transporters. AB - The silicic acid uptake kinetics of diatoms were studied to provide a mechanistic explanation for previous work demonstrating both nonsaturable and Michaelis Menten-type saturable uptake. Using (68)Ge(OH)(4) as a radiotracer for Si(OH)(4), we showed a time-dependent transition from nonsaturable to saturable uptake kinetics in multiple diatom species. In cells grown under silicon (Si)-replete conditions, Si(OH)(4) uptake was initially nonsaturable but became saturable over time. Cells prestarved for Si for 24 h exhibited immediate saturable kinetics. Data suggest nonsaturability was due to surge uptake when intracellular Si pool capacity was high, and saturability occurred when equilibrium was achieved between pool capacity and cell wall silica incorporation. In Thalassiosira pseudonana at low Si(OH)(4) concentrations, uptake followed sigmoidal kinetics, indicating regulation by an allosteric mechanism. Competition of Si(OH)(4) uptake with Ge(OH)(4) suggested uptake at low Si(OH)(4) concentrations was mediated by Si transporters. At high Si(OH)(4), competition experiments and nonsaturability indicated uptake was not carrier mediated and occurred by diffusion. Zinc did not appear to be directly involved in Si(OH)(4) uptake, in contrast to a previous suggestion. A model for Si(OH)(4) uptake in diatoms is presented that proposes two control mechanisms: active transport by Si transporters at low Si(OH)(4) and diffusional transport controlled by the capacity of intracellular pools in relation to cell wall silica incorporation at high Si(OH)(4). The model integrates kinetic and equilibrium components of diatom Si(OH)(4) uptake and consistently explains results in this and previous investigations. PMID- 18162599 TI - Mechanism-based pharmacodynamic modeling of S(-)-atenolol: estimation of in vivo affinity for the beta1-adrenoceptor with an agonist-antagonist interaction model. AB - The aim of this study was the development of an agonist-antagonist interaction model to estimate the in vivo affinity of S(-)-atenolol for the beta(1) adrenoreceptor. Male Wistar-Kyoto (WKY) rats were used to characterize the interaction between the model drugs isoprenaline (to induce tachycardia) and S(-) atenolol. Blood samples were taken to determine plasma pharmacokinetics. Reduction of isoprenaline-induced tachycardia was used as a pharmacodynamic endpoint. The pharmacokinetic-pharmacodynamic relationship of isoprenaline was first characterized with the operational model of agonism using the literature value for the affinity (K(A)) of isoprenaline (3.2 x 10(-8) M; left atria WKY rats). Resulting estimates for baseline (E(0)), maximal effect (E(max)), and efficacy (tau) were 374 (1.9%), 130 (5.9%), and 247 (33%) beats per minute, respectively. In addition, the interaction between isoprenaline and S(-)-atenolol was characterized using a pharmacodynamic interaction model based on the operational model of agonism that describes the heart rate response based on the affinity of the agonist (K(A)), the affinity of the antagonist (K(B)), the efficacy (tau), the maximal effect (E(max)), the Hill coefficient (n(H)), the concentrations of isoprenaline and atenolol, and the displacement of the endogenous agonist adrenaline. The estimated in vivo affinity (K(B)) of S(-) atenolol for the beta(1) -receptor was 4.6 x 10(-8) M. The obtained estimate for in vivo affinity of S(-)-atenolol (4.6 x 10(-8) M) is comparable to literature values for the in vitro affinity in functional assays. In conclusion, a meaningful estimate of in vivo affinity for S(-)-atenolol could be obtained using a mechanism-based pharmacodynamic modeling approach. PMID- 18162600 TI - Amiloride-insensitive Na+ and fluid absorption in the mammalian distal lung. AB - The ability of the distal lung epithelia to actively transport Na+, with Cl- and water following, from the alveolar spaces inversely correlates with morbidity and mortality of infants, children, and adults with alveolar pulmonary edema. It is now recognized, in contrast to many other Na+ transporting epithelia, that at least half of this active transport is not sensitive to amiloride, which inhibits the epithelial Na+ channel. This paper reviews amiloride-insensitive Na+ and fluid transport in the mammalian distal lung unit under basal conditions and speculates on potential explanations for this amiloride-insensitive transport. It also provides new information, using primary cultures of rat fetal distal lung epithelia and alveolar type II cells grown under submersion and air-liquid interface culture conditions, regarding putative blockers of this transport. PMID- 18162601 TI - Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells. AB - Nuclear erythroid-related factor 2 (Nrf2), a redox-sensitive transcription factor, is involved in transcriptional regulation of many antioxidant genes, including glutamate-cysteine ligase (GCL). Cigarette smoke (CS) is known to cause oxidative stress and deplete glutathione (GSH) levels in alveolar epithelial cells. We hypothesized that resveratrol, a polyphenolic phytoalexin, has antioxidant signaling properties by inducing GSH biosynthesis via the activation of Nrf2 and protects lung epithelial cells against CS-mediated oxidative stress. Treatment of human primary small airway epithelial and human alveolar epithelial (A549) cells with CS extract (CSE) dose dependently decreased GSH levels and GCL activity, effects that were associated with enhanced production of reactive oxygen species. Resveratrol restored CSE-depleted GSH levels by upregulation of GCL via activation of Nrf2 and also quenched CSE-induced release of reactive oxygen species. Interestingly, CSE failed to induce nuclear translocation of Nrf2 in A549 and small airway epithelial cells. On the contrary, Nrf2 was localized in the cytosol of alveolar and airway epithelial cells due to CSE-mediated posttranslational modifications such as aldehyde/carbonyl adduct formation and nitration. On the other hand, resveratrol attenuated CSE-mediated Nrf2 modifications, thereby inducing its nuclear translocation associated with GCL gene transcription, as demonstrated by GCL-promoter reporter and Nrf2 small interfering RNA approaches. Thus resveratrol attenuates CSE-mediated GSH depletion by inducing GSH synthesis and protects epithelial cells by reversing CSE-induced posttranslational modifications of Nrf2. These data may have implications in dietary modulation of antioxidants in treatment of chronic obstructive pulmonary disease. PMID- 18162603 TI - Animal models of human pneumonia. AB - Pneumonia is a medical and public health priority, and advances against this disease will require improved knowledge of biological mechanisms. Human pneumonia is modeled with experimental infections of animals, most frequently mice. Mouse models are leading to important discoveries relevant to pneumonia, but their limitations must be carefully considered. Several approaches to establishing pneumonia in mice have been developed, and each has specific strengths and weaknesses. Similarly, procedures for characterizing microbial and host responses to infection have unique advantages and disadvantages. Mice are not small humans, and the applicability of results from murine models to human disease depends on understanding the similarities and differences between species. Additional considerations such as mouse strain, microbe strain, and prior mouse-microbe interactions also influence the design and interpretation of experiments. Results from studies of pneumonia in animals, combined with complementary basic and translational studies, are elucidating mechanisms responsible for susceptibility to and pathophysiology of lung infection. PMID- 18162602 TI - PPAR-gamma agonists inhibit profibrotic phenotypes in human lung fibroblasts and bleomycin-induced pulmonary fibrosis. AB - Pulmonary fibrosis is characterized by alterations in fibroblast phenotypes resulting in excessive extracellular matrix accumulation and anatomic remodeling. Current therapies for this condition are largely ineffective. Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a member of the nuclear hormone receptor superfamily, the activation of which produces a number of biological effects, including alterations in metabolic and inflammatory responses. The role of PPAR-gamma as a potential therapeutic target for fibrotic lung diseases remains undefined. In the present study, we show expression of PPAR gamma in fibroblasts obtained from normal human lungs and lungs of patients with idiopathic interstitial pneumonias. Treatment of lung fibroblasts and myofibroblasts with PPAR-gamma agonists results in inhibition of proliferative responses and induces cell cycle arrest. In addition, PPAR-gamma agonists, including a constitutively active PPAR-gamma construct (VP16-PPAR-gamma), inhibit the ability of transforming growth factor-beta1 to induce myofibroblast differentiation and collagen secretion. PPAR-gamma agonists also inhibit fibrosis in a murine model, even when administration is delayed until after the initial inflammation has largely resolved. These observations indicate that PPAR-gamma is an important regulator of fibroblast/myofibroblast activation and suggest a role for PPAR-gamma ligands as novel therapeutic agents for fibrotic lung diseases. PMID- 18162604 TI - A single amino acid difference between ether-a-go-go- related gene channel subtypes determines differential sensitivity to a small molecule activator. AB - Activators of human ether-a-go-go-related gene 1 (hERG1) channels, such as (3R,4R)-4-[3-(6-methoxy-quinolin-4-yl)-3-oxo-propyl]-1-[3-(2,3,5-trifluoro phenyl)-prop-2-ynyl]-piperidine-3-carboxylic acid (RPR260243), reverse the effect of hERG1 blockers and shorten the duration of cardiac action potentials. RPR260243 (RPR) slows the rate of deactivation and shifts the voltage dependence of channel inactivation to more positive potentials. We recently mapped the binding site for RPR to several residues located near the cytoplasmic ends of the S5 and S6 helices of the hERG1 subunit. These residues are conserved in the highly homologous ether-a-go-go-related gene 3 (ERG3) subunit; however, RPR blocks ERG3 channels. Here, we compare hERG1 and rat ERG3 (rERG3) channels to explore the molecular basis for differential channel sensitivity to RPR. Channels were heterologously expressed in Xenopus laevis oocytes, and currents were recorded using the two-electrode voltage-clamp technique. Site-directed mutagenesis was used to swap the two residues within the putative binding domain that differed between hERG1 and rERG3. The differential sensitivity of hERG1 and rERG3 channels to the agonist effect of RPR could be accounted for by a single S5 residue (Thr556 in hERG1, Ile558 in rERG3). A Thr in this position favors agonist activity, whereas an Ile reveals a secondary blocking effect of RPR. PMID- 18162605 TI - Alpha2 subunit specificity of cyclothiazide inhibition on glycine receptors. AB - In the mammalian cortex, alpha2 subunit-containing glycine receptors (GlyRs) mediate tonic inhibition, but the precise functional role of this type of GlyRs is difficult to establish because of the lack of subtype-selective antagonist. In this study, we found that cyclothiazide (CTZ), an epileptogenic agent, potently inhibited GlyR-mediated current (I(Gly)) in cultured rat hippocampal neurons. The inhibition was glycine concentration-dependent, suggesting a competitive mechanism. Note that GlyRs containing the alpha2 but not alpha1 or alpha3 subunits, when being heterologously expressed in human embryonic kidney 293T cells, were inhibited by CTZ, indicating subunit specificity of CTZ action. In addition, the degree of CTZ inhibition on I(Gly) in rat spinal neurons declined with time in culture, in parallel with a decline of alpha2 subunit expression, which is known to occur during spinal cord development. Furthermore, site directed mutagenesis indicates that a single-amino acid threonine at position 59 near the N terminus of the alpha2 subunit confers the specificity of CTZ action. Thus, CTZ is a potent and selective inhibitor of alpha2-GlyRs, and threonine at position 59 plays a critical role in the susceptibility of GlyR to CTZ inhibition. PMID- 18162606 TI - Critical role of lipid raft redox signaling platforms in endostatin-induced coronary endothelial dysfunction. AB - OBJECTIVE: Endostatin (EST) was found to initiate a redox signaling cascade associated with activation of NADPH oxidase in endothelial cells (ECs). The present study tested whether EST stimulates clustering of ceramide-enriched lipid rafts (LRs), which assembles and activates NADPH oxidase to form redox signaling platforms. METHODS AND RESULTS: Using confocal microscopy, we first demonstrated a colocalization of LR clusters with NADPH oxidase subunits, gp91(phox) and p47(phox) in the ECs membrane on EST stimulation. Immunoblot analysis of floated detergent-resistant membrane fractions found that in LR fractions NADPH oxidase subunits gp91(phox) and p47(phox) are enriched and that the activity of this enzyme increased dramatically, as measured by electron spin resonance (ESR) spectrometry. This EST-increased LR platform formation was shown to be attenuated by inhibition or RNA interference of acid sphingomyelinase (A-SMase). Functionally, EST pretreatment significantly impaired bradykinin or A23187 induced vasodilation in isolated small coronary arteries, which could be partially reversed by LR disruptors. CONCLUSIONS: The early injury effect of EST on the vascular endothelium is associated with the formation of redox signaling platforms via lipid raft clustering. PMID- 18162607 TI - Transcriptional inhibition of protease-activated receptor-1 expression by prostacyclin in human vascular smooth muscle cells. AB - OBJECTIVE: Stimulation of protease-activated receptor-1 (PAR-1) by thrombin causes vascular smooth muscle cell (SMC) mitogenesis and has been implicated in the vascular response to injury. Vascular injury is also associated with enhanced formation of PGE2 and PGI2 (prostacyclin). This study investigates whether PGI2 and PGE2 modify the expression of PAR-1 and the cellular response to thrombin in human SMC. METHODS AND RESULTS: The PGI2-mimetic iloprost (1 to 100 nmol/L) attenuated mRNA, total protein, and cell surface expression of PAR-1. This was associated with inhibition of thrombin-induced mitogenesis and migration. Comparable inhibition of PAR-1 expression was observed with the selective IP receptor agonist cicaprost, the adenylyl cyclase activator forskolin, the phosphodiesterase inhibitor isobutylmethylxanthine and the PKA activator dibutyryl-cAMP. Similar effects of PGE2 required micromolar concentrations. The specific PKA-inhibitor Myr-PKI prevented PAR-1 downregulation by iloprost. The potential role of Rho family GTPases in PAR-1 regulation was also investigated. Iloprost decreased Rac1 mRNA and the Rac1 inhibitor NSC23766 mimicked the inhibitory effects of iloprost on PAR-1 protein--but not mRNA. The Rho kinase inhibitor Y27632 did not influence PAR-1 expression. CONCLUSIONS: IP-receptor agonists may limit the mitogenic actions of thrombin in human SMC by downregulating PAR-1 via modulation of cAMP-/PKA- and Rac1-dependent signaling pathways. PMID- 18162608 TI - Upregulation of pentraxin-3 in human endothelial cells after lysophosphatidic acid exposure. AB - OBJECTIVE: The earliest event in atherogenesis appears to be endothelium dysfunction. Lysophosphatidic acid (LPA), one of the major bioactive lipid components of oxidized low-density lipoproteins (oxLDL), can cause the activation of endothelial cells (ECs), which start to secrete multiple proinflammatory polypeptides/proteins. The purpose of this study was to better document the proatherogenic properties of LPA using a subproteomic approach focused on the secretome of LPA-treated ECs. METHODS AND RESULTS: The secretome of LPA-treated ECs was analyzed using the 2D-DIGE approach. Among the 20 spots displaying significant variations of abundance compared with the control cells, we identified pentraxin-3 by mass spectrometry. Pentraxin-3 upregulation was confirmed at the mRNA and protein level, both on immortalized and primary ECs. LPA- but also oxLDL-induced pentraxin-3 upregulation was reduced in the presence of an antagonist of the LPA-receptors and largely dependent on NFkappaB activation. Finally, we demonstrated, for the first time, the chemotactic activity of pentraxin-3 on human THP-1 monocytes by using a chemotaxis assay. CONCLUSIONS: Our findings favor the proatherogenic role of LPA, a bioactive lipid produced by activated platelets and present in oxLDL, because it enhances pentraxin-3 secretion that could contribute to the accumulation of monocytes in the atherosclerotic lesion. PMID- 18162609 TI - VE-cadherin: the major endothelial adhesion molecule controlling cellular junctions and blood vessel formation. AB - Vascular endothelial (VE)-cadherin is a strictly endothelial specific adhesion molecule located at junctions between endothelial cells. In analogy of the role of E-cadherin as major determinant for epithelial cell contact integrity, VE cadherin is of vital importance for the maintenance and control of endothelial cell contacts. Mechanisms that regulate VE-cadherin-mediated adhesion are important for the control of vascular permeability and leukocyte extravasation. In addition to its adhesive functions, VE-cadherin regulates various cellular processes such as cell proliferation and apoptosis and modulates vascular endothelial growth factor receptor functions. Consequently, VE-cadherin is essential during embryonic angiogenesis. This review will focus on recent new developments in understanding the role of VE-cadherin in controlling endothelial cell contacts and influencing endothelial cell behavior by various outside-in signaling processes. PMID- 18162610 TI - Oxidant generation predominates around calcifying foci and enhances progression of aortic valve calcification. AB - OBJECTIVE: We hypothesized that reactive oxygen species (ROS) contribute to progression of aortic valve (AV) calcification/stenosis. METHODS AND RESULTS: We investigated ROS production and effects of antioxidants tempol and lipoic acid (LA) in calcification progression in rabbits given 0.5% cholesterol diet +10(4) IU/d Vit.D2 for 12 weeks. Superoxide and H2O2 microfluorotopography and 3 nitrotyrosine immunoreactivity showed increased signals not only in macrophages but preferentially around calcifying foci, in cells expressing osteoblast/osteoclast, but not macrophage markers. Such cells also showed increased expression of NAD(P)H oxidase subunits Nox2, p22phox, and protein disulfide isomerase. Nox4, but not Nox1 mRNA, was increased. Tempol augmented whereas LA decreased H2O2 signals. Importantly, AV calcification, assessed by echocardiography and histomorphometry, decreased 43% to 70% with LA, but increased with tempol (P < or = 0.05). Tempol further enhanced apoptosis and Nox4 expression. In human sclerotic or stenotic AV, we found analogous increases in ROS production and NAD(P)H oxidase expression around calcifying foci. An in vitro vascular smooth muscle cell (VSMC) calcification model also exhibited increased, catalase-inhibitable, calcium deposit with tempol, but not with LA. CONCLUSIONS: Our data provide evidence that ROS, particularly hydrogen peroxide, potentiate AV calcification progression. However, tempol exhibited a paradoxical effect, exacerbating AV/vascular calcification, likely because of its induced increase in peroxide generation. PMID- 18162611 TI - Final common molecular pathways of aging and cardiovascular disease: role of the p66Shc protein. AB - Oxidative stress affects the availability of key-regulators of vascular homeostasis and controls a number of signaling pathways relevant to myocardial and vascular disease. Reactive oxygen species are generated by different intracellular molecular pathways principally located in mitochondria. The notion that mice carrying a targeted mutation of the p66(Shc) gene display prolonged lifespan, reduced production of intracellular oxidants, and increased resistance to oxidative stress-induced apoptosis prompted a series of studies aimed at defining the biochemical function of p66(Shc) and its possible implication in cardiovascular diseases. Indeed, p66(Shc-/-) mice are protected against vascular, cardiac, and renal impairment attributable to hypercholesterolemia, aging, diabetes, and ischemia/reperfusion. The present review focuses on the biochemical and physiological function of the p66(Shc) adaptor protein as well as on the mechanisms linking p66(Shc)-associated generation of free radicals to the pathophysiology of aging and cardiovascular disease. On the whole, the evidence so far reported and here discussed supports the concept that pharmacological modulation of p66(Shc) expression and activity may be a novel and effective target for the treatment of atherosclerotic vascular disease as well as myocardial adaptation to hypertrophic, inflammatory and neuro-hormonal stimuli in the overloaded heart. PMID- 18162612 TI - p38 MAPK inhibition reduces aortic ultrasmall superparamagnetic iron oxide uptake in a mouse model of atherosclerosis: MRI assessment. AB - OBJECTIVE: Ultrasmall superparamagnetic iron oxide (USPIO) contrast agents have been used for noninvasive MRI assessment of atherosclerotic plaque inflammation. The purpose of this study was to noninvasively evaluate USPIO uptake in aorta of apoE-/- mice and to determine the effects of Angiotensin II (Ang II) infusion and chronic antiinflammatory treatment with a p38 MAPK inhibitor on this uptake. METHODS AND RESULTS: ApoE-/- mice were administered saline or Ang II (1.44 mg/kg/d) for 21 days. In vivo MRI assessment of USPIO uptake in the aortic arch was observed in all animals. However, although the Ang II group had significantly higher absolute iron content (increased 103%, P<0.001) in the aortic arch compared with the saline group, the p38 MAPK inhibitor (SB-239063, 150 mg/kg/d) treatment group did not (increased 6%, NS). The in vivo MRI signal intensity was significantly correlated to the absolute iron content in the aortic arch. Histological evaluation of the aortic root lesion area showed colocalization of USPIO with macrophages and a reduction in USPIO but not macrophage content with SB-239063 treatment. CONCLUSIONS: The present study demonstrates that noninvasive assessment of USPIO uptake, as a marker for inflammation in murine atherosclerotic plaque, is feasible and that p38 MAPK inhibition attenuates the uptake of USPIO in aorta of Ang II-infused apoE-/- mice. PMID- 18162613 TI - To FAST or not to FAST: SHOUT-FAST! PMID- 18162614 TI - Perinatal stroke with no obvious cause. PMID- 18162615 TI - Mobile message for a better stroke recognition: the new concept of national campaign. PMID- 18162616 TI - The impact of intensity of aphasia therapy on recovery. PMID- 18162617 TI - Vertebral artery occlusion after chemotherapy. PMID- 18162618 TI - Can patients with atrial fibrillation be optimally risk stratified for stroke and thromboembolism? PMID- 18162619 TI - Stent-assisted endovascular thrombolysis versus intravenous thrombolysis in internal carotid artery dissection with tandem internal carotid and middle cerebral artery occlusion. PMID- 18162620 TI - Transforming guidelines in routine practice. PMID- 18162621 TI - Diffusion-weighted imaging in stroke attributable to internal carotid artery dissection: the significance of vessel patency. AB - BACKGROUND AND PURPOSE: In stroke attributable to spontaneous dissection of the internal carotid artery (sICAD), arterial patency may influence the pattern and extent of cerebral ischemia. METHODS: In 40 consecutive patients with stroke caused by sICAD, we compared the number, size and pattern of cerebral diffusion weighted imaging lesions between patients with stenotic sICAD (n=15) and occlusive sICAD (n=25). RESULTS: Patients with stenotic sICAD had more ischemic lesions (median 5, interquartile range 1 to 10) than patients with occlusive sICAD (2, 1 to 3; P=0.014). Lesion diameters were larger in occlusive sICAD (62, 50 to 99 mm) than in stenotic sICAD (25, 10 to 50 mm; P=0.007). Border-zone infarction occurred only in stenotic sICAD (7/15, 47%). Most patients with occlusive sICAD had territorial infarcts (22/25, 88%). CONCLUSIONS: In stroke attributable to sICAD, diffusion-weighted imaging characteristics may be influenced by the patency of the carotid artery. Differences in the pathogenesis of cerebral ischemia might exist between patients with stenotic and those with occlusive sICAD. PMID- 18162622 TI - Tight link between our sense of limb ownership and self-awareness of actions. AB - BACKGROUND AND PURPOSE: Hemiparetic stroke patients with disturbed awareness for their motor weakness (anosognosia for hemiparesis/-plegia [AHP]) may exhibit further abnormal attitudes toward or perceptions of the affected limb(s). The present study investigated the clinical relationship and the anatomy of such abnormal attitudes and AHP. METHODS: In a new series of 79 consecutively admitted acute stroke patients with right brain damage and hemiparesis/-plegia, different types of abnormal attitudes toward the hemiparetic/plegic limb (asomatognosia, somatoparaphrenia, anosodiaphoria, misoplegia, personification, kinaesthetic hallucinations, supernumerary phantom limb) were investigated. RESULTS: Ninty-two percent of the patients with AHP showed additional "disturbed sensation of limb ownership" (DSO) for the paretic/plegic limb. The patients had the feeling that their contralesional limb(s) do not belong to their body or even belong to another person. Analysis of lesion location revealed that the right posterior insula is a crucial structure involved in these phenomena. CONCLUSIONS: DSO for hemiparetic/-plegic limbs and AHP are tightly linked both clinically and anatomically. The right posterior insula seems to be a crucial structure involved in the genesis of our sense of limb ownership and self-awareness of actions. PMID- 18162623 TI - Predicting stroke risk in hypertensive patients with coronary artery disease: a report from the INVEST. AB - BACKGROUND AND PURPOSE: Our understanding of factors influencing stroke risk among patients with coronary artery disease is incomplete. Accordingly, factors predicting stroke risk in hypertensive, clinically stable coronary artery disease patients were determined with data from the INternational VErapamil SR trandolapril STudy (INVEST). METHODS: The effect of baseline characteristics and on-treatment blood pressure (BP) were analyzed to determine the risk of stroke (fatal or nonfatal) among the 22 576 patients enrolled. Cox proportional-hazards models (unadjusted, adjusted, and time dependent) were used to identify predictors of stroke among subgroups with these characteristics present at entry and on-treatment BP. RESULTS: Excellent BP control (at 24 months, >70% <140/90 mm Hg) was achieved during 61 835 patient-years of follow-up, as 377 patients had a stroke (6.1 strokes/1000 patient-years) and 28% of those patients had a fatal stroke. Increased age, black race, US residency, and history of prior myocardial infarction, smoking, stroke/transient ischemic attack, arrhythmia, diabetes, and coronary bypass surgery were associated with an increased risk of stroke. Achieving a systolic BP <140 mm Hg and a diastolic BP <90 mm Hg was associated with a decreased risk of stroke. There was no statistically significant difference in stroke risk comparing the verapamil SR-based with the atenolol based treatment strategy (adjusted hazard ratio=0.87; 95% CI, 0.71 to 1.06; P=0.17). CONCLUSIONS: Among hypertensive patients with chronic coronary artery disease, stroke was an important complication associated with significant mortality. Black race, US residency, and conditions associated with increased vascular disease severity and arrhythmia predicted increased stroke risk, whereas achieving a BP <140/90 mm Hg on treatment predicted a reduced stroke risk. PMID- 18162624 TI - Stroke awareness in Brazil: alarming results in a community-based study. AB - BACKGROUND AND PURPOSE: Stroke is the leading cause of death in Brazil. This community-based study assessed lay knowledge about stroke recognition and treatment and risk factors for cerebrovascular diseases and activation of emergency medical services in Brazil. METHODS: The study was conducted between July 2004 and December 2005. Subjects were selected from the urban population in transit about public places of 4 major Brazilian cities: Sao Paulo, Salvador, Fortaleza, and Ribeirao Preto. Trained medical students, residents, and neurologists interviewed subjects using a structured, open-ended questionnaire in Portuguese based on a case presentation of a typical patient with acute stroke at home. RESULTS: Eight hundred fourteen subjects were interviewed during the study period (53.9% women; mean age, 39.2 years; age range, 18 to 80 years). There were 28 different Portuguese terms to name stroke. Twenty-two percent did not recognize any warning signs of stroke. Only 34.6% of subjects answered the correct nationwide emergency telephone number in Brazil (#192). Only 51.4% of subjects would call emergency medical services for a relative with symptoms of stroke. In a multivariate analysis, individuals with higher education called emergency medical services (P=0.038, OR=1.5, 95%, CI: 1.02 to 2.2) and knew at least one risk factor for stroke (P<0.05, OR=2.0, 95% CI: 1.2 to 3.2) more often than those with lower education. CONCLUSIONS: Our study discloses alarming lack of knowledge about activation of emergency medical services and availability of acute stroke treatment in Brazil. These findings have implications for public health initiatives in the treatment of stroke and other cardiovascular emergencies. PMID- 18162625 TI - Intravenous rosuvastatin for acute stroke treatment: an animal study. AB - BACKGROUND AND PURPOSE: Statins exert rapid cholesterol-independent vasoprotective effects. Here, we tested whether postevent treatment with intravenously (i.v.) administered rosuvastatin improves acute stroke outcome in mice. METHODS: 129/SV wild-type mice were subjected to 1-hour filamentous middle cerebral artery occlusion (MCAo), followed by reperfusion, and were postevent treated with i.v. or intraperitoneal (i.p.) rosuvastatin given up to 6 hours after MCAo (dose range 0.02 to 20 mg kg(-1) body weight). RESULTS: Rosuvastatin, when administered i.v., significantly reduced lesion size when given up to 4 hours after MCAo and in doses as low as 0.2 mg kg(-1). In contrast, i.p. administration provided protection only when given directly on reperfusion at a dose of 20 mg kg(-1) but not at lower doses or later time points. Lesion protection was evident as late as 5 days after brain ischemia and was associated with functional improvements in the pole-test and wire-hanging test (2.0 mg kg( 1) dose). Neuroprotection with i.v. rosuvastatin was achieved with peak plasma concentrations <0.5 ng ml(-1) (ie, with 0.2 mg kg(-1)) and was associated with increased levels of phosphorylated Akt kinase and endothelial nitric oxide synthase in the vasculature. CONCLUSIONS: Rosuvastatin, given intravenously at pharmacologically relevant concentrations, protects from focal brain ischemia up to 4 hours after an event. In our opinion, the development of an intravenous statin formulation is warranted for acute stroke trials with statins in humans. PMID- 18162626 TI - Early neutrophilia is associated with volume of ischemic tissue in acute stroke. AB - BACKGROUND AND PURPOSE: Few data exist on the relationship between differential subpopulations of peripheral leukocytes and early cerebral infarct size in ischemic stroke. Using diffusion-weighted MR imaging (DWI), we assessed the relationship of early total and differential peripheral leukocyte counts and volume of ischemic tissue in acute stroke. METHODS: All included patents had laboratory investigations and neuroimaging collected within 24 hours of stroke onset. Total peripheral leukocyte counts and differential counts were analyzed individually and by quartiles. DWI lesions were outlined using a semiautomated threshold technique. The relationship between leukocyte quartiles and DWI infarct volumes was examined using multivariate quartile regression. RESULTS: 173 patients met study inclusion criteria. Median age was 73 years. Total leukocyte counts and DWI volumes showed a strong correlation (Spearman rho=0.371, P<000.1). Median DWI volumes (mL) for successive neutrophil quartiles were: 1.3, 1.3, 3.2, and 20.4 (P for trend <0.001). Median DWI volumes (mL) for successive lymphocyte quartiles were: 3.2, 8.1, 1.3, and 1.5 (P=0.004). After multivariate analysis, larger DWI volume remained strongly associated with higher total leukocyte and neutrophil counts (both probability values <0.001), but not with lymphocyte count (P=0.4971). Compared with the lowest quartiles, DWI volumes were 8.7 mL and 12.9 mL larger in the highest quartiles of leukocyte and neutrophil counts, respectively. CONCLUSIONS: Higher peripheral leukocyte and neutrophil counts, but not lymphocyte counts, are associated with larger infarct volumes in acute ischemic stroke. Attenuating neutrophilic response early after ischemic stroke may be a viable therapeutic strategy and warrants further study. PMID- 18162627 TI - Cognitive outcome at early school age in term-born children with perinatally acquired middle cerebral artery territory infarction. AB - BACKGROUND AND PURPOSE: To assess cognitive outcome at early school age in term born children with middle cerebral arterial (MCA) territory infarction of perinatal onset and examine the correlation between cognitive abilities and the extent of lesions as seen on neonatal MRI, epilepsy, and hemiplegia. METHODS: Thirty-one children were seen as newborns with an acutely evolving MCA territory infarction documented on neonatal MRI scan. IQ was assessed (WIPPSI/WISC where appropriate) and they had a standardized neurological examination at early school age. Lesion(s) site was recorded from the neonatal images. RESULTS: Twenty-eight of 31 children were assessed (median age 5.75 range 5.33 to 10.33 years); 1 child died and 2 were abroad. IQ was within the normal range (mean 104, range 82 to 144) in 21 (78%); 1 child did not complete all tests but had a normal PIQ; 3 had a low and 3 an exceptionally low IQ. Verbal IQs were more varied and lower than performance IQs especially in children from multilingual backgrounds. There was no consistent association between cognitive impairment, side, or extent of the MCA lesion. Cognitive impairments were more frequent in children with seizures or hemiplegia. All 6 children with low IQ also had behavioral problems or unusual associated clinical or scan features. CONCLUSIONS: In our cohort a low IQ at early school age did not occur in children with the common presentation of neonatal unilateral MCA territory infarction. Cognitive impairment appeared more frequently when an MCA arterial territory infarction, even if relatively small, was associated with other risk factors. PMID- 18162628 TI - Cell growth and differentiation in Arabidopsis epidermal cells. AB - Plant epidermal cells are morphologically diverse, differing in size, shape, and function. Their unique morphologies reflect the integral function each cell performs in the organ to which it belongs. Cell morphogenesis involves multiple cellular processes acting in concert to create specialized shapes. The Arabidopsis epidermis contains numerous cell types greatly differing in shape, size, and function. Work on three types of epidermal cells, namely trichomes, root hairs, and pavement cells, has made significant progress towards understanding how plant cells reach their final morphology. These three cell types have highly distinct morphologies and each has become a model cell for the study of morphological processes. A growing body of knowledge is creating a picture of how endoreduplication, cytoskeletal dynamics, vesicle transport, and small GTPase signalling, work in concert to create specialized shapes. Similar mechanisms that determine cell shape and polarity are shared between these cell types, while certain mechanisms remain specific to each. PMID- 18162629 TI - Expression of multiple forms of ferredoxin NADP+ oxidoreductase in wheat leaves. AB - In higher plants there are two forms of ferredoxin NADP(+) oxidoreductase (FNR), a photosynthetic pFNR primarily required for the photoreduction of NADP(+), and a heterotrophic hFNR which generates reduced ferredoxin by utilizing electrons from NADPH produced during carbohydrate oxidation. The aim of this study was to investigate the presence of multiple forms of FNR in wheat leaves and the capacity of FNR isoforms to respond to changes in reductant demand through varied expression and N-terminal processing. Two forms of pFNR mRNA (pFNRI and pFNRII) were expressed in a similar pattern along the 12 cm developing primary wheat leaf, with the highest levels observed in plants grown continuously in the dark in the presence (pFNRI) or absence (pFNRII) of nitrate respectively. pFNR protein increased from the leaf base to tip. hFNR mRNA and protein was in the basal part of the leaf in plants grown in the presence of nitrate. FNR activity in plants grown in a light/dark cycle without nitrate was mainly due to pFNR, whilst hFNR contributed significantly in nitrate-fed plants. The potential role of distinct forms of FNR in meeting the changing metabolic capacity and reductant demands along the linear gradient of developing cells of the leaf are discussed. Furthermore, evidence for alternative N-terminal cleavage sites of pFNR acting as a means of discriminating between ferredoxins and the implications of this in providing a more effective flow of electrons through a particular pathway in vivo is considered. PMID- 18162630 TI - Antisense-mediated suppression of C-hordein biosynthesis in the barley grain results in correlated changes in the transcriptome, protein profile, and amino acid composition. AB - Antisense- or RNAi-mediated suppression of the biosynthesis of nutritionally inferior storage proteins is a promising strategy for improving the amino acid profile of seeds. However, the potential pleiotropic effects of this on interconnected pathways and the agronomic quality traits need to be addressed. In the current study, a transcriptomic analysis of an antisense C-hordein line of barley was performed, using a grain-specific cDNA array. The C-hordein antisense line is characterized by marked changes in storage protein and amino acid profiles, while the seed weight is within the normal range and no external morphological irregularities were observed. The results of the transcriptome analysis showed excellent correlation with data on changes in the relative proportions of storage proteins and amino acid composition. The antisense line had a lower C-hordein level and down-regulated transcript encoding C-hordein. The production of the S-rich B/gamma- and D-hordeins was increased and significantly higher steady-state expression levels of the corresponding genes were observed. The increased synthesis of S-rich hordeins appeared to increase the demand for sulphur and the S-rich amino acids (cysteine and methionine), resulting in an up regulation of key genes in the appropriate biosynthetic pathways. This study demonstrated the utility of the grain-specific cDNA microarray analysis to detect perturbations induced by antisense suppression of plant processes. PMID- 18162633 TI - Special section: policy perspectives. PMID- 18162634 TI - The production of the "battered immigrant" in public policy and domestic violence advocacy. AB - In the context of U.S. public policy, battered immigrant signifies a person who is eligible to adjust his or her status under immigration law if he or she can demonstrate they have suffered domestic violence in the United States perpetrated by a U.S. citizen or legal permanent resident. Among community organizers, the term battered immigrant signifies a broader range of people for whom legal immigration status plays a role in their options for safety planning and/or leaving an abuser, the potential threat of deportation, and the eligibility for public benefits. Based on an ethnographic study of domestic violence advocacy with South Asian immigrants in Seattle and around the United States, this article examines how the difference in signification has direct social and political consequences with regard to who may access the benefits and protection offered to victims of domestic violence in the United States. PMID- 18162635 TI - Does using nonnumerical terms to describe risk aid violence risk communication? Clinician agreement and decision making. AB - Actuarial risk assessments yield valid numerical information about violence risk, but research suggests that forensic clinicians prefer to communicate risk using nonnumerical information (i.e., verbal terms such as high risk). In an experimental questionnaire study, 60 forensic clinicians disagreed on the interpretation of nonnumerical terms, and their nonnumerical risk estimates for one group of violent offenders were influenced by comparison with another group. Adding nonnumerical terms to numerical probability statements had no effect on hypothetical forensic decisions. These findings suggest that nonnumerical descriptive terms do not aid effective communication of violence risk and that contextual information might artificially affect estimated risk. PMID- 18162636 TI - Violence in young adolescents' relationships: a path model. AB - A structural equation model based on social cognitive theory was used to predict relationship violence from young adolescents' knowledge, self-efficacy, attitudes, and alternative conflict strategies (n = 143 male and 147 female grade 7-9 students). A direct causal effect was supported for violence-tolerant attitudes and psychologically aggressive (escalation/blame) strategies on physical violence against dating partners and friends. Knowledge and self efficacy contributed to using reasoning-based strategies, but this reduced violence only in boys' friendships. Knowledge reduced violence-tolerant attitudes, thus reducing escalation/ blame and physical violence. Attitudes toward male and female dating violence (ATMDV and ATFDV) were indicators of general attitudes toward violence among non-dating students but ATFDV affected physical violence and ATMDV affected psychological aggression for both dating boys and girls. PMID- 18162637 TI - Psychopathy and affect consciousness in young criminal offenders. AB - A key characteristic of psychopathy is the individual's problematic relation to certain affects, particularly shame. Previous research has studied relations between expressed shame and psychopathy. In this study, the author analyzes potential associations between psychopathy and consciousness of feelings (i.e., participants' ability to recognize and tolerate the feeling and describe how they believe it is expressed in their posture and verbal expressions). Psychopathy is assessed with the Psychopathy Checklist: Screening Version, and affect consciousness is assessed with a semistructured interview. Additionally, risk for criminal behavior, moral reasoning, cognitive distortions, and attachment style in the participants was assessed. The participants are 47 adolescent boys who were treated at juvenile delinquency homes. The results indicate that boys with higher ratings of psychopathy had lower consciousness of shame feelings and lower empathy scores. The results in combination with qualitative analyses of the interview answers are interpreted as indicating that consciousness of shame is specifically problematic for psychopathic adolescents. PMID- 18162638 TI - Violent men: ordinary and deviant. AB - This article deals with discourses of intimate partner violence and is based on interviews with professionals who meet violent men. The professionals emphasized the importance of men taking unreserved responsibility for their violent behavior. Intimate partner violence was viewed not only as "power and control" but as the result of complex situations and interplays. The discourses presented an ambivalent explanation of violent men as both ordinary and deviant. They were understood as having a strained background, but to be rather ordinary, often functioning well at work and in society. Yet, they have nonstandard views of women, act deviant in their communication and interplay with others, and cannot cope with certain situations in intimate relationships. Based on the interviews, men inclined to partner violence may be generalized as those who: attack immediately, explode unexpectedly, or ultimately become aggressive. The discussion challenges unreflected discourses as means for change when counteracting violence. PMID- 18162639 TI - Symptoms of trauma, perceptions of discrimination, and anger: a comparison between Australian indigenous and nonindigenous prisoners. AB - Although the need for the development and provision of culturally appropriate rehabilitation programs for offenders is widely acknowledged, there is a lack of empirical data that can be used as a basis for the development of new programs. This article reports the findings of a comparison of indigenous and nonindigenous male prisoners on a range of measures relevant to the experience of anger by indigenous prisoners in Australia. The results suggest that indigenous participants are more likely to experience symptoms of early trauma, have greater difficulties identifying and describing feelings, and perceive higher levels of discrimination than nonindigenous prisoners. The implications of this work for the development of culturally appropriate and effective anger management programs for indigenous male prisoners are discussed. PMID- 18162640 TI - Predictors of child molestation: adult attachment, cognitive distortions, and empathy. AB - A conceptual model derived from attachment theory was tested by examining adult attachment style, cognitive distortions, and both general and victim empathy in a sample of 61 paroled child molesters and 51 community controls. Results of logistic multiple regression showed that attachment anxiety, cognitive distortions, high general empathy but low victim empathy significantly increased the odds of child molester status. Findings supported theoretically based hypotheses, suggesting that attachment theory may be useful in the conceptualization and treatment of child molesters. PMID- 18162641 TI - Reasons underlying treatment preference: an exploratory study. AB - Very little is known about what factors influence women's treatment preferences after a sexual assault. To learn more about these factors, data were collected from 273 women who read a standard "if this happened to you, what would you do" scenario describing a sexual assault and subsequent trauma-related psychiatric symptoms. After reading standardized treatment options for a pharmacotherapy (sertraline) and a psychotherapy (cognitive behavioral treatment), participants made a hypothetical treatment choice and reported the main reasons for their choice. Women often cited reasons surrounding the effectiveness of a treatment as the primary reason for their treatment preference, suggesting potential masking of symptoms with the medication and more logical, long-lasting effects with the psychotherapy. Other common reasons underlying treatment preference were wariness of the medication and positive feelings about talking in psychotherapy. Better understanding factors that influence treatment preference may aid in refining psychoeducation materials regarding the psychological consequences of sexual assault and their treatment for the lay public and in helping clinicians further tailor their discussion of treatment alternatives for these women. PMID- 18162642 TI - Language intervention practices for school-age children with spoken language disorders: a systematic review. AB - PURPOSE: This systematic review focuses on peer-reviewed articles published since 1985 that assess the outcomes of language intervention practices for school-age students with spoken language disorders. METHOD: We conducted computer searches of electronic databases and hand searches of other sources for studies that used experimental designs that were considered to be reliable and valid: randomized clinical trials, nonrandomized comparison studies, and multiple-baseline single subject design studies. RESULTS: The review yielded 21 studies concerning the efficacy or effectiveness of language intervention practices with school-age children since 1985. Eleven of the studies limited participants to children in kindergarten and first grade, and no studies were located that focused on students in middle grades or high school. The relatively high quality of the studies that met our criteria, and the moderate-to-high effect sizes we calculated for the majority of studies, suggests that clinicians can have some confidence in the specific language intervention practices examined. CONCLUSION: The fact that only 21 studies met our criteria means that there is relatively little evidence supporting the language intervention practices that are currently being used with school-age children with language disorders. We outline significant gaps in the evidence and discuss the implications for clinical practice in schools. PMID- 18162644 TI - Phonological awareness intervention: beyond the basics. AB - PURPOSE: The purpose of this article is to advance practitioners' knowledge base of best practices in phonological awareness intervention to facilitate the implementation of evidence- or research-based practices in everyday clinical practice. Although most speech-language pathologists (SLPs) have a basic knowledge of phonological awareness, this article provides additional information on the variables to consider in the design and implementation of phonological awareness intervention; access to this information has a clear impact on practitioners' efforts to move research to practice. METHOD: We reviewed the relevant literature on the nature and development of phonological awareness and phonological awareness intervention to identify evidence-based intervention practices. We draw on clinical experience to supplement the research literature, particularly where the research literature provides limited direction. IMPLICATIONS: SLPs have a unique contribution to make in school-based teams' efforts to facilitate literacy development in children, particularly children who are at risk for reading disability. Research provides much information to guide clinicians in the design and implementation of phonological awareness intervention. PMID- 18162645 TI - Siblings of children with speech impairment: cavalry on the hill. AB - PURPOSE: The purpose of this article was to examine the experiences of siblings of children with speech impairment, an underresearched area of family-centered practice. METHOD: Using naturalistic inquiry, we interviewed 6 siblings and 15 significant others. Interview transcripts were analyzed for meaning statements, and meaning statements were coded and organized thematically. RESULTS: Four major themes emerged as significant in these children's experiences. The first theme described the typically positive relationship when siblings were together. The second theme related to the siblings' relationship in the context of outsiders, when the sibling of the child with speech impairment frequently undertook the roles of protector and interpreter. Exemplifying this, 1 mother described her daughter as the cavalry on the hill. In the third theme, the impact on self, siblings expressed jealousy and resentment as well as worry and concern toward the child with speech impairment. In the fourth theme, the impact on parent-child relationships, siblings expressed an awareness that they experienced less parental attention, had concerns regarding the impact of the circumstances on their parents, and took on a parent-like role toward the child with impairment, and for some, toward their parents. CONCLUSION: As part of family-centered practice, speech-language pathologists need to be aware of the important roles that siblings play and should routinely include siblings in assessment and intervention. PMID- 18162646 TI - Promoting literacy in students with ASD: the basics for the SLP. AB - PURPOSE: This article provides a tutorial for speech-language pathologists (SLPs) concerning approaches for improving the reading skills of students with autism spectrum disorder (ASD). SLPs are encouraged to modify their role to include a literacy focus, not only because of inclusion and standardized reading tests, but also because SLPs offer skills to enrich the literacy skills of students with ASD. METHOD: This article is organized to provide guidance on approaches associated with reading achievements in 3 stages of development: (a) emergent, (b) conventional, and (c) skilled reading. For each, we provide a brief overview of major achievements observed in students with typical development as well as a synopsis of what is currently known concerning the achievements of students with ASD. We then provide suggestions concerning specific approaches that can be used to further the reading and oral language skills of students with ASD within the particular stage. IMPLICATIONS: This article suggests literacy interventions that target critical oral language and literacy skills that have been well documented as areas of need among many students with ASD. SLPs can draw on this information when designing and implementing transdiciplinary literacy interventions for this growing population of students whose literacy needs are currently underserved. PMID- 18162647 TI - Empirically derived combinations of tools and clinical cutoffs: an illustrative case with a sample of culturally/linguistically diverse children. AB - PURPOSE: Using a sample of culturally/linguistically diverse children, we present data to illustrate the value of empirically derived combinations of tools and cutoffs for determining eligibility in child language impairment. METHOD: Data were from 95 4- and 6-year-olds (40 African American, 55 White; 18 with language impairment, 77 without) who lived in the rural South; they involved primarily scores from the Comprehension subtest of the Stanford-Binet Intelligence Scale Fourth Edition (CSSB; R. Thorndike, E. Hagen, & J. Sattler, 1986), but scores from an experimental nonword repetition task (NRT; C. Dollaghan & T. Campbell, 1998) were also included as supplements to these scores. RESULTS: Although the CSSB led to low fail rates in children without impairment and a statistically reliable group difference as a function of the children's clinical status but not their race, only 56% of children with impairment were accurately classified when 1 SD was employed as the cutoff. Diagnostic accuracy improved to 81% when an empirically derived cutoff of -.5 SD was used. When scores from the NRT were added to those from the CSSB, diagnostic accuracy increased to 90%. IMPLICATIONS: This illustrative case adds to the growing number of studies that call for empirically derived combinations of tools and cutoffs as one option within an evidence-based practice framework. PMID- 18162648 TI - Identifying language comprehension impairment in preschool children. AB - PURPOSE: This study examined the validity of 3 discourse comprehension measures for preschool children and the ability of a combination of them to classify children with and without language impairment. METHOD: Thirty-seven children with typical language and 12 children with language impairment completed 3 measures of oral story comprehension: the Joint Story Retell task, the Expectancy Violation Detection task, and comprehension questions. RESULTS: Discriminant analyses revealed that each measure successfully classified pre-identified groups. The clinical combination of all 3 measures resulted in 96% accurate identification of the language status of this cohort. CONCLUSION: Results support these procedures as valid measures of discourse comprehension and monitoring and provide preliminary evidence that their combination can be validly employed for identifying young children with language comprehension impairment. PMID- 18162649 TI - Training and knowledge in autism among speech-language pathologists: a survey. AB - PURPOSE: The current study was designed to answer the following questions: (a) What knowledge do school-based speech-language pathologists (SLPs) have concerning autism? (b) What educational and clinical training do SLPs receive in autism? (c) Do SLPs have confidence in their ability to provide services to children with autism and their families? METHOD: An original 52-item survey was designed to answer the research questions. Participants were recruited through e mail and were asked to respond to a Web-based survey. RESULTS: Sixty-seven school based SLPs practicing in 33 states across the United States responded to the survey. Most participants had accurate knowledge about the characteristics of children with autism; however, they had mixed perceptions of diagnostic criteria for autism. Although most participants did address autism at some level of their educational training, little time was spent discussing the topic. Additionally, some SLPs lack confidence in their abilities to provide services to children with autism. CONCLUSION: The return rate for participants was small, and it is difficult to generalize the results. However, the majority of respondents reported that they could have benefited from additional training in the area of autism. As a result, it may be necessary to consider strategies for providing this training. PMID- 18162650 TI - A qualitative examination of current guidelines for evidence-based practice in child language intervention. AB - PURPOSE: The present investigation examines the time, effort, and resources that evidence-based practice (EBP) requires, and looks at what types of results are obtained. METHOD: Principles of EBP as outlined by the American Speech-Language Hearing Association were applied to 3 treatment case studies. RESULTS: The cases took between 3 and 7 hr to complete and yielded different amounts and types of results. Individual article searches yielded better results than did searches of systematic reviews. Most evidence found was from small-group and individual case studies. CONCLUSION: EBP is a necessary part of effective and ethical treatment. The resources needed, time involved, and availability of relevant information make current guidelines for EBP challenging for most full-time practitioners. Specific recommendations to address these concerns are offered. PMID- 18162651 TI - The impact of early intervention on speech and lexical development for toddlers with cleft palate: a retrospective look at outcome. AB - PURPOSE: The purpose of the present investigation was to examine the impact of early intervention on speech and lexical measures for toddlers with cleft palate. METHOD: Speech measures of ten 27-month-old toddlers with cleft palate who had been referred for therapy at 17 months of age were compared to those of 10 toddlers with cleft palate who had been referred but did not receive therapy. Both groups were compared to 2 other groups of children who had never been referred for therapy. RESULTS: Multivariate analysis of variance revealed only 1 significant difference between the children who received therapy and those who did not receive therapy. Children who received therapy produced a significantly greater percentage of glides (p < .001) than did children who did not receive therapy. CONCLUSION: The findings of this study indicated that early intervention as conducted by the speech-language pathologists in this study was not as effective as expected for this group of toddlers with cleft palate. PMID- 18162652 TI - Absenteeism rates in students receiving services for CDs, LDs, and EDs: a macroscopic view of the consequences of disability. AB - PURPOSE: Elevated levels of absenteeism have been reported for students receiving special education services, especially students with learning disabilities (LDs) and emotional disturbances (EDs). In contrast, little is known about absenteeism rates associated with students with communication disorders (CDs). METHOD: Archival records of student attendance for the 1997-2001 academic years from a mid-sized urban school district in the Intermountain West were used to examine absenteeism rates in students receiving services for CDs, LDs, and EDs. Two cohort samples were generated (K-4th grade and 5th-9th grade). RESULTS: Students with CDs displayed absenteeism rates that were comparable to those of students receiving general education. Significant group and group x grade effects were found. Students in the LD and ED groups displayed elevated levels of absenteeism, especially at Grade 9. CONCLUSION: The outcomes of this study support the characterization of the socioemotional concomitants associated with CDs as being typologically different and exerting a weaker influence on student's health levels than those associated with either LDs or EDs. PMID- 18162653 TI - Preventing multiple-choice tests from impeding educational advancement after acquired brain injury. AB - PURPOSE: The purpose of this article is to present management strategies that can be used to breach barriers created for students with acquired brain injuries by testing the students in the multiple-choice format. METHOD: This article presents a case study of a high school student with severe hydrocephalus and difficulties with state-mandated reading comprehension tests who was denied exceptional student education services because her grades were "so good." Result Although an honor student who received academic awards, she was never taught how to pass the state reading test and was denied her diploma at graduation. IMPLICATIONS: The cognitive obstacles posed by the multiple-choice format can be specified and treated. In-service training can help school staff and officials to recognize and serve these children promptly so that academic disability can be prevented. PMID- 18162654 TI - Genes, personality, and attachment in adults: a multivariate behavioral genetic analysis. AB - Behavioral genetic methods were used to estimate genetic and environmental contributions to (a) attachment-related anxiety and avoidance and (b) the overlap between these attachment dimensions and the Big Five personality traits. Two major findings emerged from these analyses. First, individual differences in attachment-related anxiety and avoidance were heritable, and second, much of the overlap between adult attachment and the Big Five traits was due to shared genetic influences. Discussion focuses on the implications of these results for attachment theory and research and provides a speculative account of the interplay between temperamental dimensions and internal working models of relationships across the life span. PMID- 18162655 TI - What do we see in a tilted square? A validation of the figure independence scale. AB - The Figure Independence Scale (FIS) assesses people's preference for abstract figures that represent uniqueness. As psychological manifestations of cultural values cohere among each other within a cultural system, the authors argue that preference for uniqueness, as a psychological manifestation of the value for independence, can be used as an indirect measure of this value. Four studies examine the convergent, discriminant, and predictive validity of the FIS. The results indicate that liking for abstract, unique figures coheres with other specific manifestations of the value for independence (i.e., responses to explicit independence scale, need for personal control, liking for independence themes in advertisements, and use of social coping) and thus can be used as a measure of individuals' more global endorsement of this individualistic value. PMID- 18162656 TI - Longitudinal investigation of reciprocal relationship between stress reactions and hardiness. AB - Although many studies have examined hardiness as a protective factor against stress reactions, to the authors' knowledge no studies have yet examined the impact of stress reactions on hardiness. However, there is theoretical support for this notion. In this longitudinal study of 1,571 Marine recruits who participated in a highly stressful training program, the authors applied regression-based cross-lagged analyses to examine associations between stress reactions and hardiness over time for both men and women, and they investigated social support as a moderator of these relationships. Men who were hardier at Time 1 (T1) reported lower stress reactions at Time 2 (T2), and men who experienced more stress reactions at T1 were less hardy at T2. Although the negative impact of stress reactions on hardiness was strongest when social support was low for both genders, stress reactions predicted enhanced hardiness when social support was high for women only. PMID- 18162657 TI - When being a model minority is good . . . and bad: realistic threat explains negativity toward Asian Americans. AB - The current research explores the hypothesis that realistic threat is one psychological mechanism that can explain how individuals can hold positive stereotypical beliefs toward Asian Americans yet also express negative attitudes and emotions toward them. Study 1 demonstrates that in a realistic threat context, attitudes and emotions toward an anonymous group described by only positive, "model minority" attributes are significantly more negative than when the group was described using other positive attributes. Study 2 demonstrates that realistic threat significantly mediates the relationship between (a) the endorsement of the both the positive and negative stereotypes of Asian Americans and (b) subsequent negative attitudes and emotions toward them. Studies 3 and 4 conceptually replicate this effect in experimental situations involving interactions with Asian Americans in realistic threat contexts. Implications for understanding the nature of stereotyping and prejudice toward Asian Americans and other minority groups are discussed. PMID- 18162658 TI - Punishing hubris: the perils of overestimating one's status in a group. AB - Individuals engage in status self-enhancement when they form an overly positive perception of their status in a group. We argue that status self-enhancement incurs social costs and, therefore, most individuals perceive their status accurately. In contrast, theories of positive illusions suggest status self enhancement is beneficial for the individual and that most individuals overestimate their status. We found supportive evidence for our hypotheses in a social relations analysis of laboratory groups, an experiment that manipulated status self-enhancement, and a study of real-world groups. Individuals who engaged in status self-enhancement were liked less by others and paid less for their work. Moreover, individuals tended to perceive their status highly accurately. Mediation analyses showed that status self-enhancers were socially punished because they were seen as disruptive to group processes. PMID- 18162659 TI - A mile in moccasins: how situational experience diminishes dispositionism in social inference. AB - In four studies, this article investigates the impact of situational experience on social inference. Participants without firsthand experience of a situation made more extreme and erroneous inferences about the personalities of people behaving in that situation than did participants with firsthand experience. Firsthand experience, thus, appears to diminish dispositionism in social inference because it informs people about the situational constraints that guide behavior. Across all studies, participants also displayed holier-than-thou biases, overpredicting how generously they would act relative to predictions about their peers and also relative to how they actually acted when the situation came. PMID- 18162660 TI - Advantaged group's emotional reactions to intergroup inequality: the dynamics of pride, guilt, and sympathy. AB - Three studies establish intergroup inequality to investigate how it is emotionally experienced by the advantaged. Studies 1 and 2 examine psychology students' emotional experience of their unequal job situation with worse-off pedagogy students. When inequality is ingroup focused and legitimate, participants experience more pride. However, when inequality is ingroup focused and illegitimate, participants experience more guilt. Sympathy is increased when inequality is outgroup focused and illegitimate. These emotions have particular effects on behavioral tendencies. In Study 2 group-based pride predicts greater ingroup favoritism in a resource distribution task, whereas group-based sympathy predicts less ingroup favoritism. Study 3 replicates these findings in the context of students' willingness to let young immigrants take part in a university sport. Pride predicts less willingness to let immigrants take part whereas sympathy predicts greater willingness. Guilt is a weak predictor of behavioral tendencies in all studies. This shows the specificity of emotions experienced about intergroup inequality. PMID- 18162661 TI - "I don't mean to sound arrogant, but . . ." The effects of using disclaimers on person perception. AB - Limited research has examined the effects of using disclaimers on person perception, and none has examined disclaimer effectiveness. Four studies test whether disclaimers effectively ward off negative judgments regarding the specific disclaimed traits. Study 1 finds that using an arrogance disclaimer ("I don't mean to sound arrogant, but . . .") increased judgments of the speaker's arrogance but only when followed by an arrogant comment. Studies 2 and 3 yield similar findings using laziness and selfishness disclaimers. Studies 2, 3, and 4 examine five possible underlying mechanisms regarding why disclaimers can backfire. The most support was obtained for the notion that disclaimers increase an audience's expectations that the speaker will say something consistent with the unwanted trait, which then increases perceptions of that trait in the speaker. We discuss some possible moderating variables as well as some implications of these findings for general impression formation models. PMID- 18162662 TI - Are women always more interpersonally sensitive than men? Impact of goals and content domain. AB - Two studies examined motivation and content domain as possible influences on sex differences in interpersonal sensitivity. Although much research has found women to excel on tasks measuring interpersonal sensitivity, most of the tasks have measured accuracy in female-relevant domains such as emotion. The present studies measured interpersonal sensitivity, defined as accurate recall of another person, for both female-relevant and male-relevant content domains and also included motivational manipulations intended to influence men and women differently. Study 1 measured accuracy of recalling information in a written vignette about a person, and Study 2 measured accuracy of recalling details about an interaction partner. Both studies supported hypotheses about domain specificity and gender relevant motivation. However, even for male-stereotypic content and for tasks framed to favor men's motivation to perform well, men's accuracy never exceeded women's. PMID- 18162663 TI - Effects of emulsified policosanols with different chain lengths on cholesterol metabolism in heterozygous LDL receptor-deficient mice. AB - Policosanol is a mixture of long-chain primary aliphatic saturated alcohols. Previous studies in humans and animals have shown that these compounds improved lipoprotein profiles. However, more-recent placebo-controlled studies could not confirm these promising effects. Octacosanol (C28), the main component of sugarcane-derived policosanol, is assumed to be the bioactive component. This has, however, never been tested in an in vivo study that compared individual policosanol components side by side. Here we present that neither the individual policosanol components (C24, C26, C28, or C30) nor the natural policosanol mixture (all 30 mg/100 g diet) lowered serum cholesterol concentrations in LDL receptor knock-out (LDLr(+/-)) mice. Moreover, there was no effect on gene expression profiles of LDLr, ABCA1, HMG-CoA synthase 1, and apolipoprotein A-I (apoA-I) in hepatic and small intestinal tissue of female LDLr(+/-) mice after the 7 week intervention period. Finally, none of the individual policosanols or their respective long-chain fatty acids or aldehydes affected de novo apoA-I protein production in vitro in HepG2 and CaCo-2 cells. Therefore, we conclude that the evaluated individual policosanols, as well as the natural policosanol mixture, have no potential for reducing coronary heart disease risk through effects on serum lipoprotein concentrations. PMID- 18162664 TI - Editorial: medical professional liability crisis in Asia Pacific. PMID- 18162665 TI - Orthopaedic trauma surgery performed by unsupervised and supervised trainees: complication rates compared. AB - PURPOSE: To compare the complication rates associated with orthopaedic trauma surgery performed by unsupervised and supervised trainees. METHODS: In our hospital, 6361 orthopaedic trauma operations were performed between 1 January 1998 and 31 December 2002. Data pertinent to the surgeon's supervision and postoperative complications were collected. Elective operations were excluded, as consultants were almost always present. Complication rates ensuing in unsupervised and supervised groups were compared using the Chi squared test. RESULTS: Of 6361 orthopaedic trauma operations performed, 3754 (59%) were by unsupervised trainees of varying experience, whereas 2494 (39%) were by supervised trainees or consultants. In 113 (2%) of the operations, the supervision status was not recorded. The complication rate was significantly higher in the supervised than unsupervised group (5.3 vs 3.3%, Chi squared=15, df=1, p=0.0001). CONCLUSION: The complication rate was not higher for operations performed by unsupervised trainees than those performed in the presence of a consultant. PMID- 18162666 TI - Resistance of disposable drapes to bacterial penetration. AB - PURPOSE: To test the bacterial penetrability of disposable non-woven drapes used specifically for total hip arthroplasty. METHODS: 12 round agar plates were inoculated with 107 colony-forming units/ml of coagulase-negative staphylococci (CNS) and incubated in air at 37 degrees Celsius for 18 hours to obtain a semi confluent growth of organisms. Six brands of disposable drapes were tested; each was assigned to 2 plates. Each disposable drape was placed between a round agar plate and an inverted square agar plate filled with blood agar. After 30 and 90 minutes, the square agar plates were removed and incubated for 48 hours and inspected for growth of CNS. RESULTS: Bacterial penetration was time dependant. Certain brands of drapes were more impenetrable than others; none was impenetrable at all time points, but most remained so or allowed passage of fewer than 100 colony-forming units at 90 minutes. CONCLUSION: It is recommended that drapes be rigorously tested with regard to their resistance to bacterial penetration. PMID- 18162667 TI - Anti-platelet agents and surgical delay in elderly patients with hip fractures. AB - PURPOSE: To assess the risk of surgical delay in elderly hip fracture patients on anti-platelet agents. METHODS: Records of 180 patients aged over 65 years with either an intertrochanteric or femoral neck fracture were reviewed. The clopidogrel group included 10 patients on clopidogrel alone and 11 others on clopidogrel and aspirin, whereas the control group included 69 on aspirin alone and the remaining 90 not on any anti-coagulants. The 2 groups were compared with regard to time to surgery, preoperative American Society of Anesthesiologists (ASA) score, pre- and post-operative haemoglobin levels, in-patient complication rates, duration of hospital stay, and 30-day mortality. RESULTS: In the clopidogrel and control groups respectively, the mean times to surgery were 7.2 and 2.1 days (p=0.03, t-test), the mean preoperative ASA scores were 3.35 and 2.8 (p=0.29, t-test), the mean preoperative haemoglobin levels were 119 and 115 g/l (p=0.5, t-test), the mean postoperative haemoglobin levels were 98 and 96 g/l (p=0.68, t-test), the mean durations of hospital stay were 7.4 and 3.1 days (p=0.02, t-test). The 30-day mortalities were 6/21 (29%) and 6/159 (4%) [p=0.0003, Fisher's exact test]. CONCLUSION: Surgical delay in elderly patients on anti-platelet agents with hip fracture was associated with higher mortality. Despite the risk of increased blood loss, we suggest early surgery be carried out by an experienced surgeon to expedite the operating time. Pooled platelets should be given intravenously one to 2 hours preoperatively. PMID- 18162668 TI - Proximal femoral nail for treatment of trochanteric femoral fractures. AB - PURPOSE: To report outcomes of 87 consecutive patients treated with a proximal femoral nail (PFN) for trochanteric femoral fractures. METHODS: 17 men and 70 women aged 58 to 95 (mean, 85) years with trochanteric femoral fractures underwent PFN fixation using an intramedullary nail, a lag screw, and a hip pin. Fractures were classified according to the AO system; the most common fracture type was A2 (n=45), followed by A1 (n=36) and A3 (n=6). The position of the lag screw within the femoral head was measured. The lateral slide of the lag screw after fracture consolidation was measured by comparing the immediate postoperative and final anteroposterior radiographs. RESULTS: 90% of lag screws were placed in an optimal position. The length of lateral slide of the lag screw in stable A1 fractures was significantly less than that in unstable A2 fractures; it was over 10 mm in 7 of 45 patients with A2 fractures. Cut-out of lag screw did not occur, suggesting that free sliding of the lag screw facilitates direct impaction between fragments. CONCLUSION: A PFN is useful for the treatment of trochanteric femoral fractures. PMID- 18162669 TI - Intramedullary nailing versus fixed angle blade plating for subtrochanteric femoral fractures: a prospective randomised controlled trial. AB - PURPOSE: To compare closed intramedullary nailing to open reduction and internal fixation using a fixed angle blade plate for the management of subtrochanteric femoral fractures. METHODS: 58 patients were equally randomised to undergo either an intramedullary nailing (IN) or fixed angle blade plating (BP). RESULTS: There were no significant differences between the 2 groups with regard to age, time to surgery, operating time, receipt of blood transfusions, duration of hospital stay, or fracture classification. The revision rate was 28% (8/29) in the BP group and none in the IN group; the difference was statistically significant. CONCLUSION: Internal fixation using a fixed angle blade plate for subtrochanteric femoral fractures has higher implant failure and revision rates, compared to closed intramedullary nailing. PMID- 18162670 TI - The effectiveness of capsular decompression for internal fixation of intracapsular hip fractures. AB - PURPOSE: To compare the results of screw fixation plus capsular decompression versus screw fixation alone for managing intracapsular hip fractures. METHODS: Of 201 patients with intracapsular hip fractures, 99 underwent screw fixation with capsular decompression (capsular decompression group) and 102 underwent screw fixation alone (control group). The incidence and time to development of avascular necrosis of the femoral head, union rate, time to union, and other clinical parameters were compared. RESULTS: In patients with displaced fractures, the incidence of avascular necrosis was significantly higher in the control than capsular decompression group, whereas the time to development of this complication was significantly shorter. CONCLUSION: Capsular decompression did not improve the union rate and time to union in undisplaced intracapsular hip fractures, but in displaced fractures it appeared to reduce the incidence and delay the onset of avascular necrosis. PMID- 18162671 TI - Salter innominate osteotomy for treatment of developmental dysplasia of the hip. AB - PURPOSE: To compare the results of Salter innominate osteotomy (SIO) for treatment of developmental dysplasia of the hip (DDH) in children. METHODS: Between 1994 and 2002, 53 girls and 8 boys with DDH underwent open reduction and SIO; 21 were on the left side, 22 on the right side, 18 bilateral. They were divided into 2 groups: group 1 included 35 patients aged younger than 3 years (46 hips) and group 2 included 26 patients aged 3 years or older (33 hips). Clinical outcomes were assessed using the modified McKay criteria to measure pain symptoms, gait pattern, Trendelenburg sign status, and the range of hip joint movement. Radiographic outcomes were evaluated using the Severin method to measure the Sharp acetabular angle and the centre-edge angle. RESULTS: Group-1 children achieved slightly better reduction and stabilisation of the hip joint empirically, but clinical and radiographic results and complication rates in the 2 groups were not significantly different. CONCLUSION: Open reduction and SIO without preoperative traction is effective for the management of DDH in children younger than 6 years. PMID- 18162672 TI - Intramedullary nailing for pathological femoral fractures. AB - PURPOSE: To evaluate whether the operating time correlates with the survival and outcome in patients who underwent intramedullary nailing for metastatic femoral fractures. METHODS: Records of 10 men and 11 women aged 43 to 86 (mean, 66) years who underwent intramedullary nailing (4 bilaterally) for metastatic femoral fractures between 1999 and 2003 were reviewed. The patients were followed up for at least 2 years or until their death. The main outcome measure was the correlation between operating time and survival. Operating time was categorised into 5 groups from 60 to 210 minutes, with 30-minute increments. RESULTS: Operating time does not correlate with survival and outcome. The mean survival period was 9.4 months. Pain relief was achieved in 90% of the patients. There was no implant failure, but one loss of reduction. CONCLUSION: Intramedullary nailing appears safe and effective for treatment of metastatic bone disease, and confers good functional results, pain relief, and mobility. PMID- 18162673 TI - Surgical management of metastatic disease of the proximal femur. AB - PURPOSE: To review the surgical treatment for metastatic disease of the proximal femur. METHODS: Records of 8 patients who underwent endoprosthetic replacement with tumour resection (group 1) and 8 others who underwent intramedullary nailing without tumour resection (group 2) were reviewed. Treatments were based on the disease progression and patient's condition. RESULTS: In groups 1 and 2, the respective mean survival periods were 16 and 4 months. All patients in group 1 regained preoperative mobility, but only one patient in group 2 was able to walk with crutches. CONCLUSION: This was a retrospective, rather than comparative study of endoprothetic replacement and intramedullary nailing for metastatic disease of the proximal femur. Both procedures are considered palliative, and not curative. The longer survival period in group 1 was mainly due to selection of patients with better preoperative medical status. PMID- 18162674 TI - Less invasive stabilisation system plating for distal femoral fractures. AB - PURPOSE: To review the outcome of less invasive stabilisation system (LISS) plating for complex distal femoral fractures. METHODS: Records of 6 men and 11 women who underwent LISS plating for complex distal femoral fractures from September 2001 to August 2005 were reviewed. One patient who died 12 months after surgery due to a cardiac problem was excluded. The mean age of the remaining patients was 61 years and the mean follow-up period was 12 months. Four patients, 3 of whom had open fractures, had sustained high-energy trauma. According to the AO classification, 8 fractures were type 33A and 9 type 33C. RESULTS: The mean time to union was 17 weeks. Two patients with non-union underwent a second LISS plating and bone grafting, resulting in a satisfactory final outcome. Delayed radiographic union was observed in one patient, but clinically he was asymptomatic and mobile. The fracture finally united at 9 months. CONCLUSION: LISS plating is useful in treating complex distal femoral fractures, resulting in reduced blood loss and low infection rates, while achieving early mobility due to primary stability of the construct. PMID- 18162675 TI - Does flexion contracture continue to improve up to five years after total knee arthroplasty? AB - PURPOSE: To determine whether flexion contracture continues to improve up to 5 years after total knee arthroplasty (TKA). METHODS: Records of 200 women and 123 men (mean age, 72 years) with osteoarthritis and preoperative flexion contracture were reviewed. 155 (48%) involved the left knee and 168 (52%) the right knee. Patients were assessed clinically before TKA and one and 5 years later, by a single researcher using a goniometer to measure flexion contracture, range of movement, maximum flexion, and the Knee Society Score. In all patients the same prosthesis design was used. The surgical technique was standard and involved a medial parapatellar approach with bone resection. RESULTS: All variables improved significantly from pre-TKA to one-year post-TKA and from one-year post-TKA to 5 year post-TKA; respectively the mean flexion contracture decreased from 11 to 3 to 2 degrees; the mean knee range of movement increased from 90 to 103 to 107 degrees; the mean maximum flexion increased from 101 to 106 to 109 degrees; and the mean Knee Society Scores increased from 39 to 87 to 90. CONCLUSION: Functional outcome and flexion contracture may continue to improve up to 5 years after TKA. PMID- 18162676 TI - Modified Brostrom-Evans-Gould technique for recurrent lateral ankle ligament instability. AB - PURPOSE: To review the outcome of the modified Brostrom-Evans-Gould technique in patients with chronic lateral ankle ligament instability. METHODS: Between August 2003 and June 2005, 20 men aged 19 to 35 (mean, 23) years with chronic lateral instability affecting 21 ankles, underwent the modified Brostrom-Evans-Gould technique by a single surgeon. The mean follow-up period was 12 (range, 6-20) months. Patients were assessed preoperatively and postoperatively using the Kaikkonen Ankle Scoring Scale. RESULTS: Preoperatively, all patients had poor scores (<50). Postoperatively, 17 (81%) of the ankles attained excellent scores (85-100) and 4 (19%) attained good scores (70-84). CONCLUSION: The modified Brostrom-Evans-Gould technique appears effective for chronic lateral ankle ligament instability, particularly in the Asian population with a higher prevalence of generalised joint hyperlaxity. PMID- 18162677 TI - Management of acute Achilles tendon ruptures in the United Kingdom. AB - PURPOSE: To survey the management of acute Achilles tendon ruptures in the United Kingdom. METHODS: A questionnaire with 4 different Achilles tendon rupture scenarios affecting patients of different ages and activity levels was sent to orthopaedic consultants specialised in sports trauma, foot and ankle surgery. Their treatment methods including surgical techniques, immobilisation and rehabilitation regimens were surveyed. RESULTS: The response rate was 22% (51/231). Among the 51 respondents, 25 had managed less than 6 such patients in the preceding year. 26 (51%) used clinical examination (e.g. calf squeeze test) to make the diagnosis, 16 (31%) used ultrasonography, and 4 (8%) used magnetic resonance imaging. Surgical management was preferred for younger and more functionally demanding individuals. Open repairs were used more often than percutaneous repairs (72 vs 19%). Across the 4 scenarios, the mean time to return to full activity was 17 (range, 12-32) weeks for conservative to 19 (range, 10 40) weeks for surgical management. CONCLUSION: Variation in opinion among respondents was wide. Randomised controlled trials are needed to assess optimal treatment. PMID- 18162678 TI - Coracoclavicular ligament reconstruction using a gracilis tendon graft for acute type-III acromioclavicular dislocation. AB - PURPOSE: To review the functional and radiological results of patients after coracoclavicular ligament reconstruction. METHODS: Five patients aged 21 to 50 (mean, 37) years with acute Rockwood type-III acromioclavicular dislocation underwent coracoclavicular ligament reconstruction with autogenous gracilis tendon grafts. Patients were either active in sports or heavy manual workers. Assessments on shoulder function (using the Constant score), wound size, pain (using a visual analogue scale), and reduction (using radiographs of both acromioclavicular joints) were made. RESULTS: The mean follow-up period was 26 (range, 15-43) months; the mean time to return to work or sports was 14 (range, 12-20) weeks. The mean Constant score was 94 (range, 90-98). The mean donor-site scar size was 3 cm and the mean pain score was 0. No major complication or donor site morbidity was noted. There was one subluxation. CONCLUSION: Coracoclavicular ligament reconstruction using an autogenous gracilis tendon graft was safe in physically active patients having acute type-III acromioclavicular dislocation. PMID- 18162679 TI - Intramedullary fixation using multiple Kirschner wires for forearm fractures: a developing country perspective. AB - PURPOSE: To evaluate treatment results of intramedullary fixation using multiple Kirschner wires for diaphyseal forearm fractures. METHODS: Between January 2001 and January 2004, 184 patients (122 men and 62 women) with 288 displaced diaphyseal forearm fractures underwent intramedullary fixation using multiple Kirschner wires. The wires were inserted by opening the fracture site because radiographic facility for closed pinning was unavailable. The time to union, functional recovery, and the complication rates were assessed. RESULTS: 11 patients were lost to follow-up, leaving 173 whose data was analysed. In 167 (97%) of the patients the fractures united, and in 14 (8%) of them union was delayed. The mean time to union for closed fractures was 13 (range, 7-16) weeks and for open fractures it was 15 (range, 12-22) weeks. The mean time in cast was 6 (range, 3-14) weeks. In all, 34/173 (20%) had complications: superficial infection (n=13), deep infection (n=4), cross-union between bones (n=4), open fracture 'needing' a skin graft (n=2), radial nerve palsy (n=3), paraesthesia (n=1), and non-union (n=7). Among the 173 analysed patients, the infection rate was 10% and the non-union rate was 4%. 130 (75%) of the patients had the wires removed; no re-fracture occurred after wire removal. Based on the Anderson criteria, 47 (27%) of the patients attained excellent, 78 (45%) satisfactory, and 39 (23%) unsatisfactory results. In 9 (5%) of the patients, union failed (7 plain non-union and 2 due to chronic osteomyelitis). CONCLUSION: Kirschner wires are much cheaper than plates and screws, and require minimal expertise for insertion and removal. They remain acceptable for stabilising displaced diaphyseal forearm fractures in developing countries. PMID- 18162680 TI - Neurological recovery after occipitocervical fixation. AB - PURPOSE: To report on 14 consecutive cases of occipitocervical fixation. METHODS: Records of 8 men and 6 women aged 40 to 81 (mean, 57) years who underwent occipitocervical fixation and were followed up for a minimum of 2 years were retrospectively reviewed. Neurological grading was assessed before and after surgery using the Ranawat grade. Intra-operative somatosensory evoked potentials were monitored. RESULTS: The main indications for surgery were rheumatoid arthritis (n=6) and cervical metastasis (n=4). 77% of the patients demonstrated neurological improvement. Four out of the 5 non-ambulatory patients (Ranawat grade IIIB) regained ambulatory status postoperatively. No patient had neurological deterioration or evidence of vertebral artery or spinal cord injury. One endured a superficial wound infection and 2 had implant breakage. CONCLUSION: Although occipitocervical fixation is technically challenging and there are risks of serious neurologic or vascular complications, it remains a viable option with favourable results in patients requiring stabilisation of the craniocervical junction. PMID- 18162681 TI - Idiopathic scoliosis and pineal lesions in Australian children. AB - PURPOSE: To determine whether treatment of pineal lesions in children is associated with development of idiopathic scoliosis. METHODS: 38 boys and 10 girls with pineal lesions were identified. Their mean age at presentation was 10 years. The pineal pathology varied from cysts and epidermoid to teratoma, germinoma, pineocytoma, and glioblastoma. Treatment ranged from biopsy/extirpation to radiotherapy. RESULTS: 12 patients died. No scoliosis was found in any females or any of the deceased. Two boys had scoliosis: one had a 12 degree right upper thoracic curve with 32-degree kyphosis and the other had a 60 degree right thoracolumbar idiopathic curve, requiring a 2-stage arthrodesis. CONCLUSION: Pineal ablation is not related to the development of idiopathic scoliosis in humans. PMID- 18162682 TI - Management outcome and the role of manipulation in slipped capital femoral epiphysis. AB - PURPOSE: To examine and compare treatment outcomes for slipped capital femoral epiphysis (SCFE). METHODS: 24 patients with acute or acute-on-chronic SCFE underwent preoperative traction, 13 of them followed by in situ screw fixation (group A), and the remaining 11, manipulation with screw fixation (group B). Their treatment outcomes (according to the Aadalen criteria) and complications were compared. RESULTS: There were 2 treatment failures and one poor outcome in patients with acute SCFE (2 had avascular necrosis and one had slight pain with mild limitation of hip movement). The risk of developing avascular necrosis was significantly higher in patients with acute rather than acute-on-chronic or chronic slips. No significant differences were evident between groups A and B with regard to age, sex, ethnicity, outcomes, and preoperative and postoperative changes in the Southwick headshaft angle. CONCLUSION: Patients with acute or acute-on-chronic SCFE can be safely managed with straight-leg traction for up to 6 days, followed by in situ screw fixation without manipulation. Manipulation is not recommended in such slips, due to a higher avascular necrosis risk. PMID- 18162683 TI - Sacral insufficiency fractures. AB - PURPOSE: To highlight difficulties in the diagnostic process and the validity of imaging techniques for sacral insufficiency fractures. METHODS: Records of 25 women aged 68 to 95 years with sacral insufficiency fractures were reviewed. Baseline blood biochemistry and haematology test results were obtained. Pelvic anterior/posterior radiography was undertaken for all patients; additional computed tomography, technetium bone scanning, and magnetic resonance imaging were used in some. Treatments were based on the severity of the injury and the patient's mobility and cooperativeness. RESULTS: Among the 25 women, 11 had bilateral and 14 had unilateral vertical sacral fractures. Associated fractures included horizontal sacral fracture, fractures of the os pubis and ilium. Symptoms included lower back or buttock pain, abdominal pain, and those emulating radiculopathy and myelopathy, including leg weakness, sciatica, and urinary retention. The mean delay in diagnosis was 9 (range, 1-28) days. The mean recovery time between bilateral and unilateral fractures was significantly different (22 [range, 12-33] vs 14 [range, 8-36] weeks, p=0.01). No patient with bilateral fractures regained her pre-injury mobility, compared to 43% among those with unilateral fractures (p=0.02, Fisher's exact test). Computed tomography was the most reliable imaging technique; technetium bone scanning was highly sensitive but non-specific; magnetic resonance images of the fractures may mimic metastatic disease. CONCLUSION: With the increase in the elderly population, sacral insufficiency fractures may become epidemic in future. Primary and secondary osteoporoses are common causes. Once a diagnosis is established, in most cases treatment is simple but recovery may be protracted and full mobility curtailed. PMID- 18162684 TI - Combined pelvic osteotomy for deformed dysplastic acetabula: a 5-year prospective study. AB - PURPOSE: To assess clinical and radiological results of combined pelvic osteotomy (CPO) for deformed dysplastic acetabula. METHODS: Nine girls and 3 boys, aged 28 to 54 months, underwent CPO (combination of innominate osteotomy and Lance acetabuloplasty) to restore both the original shape and the mal-alignment of the acetabulum. They were followed up for at least 5 years. RESULTS: All patients attained concentric stable reduction with good acetabular cover. CONCLUSION: CPO corrected both deformed and dysplastic acetabula and prevented postoperative subluxation. PMID- 18162685 TI - Limb salvage surgery complimented by customised mega prostheses for malignant fibrous histiocytomas of bone. AB - PURPOSE: To assess functional and oncological outcomes of patients with malignant fibrous histiocytomas of bone, after limb salvage surgery complimented by a customised prosthesis. METHODS: Between May 1991 and December 2002, 15 men and 5 women (mean age, 42 years) with histologically proven malignant fibrous histiocytoma of bone underwent treatment involving limb salvage surgery complimented by a customised mega prosthesis. Most of the tumours were stage II according to the Enneking system, and located around the knee. Wide resection margins were achieved in 18 patients. RESULTS: Following a mean follow-up of 58 months, 4 patients underwent amputation for local recurrence and 5 died of the disease. Two patients had prosthesis fractures; revision of the prosthesis was carried out in one. The functional result was excellent in 5 and good in 9 patients. The Kaplan-Meier 5-year survival rates of the patients treated without chemotherapy and with chemotherapy were 50% and 76%, respectively. CONCLUSION: Limb salvage surgery with chemotherapy is a viable treatment option for patients with malignant fibrous histiocytoma of bone. It achieves higher survival rates than resection alone. Such therapy improves quality of life and provides a useful and functional limb. PMID- 18162686 TI - Congenital insensitivity to pain with anhydrosis in a Malaysian family: a genetic analysis. AB - A Malaysian family with congenital insensitivity to pain with anhydrosis was diagnosed based on clinical symptoms of chronic ulcers, joint deformities, malunited fractures, anhydrosis, and learning disabilities. We detected a compound heterozygous mutation in exon 16: V709L from the mother and G718S from the father. Two novel mutations were identified: at amino acid 709, a change of G to C at nucleotide 2209 (approximately 2209G to C) causing a valine to leucine substitution (V709L), and at amino acid 718, a change of G to A at nucleotide 2236 (approximately 2236G to A) causing a glycine to serine substitution (G718S). Polymorphisms identified were at nucleotides approximately 2113G to C and approximately 2176T to C. PMID- 18162687 TI - Anodyne therapy for recalcitrant diabetic foot ulcers: a report of four cases. AB - Four patients with diabetic foot ulcers that failed to heal after one month of treatment underwent anodyne therapy. Each therapy session lasted half an hour and was conducted 3 times a week for patients at home or daily for patients in hospital, for one to 2 months. The wound sizes and depths were graded according to the Wagner classification. Pictures were taken to evaluate the effects of anodyne therapy. All 4 patients had good healing of their foot ulcers. Anodyne therapy augments the effects of conservative treatment. Proper wound care and appropriate antibiotic coverage remain the basis of treatment. PMID- 18162688 TI - Implant infection caused by dermatitis: a report of two cases. AB - We report 2 cases of implant infection with Staphylococcus aureus associated with dermatitis. In both cases, the skin at the surgical site was normal and full antiseptic measures were taken. One patient had a history of chronic asteatotic eczema complicated by cellulitis; the other had generalised exfoliative dermatitis with an atopic aetiology. Dermatitis at sites remote from the operative site may be a contributing factor in implant infection. PMID- 18162689 TI - Gamma nail breakage: a report of four cases. AB - Gamma nails have been used extensively for the treatment of proximal femoral fractures. Nail breakage at the level of the aperture of the lag screw is rare. We report 4 such cases mainly associated with a large posteromedial cortex gap and nonunion. The need for adequate reduction to avoid such a complication is emphasised. PMID- 18162690 TI - Spontaneous spinal epidural haematoma in a 15-month-old boy presenting with a wry neck: a case report. AB - A 15-month-old boy presented with a 2-day history of a wry neck (bent to the left side) with no definite trauma. He had bilateral upper limb weakness and was afebrile, conscious, and stable. There was no spontaneous movement in both upper limbs. Magnetic resonance imaging of the cervical and thoracic spine demonstrated an extensive spontaneous spinal epidural haematoma from C3 to T8. 23 hours after admission, the patient underwent an emergency right-sided C3 to T8 hemi laminectomy and haematoma evacuation. The patent's strength gradually recovered and he attained full power 3 weeks after operation. Spontaneous spinal epidural haematoma is a rare disease in children. A high index of suspicion is essential for its effective management as the interval to operation is the most important prognostic factor. PMID- 18162691 TI - Discal cysts of the lumbar spine: a case report. AB - Discal cysts--intraspinal cysts communicating with an adjacent intervertebral disc--are an uncommon cause of lumbar radiculopathy. We report a case of discal cyst of the lumbar spine. The cyst contents were bloody and clotted rapidly; no disc materials were seen. Communication between the cyst and the intervertebral disc was detected. Histopathology of the cyst wall revealed fibrous connective tissue without synovial lining cells. We hypothesise that the discal cyst was formed by haemorrhage of the epidural venous plexus caused by separation of the peridural membrane by mechanical force transmitted by an annulus fibrosis fissure. The minute segmental motion caused by the affected disc may have stimulated continuous bleeding. PMID- 18162692 TI - Ilizarov external fixator for burst fracture of the lumbar spine: a case report. AB - A 50-year-old man presented with severe back pain and tenderness throughout the lumbar area after falling from a ladder. He had an unstable type-B burst fracture, with a spinal canal narrowing of 36% and an anterior height loss of 65%. His lower-limb neurological function was intact. An Ilizarov external spinal fixator was used; the pedicular half pins were inserted into the bilateral T11, T12, L2, and L3 pedicles; bilateral pedicular half pins were fixed at each level with external plates and rods. Postoperatively, the patient had a lordosis of 2 degrees and was able to walk 7 days later. The external fixator was removed at 10 weeks. Six years and 10 months after surgery, the patient had a kyphosis of 19 degrees that did not affect his activities of daily living. PMID- 18162693 TI - Gouty synovitis after total knee arthroplasty: a case report. AB - We report a case of acute gouty synovitis after total knee arthroplasty (TKA) in a patient with no history of gout. The diagnosis was confirmed by the presence of urate crystals in the synovial fluid. Acute gouty synovitis, though rare, should be considered in the differential diagnosis of an inflammed knee after TKA to avoid unnecessary surgical revision. PMID- 18162694 TI - Pseudoaneurysm of the geniculate artery following total knee arthroplasty: a report of two cases. AB - We report 2 cases of pseudoaneurysm of the geniculate artery after a total knee arthroplasty (TKA). The first one was located in the superomedial geniculate artery and resolved after compression dressing and warfarin cessation. The second one was located in the superolateral geniculate artery and was successfully treated with transarterial embolisation. There was no recurrence in both cases. The rare occurrence and delayed presentation of pseudoaneurysms pose a diagnostic challenge. A high index of suspicion is required to make the diagnosis. Early duplex Doppler ultrasonography is recommended when patients present with atypical knee pain and prolonged swelling after a TKA. PMID- 18162695 TI - Nocardia nova septic arthritis following total knee replacement: a case report. AB - We describe a case of Nocardia nova septic arthritis following a total knee replacement. A 55-year-old obese woman was admitted to hospital 5 months after knee surgery with a 3-week history of pain, swelling, and restricted mobility in her left knee but no preceding trauma/injury. 30 ml of cloudy joint fluid was aspirated and an arthroscopic examination showed extensive fibrin formation and synovitis. An arthroscopic washout was carried out using 16 litres of saline, followed by total synovectomy and intensive antibiotic therapy (clarithromycin 500 mg twice daily and co-trimoxazole [sulphamethoxazole 400 mg and trimethoprim 80 mg] once daily and augmentin duo forte 875 mg twice daily). At 2.5 years, the patient had recovered completely with no prosthetic loosening. PMID- 18162696 TI - Letter to the Editor: Immediate interlocking nailing versus external fixation followed by delayed interlocking nailing for Gustilo type IIIB open tibial fractures. PMID- 18162697 TI - Neurodevelopmental disorders. PMID- 18162698 TI - The impact of traumatic brain injuries: a global perspective. AB - Traumatic brain injury (TBI), according to the World Health Organization, will surpass many diseases as the major cause of death and disability by the year 2020. With an estimated 10 million people affected annually by TBI, the burden of mortality and morbidity that this condition imposes on society, makes TBI a pressing public health and medical problem. The burden of TBI is manifest throughout the world, and is especially prominent in Low and Middle Income Countries which face a higher preponderance of risk factors for causes of TBI and have inadequately prepared health systems to address the associated health outcomes. Latin America and Sub Saharan Africa demonstrate a higher TBI-related incidence rate varying from 150-170 per 100,000 respectively due to RTIs compared to a global rate of 106 per 100,000. As highlighted in this global review of TBI, there is a large gap in data on incidence, risk factors, sequelae, financial costs, and social impact of TBI. This should be addressed through planning of comprehensive TBI prevention programs in LMICs through well-established surveillance systems. Greater resources for research and prioritized interventions are critical to promote evidence-based policy for TBI. PMID- 18162700 TI - Pediatric stroke: rehabilitation of focal injury in the developing brain. AB - This review provides an overview of pediatric ischemic stroke to serve as a foundation for the discussion of rehabilitation strategies following focal injury in the developing brain. Cerebrovascular disease is an important cause of acquired brain injury in neonates and children. Ischemic strokes are caused by a multitude of risk factors and advances in neuroimaging have improved diagnosis and understanding of pathophysiology. Pediatric stroke provides the ideal model for the study of injury and recovery in a plastic nervous system. Though their brains likely posses greater potential and unique reorganizational skills, most children suffer neurological morbidity after stroke. An improved understanding of these systems is helping us understand, validate, and improve traditional approaches to rehabilitation while opening the door to new opportunities to improve outcome. All aspects of a patient's function, from the physical to psychological, including issues unique to children and their families, must be addressed and are reviewed. New advances and future directions for research are highlighted. PMID- 18162699 TI - Neural substrate differences in language networks and associated language-related behavioral impairments in children with TBI: a preliminary fMRI investigation. AB - The present study examined whether functional MRI (fMRI) can identify changes in the neural substrates of language in young children following traumatic brain injury (TBI). Eight children with TBI (F/M=3/5, age (Mean +/- SD)=7.98 +/- 1 years, range = 6-9 years) and a comparison group of nine children with orthopedic injuries (OI) (F/M=4/5, age (Mean +/- SD)=7.4 +/- 1 years, range=6-9 years) participated in an fMRI study of covert verb generation (VG). Results revealed significantly different BOLD signal activation in perisylvian language areas between the groups, after accounting for potential confounders such as verbal fluency and executive function. We also found significant associations between the BOLD signal activation and performance on language-specific neuropsychological tests (NEPSY verbal fluency score, Verbal IQ) and Glasgow Coma Scale (GCS) score. This study suggests that children with TBI have significantly different brain activation patterns in language circuitry compared to children with orthopedic injuries. Although we found clear differences in brain activation between the two groups, conventional MR images showed no evidence of structural abnormalities in five of eight children with TBI. Our study demonstrates the feasibility and potential utility of fMRI as a means of quantifying changes associated with language deficits in future pediatric TBI studies. PMID- 18162701 TI - A review of the complications of intrathecal baclofen in patients with cerebral palsy. AB - Intrathecal baclofen (ITB) has become a standard treatment for spasticity in patients with cerebral palsy (CP) over the last ten years. This review summarizes and assesses the evidence for the safety of ITB in patients with CP. The literature was searched using electronic databases and reference lists from March 2000 through April 2007. There were 36 studies identified reporting on the complications of ITB in patients with CP in non-controlled case series or single case reports; about half of these report only on the issue of complications, while the other half report on treatment effects as well. The majority of the participants reported on in these studies are in the pediatric or adolescent age range, but many studies include some adult patients with CP as well as a minority of patients with diagnoses other than CP. Conclusions on the basis of these disparate studies are limited; however, the available literature suggests that a relatively high rate of complications is associated with ITB treatment and that certain complications are associated with significant morbidity. Children appear to be at higher risk for complications than adults. However, until we have consistent, reliable reports of complications, it will not be possible to perform specific risk/benefit analyses. Recommendations for improving investigation in this important area are discussed. PMID- 18162702 TI - Use of behavioral interventions and parent education to address feeding difficulties in young children with spastic diplegic cerebral palsy. AB - The purpose of this study was to assess the effectiveness of an intensive day patient pediatric feeding program using oral motor exercises, behavioral interventions, and parental education to increase the oral feeding of children with spastic diplegic cerebral palsy. Eight children between the ages of 18 months to 4.7 years participated in the feeding program for an average of 5.8 weeks. The program consisted of structured oral motor exercises, rewards for reinforcement of appropriate feeding behaviors such as accepting food, chewing, and swallowing, as well as extinction for inappropriate feeding responses. Results show improvement in mealtime skills and behaviors necessary for increasing oral intake. There was improvement in ability to open the mouth for the presentation of the food as well as improved timeliness of swallowing without gagging, expelling or holding food in the mouth. The children were able to tolerate longer meal sessions and consume a greater quantity of food resulting in greater caloric consumption. The children who were tube fed at admission were able to decrease the amount of tube supplementation due to their improved oral intake. Caregivers improved in their ability to feed their children by providing appropriate instructions, prompts and consequences (IPC) during meals. PMID- 18162703 TI - Mechanisms of epithelial-mesenchymal transition of peritoneal mesothelial cells during peritoneal dialysis. AB - A growing body of evidence indicates that epithelial-mesenchymal transition (EMT) of human peritoneal mesothelial cells (HPMC) may play an important role in the development and progression of peritoneal fibrosis during long-term peritoneal dialysis (PD) leading to failure of peritoneal membrane function. Here, we review our own observations and those of others on the mechanisms of EMT of HPMC and suggest potential therapeutic strategies to prevent EMT and peritoneal fibrosis during long-term PD. We found that high glucose and H2O2 as well as transforming growth factor-beta1 (TGF-beta1) induced EMT in HPMC and that high glucoseinduced EMT was blocked not only by inhibition of TGF-beta1 but also by antioxidants or inhibitors of mitogen-activated protein kinases (MAPK). Since MAPKs are downstream target molecules of reactive oxygen species (ROS), these data suggest that high glucose-induced generation of ROS and subsequent MAPK activation mediate high glucose-induced EMT in HPMC. We and others also observed that bone morphogenetic protein-7 (BMP-7) prevented EMT in HPMC. Glucose degradation products (GDP) were shown to play a role in inducing EMT. Involvement of a mammalian target of rapamycin (mTOR) in TGF-beta1-induced EMT has also been proposed in cultured HPMC. A better understanding of the precise mechanisms involved in EMT of HPMC may provide new therapeutic strategies for inhibiting peritoneal fibrosis in long-term PD patients. PMID- 18162704 TI - The genotype and clinical phenotype of Korean patients with familial hypokalemic periodic paralysis. AB - Familial hypokalemic periodic paralysis (HOPP) is a rare autosomal-dominant disease characterized by reversible attacks of muscle weakness occurring with episodic hypokalemia. Mutations in the skeletal muscle calcium (CACNA1S) and sodium channel (SCN4A) genes have been reported to be responsible for familial HOPP. Fifty-one HOPP patients from 20 Korean families were studied to determine the relative frequency of the known mutations and to specify the clinical features associated with the identified mutations. DNA analysis identified known mutations in 12 families: 9 (75%) were linked to the CACNA1S gene and 3 (25%) to the SCN4A gene. The Arg528His mutation in the CACNA1S gene was found to be predominant in these 12 families. Additionally, we have detected one novel silent exonic mutation (1950C>T) in the SCN4A gene. As for a SCN4A Arg669His mutation, incomplete penetrance in a woman was observed. Characteristic clinical features were observed both in patients with and without mutations. This study presents comprehensive data on the genotype and phenotype of Korean families with HOPP. PMID- 18162705 TI - Neonatal intrahepatic cholestasis caused by citrin deficiency in Korean infants. AB - Citrin is a liver-type mitochondrial aspartate-glutamate carrier encoded by the SLC25A13 gene, and its deficiency causes adult-onset type II citrullinemia and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). Here, the authors investigated clinical findings in Korean infants with NICCD and performed mutation analysis on the SLC25A13 gene. Of 47 patients with neonatal cholestasis, three infants had multiple aminoacidemia (involving citrulline, methionine, and arginine) and galactosemia, and thus were diagnosed as having NICCD. Two of these three showed failure to thrive. The laboratory findings showed hypoproteinemia and hyperammonemia, and liver biopsies revealed micro-macrovesicular fatty liver and cholestasis. The three patients each harbored compound heterozygous 1,638 1,660 dup/ S225X mutation, compound heterozygous 851del4/S225X mutation, and heterozygous 1,638-1,660 dup mutation, respectively. With nutritional manipulation, liver functions were normalized and catch-up growth was achieved. NICCD should be considered in the differential diagnosis of cholestatic jaundice in Korean infants. PMID- 18162706 TI - Herpes simplex virus type 2 seroprevalence in Korea: rapid increase of HSV-2 seroprevalence in the 30s in the southern part. AB - To determine the characteristics of seroprevalence of herpes simplex virus type 2 (HSV-2) infection among Korean people, a cross-sectional study was conducted on three groups in 2004. The three groups consisted of the general public who visited public health centers, commercial sex workers (CSWs), and human immunodeficiency virus (HIV)-infected persons. Among the general public, HSV-2 seroprevalence rates for age under the 20s, in the 20s, 30s, 40s and the above 22.6%, 32.7% and 32.3%, respectively, which showed rapid increase of the rate in the 30s (p<0.0001). In case of the above of 19 yr old, women (28.0%) was higher than men (21.7%) (p<0.0001). The rate of CSWs (81.6%) was about 10 times higher than that of general women. In case of HIV-infected men (47.6%), the figure was about 2-3 times higher than that of general men. The low rate in the teens and the 20s proved that it is essential to develop sexually transmitted infections (STIs) prevention programs of education and publicity for them as a precaution measure. This study is the first major study of its kind on HSV-2 and would provide basic data for prevention of STIs including information about target groups subject to vaccination program. PMID- 18162707 TI - Tuberculosis among dislocated North Koreans entering Republic of Korea since 1999. AB - The collapse of North Korea's public health system has increased the development of tuberculosis (TB) in its populace. This study investigated the prevalence of active and latent TB infection (LTBI) in such people who have settled in the Republic of Korea since 1999. From 1999 to August 2006, 7,722 dislocated North Koreans entered the Republic of Korea and all were screened immediately for active TB. Demographic and clinical characteristics were reviewed from the official records of the Settlement Support Office for Dislocated North Koreans, based in the Ministry of Unification. Of 7,722 participants, 87 (1.13%) were diagnosed with active TB from 1999 to August 2006. Of these, 78 (90%) had pulmonary TB. Checking for the presence of a Bacille Calmette-Guerin (BCG) scar and tuberculin skin test has been performed in all dislocated North Koreans since November 2005. Of 1,112 participants, BCG vaccination scars were found in 67.4%. The tuberculin-positive rate using two tuberculin unit doses of the purified protein derivative RT23 (> or =10mm in diameter) was 81.5%. The prevalence of active TB and LTBI in dislocated North Koreans was high. Because this group bears a disproportionate burden of TB, we need to initiate a specific control programme and to plan for the impact of this disease in the Republic of Korea. PMID- 18162708 TI - Incidence and risk factors of insulin resistance syndrome in 20-59 year-old Korean male workers. AB - We investigated the incidence of insulin resistance syndrome (IRS) according to the criteria of diagnoses suggested by the American College of Endocrinology/ American Association of Clinical Endocrinologists and the risk factors associated with the development of IRS. Among 2,048 subjects without a history of/or drug treatment for hypertension, diabetes, dyslipidemia with normal findings at baseline, 1,578 subjects aged 20-59 yr were followed prospectively for 2 yr. The incidence of IRS was 6.9 per 100 persons/year. The relative risk (RR) due to age was 1.03 (95% CI: 1.00-1.05) with every one-year increase in age. The RR associated with an abnormal waist-hip ratio group (> or =0.9) was increased by 1.74 (95% CI: 1.17-2.58) compared to the normal group (<0.9); RR associated with abnormal alanine transferase was increased (> or =35 IU/L) by 1.70 (95% CI: 1.20 2.41) compared to the normal group (<35 IU/L); and the RR associated with abnormal lowdensity lipoprotein (LDL) cholesterol was increased (> or =160 mg/L) by 1.70 (95% CI: 1.19-2.44) compared to the normal LDL cholesterol (<160 mg/L). Lastly, the RR of current smokers was increased by 1.63 (95% CI: 1.09-2.42) compared to that of non-smokers. It is necessary to develop methods of prevention and therapeutic approach to manage the integrated risk factors as opposed to individual factors. PMID- 18162709 TI - Meta-analysis of the association between HLA-DRB1 allele and rheumatoid arthritis susceptibility in Asian populations. AB - The aims of this study were to summarize results on the association of HLA-DRB1 with rheumatoid arthritis (RA) in Asians and to determine if the shared epitope (SE) hypothesis could explain the meta-analysis results. Among the papers published between January 1987 and July 2006 on RA susceptibility in Asian Mongoloid populations (Korean, Japanese, Chinese, and Thai), 12 were selected for the metaanalysis. Mongoloid-Asian patients with RA had significantly higher frequencies of HLA-DRB1*0101, *0401, *0410, and *1001 than controls (OR 1.5-2.1, p<0.05 for association). When analyses were restricted to more ethnically homogeneous populations, HLA-DRB1*0405 showed a significant susceptibility to RA in Koreans (OR 5.65, 95% CI 4.32-7.39), whereas the HLA-DRB1*0301, *0403, *0406, *0701, *1301, and *1405 alleles showed protective association with RA (OR 0.32 0.70, p<0.05 for association). In conclusion, it was found that HLA-DRB1 *0101, *0401, *0405, *0410, and *1001 are susceptible, while HLA-DRB1* 0301, *0403, *0406, *0701, *1301, and *1405 are protective in Asian-Mongoloids. All the RA associated alleles except DRB1*0301 could be explained by the structural model supporting the SE hypothesis that RA susceptibility is determined by the combination of amino acid residues at HLA-DR beta71 and beta74, not by beta71 alone. PMID- 18162710 TI - PHEX gene mutations and genotype-phenotype analysis of Korean patients with hypophosphatemic rickets. AB - X-linked hypophosphatemic rickets (XLH) results from mutations in the PHEX gene. Mutational analysis of the PHEX gene in 15 unrelated Korean patients with hypophosphatemic rickets revealed eight mutations, including five novel mutations, in nine patients: two nonsense mutations, two missense mutations, one insertion, and three splicing acceptor/donor site mutations. Of these, c.64G>T, c.1699C>T, c.466_467 insAC, c.1174-1G>A, and c.1768+5G>A were novel mutations. To analyze the correlation between genotype and phenotype, phenotypes were compared between groups with and without a mutation, in terms of mutation location, mutation type, and sex. Skeletal disease tended to be more severe in the group with a mutation in the C-terminal half of the PHEX gene, but no genotype phenotype correlation was detected in other comparisons. Further extensive studies of the PHEX gene mutations and analyses of the genotype-phenotype relationships are required to understand PHEX function and the pathogenesis of XLH. PMID- 18162711 TI - Interferon-stimulated genes response in endothelial cells following Hantaan virus infection. AB - The regulation mechanism of interferon (IFN) and IFN-stimulated genes is a very complex procedure and is dependent on cell types and virus species. We observed molecular changes related to anti-viral responses in endothelial cells during Hantaan virus (HTNV) infection. We found that there are two patterns of gene expression, the first pattern of gene expression being characterized by early induction and short action, as in that of type I IFNs,' and the other being characterized by delayed induction and long duration, as those of IRF-7, MxA, and TAP-1/2. Even though there are significant differences in their induction folds, we found that all of IFN-alpha/beta, IRF- 3/7, MxA, and TAP-1/2 mRNA expressions reached the peak when the viral replication was most active, which took place 3 days of post infection (d.p.i.). In addition, an interesting phenomenon was observed; only one gene was highly expressed in paired genes such as IFN alpha/beta' (3/277-folds), IRF-3/7 (2.2/29.4-folds), and TAP- 1/2 (26.2/6.1 folds). Therefore, IFN-beta, IRF-7, and TAP-1 seem to be more important for the anti-viral response in HTNV infection. MxA was increased to 296-folds at 3 d.p.i. and kept continuing 207-folds until 7 d.p.i.. The above results indicate that IFN beta works for an early anti-viral response, while IRF7, MxA, and TAP-1 work for prolonged anti-viral response in HTNV infection. PMID- 18162712 TI - Indexes of suspicion of typical cow's milk protein-induced enterocolitis. AB - This study was performed to identify clinical factors that facilitate the diagnosis of typical cow's milk protein-induced enterocolitis (CMPIE). Data from 142 consecutive patients (aged 15 to 45 days, cow's milk formula- or cow's milk and breast milk mixed-fed) admitted due to vomiting and/or diarrhea were retrospectively analyzed. These 142 subjects were divided into three groups: the CMPIE, infection, and non-infection group. Each group was composed of 16 (11.3%), 102 (71.8%), and 24 (16.9%) patients, respectively. On admission, poor weight gain (p=0.003), hypoalbuminemia (p=0.035), peripheral leukocytosis (p=0.012), and metabolic acidosis (p=0.015) were found to be more significant in the CMPIE group than those in other two groups. In CMPIE, serum albumin levels decreased from 3.3+/-0.9 g/dL on admission to 2.6+/-0.3 g/dL during admission (p<0.05), and methemoglobinemia was observed in 3 patients (18.8%) (p=0.012). Multiple logistic regression analysis showed that the independent predictors of CMPIE versus the infection group were failure to gain weight (OR, 10.75 [95% CI, 1.53-66.12]) (p= 0.014) and hypoalbuminemia (OR, 9.53 [95% CI, 1.62-49.01]) (p=0.010). The early recognition of indexes of suspicion for CMPIE may be of help in the diagnosis and treatment of this disorder. PMID- 18162713 TI - A familial case of Wiskott-Aldrich Syndrome with a hotspot mutation in exon 2 of the WAS Gene. AB - The Wiskott-Aldrich syndrome (WAS) is a severe X-linked disorder characterized classically by thrombocytopenia, immunodeficiency, and eczema. The phenotype observed in this syndrome is caused by mutation in the WAS gene. Peripheral blood DNAs were isolated from an 18-month-old boy with WAS and his mother, maternal uncle, and maternal grandmother. Genetic analysis for the detection of a mutation of WAS gene was performed by polymerase chain reaction-single strand conformational polymorphism analysis (PCR-SSCP) and direct sequencing of the PCR product. In PCR-SSCP, the patient and his maternal uncle had an abnormal shift band, which was not found in normal controls, and his mother and maternal grandmother showed heterozygous bands. In direct sequencing analysis, the patient with WAS had CGC-->CAC point mutation in exon 2 that resulted in an amino acid change in codon 86 (Arg86His). The present study identified a gene mutation responsible for WAS at a mutation hotspot of the WAS gene in a Korean family. PMID- 18162714 TI - The correlation between F-wave motor unit number estimation (F-MUNE) and functional recovery in stroke patients. AB - The aim of this study was to follow up the changes in the number of motor units according to the Brunnstrom stage through a motor unit number estimation of the Fwave (F-MUNE) after a stroke, and to identify the functional significance of F MUNE. Twenty-five patients (15 men, 10 women) with a first unilateral stroke were recruited. The maximal M-potential was evoked by the supramaximal stimulation of the median nerve at the wrist, and the maximal stimulation intensity was determined on both hemiplegic and unaffected hands. The reproducible all-or-none F-wave was evoked in 30% of the maximal stimulation intensity and was constantly stimulated at that level. The prototypes of the F-wave were chosen, and the values of F-MUNE were calculated by dividing the amplitude of the maximal M potential by the mean amplitude of the F-prototype. The changes in F-MUNE were compared according to the progression of the Brunnstrom stage and correlated with those of the functional scales. The mean motor unit numbers decreased significantly in the hemiplegic side compared with the unaffected side. According to the progression of the Brunnstrom stage, the values of F-MUNE were reduced significantly by increasing the amplitude and recruitment of the F-prototype, and the functional scores also improved. These results show that the F-MUNE equation did not show a functional recovery related increase in stroke patients. PMID- 18162715 TI - Self-reported sleep latency in postmenopausal women. AB - The aim of this study was to access how self-reported sleep latency (SRSL) was affected by sleep habits, mood, and circadian rhythm in postmenopausal women. Subjects (n=384, 67.9+/-7.7 yr) completed sleep and mood questionnaires, sleep log and actigraphic data. The major urinary melatonin metabolite (6 sulphatoxymelatonin, aMT6s) was assayed in fractional urine specimens for two 24 hr intervals. Although SRSL (26.5+/-24.4 min) and actigraphic sleep latency (ASL; 27.8+/-20.0 min) were correlated (r(s)=0.361, p<0.001), the short SRSLs tended to be underestimated whereas the long SRSLs tended to be overestimated as compared to ASL. SRSL was positively correlated with the scales of insomnia, mood and hot flash, hypertension, use of anti-hypertensive drugs and the acrophase and the offset of aMT6s. SRSL was negatively correlated with the global assessment of functioning scale in DSM-IV (GAF scale), and light exposure and wrist activity. Multiple linear regression analysis showed that the best-fit model to predict SRSL was light exposure, GAF scale, and use of anti-hypertensive drugs. SRSL may be determined by psychophysiological factors as well as circadian rhythm function. Therapeutic approaches suggested for trouble falling asleep might include increased daylight exposure, improvements in general health, and modification of anti-hypertensive pharmacotherapy. PMID- 18162716 TI - ERK-1/-2 and p38 kinase oppositely regulate 15-deoxy-delta(12,14) prostaglandinJ(2)-Induced PPAR-gamma activation that mediates dedifferentiation but not cyclooxygenase-2 expression in articular chondrocytes. AB - Peroxisome proliferator-activated receptor gamma (PPAR-gamma) is a ligand activated transcription factor and plays an important role in growth, differentiation, and inflammation in different tissues. In this study, we investigated the effects of 15d-PGJ(2), a high-affinity ligand of PPAR-gamma, on dedifferentiation and on inflammatory responses such as COX-2 expression and PGE(2) production in rabbit articular chondrocytes with a focus on ERK-1/-2, p38 kinase, and PPAR-gamma activation. We report here that 15d-PGJ(2) induced dedifferentiation and/or COX-2 expression and subsequent PGE(2) production. 15d PGJ(2) treatment stimulated activation of ERK-1/-2, p38 kinase, and PPAR-gamma. Inhibition of ERK-1/-2 with PD98059 recovered 15d-PGJ(2)-induced dedifferentiation and enhanced PPAR-gamma activation, whereas inhibition of p38 kinase with SB203580 potentiated dedifferentiation and partially blocked PPAR gamma activation. Inhibition of ERK-1/-2 and p38 kinase abolished 15d-PGJ(2) induced COX-2 expression and subsequent PGE(2) production. Our findings collectively suggest that ERK-1/-2 and p38 kinase oppositely regulate 15d-PGJ(2) induced dedifferentiation through a PPAR-gamma-dependent mechanism, whereas COX-2 expression and PGE(2) production is regulated by ERK-1/-2 through a PPAR-gamma independent mechanism but not p38 kinase in articular chondrocytes. Additionally, these data suggest that targeted modulation of the PPAR-gamma and mitogen activated protein kinase pathway may offer a novel approach for therapeutic inhibition of joint tissue degradation. PMID- 18162717 TI - Plasmodium falciparum cultivation using the Petri Dish: revisiting the effect of the 'age' of erythrocytes and the interval of medium change. AB - Differences in the characteristics of the culture conditions can influence the multiplication rate of Plasmodium falciparum. The Petri dish method is one of the most popular methods of cultivating this parasite. In many previous studies, ideal culture conditions of the Petri dish method were achieved by using erythrocytes collected from blood that had been stored for at least 2 weeks, with daily changes of the medium. In the present study, we studied the multiplication rate of P. falciparum in cultures containing erythrocytes of various ages together with changing the medium at various intervals of time. Our results strongly suggest that the rate of in vitro multiplication of P. falciparum was higher in freshly collected erythrocytes than in aged erythrocytes regardless of the anticoagulant and that when the parasitemia is lower than 8% with a hematocrit of 5%, the medium change interval can be as long as 48 hr without a great reduction in the rate of multiplication. PMID- 18162718 TI - Inflammatory and remodeling events in asthma with chronic exposure to house dust mites: a murine model. AB - Although animal models with ovalbumin have been used to study chronic asthma, there are difficulties in inducing recurrence as well as in maintaining chronic inflammation in this system. Using a murine model of house dust mite (HDM) induced bronchial asthma, we examined the airway remodeling process in response to the chronic exposure to HDM. During the seventh and twelfth weeks of study, HDM were inhaled through the nose for three consecutive days and airway responsiveness was measured. Twenty-four hours later, bronchoalveolar lavage and histological examination were performed. The degree of overproduction of mucus, subepithelial fibrosis, and the thickness of the peribronchial smooth muscle in the experimental group was clearly increased compared to the control group. In addition, HDM-exposed mice demonstrated severe airway hyperreactivity to methacholine. In the bronchoalveolar lavage fluid, the number of total cells and eosinophils was increased; during the twelfth week, the number of neutrophils increased in the experimental group. With regard to changes in cytokines, the concentrations of IL-4, IL- 13, and transforming growth factor-beta (TGF-beta) were increased in the experimental group. The data suggest that eosinophils, IL 4, IL-13, and TGF-beta might play an important role in the airway remodeling process and that neutrophils may be involved with increased exposure time. PMID- 18162719 TI - Altered renal sodium transporter expression in an animal model of type 2 diabetes mellitus. AB - Hemodynamic factors play an important role in the development and/or progression of diabetic nephropathy. We hypothesized that renal sodium transporter dysregulation might contribute to the hemodynamic alterations in diabetic nephropathy. Otsuka Long Evans Tokushima Fatty (OLETF) rats were used as an animal model for type 2 diabetes. Long Evans Tokushima (LETO) rats were used as controls. Renal sodium transporter regulation was investigated by semiquantitative immunoblotting and immunohistochemistry of the kidneys of 40 week-old animals. The mean serum glucose level in OLETF rats was increased to 235+/-25 mg/dL at 25 weeks, and the hyperglycemia continued up to the end of 40 weeks. Urine protein/ creatinine ratios were 10 times higher in OLETF rats than in LETO rats. At 40th week, the abundance of the epithelial sodium channel (ENaC) beta-subunit was increased in OLETF rats, but the abundance of the ENaC gamma subunit was decreased. No significant differences were observed in the ENaC alpha subunit or other major sodium transporters. Immunohistochemistry for the ENaC beta-subunit showed increased immunoreactivity in OLETF rats, whereas the ENaC gamma-subunit showed reduced immunoreactivity in these rats. In OLETF rats, ENaC beta-subunit upregulation and ENaC gamma-subunit downregulation after the development of diabetic nephropathy may reflect an abnormal sodium balance. PMID- 18162720 TI - Erythropoietin attenuates hyperoxia-induced lung injury by down-modulating inflammation in neonatal rats. AB - This study was done to determine whether recombinant human erythropoietin (rhEPO) treatment could attenuate hyperoxia-induced lung injury, and if so, whether this protective effect is mediated by the down-modulation of inflammation in neonatal rats. Newborn Sprague Dawley rat pups were subjected to 14 days of hyperoxia (>95% oxygen) within 10 hr after birth. Treatment with rhEPO significantly attenuated the mortality and reduced body weight gain caused by hyperoxia. With rhEPO treatment, given 3 unit/gm intraperitoneally at 4th, 5th, and 6th postnatal day, hyperoxia- induced alterations in lung pathology such as decreased radial alveolar count, increased mean linear intercept, and fibrosis were significantly improved, and the inflammatory changes such as myeloperoxidase activity and tumor necrosis factor-alpha expression were also significantly attenuated. In summary, rhEPO treatment significantly attenuated hyperoxia-induced lung injury by down modulating the inflammatory responses in neonatal rats. PMID- 18162721 TI - Usefulness of pain distribution pattern assessment in decision-making for the patients with lumbar zygapophyseal and sacroiliac joint arthropathy. AB - There are currently no initial guides for the diagnosis of somatic referred pain of lumbar zygapophyseal joint (LZJ) or sacroiliac joint (SIJ). We developed a classification system of LZJ and SIJ pain, the ''pain distribution pattern template (PDPT)'' depending on the pain distribution patterns from a pool of 200 patients whose spinal pain source was confirmed. We prospectively applied the PDPT to determine its contribution to clinical decision-making for 419 patients whose pain was presumed to arise from the LZJs (259 patients) or SIJs (160 patients). Forty-nine percent (128/259) of LZJ and 46% (74/160) of SIJ arthopathies diagnosed by PDPT were confirmed by nerve blocks. Diagnostic reliabilities were significantly higher in Type A and C patterns in LZJ and Type C in SIJ arthropathies, 64%, 80%, and 68.4%, respectively. For both LZJ and SIJ arthropathies, favorable outcome after radiofrequency (RF) neurotomies was similar to the rate of positive responses to diagnostic blocks in Type A to Type D, whereas the outcome was unpredictable in those with undetermined type (Type E). Considering the paucity of currently available diagnostic methods for LZJ and SIJ arthropathies, PDPT is useful in clinical decision- making as well as in predicting the treatment outcome. PMID- 18162722 TI - A prospective randomized trial of either famotidine or pantoprazole for the prevention of bleeding after endoscopic submucosal dissection. AB - Endoscopic submucosal dissection (ESD) has been reported to have a higher bleeding rate than conventional methods. However, there are few reports on whether a proton pump inhibitor or a histamine2-receptor antagonist is the more effective treatment for preventing bleeding after ESD. In a prospective trial, patients undergoing ESD due to gastric adenoma or adenocarcinoma were randomly assigned to pantoprazole or famotidine. Both drugs were given intravenously for the first 2 days, thereafter by mouth. Eighty-five in the pantoprazole group and 79 in the famotidine group were included for analysis. Primary outcome measure was the delayed bleeding rate. Clinical characteristics were not different between the two groups. The delayed bleeding rate was significantly lower in the pantoprazole group compared with the famotidine group (3.5% vs. 12.7%, p=0.031). On multivariate analysis, the preventive use of pantoprazole (relative hazard: 0.220, 95% CI: 0.051- 0.827, p=0.026) and the specimen size (> or =34 mm, relative hazard: 4.178, 95% CI: 1.229-14.197, p=0.022) were two independent factors predictive of delayed bleeding. There were no significant differences in en bloc and complete resection rate between the two groups. In conclusion, pantoprazole is more effective than famotidine for the prevention of delayed bleeding after ESD. PMID- 18162723 TI - Long-term experience with the Bjork-Shiley Monostrut tilting disc valve. AB - The Bjork-Shiley Monostrut valve is tilting disc mechanical valve prosthesis. This study was designed to present the long-term outcome of our experience. One hundred and thirty-seven Bjork-Shiley Monostrut valves were implanted in 101 consecutive patients from November 1983 to February 1990. There were 60 male and 41 female with mean age of 34.5 yr at the time of operation. Fifty-nine patients underwent single valve replacement, 38 had double valve, and 4 had triple valve replacement. There were six in-hospital deaths (5.9%): three from cardiopulmonary bypass weaning failure and one each from septic shock, sudden cardiac arrest, and uncontrollable bleeding. Mean duration of follow-up was 181.2+/-76.2 months. Overall survival was 86.2% at 15 yr and 83.1% at 20 yr. Patients with mitral valve replacement had 93.5% and 90.2% cumulative survival at 10 and 15 yr, respectively, while patients with aortic valve replacement had 91.1% and 86.5% cumulative survival at 10 and 15 yr. Two groups had no significant difference in survival. Double valve replacement patients had 92.2% and 84.0% survival at 10 and 15 yr, respectively. There were no significant differences in survival between the single and double valve replacement groups. Freedom from thromboembolism was noted in: 97.8%, 97.8%, 96.4% and 87.8% at 5, 10, 15 and 20 yr, respectively. Absence of endocarditis was noted in 98.6% and 94.8% at 15 and 20 yr. Absence of reoperation was 92.5% at 20 yr. In conclusion, the Bjork-Shiley Monostrut valve is reliable, with a similar incidence of valve-related morbidity as in other mechanical valves. PMID- 18162724 TI - Sociodemographic and smoking behavioral predictors associated with smoking cessation according to follow-up periods: a randomized, double-blind, placebo controlled trial of transdermal nicotine patches. AB - This study investigated sociodemographic and smoking behavioral factors associated with smoking cessation according to follow-up periods. In this randomized, double-blind, placebo-controlled trial of transdermal nicotine patches, subjects were a total of 118 adult male smokers, who were followed up for 12 months. Univariable logistic regression analysis and stepwise multiple logistic regression analyses were performed to identify the predictors of smoking cessation. The overall self-reported point prevalence rates of abstinence were 20% (24/118) at 12 months follow-up, and there was no significant difference in abstinence rates between placebo and nicotine patch groups. In the univariable logistic regression analysis, predictors of successful smoking cessation were the low consumption of cigarettes per day and the low Fagerstrom Test for Nicotine Dependence (FTND) scores (p<0.05) at 3, 6, and 12 months follow-up. In the stepwise multiple logistic regression analyses, predictors of successful smoking cessation, which were different according to the follow- up periods, were found to be the low consumption of cigarettes per day at the short-term and midterm follow-up (< or =6 months), older age, and the low consumption of cigarettes per day at the long-term follow-up (12 months). PMID- 18162725 TI - Paradoxical air embolism during percutaneous nephrolithotomy: a case report. AB - Air embolism is a rare complication of percutaneous nephrolithotomy. Patent foramen ovale, which is necessary in fetal circulation, is a potential route for emboli arising from the venous system to enter the systemic arterial circulation, resulting in paradoxical air embolism syndrome. A case of paradoxical air embolism during percutaneous nephrolithotomy is presented. To our knowledge, this is the first report of paradoxical air embolism associated with patent foramen ovale during percutaneous nephrolithotomy. PMID- 18162726 TI - A case of post-streptococcal glomerulonephritis with diffuse alveolar hemorrhage. AB - Acute post-streptococcal glomerulonephritis (PSGN) is characterized by an abrupt onset of edema, hypertension, and hematuria. Life-threatening diffuse alveolar hemorrhage (DAH) is rarely associated with acute PSGN. There have been only two reported cases worldwide, and no case has been reported previously in Korea. Here, we present a patient who clinically presented with pulmonary-renal syndrome; the renal histology revealed post-infectious glomerulonephritis of immune complex origin. A 59-yr-old woman was admitted with oliguria and hemoptysis two weeks after pharyngitis. Renal insufficiency rapidly progressed, and respiratory distress developed. Chest radiography showed acute progressive bilateral pulmonary infiltrates. The clinical presentation suggested DAH with PSGN. Three days after treatment with high-dose steroids, the respiratory distress and pulmonary infiltrates resolved. Electron microscopy of a renal biopsy specimen sample revealed diffuse proliferative glomerulonephritis with characteristic subendothelial deposits of immune complex (''hump''). The renal function of the patient was restored, and the serum creatinine level was normalized after treatment. PMID- 18162727 TI - Therapeutic experience of Bing-Neel Syndrome associated with Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia is an uncommon low-grade B-cell lymphoproliferative disorder in which monoclonal immunoglobulin M is produced. Neurological symptoms due to hyperviscosity are frequent manifestations of Waldenstrom's macroglobulinemia. However, central nervous system infiltration by plasmacytoid lymphocytes (Bing-Neel syndrome) has only rarely been reported. We report a case of a 51-yr-old woman suffering from Waldenstrom's macroglobulinemia who complained of persistant headache. Brain magnetic resonance imaging revealed an extra-axial soft tissue mass along the left cavernous sinus, left tentorium, right tentorium, and falx cerebri. A stereotactic biopsy of dural tissue from the falx was performed and showed plasmacytoid lymphocyte infiltration. The patient became symptom- free with irradiation of the whole brain followed by chemotherapy with fludarabine. PMID- 18162728 TI - Morganella morganii sepsis with massive hemolysis. AB - Morganella morganii is a facultative gram-negative and anaerobic rod. It may be a cause of devastating infections in neonates and immunocompromised hosts. Some bacterial infections such as Clostridium and Vibrio are associated with hemolysis. However, massive hemolysis caused by M. morganii sepsis has not yet been reported. We observed a 59-yr-old man who had chemotherapy-induced neutropenia and was found to have massive hemolysis and metabolic acidosis due to sepsis. He died 6 hr after admission in spite of aggressive treatment. Two sets of blood cultures revealed the growth of M. morganii. We report here that M. morganii sepsis can cause fatal massive hemolysis leading to death. PMID- 18162729 TI - Pituitary apoplexy complicated by chemical meningitis and cerebral infarction. AB - A 41-yr-old man was admitted with acute headache, neck stiffness, and febrile sensation. Cerebrospinal fluid examination showed pleocytosis, an increased protein level and, a decreased glucose concentration. No organisms were observed on a culture study. An imaging study revealed pituitary macroadenoma with hemorrhage. On the 7th day of the attack, confusion, dysarthria, and right-sided facial paralysis and hemiparesis were noted. Cerebral infarction on the left basal ganglia was confirmed. Neurologic deficits gradually improved after removal of the tumor by endoscopic transnasal transsphenoidal approach. It is likely that the pituitary apoplexy, aseptic chemical meningitis, and cerebral infarction are associated with each other. This rare case can serve as a prime example to clarify the chemical characteristics of pituitary apoplexy. PMID- 18162730 TI - Chronic idiopathic myelofibrosis presenting as cauda equina compression due to extramedullary hematopoiesis: a case report. AB - Extramedullary hematopoiesis (EMH) is occasionally reported in idiopathic myelofibrosis and is generally found in the liver, spleen, and lymph nodes several years after diagnosis. Myelofibrosis presenting as spinal cord compression, resulting from EMH tissue is very rare. A 39-yr-old man presented with back pain, subjective weakness and numbness in both legs. Sagittal magnetic resonance imaging showed multiple anterior epidural mass extending from L4 to S1 with compression of cauda equina and nerve root. The patient underwent gross total removal of the mass via L4, 5, and S1 laminectomy. Histological analysis showed islands of myelopoietic cells surrounded by fatty tissue, consistent with EMH, and bone marrow biopsy performed after surgery revealed hypercellular marrow and megakaryocytic hyperplasia and focal fibrosis. The final diagnosis was chronic idiopathic myelofibrosis leading to EMH in the lumbar spinal canal. Since there were no abnormal hematological findings except mild myelofibrosis, additional treatment such as radiothepary was not administered postoperatively for fear of radiotoxicity. On 6 month follow- up examination, the patient remained clinically stable without recurrence. This is the first case of chronic idiopathic myelofibrosis due to EMH tissue in the lumbar spinal canal in Korea. PMID- 18162731 TI - Amnesic syndrome in a mammillothalamic tract infarction. AB - It is controversial whether isolated lesions of mammillothalamic tract (MTT) produce significant amnesia. Since the MTT is small and adjacent to several important structures for memory, amnesia associated with isolated MTT infarction has been rarely reported. We report a patient who developed amnesia following an infarction of the left MTT that spared adjacent memory-related structures including the anterior thalamic nucleus. The patient s memory deficit was characterized by a severe anterograde encoding deficit and retrograde amnesia with a temporal gradient. In contrast, he did not show either frontal executive dysfunction or personality change that is frequently recognized in the anterior or medial thalamic lesion. We postulate that an amnesic syndrome can develop following discrete lesions of the MTT. PMID- 18162732 TI - Autosomal dominant centronuclear myopathy with unique clinical presentations. AB - Centronuclear myopathies are clinically and genetically heterogenous diseases with common histological findings, namely, centrally located nuclei in muscle fibers with a predominance and hypotrophy of type 1 fibers. We describe two cases from one family with autosomal dominant centronuclear myopathy with unusual clinical features that had initially suggested distal myopathy. Clinically, the patients presented with muscle weakness and atrophy localized mainly to the posterior compartment of the distal lower extremities. Magnetic resonance imaging revealed predominant atrophy and fatty changes of bilateral gastrocnemius and soleus muscles. This report demonstrates the expanding clinical heterogeneity of autosomal dominant centronuclear myopathy. PMID- 18162733 TI - [Antibacterial activity and pharmacological features of fluoroquinolones and considerations on their use in a nosocomial setting]. AB - The need for appropriate use of antimicrobials limiting the insurgence, selection and spread of bacterial resistance and preventing its diffusion to the community is increasingly widely felt. Based on some CDC expert recommendations, an antibiotic therapy should be driven by the following criteria: scientifically proven clinical evidence, high probability of being beneficial to patients, safety and tolerability, drug and regimen choice made according to the infection to be treated, and cost/efficacy ratio. In this article, we compare the microbiological and pharmacological characteristics of the three most frequently used fluoroquinolones in Italy - ciprofloxacin, levofloxacin, and moxifloxacin - highlighting the main differences and peculiar aspects for each of them. We describe the microbiological profile, providing details on bacterial resistance mechanisms, the pharmacological profile, giving details on tolerability, and the clinical profile, showing the individual therapeutic efficacy, underlining the main differences in their pharmacodynamic and pharmacokinetic aspects. The main national and international guidelines recommend ciprofloxacin and levofloxacin and, to a lesser extent, moxifloxacin, for treating nosocomial infections localised in different organs and apparatuses, including severe infections in critical patients. In addition, they represent a basic therapeutic option for treatment of community-acquired infections, such as urinary and respiratory tract infections. PMID- 18162734 TI - [Effect of omega-3 fatty acids on the lipid structure of HIV-positive patients on antiretroviral therapy: personal observation of 20 subjects]. AB - Hyperlipidaemia related to antiretroviral therapy is a major issue in the management of HIV infection. In HIV positive patients, interventions on their lifestyles and dietary habits are only partially effective and pharmacological support is often required. However, fibrates and statins, interacting with antiretroviral therapy, may cause toxicity. A number of clinical trials have shown the effectiveness of omega-3 fatty acids on the hyperlipidaemia of HIV patients. In this study, the effect of six-month daily administration of omega-3 fatty acids on triglycerides, total and LDL cholesterol, and CD4 cell count was evaluated in a group of 20 HIV positive patients. Triglyceride levels significantly decreased, whilst total, LDL cholesterol and CD4 cell count did not vary. Fish oil therapy was well tolerated by all patients. Omega-3 fatty acids may contribute to reduce cardiovascular risk in HIV positive patients. PMID- 18162735 TI - [Reduced incidence of Staphylococcus isolation in nosocomial setting after introducing preoperative antibiotic prophylaxis guidelines]. AB - We compare epidemiological data from two twelve-month periods, before and after the introduction of preoperative antibiotic prophylaxis guidelines at Carlo Poma hospital in Mantova, Italy, in June 2003. Considering the results from the microbiology laboratory and the data from the pharmacy, concerning the consumption of some antimicrobials, we noted a significant decrease in the incidence of methicillin-resistant Staphylococcus aureus (MRSA) in surgical wards, where the incidence of MRSA had previously exceeded that in medical wards. At the same time, analysis of antibiotic consumption revealed a considerable decrease in third and fourth-generation cephalosporins and an increasing use of cephazolin, in compliance with prophylaxis protocol rules. This trend was confirmed by analysis of the same data regarding the first six months of 2006. PMID- 18162736 TI - Time trends in health care needs of non-EU citizens from developing countries, admitted to a general hospital in northern Italy. AB - Hospitalizations of foreign patients from developing countries outside the European Union were examined for the period 1999-2004, focusing on infectious diseases and on pregnancy issues. Patients over 14 years old had 6,003 admissions, leading to 7,231 overall diagnoses. During the 6-year study period, female hospitalizations increased steadily, with a peak in 2002 (p .001). This trend was mainly due to the rise in women from Eastern Europe (p .001), which occurs at a younger mean age versus that of males (p .001). Admission of illegal immigrants, performed on an emergency basis, accounted for an average of 9.4%. This phenomenon was very frequent in 1999 (43% of admissions), but dropped sharply after 2002 (p .001), caused by changes in Italian law. The prevalent women diagnoses were ob/gyn ones: voluntary pregnancy interruption, spontaneous abortion or pregnancy complications in 30.6% of cases, and childbirths or controls of pregnancies with a favourable outcome in 18.2% of patients. These diagnoses covered nearly 50% of hospitalizations of migrant women: other admissions were due to organic, dysmetabolic, or functional disorders, while infectious diseases were less frequent (4.6%). Among men, dysmetabolic disorder and organic-degenerative diseases, or functional illnesses (36.2%), were prominent, and significantly more frequent versus women (p .001), as well as post traumatic diseases (16.5%), and infectious illnesses (12.1%; p .001). Also generic-undefined diagnoses were proportionally numerous (6.6%): cultural language deficiencies affected the physician-patient relationship. Among infectious diseases, the main causative organisms were Mycobacterium tuberculosis (14.9%), HIV (7.1%), HBV (3.3%), and HCV (2.6%). Upper-lower airways represented the most involved organ system (45% of discharges), followed by the gastroenteric tract (16.4%), and skin-soft tissues (7.4%), while systemic infectious diseases accounted for 14.9% of episodes. Such disorders predominated (up to 90% of cases) among non-regular migrants during 1999-2000, while after 2002 an increase in infectious disorders was observed among patients from Eastern Europe. From a health care-social perspective, although a reduced incidence of infectious diseases did not occur, the possibility of attributing them to individuals of ascertained identity and housing makes it possible to trace index patients, and ultimately strive towards well-planned and effective therapeutic-preventive interventions. PMID- 18162737 TI - Infected atrial myxoma: case report and literature review. AB - Myxoma is the most common type of cardiac tumour in all age groups. It may simulate infective endocarditis but is rarely infected. We describe one case of infected left atrial myxoma caused by Enterococcus faecalis. Combined therapy, consisting of surgical management and antimicrobial therapy, was used. Histological examination of the excised tumour revealed a typical myxoma with infiltrates of neutrophils. Few cases of infected atrial myxomas have been reported in the literature. PMID- 18162738 TI - [Invasive aspergillosis in an immunocompromised patient: clinical and therapeutic considerations]. AB - The paper describes a case report of a young female with invasive aspergillosis diagnosed after brief treatment with high-dose steroids for autoimmune thrombocytopenia. Early diagnosis of invasive aspergillosis was made with cultures of tracheoaspirates and bronchoalveolar lavage and was confirmed with a transbronchial biopsy. After initial ineffective treatment with liposomal amphotericin B and dissemination from pulmonary to central nervous system involvement, treatment was switched to a combination of voriconazole and caspofungin. After marked clinical and radiological improvement, treatment was switched to the orally administered formulation of voriconazole until the complete disappearance of central nervous system lesion was observed. In the discussion section we underscore the most significant data of the host susceptibility, diagnosis of invasive aspergillosis, complications and treatment. This case ably demonstrates the efficacy of new antifungal agents, even when administered orally, and underscores the variability of host susceptibility to atypical and often unexpected invasive fungal infections. PMID- 18162739 TI - [Difficulty in diagnosing pediatric tuberculosis]. AB - Tuberculosis (TB) in children is an important warning sign in a community, as it could signal recent infection of a cavitary form in an adult. Thus, while early diagnosis is crucial for effective treatment in children, it is also imperative for the control of tuberculosis at the public health level since it allows rapid identification of contagious adult cases. Here we report four cases of difficult and delayed diagnosis of TB in children. From this experience we highlight the need for an extensive medical history of the patient during diagnostic work-up. This includes: the positive history for contact with infected adults, especially for immigrant children; exclusion of TB diagnosis for persistent respiratory symptoms (2-3 weeks) after antibiotic therapy; and the need for high-definition CT scan when the radiological picture is not specific, especially for children under 5 years of age. PMID- 18162740 TI - [The Spanish influenza pandemic]. AB - The influenza pandemic of 1918-1919, so-called Spanish influenza, spread to almost all nations worldwide. This outbreak is thought to have killed 25 million people, although some have claimed that the epidemic resulted in as many as 40 million deaths. This pandemic was a particularly dramatic event, because it occurred at the end of World War I, when both armies and the civilian population, in nations involved in the war, were exhausted. In Italy 600,000 people are estimated to have died of Spanish influenza. Together with the death of 650,000 soldiers during the war, this had a major demographic impact. We describe the course of the epidemic in Italy as a whole and in Bologna in particular. In Bologna and in its province we analysed the lists drawn up at the end of the World War I by the Central Records Office in Bologna, which coordinated research into causes of death of soldiers engaged in the conflict. We also examined the trend of burials at Certosa in Bologna in the first decades of the last century in order to establish, during the two-year period 1918-1919, the impact of the epidemic upon annual mortality. In Bologna the impact of the epidemic, albeit important in comparison to other situations, was not particularly dramatic. No special preventive measures were adopted, with the exception of isolating seriously ill patients in a former school converted by the military authorities into a hospital. Family doctors worked together actively with the city's medical authorities when the epidemiological survey was carried out. PMID- 18162743 TI - Developing a novel Poverty in Healthcare curriculum for medical students at the University of Michigan Medical School. AB - Nearly 90 million Americans live below 200% of the federal poverty threshold. The links between lower socioeconomic status and poor health are clear, and all physicians face the resulting challenges in patient care. Current medical school curricula do not adequately prepare students to address this issue despite recommendations from the Association of American Medical Colleges and the Institute of Medicine. In response, students and faculty at the University of Michigan Medical Center established the Poverty in Healthcare curriculum, which encompasses required learning experiences spanning all four years of undergraduate medical education. This article describes the design and implementation of this curriculum. The authors provide thorough descriptions of the individual learning experiences, including community site visits, longitudinal cases, mini-electives, and family centered experiences. The authors also discuss the history, costs, challenges, and evaluation process related to the Poverty in Healthcare curriculum, including issues specifically related to medical students' involvement in developing and implementing the curriculum. This information may be used as a guide for other medical schools in the development of curricula to address this current gap in medical student education. PMID- 18162745 TI - Health and disease in context: a community-based social medicine curriculum. AB - Despite the increasing attention paid to the role of social forces in determining health, most physicians finish their training ill-prepared to address these issues. The authors describe their efforts to fill that training gap for internal medicine residents at Oregon Health and Science University through a community based social medicine curriculum, designed in 2006 in conjunction with community partners at Central City Concern (CCC), an organization addressing homelessness, poverty, and addiction in downtown Portland, Oregon. The challenge was to develop a curriculum that would (1) fit within the scheduling constraints of an established categorical internal medicine residency program, (2) give all internal medicine residents a chance to better understand how social forces affect health, and (3) help show how they, as health professionals, might intervene to improve health and health care. The authors maintain that by developing this curriculum with community partners--who took the lead in deciding what residents should learn about their community and how they should learn it- the residency program is providing a relatively brief but extremely rich opportunity for residents to engage the personal, social, and health-related issues experienced by clients served by CCC. The authors first provide a brief overview of the curriculum and describe how the principles and practices of community-based participatory research were used in its development. They then discuss the challenges involved in teaching medical residents about social determinants of health, how their academic-community partnership approaches those challenges, and the recently established methods of evaluating the curriculum. PMID- 18162746 TI - Integration of the biopsychosocial model: perspectives of medical students and residents. AB - PURPOSE: To examine residents' and medical students' attitudes toward the incorporation of psychosocial factors in diagnosis and treatment and to identify barriers to the integration of evidence-based, mind-body methods. METHOD: A random sample of third- and fourth-year medical students and residents was drawn from the Masterfiles of the American Medical Association. A total of 661 medical students and 550 residents completed a survey, assessing attitudes toward the role of psychosocial factors and the clinical application of behavioral/mind-body methods. RESULTS: The response rate was 40%. Whereas a majority of students and residents seem to recognize the need to address psychosocial factors, 30%-40% believe that addressing such factors leads to minimal or no improvements in outcomes. The majority of students and residents reports that their training in these areas was ineffective, yet relatively few indicate interest in receiving further training. Females are more likely to believe in the need to address psychosocial factors. Additional factors associated with greater openness to addressing psychosocial factors include (1) the perception that training in these areas was helpful, and (2) personal use of behavioral/mind-body methods to care for one's own health. CONCLUSIONS: There is a need for more comprehensive training during medical school and residency regarding both the role of psychosocial factors in health and the application of evidence-based, behavioral/mind-body methods. The current health care structure-particularly insufficient time and inadequate reimbursement for addressing psychosocial factors-may be undermining efforts to improve patient care through inconsistent or nonexistent application of the biopsychosocial model. PMID- 18162747 TI - Enhancing behavioral science education at the Ohio State University College of Medicine. AB - The social and behavioral sciences play key roles in patient health outcomes. Given this reality, successful development of social and behavioral science curricula in medical education is critically important to the quality of patients' lives and the effectiveness of health care delivery systems. The Institute of Medicine, in a recent report, recommended that medical schools enhance their curricula in these areas and identified four institutions as "exemplars" of social and behavioral science education. The authors describe an ongoing curriculum development and improvement process that produced one such exemplary program at The Ohio State University College of Medicine. The authors provide a historical perspective on behavioral science education, discuss issues that led to curricular change, and describe the principles and processes used to implement reform. Critical factors underlying positive change are addressed: increase active learning, recruit a core group of small-group facilitators who are primary care physicians, diversify teaching methods, support student-directed educational initiatives, enhance student-teacher relationships, centralize course administration, obtain funding, implement a faculty development program, and apply curriculum quality improvement methods. Outcome data from evaluations completed by both students and small-group physician faculty are presented, and future directions regarding further revision are outlined. The authors believe that the strategies they describe can be applied at other institutions and assist behavioral science educators who may experience the challenges typically encountered in this important field of medical education. PMID- 18162748 TI - Communication skills training: describing a new conceptual model. AB - Current research in communication in physician-patient consultations is multidisciplinary and multimethodological. As this research has progressed, a considerable body of evidence on the best practices in physician-patient communication has been amassed. This evidence provides a foundation for communication skills training (CST) at all levels of medical education. Although the CST literature has demonstrated that communication skills can be taught, one critique of this literature is that it is not always clear which skills are being taught and whether those skills are matched with those being assessed. The Memorial Sloan-Kettering Cancer Center Comskil Model for CST seeks to answer those critiques by explicitly defining the important components of a consultation, based on Goals, Plans, and Actions theories and sociolinguistic theory. Sequenced guidelines as a mechanism for teaching about particular communication challenges are adapted from these other methods. The authors propose that consultation communication can be guided by an overarching goal, which is achieved through the use of a set of predetermined strategies. Strategies are common in CST; however, strategies often contain embedded communication skills. These skills can exist across strategies, and the Comskil Model seeks to make them explicit in these contexts. Separate from the skills are process tasks and cognitive appraisals that need to be addressed in teaching. The authors also describe how assessment practices foster concordance between skills taught and those assessed through careful coding of trainees' communication encounters and direct feedback. PMID- 18162749 TI - The post-baccalaureate premedical certification program at the University of North Texas Health Science Center strengthens admission qualifications for entrance into medical school. AB - The Post-Baccalaureate (postbac) Premedical Certification Program at the University of North Texas Health Science Center provides an opportunity for individuals to enhance their credentials for entry into medical school by offering a challenging biomedical science core curriculum in graduate biochemistry, cell biology, physiology, and pharmacology. In addition, students (called postbacs) receive instruction in human gross anatomy, histology, and embryology with first-year medical students. More than 90% of the students accepted into the postbac program have applied to medical school previously but have been rejected by admission committees at least once, primarily because of low cognitive scores. In spring 2001, seven postbacs completed the program, of which only one was admitted into the Texas College of Osteopathic Medicine (TCOM), the medical school affiliated with the University of North Texas Health Science Center. Three postbacs went to other medical schools. Thirty-one completed the program by spring 2006, of whom 13 were admitted to TCOM, and eight to other medical schools. After six years, 101 postbacs have completed the program, and 70 have been accepted into medical schools. Postbacs admitted into TCOM have performed well compared with their medical school classmates. Overall, average scores for postbacs are above those of their medical school classmates. In addition, postbacs have taken class leadership positions, served as tutors and mentors, and have served as school ambassadors for new applicants. The postbac premedical program has proven to be very successful in preparing students for the rigors of a medical school curriculum by allowing these students to develop the skills and confidence necessary to compete. PMID- 18162751 TI - Beyond the dual degree: development of a five-year program in leadership for medical undergraduates. AB - The current state of physician leadership education consists mainly of executive degree programs designed for midcareer physicians. In 2004, the authors proposed that, by educating medical students in physician leadership and integrating this with a business management or public health degree program, graduates, health care organizations, and communities would benefit sooner. Given the lack of program models to guide program integration and development, the authors began a one-year inquiry to build a model leadership curriculum and integrate leadership education across degree programs. The qualitative inquiry resulted in several linked tasks. First, the authors identified a feasible method for concurrently delivering all three program components (MD degree, Leadership Curriculum, and MBA or MPH degree) during a five-year plan. Second, the authors chose a competency-based educational framework for leadership and then identified, adapted, and validated existing leadership competencies to their context. Third, the authors performed an extensive program alignment to identify existing overlaps and opportunities for integration within and across program components. Fourth, the authors performed a needs analysis to identify educational gaps, subsequently leading to redesigning two courses and to designing three new courses. A description of the Leadership Curriculum is also provided. This inquiry has led to the development of the Boonshoft Physician Leadership Development Program, which provides physician leadership education integrated with medical education and education in business management or public heath. Future program initiatives include developing leadership student assessment tools and testing the link between program activities and short- and long-term outcome measures of program success. PMID- 18162753 TI - Viewpoint: developing integrated clinical programs: it's what academic health centers should do better than anyone. So why don't they? AB - Few would dispute that health care should be provided in seamless, well integrated clinical care environments that bring together the various disciplines needed to provide patient-centered care, to educate trainees, and to conduct research into a particular disease or episode of care. Yet there are relatively few examples of successful or sustained clinical integration, either in the community setting or in academic health centers (AHCs). The authors draw on their experience with several AHCs and other health care settings to address why AHCs have not made better progress in developing integrated centers of clinical care. They characterize two fundamental types of integration that have evolved within the AHC setting: lateral and vertical. Lateral integration tends to occur among similarly situated specialties. It is easier to accomplish and far more common than is vertical integration, which brings together most, if not all, of the professionals and staff necessary to treat or manage many medical conditions and health problems. The vast majority of examples of clinical integration, whether lateral or vertical, fail to integrate essential administrative and financial functions, which has significant consequences for the ability of either laterally or vertically integrated centers to provide seamless, patient-centered care. The authors identify the emergence of several new examples of vertical clinical integration that also integrate administrative and financial functions as models for AHCs to follow and derive lessons and recommendations concerning how AHCs and others can address the cultural, financial, and governance issues that continue to limit the development of vertically integrated, patient-centered care. PMID- 18162754 TI - Current issues facing academic surgery departments: stakeholders' views. AB - PURPOSE: To determine the issues important to stakeholders in today's academic surgery departments, to query key stakeholders about possible solutions, and to investigate the correlation of organizational alignment among department stakeholders with department performance. METHOD: Between July 2003 and October 2005, the author designed, piloted and carried out a study in which he interviewed key stakeholders-deans, chief executive officers/chief financial officers (CEOs/CFOs) of hospitals and health system practice plans, surgery department chairs, and surgery department members-from 12 randomly selected academic surgery departments. Important issues and solutions were identified and comparisons among stakeholder groups performed. Alignment was evaluated both among and within groups and organizations. RESULTS: Stakeholders (11 deans, 9 CEO/CFOs, 12 department chairs, 10 department faculty members) identified 12 issues and offered potential solutions and responses important to today's academic surgery department. One issue identified was promotion and tenure; nearly all stakeholders stated that its current form needed to be changed. Alignment analysis was incomplete because of inconsistent outcomes reporting. CONCLUSIONS: The uniformity of issues facing academic surgery departments and the similarity of the solutions proposed to address these issues (both study findings) suggest a need to change the paradigm and think "outside the box." The study findings suggest that academic surgery departments, under strong leaders, must establish a unified culture, define a compelling vision, articulate a clear mission, and develop fully accepted values to be successful. The study findings could be useful in designing and developing academic surgery departments in today's health care environment. PMID- 18162755 TI - Where were you when I needed you? PMID- 18162756 TI - Medicine and the arts. Pesta on the Stairs by Theodor Kittelsen. Commentary. PMID- 18162757 TI - Adapting industry-style business model to academia in a system of Performance based Incentive Compensation. AB - Performance-Based Incentive Compensation (PBIC) plans currently prevail throughout industry and have repeatedly demonstrated effectiveness as powerful motivational tools for attracting and retaining top talent, enhancing key indicators, increasing employee productivity, and, ultimately, enhancing mission based parameters. The University of Arkansas for Medical Sciences (UAMS) College of Medicine introduced its PBIC plan to further the transition of the college to a high-performing academic and clinical enterprise. A forward-thinking compensation plan was progressively implemented during a three-year period. After the introduction of an aggressive five-year vision plan in 2002, the college introduced a PBIC plan designed to ensure the retention and recruitment of high quality faculty through the use of uncapped salaries that reflect each faculty member's clinical, research, and education duties. The PBIC plan was introduced with broad, schoolwide principles adaptable to each department and purposely flexible to allow for tailor-made algorithms to fit the specific approaches required by individual departments. As of July 2006, the college had begun to reap a variety of short-term benefits from Phase I of its PBIC program, including increases in revenue and faculty salaries, and increased faculty morale and satisfaction.Successful implementation of a PBIC plan depends on a host of factors, including the development of a process for evaluating performance that is considered fair and reliable to the entire faculty. The college has become more efficient and effective by adopting such a program, which has helped it to increase overall productivity. The PBIC program continues to challenge our faculty members to attain their highest potential while rewarding them accordingly. PMID- 18162758 TI - Evidence-based appointment and promotion of academic faculty at the University of Chicago. AB - The authors report how one academic medical center (AMC) and associated nonclinical departments implemented evidence-based academic criteria and an evidence-based academic vetting process, which may be models for other institutions. In 2004-2005, The University of Chicago Division of the Biological Sciences and Pritzker School of Medicine reconceptualized its appointment, promotion, and tenure criteria to recognize all forms of scholarship as equally legitimate bases for academic tenure. The revised criteria also accommodate differences in academic effort consistent with varying clinical demands. Implementation of these criteria, however, necessitated revised practices in providing objective evidence and analysis of their satisfaction. Three complementary mechanisms now yield excellent evidence and analysis. The first, electronic forms (e-forms) comprise highly specific response items with embedded instructions, advice, and rationale. The e-forms encourage candidates and departments to provide the evidence that subsequent review needs to evaluate appointment or promotion proposals. Unexpectedly, the e-forms have been coopted as effective mechanisms for faculty development. Second, a faculty dean of academic affairs, a regular faculty member, was appointed to provide robust academic authority and perspective to the process. Third, the promotion and tenure advisory committee was restricted to evaluating academic criteria, and from considerations of institutional value. This change interposed a "firewall" between academic and institutional review. These changes have attenuated dissatisfaction with the appointments and promotions process both within and outside the AMC. PMID- 18162759 TI - When should students learn essential physical examination skills? Views of internal medicine clerkship directors in North America. AB - PURPOSE: To determine whether any consensus exists among internal medicine clerkship directors regarding when students should acquire proficiency in selected physical examination (PE) skills. METHOD: In 2004, the annual survey of Clerkship Directors in Internal Medicine (CDIM) included a question about the timing of PE-skills proficiency. (CDIM members are from 123 U.S. and Canadian medical schools.) A total of 259 members (123 institutional and 136 individual members) were asked the following question about 39 common physical examination skills, selected using a consensus process among the authors and members of the CDIM Council: "When in the medical school curriculum should medical students acquire proficiency for the following skills?" RESULTS: There were 157 respondents, an overall response rate of 60%. There were 89 (72%) responding institutional members and 68 (50%) responding individual members. Respondents agreed that 31 (80%) of the skills should be learned by the end of the clerkship year. However, considerable variability existed regarding when in the curriculum those skills should be learned: for only 18 of 39 skills was there 80% agreement on skills-learning timing. CDIM members were divided on whether normal examination findings should be learned before or during the clerkships. CONCLUSIONS: Variability existed among CDIM members regarding their expectations for the timing of student physical examination learning in the undergraduate medical curriculum. Creating a common vision among clerkship directors and faculty regarding what neophyte clinicians must learn to do and when they are expected to be able to do it will help to address the issue of physical examination proficiency standards of medical students. PMID- 18162760 TI - The influence of physicians' demographic characteristics and their patients' demographic characteristics on physician practice: implications for education and research. AB - In recent years, interest in improving health care to diverse patient populations has stimulated the development of academic and clinical resources to improve physicians' cultural competence. These efforts have focused on increasing physicians' sensitivity to the roles patients' ethnicity and culture play in health care. However, the influence of physicians' own demographic characteristics on their practice of medicine is an important, yet relatively overlooked, consideration among efforts to improve cross-cultural care. There is a strong presumption in the medical literature that clinicians are neutral operators governed by objective science and are unaffected by personal variables. Yet, there is a body of research that finds physicians' practice patterns are influenced by their own demographic characteristics, and patient care is affected by the demographic concordance or discordance of the physician-patient dyad. The author discusses this existing literature to illustrate the presence and importance of the impact of physicians' demographic characteristics on the care they provide and discusses strategies to mitigate this influence. Greater attention to understanding the way in which physician demographic characteristics influence clinical care using multidisciplinary and multimodal approaches provides an opportunity to improve the quality of medical education and improve the quality and efficacy of medical care. PMID- 18162762 TI - Viewpoint: physician, know thyself: the professional culture of medicine as a framework for teaching cultural competence. AB - The need for physicians who are well equipped to treat patients of diverse social and cultural backgrounds is evident. To this end, cultural competence education programs in medical schools have proliferated. Although these programs differ in duration, setting, and content, their intentions are the same: to bolster knowledge, promote positive attitudes, and teach appropriate skills in cultural competence. However, to advance the current state of cultural competence curricula, a number of challenges have to be addressed. One challenge is overcoming learner resistance, a problem that is encountered when attempting to convey the importance of cultural competence to students who view it as a "soft science." There is also the challenge of avoiding the perpetuation of stereotypes and labeling groups as "others" in the process of teaching cultural competence. An additional challenge is that few cultural competence curricula are specifically designed to foster an awareness of the student's own cultural background. The authors propose the professional culture of medicine as a framework to cultural competence education that may help mitigate these challenges. Rather than focusing on patients as the "other" group, this framework explores the customs, languages, and beliefs systems that are shared by physicians, thus defining medicine as a culture. Focusing on the physician's culture may help to broaden students' concept of culture and may sensitize them to the importance of cultural competence. The authors conclude with suggestions on how students can explore the professional culture of medicine through the exploration of films, role-playing, and the use of written narratives. PMID- 18162763 TI - E-cadherin/catenin complex status in solid pseudopapillary tumor of the pancreas. AB - Solid pseudopapillary tumor (SPT) of the pancreas is an uncommon neoplasm of uncertain lineage. They have been shown to express nuclear beta-catenin believed to be due to mutations of the beta-catenin gene. The aim of this study was to investigate the status of the E-cadherin/catenin complex in SPTs. We studied the expression of 4 principal members of the E-cadherin/catenin complex using immunohistochemistry and the E-cadherin gene status by screening all exons of the gene for mutations, in 6 cases of SPT. In addition to the nuclear localization of beta-catenin, we found nuclear localization of E-cadherin in all tumors with complete absence of membranous and cytoplasmic localization. Nuclear localization of E-cadherin was independent of beta-catenin. No mutations were identified in the E-cadherin gene in any of the tumors. Ten cases of pancreatic adenocarcinomas and 15 neuroendocrine tumors were studied as well for comparison. The reported changes in the expression of the principal members of the E-cadherin/catenin complex were unique to SPTs. Our study shows abnormalities in the expression of 4 principal members of the E-cadherin/catenin complex in SPTs, which may help to explain the discohesive nature of the cells and the cystic changes in these tumors, and provide additional diagnostic features. PMID- 18162764 TI - The utility of fluorescence in situ hybridization (FISH) in the diagnosis of myxoid soft tissue neoplasms. AB - Diagnosing myxoid soft tissue neoplasms can be challenging because of overlapping histologic features. Distinct chromosomal translocations have been identified in several myxoid sarcomas, including t(12;16)(q13;p11) FUS-DDIT3 in myxoid liposarcoma, t(7;16)(q34;p11) FUS-CREB3L2 in low-grade fibromyxoid sarcoma, and t(9;22)(q31;q12) EWSR1-NR4A3 in extraskeletal myxoid chondrosarcoma. These recurrent chromosomal alterations are attractive targets for diagnostic studies. To that end, dual-color, break-apart fluorescence in situ hybridization (FISH) probes spanning the genomic regions of EWSR1 (22q12), DDIT3 (12q13), and FUS (16p11) (Vysis, Downer's Grove, IL) were evaluated in formalin-fixed, paraffin embedded tissues from myxoid neoplasms, including intramuscular myxoma (n=10), myxoid liposarcoma (n=18), low-grade fibromyxoid sarcoma (n=10), extraskeletal myxoid chondrosarcoma (n=13), and myxofibrosarcoma (n=8). Of the myxoid liposarcomas, 18/18 cases had a rearrangement of the DDIT3 gene, with 17/18 (94.4%) showing both DDIT3 and FUS gene rearrangements. A FUS gene rearrangement was identified in 7/10 (70%) of low-grade fibromyxoid sarcomas, with no changes involving EWSR1 or DDIT3. An EWSR1 translocation was seen in 6/13 (46.2%) of extraskeletal myxoid chondrosarcomas, without changes in DDIT3 or FUS genes. The remaining neoplasms studied showed no rearrangements involving DDIT3, FUS, or EWSR1 genes. In conclusion, interphase FISH using DDIT3 and FUS probes identifies the characteristic translocation in myxoid liposarcoma. FUS and EWSR1 probes are useful in confirming the diagnosis of low-grade fibromyxoid sarcoma and extraskeletal myxoid chondrosarcoma, respectively. The specificity of the probes is documented as none of the non-translocation-associated myxoid tumors showed genomic abnormalities with the probes tested. FISH is capable of providing specific ancillary information useful in this often difficult differential diagnosis. PMID- 18162765 TI - t(8;13)-positive bilineal lymphomas: report of 6 cases. AB - The 8p11 myeloproliferative syndrome (EMS) is a rare hematologic malignancy characterized by myeloid hyperplasia, eosinophilia, and precursor lymphoblastic lymphoma, associated with balanced translocations involving chromosome 8p11, most commonly t(8;13)(p11;q12). Approximately 75% of EMS patients present with or develop precursor T-cell lymphoblastic lymphoma, and most subsequently develop acute myeloid leukemia. Here we describe the morphologic and immunophenotypic features of 6 cases of t(8;13)-positive bilineal lymphoma of mixed T-cell and myeloid lineage, 5 in lymph nodes and 1 in breast. The patients, 3 males and 3 females, ranged in age from 6 to 19 years. Histologically, each tumor was composed of 2 distinct cellular components: small to medium-sized T cells with scant cytoplasm that resembled lymphoblasts, and larger immature-appearing cells with more abundant eosinophilic cytoplasm that resembled myeloblasts, a subset of which expressed myeloid antigens. In all cases, the latter component tended to surround residual lymphoid follicles and/or blood vessels. Numerous eosinophils and prominent high endothelial venules were present in all of the lymph node specimens. Interestingly, cells of both components expressed CD3 on immunohistochemical stains. In conclusion, EMS associated with t(8;13) should be suspected in patients with a bilineal tumor that involves lymph nodes or other extramedullary sites. We believe that these bilineal neoplasms of mixed T-cell and myeloid lineages, which present as lymphoma, are analogous to bilineal leukemias. They likely arise from an early hematopoietic cell with potential to differentiate along T-cell and myeloid pathways. PMID- 18162766 TI - Sessile serrated adenoma (SSA) vs. traditional serrated adenoma (TSA). AB - The morphologic distinction between various serrated polyps of the colorectum may be challenging. The distinction between sessile serrated adenoma (SSA) and traditional serrated adenoma (TSA) may be difficult using currently available criteria mostly based on cytologic characteristics. We have evaluated 66 serrated polyps including 29 SSA, 18 TSA, and 19 hyperplastic polyps for overall shape of the polyps, architectural features of individual crypts, the presence of eosinophilic cytoplasm, size and distribution of the proliferation and maturation zones, as well as Ki-67 and CK20 expression. The extent of the expression of CK20 and Ki-67 could not distinguish between the 3 types of serrated polyps, but the distribution of their expression was very helpful and differences were statistically significant. The distribution of Ki-67+ cells was the single most helpful distinguishing feature of the serrated polyp type (P<0.0001, chi test). Hyperplastic polyps had regular, symmetric, and increased Ki-67 expression. SSA had irregular, asymmetric, and highly variable expression of Ki-67. TSA had low Ki-67 expression, which was limited to "ectopic crypts" and admixed tubular adenomalike areas. In serrated polyps, ectopic crypt formation (ECF) defined by the presence of ectopic crypts with their bases not seated adjacent to the muscularis mucosae was nearly exclusive to TSA and was found in all cases, while the presence of cytologic atypia and eosinophilia of the cytoplasm were characteristic, but not limited to TSA. No evidence of ECF, but nevertheless abnormal distribution of proliferation zone was characteristic of SSA, whereas HP had neither. The presence of the ECF defines TSA in a more rigorous fashion than previous diagnostic criteria and also explains the biologic basis of exuberant protuberant growth associated with TSA and the lack of such growth in SSA. Recognition of this phenomenon may also help in exploring the genetic and molecular basis for differences between SSA and TSA, because these architectural abnormalities may well be a reflection of abnormalities in genetically programmed mucosal development. PMID- 18162767 TI - Sessile serrated adenoma: challenging discrimination from other serrated colonic polyps. AB - Sessile serrated adenoma (SSA) is the proposed precursor for microsatellite unstable colorectal carcinomas and some authorities recommend that SSAs should be managed similar to adenomas. The aim of our study was to determine whether serrated polyps can be classified with sufficient consistency to support current treatment recommendations. One hundred eighty-five serrated polyps were classified as hyperplastic polyp (HP), SSA, or traditional serrated adenoma (TSA) by 5 pathologists blinded to clinical data. The observers documented which histologic features they considered most helpful in reaching their diagnosis in each case. In a second round, the observers were provided with polyp site and size. After reaching a consensus on minimum criteria for SSA and TSA, the pathologists classified another set of 50 polyps. The interobserver concordance was calculated using kappa statistics. In the first round, the overall interobserver agreement was moderate (kappa=0.55). Concordance for HP and SSA was moderate whereas it was nearly perfect for TSA. In the second round, there was no improvement in the concordance. All observers relied more often on architectural features than on cytologic ones to distinguish SSA from HP and agreement was reached that architectural features should provide the basis for the diagnosis of SSA. Subsequently, interobserver concordance was slightly improved but remained moderate (kappa=0.58). Interobserver agreement for the diagnosis of serrated polyps is moderate. However, this level of variability is acceptable because the presence of SSA indicates increased risk of developing additional serrated polyps and carcinoma, and surveillance is appropriate. PMID- 18162768 TI - Atypical cellular blue nevi (cellular blue nevi with atypical features): lack of consensus for diagnosis and distinction from cellular blue nevi and malignant melanoma ("malignant blue nevus"). AB - The distinction of cellular blue nevi (CBN) with atypical features ["atypical" CBN (ACBN)] from conventional CBN and malignant melanomas related to or derived from CBN remains a difficult problem. Here, we report on the diagnosis of various cellular blue melanocytic neoplasms by 14 dermatopathologists who routinely examine melanocytic lesions. Three parameters were assessed: (1) for between rater analyses, we calculated interobserver agreement by the kappa statistic (regardless of whether the diagnosis was correct). (2) For each individual lesion, we reported whether a majority agreement (>50%) was reached and, if so, whether the majority agreed with the gold standard diagnosis, derived from standardized histopathologic criteria for melanoma, definitive outcome such as metastatic event or death of disease, or disease-free follow-up for > or =4 years. (3) For the individual pathologists, we calculated sensitivity and specificity for each type of lesion. The study set included 26 melanocytic lesions: (1) 6 malignant melanomas developing in or with attributes of CBN; (2) 11 CBN with atypical features and indeterminate biologic potential (ACBN); (3) 8 conventional CBN; and (4) 1 common BN. The kappa values for interrater agreement varied from 0.52 (95% confidence interval 0.45, 0.58) for melanoma to 0.02 (0.05, 0.08) for ACBN and 0.20 (0.13, 0.28) for CBN. The kappa for all lesions was 0.25 (0.22, 0.28). The pathologists' sensitivities were 68.6% (61.0%, 76.1%) for melanoma, 33.1% (21.0%, 45.2%) for ACBN, and 44.6% (29.0%, 60.3%) for CBN. The specificities were 65.7% (55.8%, 75.6%) for melanoma, 84.7% (77.3%, 92.2%) for ACBN, and 89.9% (82.7%, 97.1%) for CBN. Overall, greater than 50% of the pathologists agreed and were correct in their diagnosis 38.5% (10 lesions) of the time. There was a majority agreement, but with an incorrect diagnosis, another 26.9% (7 lesions) of the time. Six of the 7 majority agreements with an incorrect diagnosis were for ACBN lesions. In summary, the results of our study indicate that there is substantial confusion and disagreement among experienced histopathologists about the definitions and biologic nature of cellular blue melanocytic neoplasms particularly those thought to have atypical features ("atypical" CBN). PMID- 18162769 TI - Lymph nodes after preoperative chemoradiotherapy for rectal carcinoma: number, status, and impact on survival. AB - The number and status of lymph nodes examined is crucial for tumor staging. Impact of preoperative chemoradiotherapy on lymph nodes status and survival is still controversial in rectal carcinoma. The aim of this study was (i) to define the impact of preoperative chemoradiotherapy on the number of both retrieved and positive lymph nodes in rectal cancer specimen, (ii) to evaluate the influence of the number of lymph nodes retrieved on survival in patients treated by preoperative chemoradiotherapy. From 1994 to 2004, 495 patients underwent rectal excision for cancer, of which 332 received long course preoperative radiotherapy. Surgery and pathologic assessment were standardized. Multivariate analysis evaluated the influence of clinical and pathologic variables on the number of both retrieved and positive lymph nodes. Kaplan-Meier method and log-rank test assessed the relation between survival and the number of lymph nodes retrieved in patients treated by preoperative chemoradiotherapy. Compared with surgery alone, preoperative chemoradiotherapy decreased both the mean number of lymph nodes retrieved (17 vs. 13; P<0.001) and the mean number of positive lymph nodes (2.3 vs. 1.2; P=0.001). Multivariate analysis confirmed the independent impact of preoperative chemoradiotherapy on retrieved and positive lymph nodes. In patients treated by preoperative chemoradiotherapy, the 5-year overall (71%) and disease free (60%) survival was not associated with the number of lymph nodes retrieved. Although long course preoperative chemoradiotherapy decreases by 24%, the mean number of lymph nodes retrieved and by 48% the mean number of positive lymph nodes, survival was not influenced by the number of lymph nodes retrieved in irradiated rectal specimen. PMID- 18162770 TI - Atypical genital nevi. A clinicopathologic analysis of 56 cases. AB - Atypical genital nevi are rare melanocytic lesions that most commonly arise on the vulva of young women. They are currently regarded as nevi of special sites, in that despite histologically worrisome features, their clinical behavior is reportedly benign. However, only few studies with limited follow-up data are available. To better characterize the clinical presentation and behavior of these lesions and to further delineate their histologic features, we retrieved 56 atypical genital nevi arising in the lower female genital tract from our departmental and consultation files. The 56 lesions arose in 55 female patients with a median age of 26 years (range, 6 to 54 y). The dominant histologic feature was a lentiginous and nested junctional component composed of prominent round or fusiform nests, which often showed retraction artifact and/or cellular dyscohesion. Cytologic atypia was mild in 11 cases (20%), moderate in 34 (60%), and severe in 11 (20%). Ten cases (18%) showed focal pagetoid spread, with extension to the granular layer and stratum corneum in 1 case. The atypical junctional melanocytic proliferation was associated with a large common dermal nevus component that dominated the lesion in 26 cases (46%). Adnexal spread (46%) and nuclear atypia of melanocytes situated in the superficial dermis (39%) were relatively common, but dermal mitoses (7%) were uncommon and maturation was present in all cases. A broad zone of dense eosinophilic fibrosis within the superficial dermis was a frequent finding (41%). Clinical follow-up was available in 45 cases (80%) with a median follow-up period of 3.5 years (range, 1 to 16 y). Only 1 lesion recurred, 1.5 years after the initial excision. The original nevus in this patient had only mild cytologic atypia and was present at the margins of excision. The recurrent/persistent nevus was reexcised, and there was no further clinical recurrence in 11.5 additional years of follow-up. Our data support the hypothesis that atypical genital nevi have a benign clinical course despite their occasionally striking cytologic and architectural atypia. Awareness and recognition of this group of melanocytic lesions is important to avoid over diagnosis as melanoma with subsequent wide excision and possibly sentinel lymph node biopsy. PMID- 18162771 TI - Partial atrophy in prostate needle biopsies: a detailed analysis of its morphology, immunophenotype, and cellular kinetics. AB - We systematically analyzed 73 prospectively collected partial atrophy (PA) foci from over 185 prostate needle biopsy cases to characterize them along 3 fronts: morphologic, as it can be a mimic of prostate cancer (PCa), immunohistochemical (basal cell markers and alpha-methyl acyl-CoA racemase), as it often shares the staining characteristics of PCa, and cellular kinetics (MIB-1 proliferation marker), as it belongs to the larger group of focal atrophy, some of which have been shown to be proliferative and associated with chronic inflammation. The following morphologic features were prominent at low magnification: small to mid sized glands with circumscribed (70%) or disorganized growth pattern (30%), presence of stellate/undulated gland lumina (92%), associated few completely atrophic glands within the PA focus (97%), and scant apical but abundant lateral pale/clear cytoplasm similar to adjacent benign glands (100%). On higher magnification, 33% of foci contained micronucleoli, but all lacked nuclear enlargement (100%) or macronucleoli (100%), characteristic of PCa. No adjunctive features of PCa were seen. Patchy basal cell staining was observed in 52/71 (73%), whereas 4/71 (6%) were completely negative. alpha-methyl acyl-CoA racemase demonstrated variable expression, stronger than the benign glands in 7/72 (10%) foci. Associated pathology included PCa (42%), and complete atrophy (91%), distinct from PA foci. There was no difference between the mean proliferative index of the PA foci compared with the benign glands [5.5 (range 0 to 30) and 5.6 (range 0 to 31), respectively, P=0.97 by paired t test], as measured quantitatively by ChromaVision system. PA foci were rarely associated with inflammation (1%). Familiarity with these morphologic features and staining characteristics will allow its confident separation from cancer, especially in limited biopsy material. PA foci do not represent a spectrum of proliferative inflammatory atrophy, justifying its term. PMID- 18162772 TI - Small cell carcinoma of the prostate. A morphologic and immunohistochemical study of 95 cases. AB - Small cell carcinoma of prostate is rare, with the literature consisting of case reports and small series. The current work analyzes the morphology and immunohistochemistry of 95 cases of prostatic small cell carcinoma diagnosed at our institution. Specimens included 55 needle biopsies, 27 transurethral resections, 4 radical prostatectomies, and 9 biopsies from metastatic sites (some patients with >1 procedure). Patients ranged in age from 44 to 92 years old (mean: 69 y). Although serum prostate-specific antigen (PSA) in some cases was very high (up to 1896 ng/mL), the median value was only 4.0 ng/mL. Of cases with available information, 33/78 (42%) had a history of usual prostatic adenocarcinoma. The interval between the diagnosis of small cell carcinoma and prior usual prostatic cancer ranged from 1 to 300 months (median 25 mo). Pure small cell carcinoma was seen in 54/95 (57%) of cases with the remaining cases admixed with prostate adenocarcinoma. In cases with adenocarcinoma, there was a sharp demarcation between small cell carcinoma and adenocarcinoma in 20.5% of cases; in the remaining cases there was gradual merging of the 2 components. In mixed cases, small cell carcinoma predominated (median: 80% of the tumor); the Gleason score of the adenocarcinoma was > or =8 in 85% of these cases. In 61 cases (64%), small cell carcinoma was classic "oat cell" morphology with remaining the "intermediate cell" variant. Of the 95 cases: necrosis was seen in 40% (2% to 95% of the tumor); giant bizarre cells in 19%; Indian filing in 21%; rosette formation in 29%; focal vacuolated cytoplasm in 18%; and desmoplasia in 20%. Most (88%) of small cell carcinoma were positive for at least 1 neuroendocrine marker. In the small cell carcinoma component, 14/73 (19%) were positive for PSA, 17/61 (28%) positive for prostein (P501S), and 15/59 (25%) positive for prostate-specific membrane antigen, although often very focally. Stains for thyroid transcription factor-1 were positive in 23/44 (52.3%) cases. In this, the largest study of prostatic small cell carcinoma, we highlight the presence of morphologic features that may result in its underdiagnosis. Other more classic histologic features of small cell carcinoma along with rosettes are critical for its accurate diagnosis. P501S and prostate-specific membrane antigen were better in identifying the prostatic origin of small cell carcinoma than PSA, although the majority (60%) of prostatic small cell carcinomas were negative for all 3 markers. PMID- 18162773 TI - Sporadic cutaneous angiosarcomas: a proposal for risk stratification based on 69 cases. AB - Angiosarcomas have traditionally been considered high-grade lesions for which histologic features and grading have played no role in prognostication and, consequently, they have been excluded from the American Joint Committee on Cancer staging system. We have, therefore, analyzed 69 cutaneous angiosarcomas seen in consultation and not associated with lymphedema or prior radiation therapy to determine if a combination of histologic and clinical parameters could be used to differentiate indolent from aggressive tumors. The clinical features analyzed included patient age, location, size, depth, and focality of the lesion; the histologic features studied included pattern of growth (vasoformative vs. solid), nuclear grade (high vs. low), necrosis (present/absent), cell type (epithelioid or spindled), extent of inflammatory infiltrate (minimal vs. marked), and mitotic rate. Lesions occurred on the head/neck (49), extremities (15), and trunk (5) of adults (21 to 94 y) and predominated in males (41 males; 28 females). Tumors ranged in size from 0.3 to 15 cm (average 3.1 cm) and involved the papillary (n=9), reticular (n=16), or deep dermis/ subcutis (n=30). They could be predominantly vasoformative (n=28) to solid (greater than 50% solid, n=41). Most lesions were of high (n=65) as opposed to low (n=4) nuclear grade, were mitotically active (0 to 99/10 high power fields), and occasionally displayed necrosis (n=14) and epithelioid features (n=21). Inflammatory infiltrates were minimal in most cases. Follow-up information was obtained for all patients. Recurrences developed in 18 patients (26%) and metastasis in 15 (22%) to the following sites: lung (6), lymph node (7), liver (2), bone (2), and parotid gland (1). Forty-seven patients died (30 of disease) and 22 were alive at last known follow-up (range, 16 to 158 mo; mean, 65; median, 36). Five-year disease-specific survival was 48%. By univariate analysis only older age, anatomic site, necrosis, and epithelioid features correlated with increased mortality. Tumors were stratified into low (n=41) or high (n=28) risk groups based on necrosis and/or epithelioid features. By multivariable analysis, high-risk group (hazard ratio 4.07, P=0.0004) and age >70 (hazard ratio 2.79, P=0.012) were associated with increased mortality, and tumor depth (P=0.048) correlated with the risk of local recurrence. The high-risk group had a significantly worse prognosis than the low risk group with 3-year survival of 24% and 77%, respectively. No patients with high-risk features survived 5 years. In conclusion, we report that a combination of clinical and histologic features allows stratification of angiosarcoma patients into 2 risk groups that are strongly associated with marked differences in clinical course. These features seem to diminish in importance with increased tumor size and are probably most useful in tumors less than 5 cm in maximum dimension. PMID- 18162774 TI - Diagnostic utility of S100P and von Hippel-Lindau gene product (pVHL) in pancreatic adenocarcinoma-with implication of their roles in early tumorigenesis. AB - Recently, we demonstrated von Hippel-Lindau gene product (pVHL) was expressed in normal pancreatic ducts but absent in pancreatic ductal adenocarcinoma (PDA). Previous studies have suggested the diagnostic value of S100P, S100A4, and S100A6 in PDA. In this study, we evaluated pVHL, S100P, S100A4, and S100A6 as potential markers for PDA, pancreatic intraepithelial neoplasia (PanIN), ampullary adenocarcinoma (AAD), and cholangiocarcinoma (CC). Immunostains were performed on 56 PDA cases, 20 AAD cases, and 28 CC cases using antibodies against pVHL, S100P, S100A6, and S100A4. Western blots were also performed on 2 cases of PDA and the matching non-neoplastic pancreatic tissues. Of the 56 PDA cases, immunoreactivity for S100P, S100A6, and S100A4 was observed in 56, 55, and 41 cases, respectively. Non-neoplastic ductal epithelium was negative for S100P in all cases. Ninety percent of PanINs were also positive for S100P. pVHL was not detected in all PDAs and 96% of PanINs by immunohistochemistry. S100P, S100A4, and S100A6 were present in a significant number of AADs and CCs; and pVHL expression was observed in 25% of AADs and 21% of CCs. Our data indicate that (1) S100P and pVHL are a pair of sensitive and specific markers for identifying primary PDA and PanIN; (2) up regulation of S100P and down-regulation of pVHL may play a role in early tumorigenesis in PDA; and (3) the 4 markers studied have limited value in differentiating among PDA, AAD, and CC. PMID- 18162775 TI - Pseudocarcinomatous epithelial hyperplasia in the bladder unassociated with prior irradiation or chemotherapy. AB - Pseudocarcinomatous epithelial hyperplasia in the bladder is a little known phenomenon, recognized to be associated with prior irradiation and/or chemotherapy. Whether this process can occur outside of this setting has not been studied. We identified 8 of these cases mimicking invasive urothelial carcinoma from our consultation files from 07/04 to 07/06 with no prior history of radiation or chemotherapy. The mean age at diagnosis was 65 years (range, 42 to 81 y), with 5 of the 8 males. Seven patients had a potential etiology for these changes that could either have resulted in localized ischemia or injury to the urothelium. These included case 1: atrial fibrillation, hypertension, congestive heart failure, gastrointestinal bleeding, and coronary artery vascular disease; case 2: coronary angioplasty, atrial fibrillation, hyperlipidemia, and amputation of arm for ischemia; case 3: hypertension, uncontrolled diabetes, hyperlipidemia, and atrial fibrillation; case 4: underlying arteriovenous malformation of the bladder; cases 5 to 6: history of indwelling Foley catheter; and case 7: history of radical prostatectomy for prostate cancer but no radiation. One patient had no potential contributing factors. All 8 patients presented with gross hematuria. At cystoscopy, 7 patients had polypoid lesions with 1 appearing nonpolypoid. Histologically, all cases showed epithelial proliferation of urothelium with cells having prominent eosinophilic cytoplasm. This process that mimicked invasive cancer within the lamina propria was marked in 3 cases (38%). Moderate nuclear pleomorphism was seen in 6 cases (75%). Only 1 case revealed mitotic figures. Ulceration was seen in 1 case. All cases showed some degree of hemorrhage with hemosiderin deposition identified in 3 cases (38%). Fibrin deposition was present in 1 case within the stroma, 3 cases in the vessels, and 4 cases in both. Five cases show stromal fibrosis. Edema and vascular congestion were common features (90% and 100%, respectively). Six out of 8 cases were accompanied by moderate to marked acute and chronic inflammation. The original diagnosis included nested variant urothelial carcinoma (1 case), atypical suspicious for invasive carcinoma (5 cases), hemangioma (1 case), and eosinophilic cystitis (1 case). Patients were followed for a mean of 16.5 months (range, 10 to 34 mo), and none developed bladder cancer. As a rare response to ischemia and chronic irritation, pseudocarcinomatous epithelial proliferations in the bladder may be confused with invasive urothelial carcinoma. Pathologists must be aware of the histologic changes mimicking cancer, and recognize that it can occur outside of the setting of prior irradiation or chemotherapy. PMID- 18162776 TI - The Use of p16 in enhancing the histologic classification of uterine smooth muscle tumors. AB - BACKGROUND: Uterine smooth muscle tumors can usually be divided histologically into leiomyoma (L) and leiomyosarcoma (LMS). Occasionally, the histologic features are indeterminate and classified as smooth muscle tumor of uncertain malignant potential (STUMP). Recent gene expression studies have found p16 overexpressed in LMS when compared with normal myometrium. This study evaluated the protein expression of p16 by immunohistochemistry in LMS, L, and normal myometrium. Additionally, 8 tumors originally classified as STUMP were evaluated for p16 expression and correlated to their clinical outcome. METHODS: A tissue microarray was constructed and composed of 15 LMS, 8 STUMPs, 22 L, and 10 samples of normal myometrium. p16 expression was correlated with clinical outcome and histologic features. RESULTS: Twelve of the 15 LMS strongly and diffusely expressed p16, 3 of the L had focal p16 staining, and none of the normal myometria were p16 positive. Three of the tumors originally classified as STUMP developed metastatic disease and 2 of these tumors had strong, diffuse p16 positivity. Histologically, these 2 cases were characterized by coagulative tumor cell necrosis and only mild cytologic atypia. CONCLUSIONS: p16 is preferentially expressed in LMS with only rare L showing positivity. Histologically, tumors with coagulative tumor cell necrosis alone were clinically LMS. In those cases in which the type of necrosis is uncertain (coagulative tumor cell vs. hyalinized), the addition of p16 may aid in discerning a subset of STUMP that should be classified as LMS. PMID- 18162777 TI - Sox2 expression in brain tumors: a reflection of the neuroglial differentiation pathway. AB - Sox2 is a key transcription factor that maintains the proliferation of neuroglial stem cells and inhibits neuronal fate commitment. Moreover, it was recently found that brain tumors contain stem cells that resemble normal neuroglial stem cells in many respects. This study was undertaken to describe Sox2 expression in various brain tumors, and to determine whether Sox2 expression is a universal feature of brain tumors, or whether its expression is limited to a specific lineage of brain tumors. Sox2 immunohistochemistry was performed on 194 brain tumor tissues of various kinds. Fetal and adult normal brain tissues obtained by autopsy and brain tissues of epilepsy patients with cortical dysplasia were used as controls. Semiquantitative reverse transcription polymerase chain reaction was used to confirm the immunohistochemical results. Double immunofluorescence was performed to characterize the lineage of Sox2-positive cells. Sox2 was found to be expressed in various glial tumors, including those with astroglial, oligodendroglial, and ependymal lineages, and in the glial components of mixed neuroglial tumors, regardless of pathologic grade. In brain tumors of embryonal origin, supratentorial primitive neuroectodermal tumors showed robust Sox2 expression, whereas medulloblastomas and pineoblastomas did not. The majority of Sox2-positive tumor cells coexpressed glial fibrillary acidic protein, and most Sox2-negative cells in medulloblastomas and pineoblastomas showed neuronal differentiation. This study suggest that Sox2 may be a tumor marker of glial lineages rather than a universal brain tumor stem cell marker, because its expression pattern was found to correspond to differentiation pathways. On the other hand, the aberrant coexpressions of Sox2 and of a neuronal marker were widely observed in glioblastomas, which reflects a disorganized differentiation pattern that characterizes highly malignant tumors. PMID- 18162778 TI - Characteristic expression patterns of TCL1, CD38, and CD44 identify aggressive lymphomas harboring a MYC translocation. AB - The distinction between Burkitt (BL) or atypical Burkitt/Burkitt-like lymphomas harboring a MYC translocation (MYC+) and diffuse large B-cell lymphomas (DLBCLs) with high proliferation fractions but without a MYC translocation (MYC-) can be difficult using standard morphologic and immunohistochemical criteria. Recently, unique gene expression profiles differentiating BL and DLBCL were reported and include higher transcript levels of T-cell leukemia-1 (TCL1) and CD38 and lower transcript levels of CD44 in MYC+ BL relative to MYC- DLBCL. We examined a cohort of 67 cytogenetically defined aggressive lymphomas using immunohistochemical techniques for expression of TCL1, CD38, and CD44 and found distinct expression patterns between MYC+ and MYC- tumors. Furthermore, these markers are better predictors of MYC status than combined staining for CD10 and BCL2. Thus staining for TCL1, CD38, and CD44 are useful ancillary tests to identify B-cell tumors for which confirmatory cytogenetic and/or fluorescent in situ hybridization studies assessing the status of the MYC locus should be pursued. PMID- 18162779 TI - Podoplanin is a better immunohistochemical marker for sarcomatoid mesothelioma than calretinin. AB - Immunohistochemistry using a broad panel of markers is an invaluable tool for diagnosing sarcomatoid mesothelioma. Membranous podoplanin staining has been proposed as a specific and sensitive marker to distinguish epithelioid mesothelioma from adenocarcinoma. We found that cytoplasmic podoplanin staining was present in sarcomatoid mesotheliomas, and wanted to explore the reproducibility and specificity of this staining pattern. Immunohistochemistry for podoplanin, using 2 podoplanin antibodies (antipodoplanin and D2-40), was performed in 55 mesotheliomas (24 epithelioid, 18 sarcomatoid, and 13 biphasic), 80 pulmonary adenocarcinomas, 8 synovial sarcomas, and 16 sarcomatoid carcinomas. Expression of calretinin, vimentin, MOC31, and TTF-1 was also examined in all adenocarcinomas, sarcomatoid carcinomas, 7 synovial sarcomas, and 21 of the mesotheliomas. Calretinin staining performed previously on an additional 31 mesotheliomas was reviewed. Using membranous or cytoplasmic staining as indicative of positivity, we found that antipodoplanin and D2-40 each stained 84% of mesotheliomas (antipodoplanin: 46/55; D2-40: 38/44), including 72% of sarcomatoid mesotheliomas (antipodoplanin: 13/18; D2-40: 11/14). With antipodoplanin antibody, no staining was seen in the pulmonary adenocarcinomas (0/80, 0%) or the synovial sarcomas (0/8, 0%), and weak cytoplasmic staining was seen in only 1 sarcomatoid carcinoma (1/15, 7%). D2-40 showed similar results, staining 3% (2/80) of pulmonary adenocarcinomas, 13% (1/8) of synovial sarcomas, and 8% (1/13) of sarcomatoid carcinomas. Overall sensitivities and specificities were 84% and 99% for antipodoplanin, and 86% and 96% for D2-40. These findings suggest that cytoplasmic podoplanin expression may be useful in the diagnosis of sarcomatoid mesothelioma, although it should be used with caution on biopsy material. PMID- 18162780 TI - Distinction of primary and metastatic mucinous tumors involving the ovary: analysis of size and laterality data by primary site with reevaluation of an algorithm for tumor classification. AB - Distinction of primary ovarian mucinous tumors from metastatic/secondary mucinous tumors involving the ovaries is often challenging, not only at the time of intraoperative assessment when requested for surgical management (staging decisions) but also for final pathologic diagnosis. Previous studies have shown that a simple algorithm using tumor size and laterality (bilateral tumors of any size, or unilateral tumor <10 cm=metastatic; unilateral tumor > or =10 cm=primary) can accurately classify a substantial majority of tumors. To assess the general utility of this algorithm for distinction of primary and secondary mucinous tumors in the ovary and address the occurrence of exceptions (large unilateral metastases), analysis of tumor size and laterality data was performed using 194 tumors (52 primary tumors and 142 metastases), with metastases subclassified by primary site [colorectum (46), appendix (28 low-grade tumors, 20 carcinomas), pancreaticobiliary tract (20), small intestine (3), stomach (5), and endocervix (20)]. Performance of the algorithm was evaluated using the originally proposed method and modified size criteria were analyzed to optimize tumor classification. The original algorithm correctly classified 84% of tumors overall, including 100% of primary ovarian tumors and 77% of all metastases (colorectal: 74%; appendiceal: 79% of low-grade tumors, 100% of carcinomas; pancreaticobiliary: 95%; small intestinal: 33%; gastric: 80%; endocervical: 55%). By adjusting the size criterion to 12 cm, performance of the algorithm was both maintained for primary ovarian tumors and improved for metastases, with correct classification of 86% of tumors overall, including 100% of primary tumors and 80% of metastases. Performance was optimized at 13 cm, with correct classification of 87% of tumors overall, including 98% of primary tumors and 82% of metastases (colorectal: 80%; appendiceal: 79% of low-grade tumors, 100% of carcinomas; pancreaticobiliary: 100%; small intestinal: 33%; gastric: 100%; endocervical: 70%). Of the more common metastases, metastatic colorectal and endocervical carcinomas provided the greatest number of exceptions, even when analyzed with the optimized size criterion. Recognition that metastatic colorectal carcinomas represent the most common metastases and have a greater tendency to violate the algorithm should prompt lowering of the threshold for suggesting the possibility of metastatic colorectal carcinoma for tumors displaying any microscopic features suggestive of that diagnosis, even when a history of primary colorectal carcinoma is lacking. Use of the algorithm is intended as an adjunct to the complete clinicopathologic evaluation that ideally should occur when problematic mucinous tumors in the ovary are encountered. PMID- 18162781 TI - CD25 expression on cutaneous mast cells from adult patients presenting with urticaria pigmentosa is predictive of systemic mastocytosis. AB - Urticaria pigmentosa (UP) is a clinicopathologic term used to describe reddish brown cutaneous macules and papules, characterized histologically by mast cell infiltration of the papillary and upper reticular dermis and reactive basal hyperpigmentation of the overlying epidermis. Although typically a benign, self limited disorder of childhood, a significant proportion (up to 30%) of adolescent and adult-onset UP represents cutaneous involvement by underlying systemic mastocytosis (SM). Predicting the course of cutaneous mast cell disease has been limited by a lack of diagnostic and prognostic markers. In patients with SM, neoplastic bone marrow mast cells show aberrant surface expression of CD25. However, whether CD25 expression on cutaneous mast cells is associated with underlying SM is unknown. In this study, we performed a clinicopathologic analysis of 30 adult patients presenting with UP between 1987 and 2007. Cutaneous mast cell infiltration pattern, cytomorphology, density, and CD25 immunoreactivity were correlated with underlying or subsequent SM. On the basis of clinical and pathologic follow-up, 10 of 30 (33%) patients were diagnosed with SM and 20 of 30 (67%) with limited cutaneous mastocytosis (CM). Although cutaneous mast cell density was slightly higher in patients with SM compared to those with limited CM (P=0.047), neither mast cell cytomorphology nor infiltration pattern correlated with underlying systemic disease. However, cutaneous mast cells from all 10 patients with SM (100%) were immunoreactive for CD25, compared to only 5 of 20 (25%) with limited CM (P<0.001). Our findings suggest that immunoreactivity for CD25 in cutaneous mast cells may be useful for stratifying adult patients presenting with UP for additional clinical evaluation. PMID- 18162782 TI - Primary mediastinal seminoma: a comprehensive assessment integrated with histology, immunohistochemistry, and fluorescence in situ hybridization for chromosome 12p abnormalities in 23 cases. AB - Accurate diagnosis of mediastinal seminoma is critical because of its favorable response to radiation therapy and/or cisplatin-based chemotherapy. Immunohistochemical staining for OCT4 has recently been validated as a powerful tool for detecting gonadal seminoma. However, discrepancies between the genetic alterations and immunoprofiles of mediastinal and testicular seminomas have been reported, raising the question of whether techniques that are useful in the diagnosis of gonadal seminoma are applicable to its mediastinal counterpart. The present study was conducted to evaluate the morphologic and immunohistochemical characteristics and chromosomal abnormalities of 12p in 23 primary mediastinal seminomas and to compare their applicability as diagnostic tools. Dual-color fluorescence in situ hybridization (FISH) analyses for chromosome 12p and immunostains for OCT4, c-kit, placental-like alkaline phosphatase, CD30, and a panel of cytokeratins, including cytokeratin AE1/AE3 (AE1/3), high molecular weight cytokeratin (34betaE12, HMWCK), CAM5.2, cytokeratin 7 (CK7), cytokeratin 20 (CK20), and epithelial membrane antigen were performed. Lymphocytic infiltration was found in all 23 cases (100%). The incidence of other histologic characteristics were as follows: fibrous septa/stroma (21 cases, 91%), prominent tumor cell nucleoli (21 cases, 91%), clear tumor cell cytoplasm (20 cases, 87%), distinct tumor cell borders (20 cases, 87%), granulomatous inflammation (17 cases, 74%), cellular pleomorphism (10 cases, 43%), necrosis (8 cases, 35%), prominent cystic change (2 cases, 8%), intercellular edema (1 case, 4%), and syncytiotrophoblasts (1 case, 4%). The mean mitotic count was 4.4 (range 0 to 16) per 10 high-power fields. Moderate to strong nuclear OCT4 staining was identified in all 23 cases (100%). Seventeen tumors (74%) showed membranous expression of c kit, with variable staining intensity and percentages. Weakly to moderately intense immunostaining for placental-like alkaline phosphatase was identified in 10 cases (43%) with occasional background staining artifact. The incidences of positive staining were 43% for AE1/3, 39% for HMWCK, 48% for CAM5.2, 39% for CK7, and 9% for epithelial membrane antigen, respectively. In most cases, these epithelial markers highlighted only a small proportion of tumor cells with variable intensities. Immunostaining for CD30 and CK20 was completely negative in all seminomas. Twenty-two seminomas (96%) revealed chromosome 12p abnormalities, including 12p amplification in 20 cases (87%) or i(12p) in 15 cases (65%). Lymphocytic infiltration is the most common histologic feature observed in primary mediastinal seminoma and both OCT4 immunostain and FISH for 12p abnormalities can be very helpful in diagnosing mediastinal seminoma. The intense staining pattern of OCT4 and the high sensitivity of FISH make them superior to other auxiliary diagnostic utilities for detecting seminoma. In addition, the incidences of cytokeratin expression of primary mediastinal seminoma are similar to those of its gonadal counterpart and pathologists must exercise caution in the interpretation of epithelial markers in mediastinal neoplasms. PMID- 18162783 TI - Metastatic peritoneal neurocutaneous melanocytosis. AB - Neurocutaneous melanosis, better referred to as neurocutaneous melanocytosis (NCM), is a rare congenital disorder occurring in childhood characterized by proliferation of melanocytes in the central nervous system (CNS), associated with large congenital melanocytic nevi. The phenotype of the CNS lesions varies, ranging from that of a benign, nevuslike lesion, to one of an aggressive-looking, atypical cell proliferation; however, specific diagnostic criteria allow the distinction from CNS metastasis of a primary skin melanoma. NCM can present with severe neurologic manifestations, and usually has a relentless clinical progression whence neurologic symptoms appear. Dissemination to the peritoneal surface by ventriculo-peritoneal shunting has been exceptionally observed, and we describe 2 cases of such occurrence, one of which was associated with a "bulky perineal nevocytoma" with complex cytogenetic rearrangements. This "metastatic" spreading supports an aggressive phenotype, able to seed and establish new colonies, although only after facilitated translocation of the proliferating cells through the shunt conduit; the aggressiveness of these lesions in our cases is further supported by the histopathologic features and clinical course. The biologic features of NCM cells merit further exploration, as they may shed light on a much more frequent neoplastic neurocristopathy, namely, malignant melanoma. PMID- 18162784 TI - Choroid plexus papilloma with neuropil-like islands. AB - Choroid plexus papilloma may display unusual histologic features, but the presence of neuronal differentiation at the light microscopic level has not yet been described. We thus report a choroid plexus papilloma with neuropil-like islands located within the lateral ventricle of an 11-year-old girl. The absence of atypical histologic features associated with recurrence (particularly increased mitotic activity) and recurrence-free follow-up upon gross total resection are compatible with a diagnosis of choroid plexus papilloma (WHO grade I). This case further emphasizes the capacity of choroid plexus tumor cells toward neuronal differentiation, and expands the spectrum of tumors, which may contain neuropil-like islands. PMID- 18162785 TI - Divergent myoid, neuroendocrine, and perineural differentiation in a nasal tumor of a patient with Carney complex. AB - A 39-year-old woman with Carney complex presented with a stroke of undetermined etiology. Computed tomography showed bilateral thalamic infarctions and also an unsuspected multicompartmental cystic neoplasm that had eroded the anterior clivus and extended forward into the nasopharynx. Histologically, the mass appeared benign and was composed of spindle cells and multiple foci of striated muscle. Immunohistochemically, the spindle cells were strongly reactive for S-100 protein and to a lesser extent for CD57, collagen IV, neuron-specific enolase, smooth muscle actin, epithelial membrane antigen, and glut-1. The striated muscle cells were positive for desmin and myogenin. The MIB-1 labeling index was 0.5%. Ultrastructural examination was necessary to reveal the full extent of divergent differentiation. Ultrastructurally, the spindle cells showed divergent differentiation along several cell lines, including smooth muscle, neuroendocrine, hybrid smooth muscle-neuroendocrine, perineural-like cells, and striated muscle. The occurrence of this unique lesion in a patient with the Carney complex raises the possibility that it may be a rare component of the syndrome. PMID- 18162787 TI - HHV8 in plasmablastic lymphoma. PMID- 18162788 TI - Rhabdomyosarcoma of the adult uterus. PMID- 18162790 TI - Ambulatory Patient Groups, Version 3.0--a classification system for payment of ambulatory visits. AB - The Ambulatory Patient Groups (APGs) are a patient classification system that was designed to be used as the basis of an Outpatient Prospective Payment System (OPPS). Although 6 major non-Medicare payers had implemented an APG-based OPPS between 1995 and 2000, the implementation of the Ambulatory Payment Classification (APC)-based Medicare OPPS shifted the focus of outpatient payment reform among payers to APC-based systems. Unfortunately, the APC OPPS is not really a prospective payment system and has become essentially a variant of a fee for-service system. As a result, most major non-Medicare payers have rejected APCs as a model for outpatient payment reform and a renewed interest in the original APG OPPS design has occurred. This article reviews the basic components of an OPPS, compares and contrasts an APG- and APC-based OPPS, describes the differences between APG, Version 2.0, and APG, Version 3.0, and summarizes the key policy decisions payers will need to make in implementing an OPPS. PMID- 18162791 TI - The use of ambulatory patient groups for regulation of hospital ambulatory surgery revenue in Maryland. AB - The Maryland Health Services Cost Review Commission (HSCRC or the commission) is a government agency with the authority to establish rates for both inpatient and outpatient services for all general acute care hospitals in the state. By law and consistent with the state's unique Medicare waiver, all payers (including Medicare and Medicaid) must pay hospitals on the basis of these rates. The HSCRC has used diagnosis related groups to set case-mix-adjusted limits on the revenue per discharge for inpatient services (similar to Medicare inpatient prospective payment nationally) yet, the Maryland rate-setting system for outpatient services has not embodied incentives to control utilization of services. Beginning in the state's fiscal year 2008, the HSCRC is implementing regulation of ambulatory surgery services using ambulatory patient groups to provide better incentives to control utilization, and to facilitate comparisons of the case-mix-adjusted charges per ambulatory surgery case across hospitals. Maryland has been an innovator in the design and successful implementation of payment systems and other incentive mechanisms to constrain hospital cost, maintain payment equity, and ensure access to needed hospital care. The HSCRC's adoption of all patient refined diagnosis related groups and the hospital-specific relative value method for establishing diagnosis related group weights in 2005 was relevant to the Centers for Medicare and Medicaid Services' decision to move to Medicare severity diagnosis related groups beginning in federal fiscal year 2008, and to consider the use of hospital-specific relative value weights. The HSCRC's decision to use ambulatory patient groups for ambulatory surgery is an attempt to apply the most effective features of inpatient payment systems, prospective payment, including incentives to control service volumes. As such, it represents a radical departure from prevailing payment arrangements in that it seeks to remove the traditional distinction between inpatient and outpatient surgical services, a distinction that has blocked the development of effective and well-integrated outpatient payment systems for decades. This article describes the policy rationale for this system, the analysis that was performed, and the methods that will be used to control the revenue per case and compare the relative charges of the hospitals. PMID- 18162792 TI - Reforming the primary care physician payment system: eliminating E & M codes and creating the financial incentives for an "advanced medical home". AB - The problem faced by primary care physicians is that they can only maintain or increase their (inflation adjusted) incomes by increasing the volume of visits and associated services. The fundamental flaw in a fee-for-service system is that only paying for individual services creates incentives for more services. This article offers a very different approach to paying primary care physicians that will result in both significantly higher incomes for these underpaid professionals together with incentives for creating a medical home. PMID- 18162793 TI - Outpatient hospital services payment system. PMID- 18162794 TI - Who is in charge? Even affluent patients suffer consequences of fragmented care. PMID- 18162795 TI - The feasibility and value of new measures showing patterns of quality for patients with 3 chronic conditions. AB - The aim of this study was to create new measures of quality that combine individual service measures. Using an all-or-none approach, we identify 5 levels of care reflecting the extent to which optimal patterns of service were obtained by patients with asthma, diabetes, and heart failure. We also assess the feasibility of these levels-of-care measures and their potential value in quality improvement efforts. The study was designed to analyze claims data to reflect patterns of services used in a single metropolitan market of about 1 million residents in the northeastern United States. More than 80,000 patients insured over 4 years (1994-1997) had claims with 1 or more of 3 chronic conditions. The analysis showed that the measures discriminated effectively among groups of patients with the 3 chronic conditions and highlighted areas to target quality improvement efforts. Although the numbers vary by year, for two of the diagnoses, most patients were in the lowest categories (59%-75%), and for the third, 40% were in these categories. Few were in the highest category. Most patients were in the same category from one year to the next. The levels-of-care approach to quality measurement can help caregivers and policy makers find methods for avoiding unnecessary utilization and expenditures while raising--not lowering- the probability that utilization patterns will conform to condition-specific recommended care. PMID- 18162796 TI - The value of a drug: from innovation to the payment via Karl Marx. PMID- 18162797 TI - Commentary on Giovanni Apolone's "The Value of a Drug: From Innovation to the Payment via Karl Marx". PMID- 18162798 TI - Commentary on Giovanni Apolone's "The Value of a Drug: From Innovation to the Payment via Karl Marx". PMID- 18162799 TI - Something new every day: defining innovation and innovativeness in drug therapy. AB - The word "innovation" comes from the Latin noun innovatio, derived from the verb innovare, to introduce [something] new. It can refer either to the act of introducing something new or to the thing itself that is introduced. In terms of commerce, it is defined in the Oxford English Dictionary as "the action of introducing a new product into the market; a product newly brought on to the market," a definition that illustrates both aspects of the word's meaning. "Innovativeness" is the property of being an innovation. Here I identify several different types of innovativeness in drug therapy, including structural, pharmacological or pharmacodynamic, pharmaceutical, and pharmacokinetic innovativeness, and I stress the over-riding importance of clinical innovativeness, which should result in a better benefit to harm balance at an affordable cost. PMID- 18162800 TI - Adoption of health center performance measures and national benchmarks. AB - This study examines the adaptability of standardized performance measurement tools in 3 community-based health centers. Although health centers have considerable experience in the area of performance reporting, they do not currently participate in a national reporting system that is transparent and standardized. The analysis of the data collected from health centers indicates that not only can these safety net providers readily integrate standardized measures, the quality of care being provided compare favorably to national benchmarks. With evidence of solid performance may come the types of financial adjustments essential to permitting health centers to move more decisively into the broader private health insurance markets that may exist in their service areas. PMID- 18162801 TI - Improving patient care through health-promotion outreach. AB - Patient receipt of recommended preventive and chronic disease care is suboptimal, partly because of poorly organized clinical information and time-limited and sporadic appointments with medical care providers. To overcome these problems, we describe the features of a novel health-promotion outreach system based on electronic data queries, active patient-centered outreach, and minimizing patient barriers to adherence. We describe advantages of this system over traditional preventive care delivery systems and chronic disease management programs, and explain how it complements and helps to promote the type of clinical practice change contemplated by the chronic care model. PMID- 18162803 TI - PEPFAR reauthorization: can US aid keep up with the AIDS pandemic? PMID- 18162804 TI - Risk assessment to guide the prevention of cervical cancer. AB - Advances in screening and diagnosis make it increasingly possible to prevent cervical cancer. However, if misused or poorly understood, these new tools will only increase costs and potentially harm patients without benefit. As a framework for standardized care that maximizes patient safety and well-being, we propose that a risk model be adopted to guide clinical management now and in the future. The model would use thresholds of increasing risk for cervical precancer and treatable cancer to guide clinical decision making for screening intensity, diagnostic evaluation, or treatment. Experts would decide on these risk thresholds and stratum based on the patient risk to benefit, independent of current (e.g., cytology, carcinogenic human papillomavirus testing, and colposcopy) and future methods of measuring risk. A risk management model for cervical cancer prevention, based on appropriate clinical actions that correspond to risk stratum, can result in better allocation of resources to and increased safety for women at the greatest risk and increased well-being for women at the lowest risk. PMID- 18162805 TI - Why do human papillomavirus infections induce sharply demarcated lesions of the cervix? AB - Chronic infections with high-risk human papilloma virus (HPV) have been established to cause cervical cancer. The causal pathogenesis of cervical carcinoma is believed to be the result of the proliferation of one or, at most, a few HPV-transformed cells. So far, it has not been possible to identify and analyze these early initial lesions or cell clusters. Invasive cervical cancer arises in cervical intraepithelial neoplasia, which in turn develops preferentially in squamous metaplasia in clearly demarcated fields. These fields are the main salient morphologic characteristic in cervical intraepithelial neoplasia and can be recognized colposcopically as well as histomorphologically. The precise mechanism for the development of separate, well-delineated fields in HPV-related intraepithelial neoplasia and the variable susceptibility of stem or reserve cells for different HPV genotypes remains unsolved. In cervical carcinogenesis, the link between formal pathogenesis apparent as colposcopic and histomorphologic changes and the causal pathogenesis of HPV-induced genetic changes is still missing and an issue for future research. PMID- 18162806 TI - Should unsatisfactory colposcopy necessitate treatment of cervical intraepithelial neoplasia 1? AB - OBJECTIVE: To evaluate the prevalence of cervical intraepithelial neoplasia (CIN) 2 or worse among women undergoing cone biopsy for CIN 1, stratified by colposcopic adequacy. MATERIALS AND METHODS: A cross-sectional cohort study was performed using a colposcopic database of 3,004 women seen between August 1999 and December 2005. Data collected included demographic information, indications for treatment, adequacy of colposcopic examination, and final cone pathology. A satisfactory colposcopy was defined as being able to define the entire squamocolumnar junction and visualize all lesions in their entirety. Descriptive statistics and crude and adjusted odds ratios with 95% confidence intervals were calculated. RESULTS: Of the 440 cone biopsies, 50 (11%) were done for CIN 1, of which 9 (18%) demonstrated CIN 2 or worse, and 23 (46%) were done in the setting of unsatisfactory colposcopy. Compared with women with satisfactory colposcopy, women with unsatisfactory colposcopy were more likely to be older (median age = 33 vs 25 years, p = .01) and to have CIN 1 on endocervical sampling (5/22 [22%] vs 0, p < .01). Persistent (18 months or more) CIN 1 was more commonly encountered in women with satisfactory colposcopy (21/27 [78%] vs 7/23 [30%], p < .01). Women with unsatisfactory colposcopic examinations were at decreased risk of CIN 2+ on final pathology compared with women with satisfactory examinations (1/23 [4%] vs 8/27 [30%], odds ratios = 0.08, 95% confidence intervals = 0.01 0.95). CONCLUSIONS: Women with unsatisfactory colposcopy and CIN 1 are unlikely to have high-grade cervical neoplasia on final pathology. For this subset of women, conservative management is preferable to immediate treatment. PMID- 18162807 TI - Outcomes after diagnosis of vaginal intraepithelial neoplasia. AB - OBJECTIVE: To describe presenting characteristics and outcomes after a diagnosis of vaginal intraepithelial neoplasia (VAIN). MATERIALS AND METHODS: Histologically confirmed cases of VAIN identified colposcopically after abnormal cytology were reviewed retrospectively. VAIN 1 was managed with observation, office loop excision, or laser ablation; unifocal or clustered VAIN 2,3 was managed with loop excision; and multifocal VAIN 2,3 was managed with laser ablation. Women were followed up cytologically at 6-month intervals. Clearance was defined as 2 consecutive negative cytology results. RESULTS: Thirty-six cases were identified with adequate follow-up (median = 20 months). VAIN 1 was found in 17 women, VAIN 2 in 8, and VAIN 3 in 11. All VAIN 1 lesions treated with loop excision were cleared, as were 8 of 12 VAIN 1 cases observed without therapy, 5 of 8 cases of VAIN 2, and 7 of 11 cases of VAIN 3. One woman had persistent VAIN 2; all other cases of persistence were low grade. No cases of progression and no treatment complications were identified. CONCLUSIONS: This management strategy seemed to be safe and effective for the control of VAIN. PMID- 18162808 TI - Is this the end of the line for the moderate dyskaryotic smear? AB - OBJECTIVE: The British Society for Clinical Cytology has recently proposed that the terminology for cervical smear reporting is to be changed from a 3-tier system (mild, moderate, severe dyskaryosis) to a 2-tier system of low-grade and high-grade dyskaryosis. This modification eliminates the central category of moderate dyskaryosis which would be incorporated into the high-grade group. The aim of this study was to investigate the role of the moderate dyskaryotic smear in clinical practice. MATERIALS AND METHODS: A retrospective review of all women who were referred for colposcopy because of a moderate dyskaryotic smear was carried out for a 6-month period. Data collected included colposcopic impression, procedure performed and final histopathology. Two cytologists who were unaware of the original smear report were asked to reclassify these smears using the new 2 tier system. Their findings were compared with the documented colposcopic and histopathology results. RESULTS: One hundred women with moderate dyskaryotic smears were referred for colposcopy during the study period. Most of these were reclassified as high-grade dyskaryosis using the new system. Fifty-six (72%) of the moderate dyskaryotic smears which were correctly regraded as high grade by cytologist 1 were found to have cervical intraepithelial neoplasia 2/3 on final histopathology, whereas for cytologist 2, 66 (68%) were found to have high-grade cervical intraepithelial neoplasia. CONCLUSION: There is no clinical benefit in retaining the term moderate dyskaryosis. This study emphasizes the need for a uniform 2-tier system. PMID- 18162809 TI - Overcoming the challenging cervix: techniques to access the uterine cavity. AB - OBJECTIVE: To identify cervical stenosis and review medical, surgical, and radiological modalities to access the uterine cavity. MATERIALS AND METHODS: Computerized searches of Medline and PubMed from 1996 to 2005 were conducted using the key words "cervix," "cervical ripening," and "cervical stenosis." References from identified publications were manually searched and cross referenced to identify additional relevant articles. We review relevant techniques on how to access the uterine cavity when cervical stenosis is encountered. RESULTS: Many gynecologic procedures require uncomplicated access through the cervix to access the uterine cavity, including hysteroscopy, dilation and curettage, sonohysterogram, hysterosalpingogram, endometrial biopsy, and embryo transfer for in vitro fertilization. These procedures can be quite complicated when a cervix is obstructed. Management techniques described include the medical use of misoprostol and laminaria, intraoperative ultrasound guidance, and operative creation of a new passage. Additionally, techniques for bypassing the obstructed cervix and preventing cervical stenosis have been described. CONCLUSIONS: Cervical stenosis can result in iatrogenic complications. Preoperative identification, cervical ripening agents, osmotic dilators, and the use of ultrasound guidance are useful in overcoming cervical stenosis. It is also key to identify those at risk for cervical stenosis and implement preventative techniques as needed. PMID- 18162810 TI - Benign ductal hyperplasia and adenosis of mammary-like glands of the vulva. PMID- 18162811 TI - A rare mass formation of the vulva: giant cavernous hemangioma. AB - OBJECTIVE: Vulvar hemangioma is an extremely rare pathology in adult women causing sexual dysfunction, pain, and cosmetic problems requiring therapeutic intervention. MATERIALS AND METHODS: We present a case of an isolated giant vulvar cavernous hemangioma with a 2-year history that macroscopically seemed to be a vulvar varicosis but was actually detected at the end to be cavernous hemangioma in an adult patient with no history of other disease. RESULTS: The patient was treated with surgical excision because of progressive complaints. Histopathologic evaluation suggested cavernous hemangioma. CONCLUSION: Hemangiomas can undergo spontaneous involution; thus, treatment may be reserved for those lesions of functional or psychological concern. In this case, surgical correction was used both as a diagnostic method and as a therapeutic intervention because of progressive complaints such as pain, feeling of pressure, and sexual dysfunction. PMID- 18162812 TI - Caffeine-related genital skin pain. AB - Genital hyperesthesia is more recognized in women but may affect men. We report both a female and a male patient in whom caffeine intake was associated with genital hyperesthesia. PMID- 18162813 TI - Significance of a diagnosis of microorganisms on pap smear. AB - The Pap smear has been in use for more than half a century as the primary screening test for preinvasive and invasive lesions of the uterine cervix. Although not the primary use and an imperfect test, it can be extremely useful in the diagnosis of some microorganisms. This review focuses on the use of the Pap smear in the diagnosis of several microorganisms including Actinomyces, Chlamydia trachomatis, Candida, Trichomonas vaginalis, Leptothrix vaginalis, Herpes Simplex Virus, the causative agents of bacterial vaginosis, and other rarer organisms. The accuracy of diagnosis using the smear varies among the different organisms in question. PMID- 18162814 TI - Spectroscopic imaging as triage test for cervical disease. PMID- 18162815 TI - A 23-year-old woman has her first Pap smear, thin prep smear interpreted as "high grade squamous intraepithelial lesion". PMID- 18162822 TI - Home study course: winter 2008. AB - OBJECTIVE: The Home Study Course is intended for the practicing colposcopist or practitioner who is seeking to develop or enhance his/her colposcopic skills. The goal of the course is to present colposcopic cases that are unusual or instructive in terms of appearance, presentation, or management or that demonstrate new and important knowledge in the area of colposcopy or pathology. Participants may benefit from reading and studying the material or from testing their knowledge by answering the questions. ACCME ACCREDITATION: The American Society for Colposcopy and Cervical Pathology (ASCCP) is accredited by the Accreditation Council for Continuing Medical Education (ACCME) to provide continuing medical education for physicians. The ASCCP designates this education activity for a maximum of 1 AMA PRA Category I Credittrade mark. Physicians should only claim credit commensurate with the extent of their participation in the activity. The ASCCP also designates their educational activity for 1 Category 1 credit hour of the ASCCP's Program for Continuing Professional Development. Credit is available for those who choose to apply. The Home Study Course is planned and produced in accordance with the ACCME's Essential Areas and Elements. DISCLOSURE: The clinical history and images in the Home Study Course may represent an actual case, but not always. To improve educational quality, some gross, cytological, or histological images may come from photographic libraries. Good teaching cases are often difficult to obtain, and we encourage our readers to submit cases with high-quality images to the Home Study Course editor or executive editor to consider for publication. Lastly, faculty must disclose any significant financial interest or relationship with proprietary entities that may have a direct relationship to the subject matter. For this course, the authors had the following relationships to report: Gordon D. Davis, MD, FACOG: No such relationship to report Joseph A.D. Brooks, MD: No such relationship to report. PMID- 18162824 TI - Presentation of the 2007 Shwachman Award to Emanuel Lebenthal, M.D. PMID- 18162825 TI - Presentation of the 2007 NASPGHAN Distinguished Service Award to Susan J. Henning, PhD. PMID- 18162826 TI - Presentation of the 2007 Murray Davidson Award to Michael K. Farrell, MD. PMID- 18162827 TI - Wireless capsule endoscopy: indications, limitations, and future challenges. AB - Wireless capsule endoscopy is one of the recent inventions that has made an impact in the diagnostic work-up of gastrointestinal diseases, mainly in small intestinal pathology, the part of the gut that cannot be totally visualized by upper and lower gastrointestinal endoscopy. Since the first report documenting the use of capsule endoscopy, many adult clinical trials have taken place. Although the Food and Drug Administration approved the use of the capsule in children (ages 10-18 years) in October 2003, few small clinical trials exploring the diagnostic yield of capsule endoscopy in pediatric populations have been published. PMID- 18162828 TI - Evaluation and management of persistent problems after surgery for Hirschsprung disease in a child. AB - Hirschsprung disease occurs approximately once in every 5000 live-born infants. It is characterized by the absence of ganglion cells in the myenteric and submucosal plexuses of the intestine. Once the diagnosis of Hirschsprung disease has been made, most patients are now treated with a transanal approach to resection. We review the early and late postoperative complications. Late complications include persistent mechanical obstruction, recurrent or acquired aganglionosis, disordered motility in the proximal colon or small bowel, internal sphincter achalasia, or functional megacolon caused by stool-holding behavior. These children require complex interdisciplinary care to ensure an adequate quality of life. PMID- 18162829 TI - GLP-2 administration results in increased proliferation but paradoxically an adverse outcome in a juvenile piglet model of short bowel syndrome. AB - OBJECTIVE: The objective of the present study was to examine the effect of glucagon-like peptide-2 (GLP-2) administration in a piglet, juvenile model of short bowel syndrome. MATERIALS AND METHODS: Four-week-old piglets underwent either a sham operation or 75% small bowel resection. Postoperatively, piglets received either polymeric infant formula diet or the diet and subcutaneous human recombinant GLP-2 (1600 microg/day for 7 days, 800 microg/day thereafter). Food intake was monitored throughout the experiment, and stool and serum samples obtained fortnightly. After the piglets were killed, tissues were obtained from the duodenum, jejunum, ileum, and terminal ileum, and used for morphological and functional analysis. RESULTS: Treatment with GLP-2 resulted in significantly increased numbers of proliferating and apoptotic cells in the ileum of sham and small bowel resection piglets (P < 0.05). GLP-2 administration resulted in decreased weight gain, serum albumin, and disaccharidases in both sham and small bowel resection piglets (P < 0.001 compared with polymeric infant formula diet alone). CONCLUSIONS: This is the first study to our knowledge to examine the effect of GLP-2 administration in a juvenile short bowel syndrome model. Contrary to adult rodent studies, administration of GLP-2 resulted in adverse outcomes including reduced ability to gain weight; decreased serum albumin, tissue maltase, and sucrase; and villous atrophy. We anticipate this information will have important implications for future paediatric clinical trials. PMID- 18162830 TI - Protective effects of L-arginine on rat terminal ileum subjected to ischemia/reperfusion. AB - OBJECTIVES: Studies have shown that nitric oxide (NO) may play a major role in sustaining mucosal integrity; however, NO has been also implicated in the pathogenesis of ischemia/reperfusion (I/R)-related tissue injury. We investigated the effects of L-arginine and NG-nitro L-arginine methyl ester (L-NAME) on the acetylcholine-induced contractile response of ileum and the levels of malondialdehyde (MDA) and reduced glutathione (GSH). Histopathological changes were also evaluated in ileal preparations. MATERIALS AND METHODS: Male Wistar Albino rats were subjected to mesenteric ischemia (30 min) followed by reperfusion (3 hours). Four groups were designed: sham-operated control; I/R; I/R and L-arginine pretreatment; and I/R and L-NAME pretreatment. After reperfusion, ileum specimens were collected to determine the parameters mentioned above. RESULTS: Following reperfusion, a significant decrease in acetylcholine-induced contractile response, an increase in lipid peroxidation, a decrease in GSH content, and mucosal damage of the ileal preparations were observed. We showed that decreased contractility, increased lipid peroxidation, and reduced GSH content have been reversed by L-arginine but not by L-NAME. Mucosal injury was significantly lowered in the L-arginine group. CONCLUSIONS: Treatment with L arginine exerted a protective effect in intestinal I/R injury, which was mediated in part by regulating MDA and GSH levels, consequently ameliorating impaired contractile response and mucosal injury. PMID- 18162831 TI - Mutation analysis of endothelin-B receptor gene in patients with Hirschsprung disease in Taiwan. AB - BACKGROUND: Endothelin-B receptor (EDNRB) signaling pathway is associated for Hirschsprung disease (HSCR). The aim of this study was to investigate the EDNRB gene mutation in patients with HSCR in Taiwan and correlate the genotype and phenotype. PATIENTS AND METHODS: Using polymerase chain reaction amplification and direct sequencing, we screened for mutations in the coding regions and intron/exon boundaries of the EDNRB gene in 39 isolated HSCR cases and compared them with those in 400 control chromosomes. RESULTS: In 3 cases, heterozygous variations in exon 1 and 2 of the EDNRB gene predicted missense mutations of the first cytosolic (M132I), second transmembrane (I157V), second exoplasmic (M173T), and third transmembrane (V185M) domains of the EDNRB protein. Three of the 4 mutations in our study have not been reported previously. For total 39 unrelated cases, the mutation rates were estimated to be 10% (3 of 30) for short-segment HSCR and 7.7% (3 of 39) for all HSCR cases. CONCLUSIONS: We did not detect a significant genotype-phenotype correlation. In conclusion, this study identified 4 mutations within the EDNRB gene associated with HSCR. Because HSCR is a multifactorial and multigene disorder, the higher mutation rate of 10% for short segment HSCR suggests the important role that the EDNRB gene plays in the pathogenesis of short-segment HSCR in Taiwan. PMID- 18162832 TI - Intestinal permeability and effects of Lactobacillus rhamnosus therapy in children with short bowel syndrome. AB - BACKGROUND: The present study was conducted to investigate the effects of Lactobacillus rhamnosus (also known as LGG) on intestinal permeability (IP) in children with short bowel syndrome (SBS). PATIENTS AND METHODS: In a double blind, placebo-controlled crossover clinical trial, baseline IP (ie, lactulose-to mannitol ratio) was measured in subjects with SBS and healthy control subjects. Subjects with SBS received LGG or placebo for 4 weeks, followed by a 3-week washout before therapy was crossed over for another 4 weeks. IP, quantitative fecal cultures for Lactobacillus species (in colony-forming units [cfu] per gram of stool) and hydrogen breath test (HBT) were performed during LGG and placebo phases of therapy. RESULTS: Twenty-one children (SBS, n = 9; control, n = 12) with a median age of 4.5 years (range 1.6-16.4 years) enrolled. Baseline IP measurements were similar in patients with SBS and control subjects: 0.08 +/- 0.06 (mean +/- SD) versus 0.07 +/- 0.05 (P = 1.0). IP was correlated with age in control subjects (r = -0.83, P = 0.001) but not among patients with SBS (r = 0.55, P = 0.16). Fecal colonization with Lactobacillus species did not differ during LGG versus placebo therapy (median 1.4 x 10(9) cfu/g [range 4.0 x 10(5) to 4.0 x 10(9) cfu/g] vs 6.0 x 10(9) cfu/g [1.0 x 10(3) to 1.0 x 10(10) cfu/g], respectively; P = 0.83). LGG therapy had no consistent effects on IP (P = 0.58) or its relationship with age (r = -0.40, P = 0.29), and was associated with conversion to positive HBT results in 1 subject. CONCLUSIONS: In this sample of children with SBS, the IP was within normal limits but did not correlate with age. LGG therapy had no consistent effects on IP. These findings do not support empiric LGG therapy to enhance IP in children with SBS. PMID- 18162833 TI - Significance of bowel wall abnormalities at ultrasound in Henoch-Schonlein purpura. AB - OBJECTIVES: To describe the polymorphism of intestinal ultrasound findings in children with gastrointestinal symptoms of Henoch Schonlein purpura (HSP) and to investigate the clinical relevance of these findings in terms of symptoms and length of hospitalization. PATIENTS AND METHODS: Clinical and ultrasound data from 43 consecutive children with HSP (36 with and 7 without abdominal symptoms) were reviewed. Patients with abdominal symptoms of HSP were divided into 4 groups (0-III) representing at ultrasound normal appearance and differentiated, pseudodifferentiated, and dedifferentiated bowel wall thickening, respectively. The diagnostic value of ultrasound in diagnosing gastrointestinal involvement of HSP (grades I-III) was calculated using as the standard of reference the absence or presence of clinical symptoms. Average duration of symptoms and hospitalization in the 4 groups was compared. RESULTS: The respective sensitivity, specificity, positive predictive value, and negative predictive value of ultrasound for the diagnosis of gastrointestinal involvement of HSP was 83.3%, 100%, 100%, and 53.8%. Groups 0 to III contained, respectively, 5/36 (13.9%), 6/36 (16.7%), 22/36 (61.1%), and 3/36 (8.3%) patients. The groups' average duration of symptoms, respectively, was 2.20 +/- 2.06, 5.67 +/- 1.88, 6.29 +/- .94, and 17.67 +/- 2.66 days, whereas the length of hospitalization, respectively, was 4.80 +/- 2.96, 9.17 +/- 2.70, 11.46 +/- 1.35, and 24.67 +/- 3.82 days. The duration of both symptoms and hospitalization was significantly higher in group III than in the other groups (P < 0.05). CONCLUSIONS: In children with gastrointestinal involvement of HSP, dedifferentiated wall thickening as shown by ultrasound is associated with a poor clinical prognosis. PMID- 18162834 TI - Tegaserod use in children: a single-center experience. AB - BACKGROUND: Tegaserod is increasingly prescribed by pediatric gastroenterologists even though there are few published data concerning its use in children. The aim of this study was to describe the authors' experience with tegaserod in children. PATIENTS AND METHODS: Patients treated with tegaserod from 2004 through 2006 were included in this study. Defecation and fecal incontinence frequency and global assessment of relief of symptoms were assessed. RESULTS: Seventy-two patients (44 girls) ranging in age from 1.1 to 18.3 years constitute the patient sample of this report. The median age was 10 years and the median follow-up after initiation of tegaserod treatment was 11.3 months (range 2.3-45.2 months). Indications to prescribe tegaserod were constipation (58%) and a variety of other conditions including functional dyspepsia or inflammatory bowel disease (42%). Defecation frequency increased after tegaserod use (1 vs 7/week, P < 0.001) and presence of fecal incontinence decreased (47% vs 23%, P < 0.001) in the constipation group. Parents rated relief of constipation as moderate or significant in 71% of cases in the constipation group. In the group with other indications to start tegaserod therapy, moderate or significant relief of abdominal pain and bloating was noted in 64% and 68% of patients, respectively. The median dose of tegaserod prescribed was 0.22 mg x kg x day (range 0.05-0.87 mg x kg(-1) x day(-1)). Adverse events were observed in 32% of the patients. The most common side effects were self-limiting diarrhea (20%) and abdominal pain (8%). Only one patient discontinued tegaserod because of side effects; this patient experienced pain at his cecostomy site. CONCLUSIONS: Tegaserod seems to relieve a variety of functional gastrointestinal symptoms in children. Further randomized controlled studies are needed to support the specific pediatric target of prescribing tegaserod. PMID- 18162835 TI - A retrospective assessment of magnetic resonance cholangiopancreatography in children. AB - OBJECTIVE: To evaluate the utility of magnetic resonance cholangiopancreatography (MRCP) in children and to compare MRCP with direct cholangiopancreatography (CP). MATERIALS AND METHODS: We performed an unblinded, retrospective chart review of 32 children (ages 0-18 years, 17 male) who underwent MRCP between January 2002 and June 2005. MRCP, endoscopic retrograde cholangiopancreatography (ERCP), percutaneous fluoroscopic or intraoperative studies of the pancreatobiliary tree, and clinical outcomes were evaluated. RESULTS: Seventeen (52%) children had MRCP alone, 15 (48%) had both MR and direct CP. MRCP results correlated with other evaluative methods in 14/15 (93%) cases. There was 1 false positive (anomalous pancreatic duct union) and 0 false negatives for anatomic abnormalities. Therapeutic intervention was performed in 7 of 28 children initially evaluated by MRCP (2 sphincter of Oddi dysfunction, 2 choledocholithiasis, 2 primary sclerosing cholangitis, 1 congenital hepatic cysts) and 1 of 4 children initially evaluated by ERCP (primary sclerosing cholangitis). All 17 children initially evaluated by MRCP had no change in clinical status to suggest a missed anatomic lesion or therapeutic opportunity. CONCLUSIONS: In this retrospective study, MRCP was sensitive and specific in identifying anatomic abnormalities of the pancreatobiliary tree in children. MRCP should be considered before direct CP to evaluate anatomic abnormalities of the pancreatobiliary tree. PMID- 18162836 TI - Effect of treatment of hepatic histopathology in children and adolescents with autoimmune hepatitis. AB - OBJECTIVE: To evaluate the effects of treatment on liver histopathology of children and adolescents diagnosed with autoimmune hepatitis. PATIENTS AND METHODS: Histopathological evaluation of liver biopsies performed at diagnosis or early during treatment, compared with biopsies after immunosuppressive treatment of 20 children and adolescents diagnosed with autoimmune hepatitis who presented clinical and biochemical remission for at least 24 months. Liver histopathology was assessed by 2 pathologists. Agreement between their interpretations was analyzed with kappa statistics. RESULTS: The age at diagnosis of the 20 patients varied from 1.7 years to 11.6 years (median = 6.6 years); 18 were females. The mean duration of clinical and biochemical remission, up to performance of liver biopsy, was 4.1 years. The mean interval between the initial and posttreatment biopsies was 4.6 years (range 2-9.7 years). Assessing the necroinflammatory activity, a mean reduction by 6.3 (observer 1) and 7.3 (observer 2) in the grading score was observed (P = 0.00000). When staging was assessed, a mean reduction by 1.8 (observers 1 and 2) in the score was observed after treatment (P = 0.00008), and there was no progression toward cirrhosis. No regression of cirrhosis was observed. CONCLUSIONS: On histopathological evaluation, the immunosuppressive treatment improved the fibrosis scores, with an arrest in its progression and no development into cirrhosis. Fibrosis control is mainly associated with regression of necroinflammatory activity, which is the main treatment component in autoimmune hepatitis. PMID- 18162837 TI - Na+-dependent neutral amino acid transporter ASCT2 is downregulated in seriously traumatized human intestinal epithelial cells. AB - OBJECTIVE: Serious trauma to the body often is associated with changes in protein metabolism in multiple organs and tissues. Clinically, the catabolic response results in a generalized negative nitrogen balance. Nutrition support has been an important component of the care of seriously traumatized patients. However, during states of severe trauma, enterocyte transport function remains unclear. This study aims to quantitate the Na+-dependent neutral amino acid transport and expression of its transporter in traumatically injured Caco-2 cell lines. MATERIALS AND METHODS: Transport and transporter of Na+-dependent neutral amino acid in Caco-2 cell lines were characterized. Then the cell lines were cultured under hypoxic, nutrient-deprived, and ischemic conditions for 1, 2, 4, and 6 hours. After severe trauma was performed, we investigated the transport of Na+ dependent neutral amino acids and the expression of transporter protein and mRNA in apical membrane vesicles. RESULTS: Among the neutral amino acid transporters, only ASCT2 mRNA was amplified successfully. Under nutrient-deprived and ischemic conditions, transport of L-alanine and L-glutamine decreased significantly compared with control (P < 0.01), whereas hypoxia had no significant effect. The changes were associated with a decrease in maximum transport velocity without an influence on transport affinity. Expression of relative transporter proteins and mRNA decreased significantly compared with control (P < 0.01). CONCLUSIONS: Na+ dependent neutral amino acid transport and its key transporter are differently regulated during state of traumatic injury. It may be of use to provide some strategies targeting the special nutrient requirements and transport capabilities of seriously traumatized patients. PMID- 18162838 TI - Critical analysis of bariatric procedures in Prader-Willi syndrome. AB - Prader-Willi syndrome (PWS) is a complex genetic disorder localized to chromosome 15 and is considered the most common genetic cause of the development of life threatening obesity. Although some morbidities associated with PWS, including respiratory disturbance/hypoventilation, diabetes, and stroke, are commonly seen in obesity, others such as osteoporosis, growth hormone deficiency, and hypogonadism, and also altered pain threshold and inability to vomit, pose unique issues. Various bariatric procedures have been used to cause gastric stasis, decrease gastric volume, and induce malabsorption, with poor results in PWS patients in comparison with normal obese individuals. PMID- 18162839 TI - Salmonella gastroenteritis mimicking onset of inflammatory bowel disease in children. PMID- 18162840 TI - Videomanometric evaluation of pharyngo-oesophageal dysmotility in children with velocardiofacial syndrome. PMID- 18162841 TI - Superior mesenteric artery syndrome: an uncommon cause of feeding intolerance in infancy. PMID- 18162843 TI - Gastroesophageal reflux in preterm infants: how acid should it be? PMID- 18162844 TI - Complementary feeding: a commentary by the ESPGHAN Committee on Nutrition. AB - This position paper on complementary feeding summarizes evidence for health effects of complementary foods. It focuses on healthy infants in Europe. After reviewing current knowledge and practices, we have formulated these conclusions: Exclusive or full breast-feeding for about 6 months is a desirable goal. Complementary feeding (ie, solid foods and liquids other than breast milk or infant formula and follow-on formula) should not be introduced before 17 weeks and not later than 26 weeks. There is no convincing scientific evidence that avoidance or delayed introduction of potentially allergenic foods, such as fish and eggs, reduces allergies, either in infants considered at increased risk for the development of allergy or in those not considered to be at increased risk. During the complementary feeding period, >90% of the iron requirements of a breast-fed infant must be met by complementary foods, which should provide sufficient bioavailable iron. Cow's milk is a poor source of iron and should not be used as the main drink before 12 months, although small volumes may be added to complementary foods. It is prudent to avoid both early (<4 months) and late (>or=7 months) introduction of gluten, and to introduce gluten gradually while the infant is still breast-fed, inasmuch as this may reduce the risk of celiac disease, type 1 diabetes mellitus, and wheat allergy. Infants and young children receiving a vegetarian diet should receive a sufficient amount ( approximately 500 mL) of breast milk or formula and dairy products. Infants and young children should not be fed a vegan diet. PMID- 18162845 TI - CFC1 gene involvement in biliary atresia with polysplenia syndrome. AB - The present report describes CFC1 gene analysis in 10 patients with polysplenia syndrome. The heterozygous transition c.433G>A (Ala145Thr) located in exon 5 was identified in 5 patients, with a twice-higher frequency than in control patients. These results suggest that heterozygous CFC1 mutation may represent a genetic predisposition to biliary atresia splenic malformation syndrome. PMID- 18162846 TI - Very low birth weight infants as a model to study genetic influences on neonatal weight gain. AB - In a cohort of 829 preterm infants (birth weight below 1500 g) we identified 13 monozygotic, 10 same-sex dizygotic, and 12 same-sex matched singleton pairs. The difference in daily weight gain within pairs was significantly lower in monozygotic twins compared with dizygotic twins or matched singleton pairs. Our data support a strong genetic influence on postnatal growth in preterm infants. Therefore, weight gain of preterm infants may be an interesting model to study polymorphic variants of genes regulating neonatal resorption, metabolism, or energy expenditure, and their influence on weight gain in preterm infants. PMID- 18162847 TI - Pilot evaluation of an instrument to measure quality of life in British children with inflammatory bowel disease. AB - The IMPACT questionnaire was developed in Canada to measure quality of life in children with inflammatory bowel disease (IBD). In the present study, 20 children with IBD completed 2 versions of the IMPACT questionnaire with a Likert scale or visual analog scale (VAS), 5 of whom expressed problems with language or phrasing. Difficult words included "restrictions," "moderate," "diarrhea," "school break," and the abbreviation "IBD." Fifteen children preferred the Likert scale to the VAS (chi = 20, P < 0.01). Rewording the difficult words and using a Likert scale should facilitate completion of the IMPACT questionnaire in the United Kingdom. Further validation is needed to ensure that the instrument is reliable and valid. PMID- 18162848 TI - Evidence-based health policy supports single-payer health insurance reform. PMID- 18162849 TI - Uninsured but eligible children: are their parents insured? Recent findings from Oregon. AB - BACKGROUND: Despite expansions in public health insurance programs, millions of US children lack coverage. Nearly two-thirds of Oregon's uninsured children seem to be eligible for public insurance. OBJECTIVES: We sought to identify uninsured but eligible children and to examine how parental coverage affects children's insurance status. METHODS: We collected primary data from families enrolled in Oregon's food stamp program, which has similar eligibility requirements to public health insurance in Oregon. In this cross-sectional, multivariable analysis, results from 2861 surveys were weighted back to a population of 84,087 with nonresponse adjustment. Key predictor variables were parental insurance status and type of insurance; the outcome variable was children's insurance status. RESULTS: Nearly 11% of children, presumed eligible for public insurance, were uninsured. Uninsurance among children was associated with being Hispanic, having an employed parent, and higher household earnings (133-185% of the federal poverty level). Children with an uninsured parent were more likely to be uninsured, compared with those who had insured parents (adjusted odds ratio 14.21, 95% confidence interval 9.23-20.34). More surprisingly, there was a higher rate of uninsured children among privately-insured parents, compared with parents covered by public insurance (adjusted odds ratio 4.39, 95% confidence interval 2.00-9.66). CONCLUSIONS: Low-income Oregon parents at the higher end of the public insurance income threshold and those with private insurance were having the most difficulty keeping their children insured. These findings suggest that when parents succeed in pulling themselves out of poverty and gaining employment with private health insurance coverage, children may be getting left behind. PMID- 18162850 TI - Medicaid preferred drug lists' costs to physicians. AB - BACKGROUND: Medicaid preferred drug lists (PDLs) might reduce costs for Medicaid programs while creating costs to physicians. OBJECTIVES: To measure the costs from complying with Medicaid PDLs for primary care physicians and cardiologists, and to quantify the costs of a hypothetical PDL for Medicare Part D. RESEARCH DESIGN: We analyzed cardiologists' and primary care physicians' experiences with Medicaid PDLs for antihypertensives and statins in 9 states. Physicians' prescribing volumes and PDL compliance were generated by combining pharmacy claims data from Wolters Kluwer Health with the state PDLs. These data were augmented with a survey of physicians. A Monte Carlo simulation was used to randomly assign each relevant physician in the state to a survey response. Estimates of the cost of a potential Part D PDL relied on the volume of Part D claims reported by Centers for Medicare and Medicaid Services (CMS) through May 2006. RESULTS: Physicians' PDL-related costs averaged $8.02 [95% confidence interval (CI): $7.25-$8.78] per prescription. Average cost per prescription not covered by the PDL was $14.41 (95% CI: $13.29-$15.53), and average cost per prescription covered by the PDL was $6.59 (95% CI: $5.91-$7.28). Medicaid PDL costs per physician averaged $1110 (95% CI: $1061-$1161) annually for statins and antihypertensives alone, and this varied across states. Similar restrictions under Medicare Part D across all therapeutic classes would have cost physicians $3.18 billion (95% CI: $2.88-$3.49 billion) in 2006. CONCLUSIONS: Medicaid PDLs generate considerable costs to physicians, as would adoption of PDLs in Medicare Part D. Policymakers should weigh these and other costs against the benefits of PDLs. PMID- 18162851 TI - Costs of intravenous adverse drug events in academic and nonacademic intensive care units. AB - BACKGROUND: Adverse drug events (ADEs), particularly those involving intravenous medications (IV-ADEs), are common among intensive care unit (ICU) patients and may increase hospitalization costs. Precise cost estimates have not been reported for academic ICUs, and no studies have included nonacademic ICUs. OBJECTIVES: To estimate increases in costs and length of stay after IV-ADEs at an academic and a nonacademic hospital. RESEARCH DESIGN: This study reviewed medical records to identify IV-ADEs, and then, using a nested case-control design with propensity score matching, assessed differences in costs and length of stay between cases and controls. SUBJECTS: : A total of 4604 adult ICU patients in 3 ICUs at an academic hospital and 2 ICUs at a nonacademic hospital in 2003 and 2004. MEASURES: Increased cost and length of stay associated with IV-ADEs. RESULTS: : Three hundred ninety-seven IV-ADEs were identified: 79% temporary physical injuries, 0% permanent physical injuries, 20% interventions to sustain life, and 2% in-hospital deaths. In the academic ICUs, patients with IV-ADEs had $6647 greater costs (P < 0.0001) and 4.8-day longer stays (P = 0.0003) compared with controls. In the nonacademic ICUs, IV-ADEs were not associated with greater costs ($188, P = 0.4236) or lengths of stay (-0.3 days, P = 0.8016). Cost and length-of stay differences between the hospitals were statistically significant (P = 0.0012). However, there were no differences in IV-ADE severity or preventability, and the characteristics of patients experiencing IV-ADEs differed only modestly. CONCLUSIONS: IV-ADEs substantially increased hospitalization costs and length of stay in ICUs at an academic hospital but not at a nonacademic hospital, likely because of differences in practices after IV-ADEs occurred. PMID- 18162852 TI - Do postoperative complications vary by hospital teaching status? AB - OBJECTIVE: The purpose of this paper is to assess postoperative patient safety outcomes across teaching and nonteaching hospitals and to examine the relation of hospital and patient factors to patient safety outcomes. RESEARCH DESIGN AND METHODS: The Nationwide Inpatient Sample and American Hospital Association annual survey data were used for analyses. Patient safety indicators (PSIs) developed by the Agency for Healthcare Research and Quality (AHRQ) were used to identify 6 postoperative PSIs. The study sample consisted of 646 acute care hospitals, divided into nonteaching (n = 400), minor teaching (n = 207), and major teaching hospitals (n = 39). The unit of analysis was the patient. Associations between hospital teaching status and patient and hospital characteristics were determined using one-way analysis of variance and Pearson chi test. Multivariable analysis using generalized estimating equation regression models assessed the relationship between teaching status and PSIs. RESULTS: Bivariate results showed higher observed PSI rates at major teaching hospitals. Results from multivariable analyses, after adjusting for hospital size, staffing variables, patient case mix, and other risk factors, showed that major teaching hospitals had significantly higher odds of postoperative pulmonary embolism or deep vein thrombosis and postoperative sepsis, lower odds of postoperative respiratory failure, and showed no difference for postoperative hip fracture, postoperative hematoma or hemorrhage, and postoperative physio-metabolic derangement. CONCLUSIONS: The present analysis found an inconsistent relationship between teaching status and postoperative patient safety event rates. Teaching status of the hospital was associated with numerous hospital and patient characteristics which mediate the relationship between teaching status and PSIs. PMID- 18162853 TI - Medicaid payment rates, case-mix reimbursement, and nursing home staffing--1996 2004. AB - OBJECTIVE: We examined the impact of state Medicaid payment rates and case-mix reimbursement on direct care staffing levels in US nursing homes. METHODS: We used a recent time series of national nursing home data from the Online Survey Certification and Reporting system for 1996-2004, merged with annual state Medicaid payment rates and case-mix reimbursement information. A 5-category response measure of total staffing levels was defined according to expert recommended thresholds, and examined in a multinomial logistic regression model. Facility fixed-effects models were estimated separately for Registered Nurse (RN), Licensed Practical Nurse (LPN), and Certified Nurse Aide (CNA) staffing levels measured as average hours per resident day. RESULTS: Higher Medicaid payment rates were associated with increases in total staffing levels to meet a higher recommended threshold. However, these gains in overall staffing were accompanied by a reduction of RN staffing and an increase in both LPN and CNA staffing levels. Under case-mix reimbursement, the likelihood of nursing homes achieving higher recommended staffing thresholds decreased, as did levels of professional staffing. Independent of the effects of state, market, and facility characteristics, there was a significant downward trend in RN staffing and an upward trend in both LPN and CNA staffing. CONCLUSIONS: Although overall staffing may increase in response to more generous Medicaid reimbursement, it may not translate into improvements in the skill mix of staff. Adjusting for reimbursement levels and resident acuity, total staffing has not increased after the implementation of case-mix reimbursement. PMID- 18162854 TI - The effect of acupuncture utilization on healthcare utilization. AB - OBJECTIVE: To determine whether acupuncture is a complement to or substitute for various medical services. DATA SOURCE: This study used managed care claims data from a midsize metropolitan insurance company from 2002. Zip code level data from the 2000 US Census was also incorporated. The original dataset contained medical and drug claims data for every eligible acupuncture user (n = 1688) and every 18th eligible nonacupuncture user (n = 16,282) covered by the data provider. STUDY DESIGN: Simultaneous equations models with an exclusion restriction were used in this cross-sectional study. The influence of acupuncture utilization was assessed independently on each conventional service of interest, controlling for numerous clinical and demographic characteristics. Bivariate probit models were estimated using distance to the nearest acupuncturist as the exclusion restriction. RESULTS: Acupuncture was a statistically significant (P < 0.05) substitute for primary care, all outpatient services, pathology services, all surgery, and gastrointestinal medications. Acupuncture seemed to complement numerous therapies, particularly chiropractic and physical therapy; however, acupuncture did not statistically significantly complement any therapies after controlling for unobservable characteristics that influence the use of acupuncture and/or conventional medicine. CONCLUSIONS: Acupuncture is an economic substitute for some medical services and pharmaceuticals, a finding of some importance to insurers, healthcare practitioners, and policy makers. The fact that acupuncture has an effect on other medical services needs to be explored more fully with an emphasis on how this substitution impacts patient health. PMID- 18162855 TI - Disenrollment from Medicaid after recent cancer diagnosis. AB - OBJECTIVES: We examine the frequency with which newly diagnosed cancer patients are covered by Medicaid in Washington State and the duration of coverage. METHODS: Medicaid enrollment and claims files were linked to the Washington State Cancer Registry to identify all Medicaid enrollees with breast, cervical, lung, colorectal, and prostate cancer between 1997 and 2002. RESULTS: We identified 5009 newly diagnosed cancer patients covered by Medicaid, approximately 13% of the total cases diagnosed in subjects less than 65 years of age in Washington State. The majority, 2866 (57%), enrolled in Medicaid around the time of diagnosis; the remainder had been enrolled at least 3 months before diagnosis. Persons enrolled at diagnosis had later-stage cancer; those enrolled before diagnosis had more noncancer comorbidities. Overall, 18% had disenrolled by 6 months after diagnosis; 34% by 1 year; and 54% by 2 years. CONCLUSIONS: Medicaid patients with cancer in Washington State experience a high rate of disenrollment within 1 year after diagnosis. Further research is needed to determine whether disenrollment compromises initial therapy or follow-up care. PMID- 18162856 TI - Continuity of care and cancer screening among health plan enrollees. AB - CONTEXT: Although having a usual source of care has been associated with cancer screening, whether there is additional benefit from continuity with a specific physician is uncertain. In addition, little is known about the relationship between continuity of care and receipt of colorectal and prostate cancer screening. METHODS: Subjects were enrolled in a Washington State health plan that operates an integrated delivery system that emphasizes access to primary care. Among patients age 50-78 years old with 2 or more primary care visits in 2002 2003 (N = 67,633), we determined whether higher continuity (>/=50% of visits with the most visited primary care provider) was associated with colorectal, breast, and prostate cancer screening. Random-effects logistic regression estimated adjusted percentages of patients who received fecal occult blood testing, lower endoscopy (sigmoidoscopy or colonoscopy), screening mammography, and prostate specific antigen (PSA) testing. RESULTS: Patients with higher continuity were more likely to receive fecal occult blood testing than patients with lower continuity (28.9% vs. 26.8%; P < 0.001) but less likely to receive lower endoscopy (12.9% vs. 14.3%; P < 0.001). Although higher continuity was not significantly associated with screening mammography (P = 0.38), men with higher continuity were more likely to receive PSA testing than men with lower continuity (39.4% vs. 37.4%; P = 0.008). CONCLUSIONS: In an insured population with a high degree of primary care access, continuity with a specific primary care physician was associated with the selection of less invasive colorectal cancer screening tests by patients and physicians and greater likelihood of PSA testing. PMID- 18162857 TI - Quality of care and mortality among patients with stroke: a nationwide follow-up study. AB - BACKGROUND: The relationship between process and outcome measures among patients with stroke is unclear. OBJECTIVES: To examine the association between quality of care and mortality among patients with stroke in a nationwide population-based follow-up study. METHODS: Using data from The Danish National Indicator Project, a quality improvement initiative with participation of all Danish hospital departments caring for patients with stroke, we identified 29,573 patients hospitalized with stroke between January 13, 2003 and October 31, 2005. Quality of care was measured in terms of 7 specific criteria: early admission to a stroke unit, early initiation of antiplatelet or oral anticoagulant therapy, early examination with computed tomography/magnetic resonance imaging scan, and early assessment by a physiotherapist, an occupational therapist, and of nutritional risk. Data on 30- and 90-day mortality rates were obtained through the Danish Civil Registration System. RESULTS: Six of 7 of these criteria were associated with lower 30- and 90-day mortality rates. Adjusted mortality rate ratios corrected for clustering by department ranged from 0.41 to 0.83. We found indication of an inverse dose-response relationship between the number of quality of care criteria met and mortality; the lowest mortality rate was found among patients whose care met all criteria compared with patients whose care failed to meet any criteria (ie, adjusted 30-day mortality rate ratios: 0.45, 95% confidence interval: 0.24-0.66). When analyses were stratified by age and sex, the dose-response relationship was found in all subgroups. CONCLUSIONS: Higher quality of care during the early phase of stroke was associated with substantially lower mortality rates. PMID- 18162858 TI - Hospital variation in use of secondary preventive medicine after discharge for first acute myocardial infarction during 1995-2004. AB - OBJECTIVE: To examine temporal trends in hospital use of secondary preventive medicine after discharge for first acute myocardial infarction (AMI) in Denmark. DESIGN: Observational study from national administrative databases of 60,339 patients who survived a first AMI at 73 acute-care hospitals during 1995-2004. OUTCOME MEASURES: At least 1 prescription claim for angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, or statins within 90 days of discharge for AMI. FINDINGS: The odds ratios between hospitals in the highest and lowest deciles, adjusted for age, gender, period, income, comorbidity, concomitant, and prior pharmaceutical therapy, in 1995 were 8.5 [95% confidence interval (CI), 5.5-12.2] for beta-blockers, 3.0 (2.3-3.7) for ACE inhibitors, and 6.2 (4.1-8.8) for statins. By 2004, the hospital variation had decreased for beta-blockers (3.2; 2.3-4.0) and statins (4.2; 3.0-5.5) but had increased for ACE inhibitors (3.8; 2.7-4.9). All the changes over time were significant (P < 0.001). Geographical characteristics of the hospital explained 32% of the variation in use of beta blockers in 2004 and 27% in 1995, 39% of the variation in use of ACE inhibitors in 2004 and 3% in 1995, and 29% of the variation in use of statins and 19% in 1995. CONCLUSIONS: Hospital use of secondary preventive medicine after discharge for AMI varied substantially. Hospital variation in use of beta-blockers and statins decreased with time whereas variation in use of ACE inhibitors increased. This may be attributed to gradually better agreement for the use of beta-blockers and statins and lesser agreement for the use of ACE inhibitors. PMID- 18162859 TI - How a therapy-based quality improvement intervention for depression affected life events and psychological well-being over time: a 9-year longitudinal analysis. AB - BACKGROUND: Short-term quality improvement (QI) interventions for depression can improve long-term mental health but mechanisms are unknown. We hypothesized that 1 pathway for such health benefits was an indirect effect with QI reducing risk factors for depression such as stressful life events. OBJECTIVE: To determine whether 6-12 month QI programs for depression reduce negative life events at 5 year follow-up and to model the relationship between program implementation, life events and mental health over 9 years. DESIGN: Forty-six primary care clinics in 6 managed care organizations were randomized to usual care or 1 of 2 QI interventions. We focus on the intervention that provided resources to assess and manage depression while particularly facilitating access to evidence-based psychotherapy ("QI-Therapy"). SUBJECTS: A total of 1300 enrolled patients with current depressive symptoms, who had data at any of 4 data points: baseline, or follow-up year 1, 5, or 9. MEASURES: Total and negatively-evaluated life events and psychologic well-being. RESULTS: A path model showed that QI-Therapy, in addition to improving psychologic well-being at year 1 (P = 0.0033), reduced negative life events at year 5 (P = 0.0033). This effect was not fully explained by improved psychologic well-being. Better mental health (P < 0.0001) and fewer negative life events (P = 0.0013) at year 5 were associated with improved psychologic well-being at 9 years. CONCLUSIONS: Depression QI programs that include resources for psychotherapy can reduce occurrence of life events, further protecting subsequent mental health. Implications for the design of QI programs and development of prevention interventions are discussed. PMID- 18162860 TI - Spiritual care at the end of life in long-term care. AB - BACKGROUND: There is growing attention given to the spiritual needs of dying patients and long-term care (LTC) facilities are common settings in which patients receive care as they approach death. OBJECTIVES: To describe the sources of support, the structure and processes of spiritual care in LTC, and examine the relationship between these components and family ratings of overall care. RESEARCH DESIGN: After-death interviews of family members of decedents. SUBJECTS: Family members of 284 decedent residents from a stratified sample of 100 residential care/assisted living facilities and nursing homes in Florida, Maryland, New Jersey, and North Carolina. MEASURES: : Interview items included sources of spiritual support, processes of spiritual care, and the impression of overall care (4 = very good, 3 = good, 2 = fair, 1 = poor) for decedents. Facility-level data included demographics, counseling by clergy, on-site religious services, hospice services, and hospice unit. RESULTS: Most decedents (87%) received assistance with their spiritual needs and those who received spiritual care were perceived by family members to have had better overall care (3.59 vs. 3.25, P = 0.002). Family ratings of care ratings were higher for those who received spiritual support or care from facility staff when compared with those who did not (3.76 vs. 3.49, P < 0.001) and better care was associated with the facilitation of individual devotional activities (3.87 vs. 3.53, P = 0.001). CONCLUSIONS: Spiritual support and care are associated with better overall care at the end of life for LTC residents, and interventions to improve this type of care may best target interactions between residents and facility staff. PMID- 18162861 TI - Chronic disease and severe disability among working-age populations. AB - BACKGROUND: Recent work has shown that rates of severe disability, measured by the inability to perform basic activities of daily living, have been rising in working age populations. At the same time, the prevalence of important chronic diseases has been rising, while others falling, among working age populations. Chronically ill individuals are more likely than others to have activity of daily living limitations. OBJECTIVE: We examine the extent to which chronic disease trends can explain these disability trends. DATA: We use nationally representative survey data from the 1984-1996 National Interview Survey, which posed a consistent set of questions regarding limitations in activities of daily living over that period. METHODS: We decompose trends in disability into 2 parts 1 part due to trends in the prevalence of chronic disease and the other due to trends in disability prevalence among those with chronic disease. RESULTS: : Our primary findings are that for working age populations between 1984 and 1996: (1) disability prevalence fell dramatically among the nonchronically ill; (2) rising obesity prevalence explains about 40% of the rise in disability attributable to trends chronic illness; and (3) rising disability prevalence among the chronically ill explains about 60% of the rise in disability attributable to trends in chronic illness. CONCLUSIONS: Disability prevention efforts in working age populations should focus on reductions in obesity prevalence and limiting disability among chronically ill populations. Given the rise in disability among these population subgroups, it is unclear whether further substantial declines in elderly disability can be expected. PMID- 18162862 TI - The costs of nosocomial infections. AB - BACKGROUND: Although nosocomial infections (NIs) are widely regarded as expensive complications of healthcare delivery, their costs have not been rigorously quantified in large-scale studies. Additionally, problems that can bias cost estimates have often gone unaddressed. For example, are NIs more likely to cause significant extra length of stay (LOS) and costs, or are they more likely to be relatively inexpensive and inevitable consequences of long and expensive hospitalizations? This study is the largest of its kind to provide a rigorous analysis of the costs of NIs. OBJECTIVE: To precisely bound the attributable costs of a NI using large-scale data and to determine the effects of endogeneity between NIs and LOS on cost estimates. DESIGN, SETTING AND PATIENTS: Discharge diagnoses, cost, LOS, and NI data were collected for 1,355,347 admissions from March 30, 2001 to January 31, 2006 in 55 hospitals. MAIN OUTCOME MEASURES: The cost effects of NIs (in 2007 $) were estimated using multivariable regression models. Restricted models were applied to determine how cost estimates are confounded by disease severity and LOS. RESULTS: NIs are associated with $12,197 (95% CI, $4862-$19,533, P < 0.001) in incremental cost. A lower bound estimate of infection cost, controlling for LOS, is $4644 (95% CI, $1266-$7391). CONCLUSIONS: NIs are associated with substantial increases in the costs of inpatient care, even when estimates are corrected for potential endogenous confounding. PMID- 18162864 TI - Intraventricular dysembryoplastic neuroepithelial tumor: case report. AB - OBJECTIVE: The most common localization of dysembryoplastic neuroepithelial tumors (DNTs) is the supratentorial cortex, often in the temporal lobe. However, intraventricular localization of a DNT is extremely rare. CLINICAL PRESENTATION: A 30-year-old woman presented with a 1-year history of epileptic seizures. The seizures had not been controlled despite standard doses of antiepileptics. INTERVENTION: Neuroimaging results demonstrated a lesion located in the occipital horn of the right lateral ventricle. The lesion was totally removed. Based on histopathological and immunohistochemical evaluation, a DNT was diagnosed. Over the course of the next 8 months, the patient's epileptic seizures were under control. The most recent neuroimaging examinations revealed neither residual nor recurrent tumor. CONCLUSION: Because DNTs are surgically curable and neither radiotherapy nor chemotherapy is required after surgery, recognition of an intraventricular DNT in this location is extremely important. PMID- 18162865 TI - Malignant peripheral nerve sheath tumor of the occipital region: case report. AB - OBJECTIVE: A rare case of a malignant peripheral nerve sheath tumor of the occipital region is presented. The role of postoperative radiotherapy in such cases is reviewed. CLINICAL PRESENTATION: A 36-year-old man presented with a small spongy swelling in the posterior aspect of the cranium since childhood. The swelling gradually increased for 2 years and then rapidly over the next 7 months. There were no associated symptoms of headache, vomiting, vertigo, focal weakness, or visual impairment. INTERVENTION: A small occipital craniectomy for total excision of the tumor was attempted. However, as a result of intracranial extension to the transverse sinus, the tumor could not be completely excised. Local radiotherapy to a dose of 59.4 Gy in 33 fractions over 6.5 weeks was delivered to the residual microscopic disease in the posterior part of the cranium. The patient has been followed up regularly for the past 28 months and continues to remain asymptomatic. CONCLUSION: An aggressive malignant peripheral nerve sheath tumor of an atypical site such as the scalp, in which complete surgery may not always be possible, could benefit from postoperative radiotherapy. A dose of approximately 60 Gy could be effectively delivered with a satisfactory outcome. PMID- 18162866 TI - Subpial hemangiopericytoma with marked extramedullary growth: case report. AB - OBJECTIVE: Hemangiopericytomas, vascular tumors arising in soft tissue, are relatively rare in the central nervous system; they comprise less than 1% of all hemangiopericytomas. Central nervous system hemangiopericytomas occur primarily in the epidural space of the brain and spinal cord. There are no previous reports of subpial, extramedullary growing central nervous system hemangiopericytomas. CLINICAL PRESENTATION: We document the first case of a subpial hemangiopericytoma with extramedullary growth in the thoracic spine. The patient was a 31-year-old man who developed progressively worsening left lower limb numbness that was followed by gait disturbance over the course of 4 months. INTERVENTION: Magnetic resonance imaging revealed an intradural tumor at the T4-T6 level of the thoracic spine. Because the patient's symptoms progressed, he underwent resection of the tumor, which had arisen in the spinal cord subpially without attachment to the dura mater. CONCLUSION: The pathological diagnosis was hemangiopericytoma. Differential diagnoses include hemangioblastoma, meningioma, schwannoma, and solitary fibrous tumor, the clinical course and prognosis of which are different from hemangiopericytoma. Our experience indicates that hemangiopericytomas can occur as intradural tumors arising from the subpial portion. PMID- 18162867 TI - A case of cranial fasciitis originating within the diploic space of an adult: case report. AB - OBJECTIVE: Cranial fasciitis rarely occurs as a circumscribed lesion in the diploic space without soft tissue involvement. It is even more rare in adults. We discuss the radiographic and pathological differential diagnosis for cranial fasciitis and review the literature regarding clinical presentation and prognosis. CLINICAL PRESENTATION: A 27-year-old woman presented with scalp pain and headaches for several months. On physical examination, she had a raised, firm, tender mass above the right ear that was 3 cm in diameter over which there was mild alopecia. Magnetic resonance imaging scans showed a well circumscribed contrast enhancing mass within the diploic space. INTERVENTION: Because of the patient's symptoms and the lack of a diagnosis, magnetic resonance imaging-guided surgical resection of the cranial lesion followed by placement of a titanium cranioplasty was recommended. CONCLUSION: A nonspecific radiographic appearance warrants biopsy. Complete excision is the optimal treatment choice when the possibility of malignancy cannot be ruled out. PMID- 18162868 TI - Outcome of brain abscess by Clostridium perfringens. AB - OBJECTIVE: Despite the aggressive infection of soft tissue caused by Clostridium perfringens (gas gangrene-necrotizing fasciitis), a brain abscess with this bacteria treated by early surgical excision, debridement of necrotic tissue, and antibiotic coverage may be expected to have a good recovery. Long-term follow-up has not been well established in this group of patients. We report this case to show the outcome at 3 years post surgical and antibiotic treatment for C. perfringens brain abscess and stress the need for urgent intervention to achieve good outcome. We also present a literature review of Clostridial brain abscesses since the 1960s. CLINICAL PRESENTATION: A 53-year-old man was brought to the emergency room after having a witnessed seizure status postassault 3 days before admission. On presentation, he was febrile, disoriented, lethargic, and demonstrated right upper extremity weakness. A computed tomographic scan of the head showed a left frontoparietal depressed cranial fracture complicated with gas and intraparenchymal air fluid level cavity. INTERVENTION: Emergent surgery for debridement and excision of necrotic tissue was performed. Empiric intravenous antibiotic therapy was started and penicillin G was added for 6 weeks after C. perfringens was demonstrated. CONCLUSION: Despite the severe infection and effect of C. perfringens in soft tissues in the brain, it appears that emergent surgical debridement and antibiotic coverage will yield an excellent outcome for these patients. PMID- 18162870 TI - Athletic concussion: current understanding as of 2007. PMID- 18162873 TI - Passion for one's work. PMID- 18162872 TI - The combined transmastoid retro- and infralabyrinthine transjugular transcondylar transtubercular high cervical approach for resection of glomus jugulare tumors. PMID- 18162875 TI - Three-day phenytoin prophylaxis is adequate after subarachnoid hemorrhage. PMID- 18162877 TI - Synthetic nerve guide implants in humans: a comprehensive survey. PMID- 18162878 TI - Brain computed tomography angiographic scans as the sole diagnostic examination for excluding aneurysms in patients with perimesencephalic subarachnoid hemorrhage. PMID- 18162879 TI - When the bone flap hits the floor. PMID- 18162880 TI - Preserving the ligamentum flavum in lumbar discectomy: a new technique that prevents scar tissue formation in the first 6 months postsurgery. PMID- 18162881 TI - Minimally invasive atlantoaxial fixation with a polyaxial screw-rod construct: technical case report. PMID- 18162882 TI - Mapping of visuospatial functions during brain surgery: a new tool to prevent unilateral spatial neglect. PMID- 18162883 TI - Hypertension, age, and location predict rupture of small intracranial aneurysms. PMID- 18162886 TI - History of endovascular surgery: personal accounts of the evolution. PMID- 18162890 TI - Chained lightning: part III--Emerging technology, novel therapeutic strategies, and new energy modalities for radiosurgery. AB - Radiosurgery is fundamentally the harnessing of energy and delivering it to a focal target for a therapeutic effect. The evolution of radiosurgical technology and practice has served toward refining methodologies for better conformal energy delivery. In the past, this has resulted in developing strategies for improved beam generation and delivery. Ultimately, however, our current instrumentation and treatment modalities may be approaching a practical limit with regard to further optimizing energy containment. In looking forward, several strategies are emerging to circumvent these limitations and improve conformal radiosurgery. Refinement of imaging techniques through functional imaging and nanoprobes for cancer detection may benefit lesion localization and targeting. Methods for enhancing the biological effect while reducing radiation-induced changes are being examined through dose fractionation schedules. Radiosensitizers and photosensitizers are being investigated as agents for modulating the biological response of tissues to radiation and alternative energy forms. Discovery of new energy modalities is being pursued through development of microplanar beams, free electron lasers, and high-intensity focused ultrasound. The exploration of these future possibilities will provide the tools for radiosurgical treatment of a broader spectrum of diseases for the next generation. PMID- 18162891 TI - Characteristics of nontraumatic subarachnoid hemorrhage in the United States in 2003. AB - OBJECTIVE: Substantial progress has been made in the diagnosis and treatment of subarachnoid hemorrhage (SAH). However, studies of SAH in the United States do not include information more recent than 2001, precluding analysis of shifts in treatment methods. We examined the epidemiology and in-hospital outcomes of nontraumatic SAH in the United States. METHODS: We analyzed nationally representative data from the 2003 Nationwide Inpatient Sample of the Healthcare Cost and Utilization Project to determine demographic and hospital characteristics, treatments, and in-hospital outcomes of patients with nontraumatic SAH. RESULTS: In 2003, there were an estimated 31,476 discharges for nontraumatic SAH among patients aged 17 years or older, or 14.5 discharges per 100,000 adults. The in-hospital mortality rate was 25.3%. Microvascular clipping was performed in 7513 discharges, or 23.9% of inpatients with nontraumatic SAH; endovascular coiling was performed in 2849 discharges (9.1%). Adjusted odds of treatment with either procedure were significantly higher in urban teaching hospitals compared with urban nonteaching hospitals (odds ratio, 1.62; 95% confidence interval, 1.00-2.62) or rural hospitals (odds ratio, 3.08; 95% confidence interval, 1.93-4.91). CONCLUSION: The in-hospital mortality rate associated with nontraumatic SAH continues to exceed 25%. Although it is unclear how many patients with nontraumatic SAH were actually diagnosed with a cerebral aneurysm, this study suggests that less than one-third of patients hospitalized for SAH receive surgical or endovascular treatment. Prospective studies are needed to elucidate either what systematic coding error is occurring in the national database or why patients may not receive treatment to secure a ruptured aneurysm. PMID- 18162892 TI - Evaluation of patients with spontaneous subarachnoid hemorrhage and negative angiography. AB - OBJECTIVE: The evaluation of patients with subarachnoid hemorrhage (SAH) with negative initial catheter-based angiography is a diagnostic challenge. Better diagnostic strategies based on hemorrhage patterns are needed. METHODS: We retrospectively investigated the yield of focused history taking, magnetic resonance imaging of the brain and cervical spine, follow-up vascular imaging, laboratory investigations, and craniotomy for vessel exploration in 100 patients with SAH and negative initial catheter-based angiography. RESULTS: The most common distribution of hemorrhage was in a "classic" aneurysmal pattern filling the basal cisterns or posterior fossa (44 patients). A cause was determined in 13 patients (13%), the most common of which was aneurysm (7 patients). Repeat angiography was the most useful diagnostic modality, detecting seven lesions. The yield of the second angiogram was best in patients with a classic hemorrhage pattern (10%) and worse in patients with a negative computed tomographic scan and positive lumbar puncture (0%). The most common reason that a lesion was not detected on initial angiography was aneurysmal thrombosis (five patients). Magnetic resonance imaging of the brain and cervical spine detected one cervical ependymoma. Factors that may contribute to SAH, such as antiplatelet agent use and drug use, were found in 13 patients (13%). Adjunctive laboratory studies alerted practitioners to modifiable risk factors. CONCLUSION: These data suggest useful modifications to current diagnostic paradigms for patients with angiographically negative spontaneous SAH. PMID- 18162893 TI - Admission angiographic cerebral circulation time may predict subsequent angiographic vasospasm after aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Angiographic cerebral vasospasm occurs in approximately 70% of patients hospitalized after aneurysmal subarachnoid hemorrhage (SAH) and is associated with poor outcome. In this study, we examined whether or not cerebral circulation time (CCT) measured with digital subtraction angiography was associated with angiographic vasospasm. METHODS: Patients who underwent cerebral angiography within 24 hours of SAH were analyzed. Contrast dye transit time from the arterial to the venous phase was measured to obtain CCT (supraclinoid internal carotid artery to parietal cortical veins) and microvascular CCT (cortical middle cerebral artery to parietal cortical veins). Patients with ruptured anterior circulation aneurysms and vasospasm on follow-up angiography (Group A) were compared with patients with SAH without vasospasm (Group B) and with normal control subjects (Group C). RESULTS: There were 20 patients in Group A (mean age, 51 +/- 13 yr), 17 patients in Group B (56 +/- 12 yr), and 98 patients in Group C (52 +/- 12 yr). CCT in patients in Group A (7.7 +/- 1.9 s) was significantly longer than those in Groups B (6.6 +/- 1.2 s; P = 0.005) and C (5.9 +/- 1 s; P < 0.001). Microvascular CCT in patients in Group A (7.1 +/- 1.8 s) was significantly longer than those in Groups B (6.1 +/- 1.2 s; P = 0.003) and C (5.4 +/- 0.9 s; P < 0.001). CONCLUSION: Prolonged CCT, a measurement of increased small vessel resistance, can be identified within 24 hours after SAH and is associated with subsequent angiographic vasospasm. These results suggest that microcirculation changes may be involved in vasospasm. PMID- 18162894 TI - Completion angiography for surgically treated cerebral aneurysms: an economic analysis. AB - OBJECTIVE: To compare cost-effectiveness of different strategies for completion angiography after cerebral aneurysm clipping. METHODS: A literature search was used to determine the outcome probabilities and costs of various strategies. The pooled results were used in a Markov cost-effectiveness model to compare quality adjusted life-years and costs of each strategy. Sensitivity (threshold) analyses and Monte Carlo simulation were used to test variation in the model. RESULTS: Routine (for all cases) intraoperative angiography proved to be slightly more cost-effective than selective (only for cases deemed "high risk") intraoperative angiography, being both less costly and more effective. Routine postoperative angiography was the least cost-effective. However, in centers whose rates of clip induced arterial compromise are much lower than the averages reported in the literature, selective angiography might be warranted. CONCLUSION: Routine intraoperative angiography remains the most cost-effective form of completion angiography after aneurysm clipping, at least at our present state of technology. PMID- 18162895 TI - Selective versus routine intraoperative shunting during carotid endarterectomy: a multivariate outcome analysis. AB - OBJECTIVE: The use of intraluminal shunting during carotid endarterectomy (CEA) remains controversial. Over the years, different shunting strategies have been used. More recently, the use of intraoperative electroencephalography and somatosensory evoked potential monitoring with selective intraluminal shunting has been explored. No studies have assessed the independent association of selective versus routine intraluminal shunting to outcomes after CEA. METHODS: The clinical and radiological records of all patients undergoing CEA from 1994 to 2006 at an academic institution were reviewed retrospectively to assess outcomes at 72 hours. The independent association of selective intraluminal carotid artery shunting during CEA and perioperative stroke within 72 hours was assessed through multivariate logistic regression analysis. RESULTS: In 1411 patients with both symptomatic and asymptomatic extracranial carotid artery disease, there were a total of 49 (3.5%) perioperative strokes after CEA. There were two (1%) cases of perioperative strokes among 194 patients in the selective shunting group compared with 47 out of 1217 (4%) in the routine shunting group (P = 0.04). Symptomatic carotid artery disease was associated with a twofold increase in the odds of experiencing perioperative stroke (odds ratio, 1.95; 95% confidence interval, 1.08-3.52; P = 0.03). Patients undergoing electrophysiological monitoring with selective intraluminal carotid artery shunting were more than seven times less likely to experience a perioperative stroke (odds ratio, 0.05; 95% confidence interval, 0.01-0.40; P < 0.01). Increasing cumulative surgical volume, particularly more than 200 total cases, was associated with more than a twofold decrease in perioperative stroke (odds ratio, 0.38; 95% confidence interval, 0.20 0.74; P < 0.01). CONCLUSION: Regardless of symptomatic carotid artery disease or cumulative surgical volume, patients undergoing CEA with intraoperative electroencephalography and somatosensory evoked potential monitoring with selective intraluminal carotid artery shunting had a stroke rate lower than that of the routine shunting group. Selective shunting based on electroencephalography and somatosensory evoked potential monitoring may be superior to the nonselective strategy. PMID- 18162896 TI - Prognostic significance of transdural invasion of cranial base malignancies in patients undergoing craniofacial resection. AB - OBJECTIVE: Invasion of the brain and/or dura is a known negative prognostic factor for patients undergoing craniofacial resection for cranial base malignancy. However, an evaluation of factors that may affect prognosis in this patient subgroup has not been undertaken. METHODS: Between 1993 and 2003, 212 patients underwent craniofacial resection for primary malignancy of the cranial base at the University of Texas M.D. Anderson Cancer Center. Twenty-eight patients (eight women, 20 men; median age, 52 yr; age range, 26-76 yr) had evidence of transdural spread (subdural tumor or brain invasion) of malignancy. These patients were identified and a retrospective review of prospectively collected data was undertaken. RESULTS: Subdural tumors were found in 16 of these patients, and brain invasion was detected in 12. Gross total resections were achieved in 22 patients: 13 with microscopically negative margins, eight with positive margins, and one with unspecified margins. Surgical complications occurred in six patients. There was no surgical mortality. The 5-year actuarial overall survival (OS) was 58%. Eleven patients had no evidence of disease, 11 died of disease, and six were alive with disease at the end of the follow-up period. The median actuarial progression-free survival (PFS) was 38 months (95% confidence interval, 4-72 mo). Gross total resection with negative margins was the key positive predictor of OS and PFS. Brain invasion was a negative predictor of survival (significant for PFS; trend only for OS). There was a trend for shorter OS and PFS in patients with high-grade tumors. CONCLUSION: Overall OS and PFS in highly selected patients with transdural invasion of cranial base malignancy is similar to what has been historically reported for patients without such invasion. The most important variables positively affecting OS and PFS seem to be the ability to achieve a microscopically margins-negative resection followed by absence of brain invasion. Performing this resection in a piecemeal fashion does not seem to affect survival outcomes. PMID- 18162897 TI - Etiopathological factors related to hydrocephalus associated with vestibular schwannoma. AB - OBJECTIVE: We retrospectively analyzed various clinical factors to determine whether or not these factors are etiopathologically related to the development of hydrocephalus in patients with vestibular schwannomas. METHODS: There were 68 patients (29 men, 39 women) in this study who underwent resection of a vestibular schwannoma. The age at the time of surgery ranged from 19 to 76 years (mean age, 51.4 yr). The maximum diameter of the tumor in the cerebellopontine cistern ranged from 0 (localized within the internal auditory canal) to 56 mm (mean, 32.0 +/- 12.9 mm). Cerebrospinal fluid (CSF) protein concentration in the cerebellomedullary cistern was measured intraoperatively in all patients. RESULTS: Sixteen (23.5%) of the 68 patients exhibited radiographic evidence of hydrocephalus. Univariate analysis of various factors revealed that both tumor size and CSF protein concentration were positively related to development of hydrocephalus (P < 0.05 and P < 0.01, respectively). However, in multiple logistic regression analysis, only the CSF protein concentration was predictive for development of hydrocephalus (P = 0.022). There was a trend toward increased CSF protein concentration in patients with a large tumor (> or = 40 mm) compared with those with a small tumor (< 40 mm) (P = 0.06). CONCLUSION: A high CSF protein concentration in fluid from the cerebellomedullary cistern is one of the most important factors contributing to hydrocephalus associated with vestibular schwannoma. It is important to judge whether or not any further treatment is required for hydrocephalus, in addition to tumor resection, especially in patients with communicating hydrocephalus. PMID- 18162898 TI - World Health Organization Grades II and III meningiomas are rare in the cranial base and spine. AB - OBJECTIVE: This study was undertaken to assess a possible relationship between the tumor location and the incidence of World Health Organization (WHO) Grades II and III meningiomas. METHODS: A retrospective review of 794 consecutive patients who underwent meningioma resection between January 1991 and March 2004 was conducted. Among these, 47 patients (5.9%) with WHO Grade II meningiomas and 16 patients (2%) with Grade III meningiomas were further analyzed. Tumor location was assessed using preoperative magnetic resonance imaging scans and/or operative reports. Histological grading was done according to the WHO 2000 Classification scheme. RESULTS: WHO Grade II tumors were found in eight out of 289 (2.8%) cranial base meningiomas and in zero spinal meningiomas, compared with 39 out of 429 (9.1%) non-cranial base meningiomas. Grade III histology was encountered in two (0.7%) cranial base tumors and in one out of 76 (1.3%) spinal tumors, compared with 13 (3%) non-cranial base tumors. The combined incidence of Grades II and III meningiomas was significantly lower in the cranial base (3.5%) and spinal (1.3%) locations compared with non-cranial base locations (12.1%) (P < 0.001). CONCLUSION: WHO Grades II and III meningiomas occur far less frequently in the cranial base and spinal locations. Tumors arising from these locations may have different mechanisms of tumorigenesis and/or progression compared with meningiomas arising from other (non-cranial base) regions. PMID- 18162899 TI - Brainstem corticospinal tract diffusion tensor imaging in patients with primary posterior fossa neoplasms stratified by tumor type: a study of association with motor weakness and outcome. AB - OBJECTIVE: Diffusion tensor imaging (DTI) allows in vivo delineation of brainstem white matter tracts. The purpose of this study was to determine whether or not abnormalities of DTI metrics and fiber tractography correlate with neurological deficits and clinical status in patients with primary posterior fossa tumors. METHODS: A review of patients with primary posterior fossa tumors who underwent magnetic resonance imaging with DTI was performed. Patients were stratified by tumor type (well-circumscribed or infiltrating lesions). Fractional anisotropy (FA) color maps were used to localize the corticospinal tracts within the brainstem. FA, mean diffusivity, and eigenvalues were measured. Tractography was performed. Correlations between DTI metrics and clinical status and between DTI metrics and neurological examination findings were assessed within each patient group using Bonferroni correction for multiple comparisons. Comparisons of DTI metrics were also made between patient groups (infiltrating lesions versus well circumscribed lesions). RESULTS: Thirty patients were studied (mean age, 14.1 yr; 16 male, 14 female). Eighteen patients had infiltrating lesions and 12 had well circumscribed lesions. Twelve patients (four well-circumscribed and eight infiltrating) demonstrated motor weakness on physical examination (four right, three left, five bilateral). Patients with well-circumscribed lesions and weakness had higher mean diffusivity and lower FA in the contralateral corticospinal tract (P < 0.05). No such association was seen in patients with infiltrating tumors. In 102 total patient-years of follow-up (average follow-up period, 4.2 yr), 17 patients (six well-circumscribed and 11 infiltrating lesions) demonstrated complete response or stable disease and six patients (three well circumscribed and three infiltrating lesions) demonstrated progressive disease or death. No differences were seen in terms of DTI metrics between patients with infiltrating lesions and those with well-circumscribed lesions. Patients with well-circumscribed tumors and a bad outcome had significantly lower transverse eigenvalue measures in the corticospinal tracts compared with those with a more favorable clinical status (P < 0.05). CONCLUSION: In patients with well circumscribed primary posterior fossa masses, higher mean diffusivity and lower FA in the brainstem corticospinal tract are associated with contralateral motor deficits; lower transverse eigenvalue may be observed with an unfavorable clinical outcome. PMID- 18162900 TI - Magnetoencephalographic study of posterior tibial nerve stimulation in patients with intracranial lesions around the central sulcus. AB - OBJECTIVE: To study interhemispheric differences of somatosensory evoked field (SEF) characteristics and the spatial distribution of equivalent current dipole sources in patients with unilateral hemispheric lesions around the central sulcus region. METHODS: In 17 patients with perirolandic lesions, averaged somatosensory responses after posterior tibial nerve stimulation at the ankle were recorded with magnetoencephalography. Dipole source solutions in the affected (AH) and unaffected (UH) hemispheres were analyzed and compared for latency, equivalent current dipole strength, root mean square, and spatial distribution in relation to clinical findings. RESULTS: Three main SEF components, P45m, N60m, and P75m, were identified in the hemisphere contralateral to the stimulated nerve. Dipole strength for the P45m component was significantly higher in the AH compared with the UH. SEF characteristics in the AH and UH showed no significant differences with respect to component latency or dipole strength of the N60m and P75m components. Interdipole location asymmetries exceeded 1.0 cm in 71% of the patients. Comparison of the posterior tibial nerve evoked responses (P45m and N60m) in patients with motor deficits and patients without deficits showed that these responses are enlarged in the AH when perirolandic lesions are present. Patients with motor deficits also showed an increased response for P45m in the UH. CONCLUSION: The results of posterior tibial nerve SEFs suggest spatial and functional changes in the somatosensory network as a result of perirolandic lesions with a possible relationship with clinical symptoms. The results can provide further basis for the evaluation of cortical changes in the presence of perirolandic lesions. PMID- 18162901 TI - The neuropsychology of patients with clinically diagnosed idiopathic normal pressure hydrocephalus. AB - OBJECTIVE: To compare the neuropsychological performance of patients with idiopathic normal pressure hydrocephalus (INPH) with that of healthy individuals and to examine its relation to neurological signs, vascular comorbidity, and background factors. METHODS: A consecutive series of 58 patients with INPH underwent neurological, neuroradiological, and neuropsychological examinations. The neuropsychological tests, measuring vigilance, fine movements of the hands, learning, working memory, and aspects of executive functioning, were also administered to 108 healthy individuals. RESULTS: Patients performed worse than healthy individuals on all included tests. Significant correlations between test results were more frequent and stronger among patients and the degree of neuropsychological impairment was related to the severity of other signs of INPH. Patients with vascular risk factors performed worse than those without. CONCLUSION: The neuropsychological deficits in INPH are widely distributed, interrelated, associated with neurological signs, and aggravated by vascular comorbidity. PMID- 18162902 TI - Measurement of head impacts in collegiate football players: an investigation of positional and event-type differences. AB - OBJECTIVE: There exists a need to better understand the biomechanical forces associated with head impacts in American football. The purpose of this study was to investigate whether or not differences in head accelerations existed between different player positions and different event types in collegiate football. We also sought to identify whether or not any associations existed between high magnitude impacts and location of head impacts. METHODS: We conducted a prospective field study in which accelerometers were embedded in the football helmets of 72 collegiate football players. Linear accelerations of all head impacts sustained over the course of the 2005 and 2006 National Collegiate Athletic Association football seasons were collected. One-way analyses of variance and chi tests of association assessed positional, event type, and location of head impact differences. RESULTS: Football players consistently sustained head impacts between 21 and 23 g. Positional differences were identified within our sample. Impacts sustained during helmets-only practices were greater than those sustained in games or scrimmages. There was an association between position and high-magnitude impacts, as well as between high magnitude impacts and location of head impact, with the likelihood of impacts to the top of the head much higher than those to the front, back, left, and right sides. CONCLUSION: Less than 0.35% of impacts exceeding theoretical injury thresholds resulted in concussion. More injury data are required before any theoretical thresholds for injury can be confirmed. Coaches and sports medicine professionals should recognize that head impacts sustained in helmets-only practices are as severe as games or scrimmages; there seem to be no "light" days for football players. PMID- 18162903 TI - Measurement of head impacts in collegiate football players: clinical measures of concussion after high- and low-magnitude impacts. AB - OBJECTIVE: It has been speculated that a theoretical injury threshold of 70 to 75 g may exist for concussions in football players. We aimed to investigate acute balance and neurocognitive performance after head impacts exceeding a theoretical injury threshold in the absence of both self-reported symptoms and a concussion diagnosis 24 hours before testing. METHODS: Forty-three Division I collegiate football players participated in this double-blind, repeated-measures study. Subjects participated in three test sessions (baseline, low impact, and high impact) separated by at least 2 weeks. The Head Impact Telemetry System (Simbex, Lebanon, NH) recorded real-time head impacts sustained during practices and games. The Automated Neuropsychological Assessment Metrics assessed neurocognitive performance. The NeuroCom Sensory Organization Test (NeuroCom International Inc., Clackamas, OR) assessed postural stability. The Graded Symptom Checklist evaluated symptom presence and severity in our participants. RESULTS: After the low-impact test session (<60 g), we observed improvements in the Math Processing (F(1, 26) = 9.797; P = 0.004), Matching to Sample (F(1, 26) = 6.504; P = 0.017), and Sternberg Procedure (F(1, 26) = 5.323; P = 0.030) Automated Neuropsychological Assessment Metrics test modules. Statistically significant differences were also observed after the high-impact test session (>90 g) with improvements in Math Processing (F(1, 22) = 16.629; P < 0.001), Procedural Reaction Time (F(1, 22) = 14.668; P < 0.001), and the total number of symptoms reported (F(1, 22) = 10.267; P = 0.004). Neurocognitive improvements were likely attributed to a learning effect. CONCLUSION: Our findings suggest that sustaining an impact greater than 90 g does not result in acute observable balance and neurocognitive deficits within 24 hours of sustaining the impact. Although previous studies have suggested a theoretical injury threshold, none have been founded on empirical data collected on the playing field in real-time. Future studies should consider the cumulative effects of impacts of varying magnitudes. PMID- 18162904 TI - Measurement of head impacts in collegiate football players: relationship between head impact biomechanics and acute clinical outcome after concussion. AB - OBJECTIVE: To determine the relationship between recorded head accelerations and impact locations and acute clinical outcome of symptomatology, neuropsychological, and postural stability tests after cerebral concussion in Division I collegiate football players. METHODS: A prospective field study was used in which accelerometers were embedded in the football helmets of 88 collegiate football players. Linear and rotational accelerations of all head impacts sustained over the course of 2004 to 2006 National Collegiate Athletic Association football seasons were collected in real-time. Change scores were calculated on clinical measures from the players' preseason baseline to postinjury (within 48 h) and regressed against the recorded linear and rotational accelerations of the head at the time of the concussion. RESULTS: Thirteen concussions were recorded ranging in impact magnitudes of 60.51 to 168.71 g. Linear regression showed no significant relationships between impact magnitude (linear or rotational acceleration) or impact location and change scores for symptom severity, postural stability, or neurocognitive function (P > 0.05). CONCLUSION: Our findings suggest that football players are concussed by impacts to the head that occur at a wide range of magnitudes and that clinical measures of acute symptom severity, postural stability, and neuropsychological function all appear to be largely independent of impact magnitude and location. Because of the varying magnitudes and locations of impacts resulting in concussion as well as other factors such as the frequency of subconcussive impacts and number of previous concussions, it may be difficult to establish a threshold for concussive injury that can be applied to all football players. PMID- 18162905 TI - Gamma knife radiosurgery for trigeminal neuralgia: results and potentially predictive parameters--part I: Idiopathic trigeminal neuralgia. AB - OBJECTIVE: Gamma knife radiosurgery (GKR) is an increasingly used, minimally invasive treatment option for patients with trigeminal neuralgia (TN) refractory to medical therapy. This retrospective study evaluates the long-term results and side effects of GKR in the treatment of TN focusing on potentially predictive factors. METHODS: One hundred sixty patients with TN were included in this study (minimum follow-up, >6 mo; mean, 37.4 mo; range, 6-144 mo). In 92 patients, GKR represented the first nonmedical option ("primary GKR"). In 68 patients, invasive treatments had been previously attempted. All patients were treated using a single 4-mm collimator shot targeting the pontine trigeminal root entry zone with a maximal dose of 75 to 95 Gy. Brainstem dose exposure never exceeded 15 Gy. Treatment outcome results were classified as Grade I (pain-free with no pharmacological treatment), Grade II (pain-free with pharmacological treatment), and Grade III (no result). Data were analyzed using the log-rank test for univariate analysis and the ordered logit model for multivariate analysis. RESULTS: In the overall series, 98 (61%) out of 160 patients reached a Grade I outcome, 45 (29%) reached a Grade II outcome, and 17 (10%) patients had no results from GKR. These results were encouraging for patients with typical facial pain features and for patients treated by a "primary" gamma knife. Considering the global outcome, the most effective and safest dose was found to be in the 80 to 90 Gy range. CONCLUSION: According to our experience, GKR represents a reliable second-line therapeutic approach for TN after pharmacological failure. Favorable prognostic factors include "primary GKR" and maximal GKR dose ranging between 80 and 90 Gy. PMID- 18162906 TI - Stab injuries to the spinal cord: a retrospective study on clinical findings and magnetic resonance imaging changes. AB - OBJECTIVE: This study was undertaken to document changes on magnetic resonance imaging (MRI) scans after a stab to the spinal cord. The aim of the study was to determine if routine MRI scans for this type of injury would lead to a change in management. METHOD: All patients with a stab wound to the spinal cord sustained between November 2004 and July 2005 were retrospectively enrolled. All of the patients were examined, a data form was completed, and MRI was performed within 72 hours on arrival at Groote Schuur Hospital. RESULTS: Twenty-two patients were imaged during the study period. MRI results were reported by a neuroradiologist. The most common change seen on MRI scans was a spinal cord tract and cord signal change. Other imaging findings that were documented included cord swelling and extra-axial collections. Progressive neurological deficits developed in two patients, both as a result of sepsis. An intramedullary abscess was found on contrast follow-up imaging in only one of the two patients, and this was surgically drained. Five patients had extradural collections visible on MRI scans; however, all of these patients either had complete cord transection on MRI scans at the time of injury or in cases of incomplete injury (Brown Sequard syndrome), a tract was visibly affecting the hemicord that explained the injury and there was no mass effect from the extradural collection. CONCLUSION: Routine MRI scans performed on admission of patients with stab injuries to the spinal cord did not lead to surgical intervention unless there was progressive neurological deficit. In such cases, a contrast MRI scan should be performed to rule out sepsis as the most likely cause of deterioration. PMID- 18162907 TI - Nerve regeneration through nerve autografts after local administration of brain derived neurotrophic factor with osmotic pumps. AB - OBJECTIVE: To determine whether or not administration of brain-derived neurotrophic factor (BDNF) with osmotic pumps at the site of the proximal stump of a peripheral nerve autograft can improve peripheral nerve regeneration. METHODS: The tibialis branch of the sciatic nerve was transected and grafted with a 20-mm nerve autograft. Wistar rats (Harlan iberica, Barcelona, Spain) (n = 70) were divided into four groups: a nongrafted control group (Group I, n = 10), a grafted but nontreated control group (Group II, n = 20), a grafted saline-treated group (Group III, n = 20), and a grafted and BDNF-treated group (Group IV, n = 20). BDNF was delivered at a rate of 6 microg/day for 2 weeks after nerve repair using osmotic pumps subcutaneously implanted with a connecting tube, the distal end of which faced the proximal stump of the nerve graft. The animals were euthanized at 6 weeks. Spinal motoneurons were quantified as well as axons at the tibialis branch 5 mm distal to the distal nerve repair site. Neuron size was categorized as large (>25 microm) or small (<25 microm). RESULTS: The statistical comparisons between the mean number of neurons in Groups II and III showed no statistical differences (P = 0.27), but there were statistically significant differences between Groups II and IV (P = 0.02) and III and IV (P = 0.02). Labeling of neurons in the group treated with BDNF represents 76% of neurons found on the nonoperated control Group I, which, in turn, is superior to the 51% of neurons found in the nontreated autograft Groups II and III. Regarding the size of motoneurons, there were no statistically significant differences between groups (P > 0.1). Finally, there were no statistically significant differences among Groups II, III, and IV regarding the number of distal axons. CONCLUSION: BDNF delivered through osmotic pumps was found to have a significant capacity for improving the presence of motoneurons in the ventral spinal horn and, thus, capacity to improve nerve regeneration through nerve autografts. However, in this study, BDNF did not specifically protect against injury to motoneurons, depending on the soma size. PMID- 18162908 TI - Impact of different types of facial nerve reconstruction on the recovery of motor function: an experimental study in adult rats. AB - OBJECTIVE: Poor functional recovery after facial nerve reconstruction is characterized by mass movements and synkinesis. Major reasons are axonal sprouting from the regenerating axons leading to misdirected reinnervation and hyperinnervation as well as polyinnervation of the mimic muscle end plates. We analyzed whether or not the type of nerve reconstruction influenced these pathological phenomena. METHODS: The experiments were performed on 48 adult rats divided into four groups. One group served as an intact control and the experimental groups were subjected to facial-facial nerve repair (FFN), facial nerve interpositional grafting, and hypoglossal-facial nerve repair (HFN), with 12 subjects in each group. Two months later, functional recovery was measured by biometrical motion analysis of whisking. Retrograde fluorescence labeling of the brainstem motoneurons was used to quantify the degree of collateral axonal branching at the lesion site. Fluorescence histochemistry of sections through the levator labii superioris muscle was performed to quantify the degree of polyinnervation after surgery. RESULTS: The type of nerve reconstruction significantly influenced the regeneration. The whisking amplitude did not recover completely regardless of the type of reconstruction. The angular velocity and angular acceleration of the vibrissal hairs showed a full recovery after facial nerve interpositional grafting and HFN, whereas these parameters remained decreased after FFN. Significantly less collateral branching and polyinnervation of the end plates were determined after grafting and HFN than after FFN. CONCLUSION: No type of immediate facial nerve reconstruction results in a full recovery in the rat. However, the morphological and functional recovery was significantly better after grafting and HFN than after FFN. PMID- 18162909 TI - Genetic modification of human sural nerve segments by a lentiviral vector encoding nerve growth factor. AB - OBJECTIVE: Autologous nerve grafts are used to treat severe peripheral nerve injury, but recovery of nerve function after grafting is rarely complete. Exogenous application of neurotrophic factors may enhance regeneration, but thus far the application of neurotrophic factors has been hampered by fast degradation after local application and unwanted side effects after systemic application. These problems may be overcome with the use of lentiviral (LV) vectors that direct sustained local transgene expression in cells. METHODS: Human sural nerve segments were either submerged in or injected with LV vectors encoding green fluorescent protein and cultured in vitro. Production of nerve growth factor (NGF) by nerve segments after injection of LV-NGF was quantified. The effect of NGF produced by LV-transduced fibroblasts derived from human sural nerve segments was assessed on neurite outgrowth in vitro. RESULTS: The injection of vector into nerve segments is a more effective way to deliver the vector than submersion of the nerve in vector-containing medium, leading to large numbers of transduced fibroblasts over a significant extent inside the nerve. The injection of LV-NGF leads to a gradual increase of NGF production, reaching a plateau after 4 days. LV-NGF-transduced human fibroblasts promote neurite outgrowth in vitro. CONCLUSION: We have developed a method to transduce cells in human sural nerve segments with LV vector. This approach holds promise as a powerful novel adjuvant therapy for peripheral nerve surgery and can be performed without changing the routine practice of nerve grafting. PMID- 18162910 TI - O-desulfated heparin improves outcome after rat cerebral ischemia/reperfusion injury. AB - OBJECTIVE: Inflammatory cascades play a significant role in progressive neurological injury after transient cerebral ischemia. It has been demonstrated that heparin, a potent anticoagulant, also possesses anti-inflammatory properties that diminish postreperfusion damage after stroke. However, the potential for heparin to induce hemorrhagic transformation of an infarct has deterred its use in cases of focal cerebral ischemia. In this study, we examined whether or not administration of a novel O-desulfated heparin (ODSH), with significantly decreased anticoagulant activity but active anti-inflammatory effects, would ameliorate inflammatory neurological injury without increasing intracerebral hemorrhage in a rat model of transient middle cerebral artery occlusion. METHODS: Rats were injected immediately before ischemia with phosphate-buffered saline or ODSH (5 mg/kg, intravenously) and then every 12 hours (15 mg/kg, subcutaneously) for 72 hours. The animals were assessed for neurological function using a foot fault test and modified Bederson scale on Days 1, 2, and 5; plasma samples were analyzed for activated clotting time at multiple time points after the initial ODSH dose. After sacrifice on Day 5, infarct volume was determined and brain tissue was examined for evidence of hemorrhage both grossly and using a previously validated spectrophotometric hemoglobin assay. RESULTS: ODSH-treated animals demonstrated significantly improved foot fault performance (P = 0.03) on Day 5 and reduced stroke volumes (P = 0.03) relative to controls. Although the brains of ODSH-treated rats exhibited significantly higher hemoglobin levels in a standardized assay (P = 0.01), there were no incidences of gross hemorrhage observed in either group, and activated clotting time measurements for the treated animals were not significantly elevated over baseline at any time point. CONCLUSION: Our findings indicate that ODSH can be administered at a dose that provides postischemic anti-inflammatory neuroprotection without an increased risk of intracerebral hemorrhage. PMID- 18162911 TI - Quantification of hemodynamic changes induced by virtual placement of multiple stents across a wide-necked basilar trunk aneurysm. AB - OBJECTIVE: The porous intravascular stents that are currently available may not cause complete aneurysm thrombosis and may therefore fail to provide durable protection against aneurysm rupture when used as a sole treatment modality. The goal of this study was to quantify the effects of porous stents on aneurysm hemodynamics using computational fluid dynamics. METHODS: The geometry of a wide necked saccular basilar trunk aneurysm was reconstructed from a patient's computed tomographic angiography images. Three commercial stents (Neuroform2; Boston Scientific/Target, San Leandro, CA; Wingspan; Boston Scientific, Fremont, CA; and Vision; Guidant Corp., Santa Clara, CA) were modeled. Various combinations of one to three stents were virtually conformed to fit into the vessel lumen and placed across the aneurysm orifice. An unstented aneurysm served as a control. Computational fluid dynamics analysis was performed to calculate the hemodynamic parameters considered important in aneurysm pathogenesis and thrombosis for each of the models. RESULTS: The complex flow pattern observed in the unstented aneurysm was suppressed by stenting. Stent placement lowered the wall shear stress in the aneurysm, and this effect was increased by additional stent deployment. Turnover time was moderately increased after single- and double stent placement and markedly increased after three stents were placed. The influence of stent design on hemodynamic parameters was more significant in double-stented models than in other models. CONCLUSION: Aneurysm hemodynamic parameters were significantly modified by placement of multiple stents. Because the associated modifications may be helpful as well as harmful in terms of rupture risk, use of this technique requires careful consideration. PMID- 18162912 TI - Challenges and opportunities for recruiting a new generation of neurosurgeons. AB - Several factors have converged to raise concern among program directors about attracting and training the next generation of neurosurgeons. These include the relatively new duty-hour regulations, the projected physician shortage, and the preference of many current medical students for controllable lifestyles. Attracting top talent into training programs may require innovations geared to Generation X such as policies supporting work-life balance, flexible work options, lots of feedback, mentoring programs, talented leadership, and standardized communication strategies during patient handoffs. Larger programmatic changes may also be needed such as "competency-based" training and additional years of training for mastery of highly specialized procedures. PMID- 18162913 TI - The fiancee with a blue face by Marc Chagall (1887-1985). PMID- 18162914 TI - Anatomy of the cranial nerves in medieval Persian literature: Esmail Jorjani (AD 1042-1137) and The treasure of the Khwarazm shah. AB - Esmail Jorjani was an influential Persian physician and anatomist of the 12th century who did most of his writing after his seventh decade of life. Jorjani's comprehensive textbook of medicine, Zakhirey-e Khwarazmshahi (The Treasure of the Khwarazm Shah) was written in approximately AD 1112 and is considered to be the oldest medical encyclopedia written in Persian. This was an essential textbook for those studying medicine during this time. We describe the life and times of Jorjani and provide a translation and interpretations of his detailed descriptions of the cranial nerves, which were written almost a millennium ago. Medieval Persian and Muslim scholars have contributed to our current knowledge of the cranial nerves. Some of these descriptions, such as the eloquent ones provided by Jorjani, were original and have gone mostly unknown to post-Vesalian European scholars. PMID- 18162916 TI - Know your targets, then align your goals. PMID- 18162918 TI - Physician involvement? Plan on it. PMID- 18162920 TI - Nursing management's visionary leader 2007. PMID- 18162921 TI - And two become one: unit integration. AB - Time and space must take on a new dimension for nursing care as health systems evolve into expanded networks. PMID- 18162922 TI - On the case: effective care transitions. AB - In this new era of patient safety, the case manager, as an advocate and facilitator of care, has a pivotal role on the front line of healthcare delivery. PMID- 18162924 TI - How reflective is our practice? AB - Awareness education helps reduce dysfunctional nurse-to-nurse interaction. PMID- 18162925 TI - ICU survival guide. AB - A nurse manager, preceptor, and novice explore what it takes to cut it in critical care. PMID- 18162926 TI - Ultrafiltration: an alternative for select heart failure patients. PMID- 18162927 TI - Make transformational leadership work for you. PMID- 18162929 TI - A review of community-associated methicillin-resistant Staphylococcus aureus skin and soft tissue infections. AB - Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as a cause of infection among otherwise healthy children and adults in the community. Skin and soft tissue infections are most common, but invasive manifestations also occur. A limited number of strains that may possess unique virulence or transmissibility factors have accounted for the majority of these infections. These strains emerged in the community but now are being transmitted in both community and healthcare settings. Incision and drainage remains the primary treatment for skin abscesses. Strains of MRSA circulating in the community generally are susceptible to a number of nonbeta-lactam antimicrobial agents, although resistance patterns may vary temporally and geographically. Educating patients on strategies to prevent further transmission is a critical component of case management. More data are needed to determine optimal strategies for management and prevention of MRSA skin infections in the community. PMID- 18162930 TI - Symptom profile of common colds in school-aged children. AB - BACKGROUND: Signs and symptoms of a common cold reported in young children are those perceived by caretakers. Objective signs include cough, fever, and sneezing. Subjective symptoms include nasal congestion, feverishness, headache, and sore throat. School-aged children may provide a more accurate picture of the symptom profile during colds because they can self-report. METHODS: Using preprinted diary sheets listing common signs and symptoms, diaries were kept for school-aged children for 10 days after onset of a cold. Nasopharyngeal aspirates were analyzed for respiratory viruses and potential bacterial pathogens. RESULTS: Out of 81 colds studied, the most common signs were cough and sneezing, although the most common symptoms were nasal congestion and runny nose. Other symptoms, including feverishness and headache, were each reported in 15% of children at onset. The majority of children (73%) continued to be symptomatic 10 days after onset. Rhinovirus was detected in 46% and 1 or more potential bacterial pathogens (Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis) in 29% of episodes. Symptom profiles for rhinovirus illnesses and those in which potential pathogenic bacteria were detected were not different from the rest. CONCLUSION: The common cold in school-aged children is characterized by nasal congestion, cough, and runny nose. Signs and symptoms usually continue for at least 10 days. PMID- 18162931 TI - Seasonal distribution of otitis media pathogens among Costa Rican children. AB - BACKGROUND: Otitis media is an important cause of pediatric consultation, and knowledge of yearly pathogen distribution might improve antimicrobial selection. OBJECTIVES: To determine the seasonal pathogen and antimicrobial resistance distribution among Costa Rican children with otitis media. METHODS: Between 1999 and 2004, 952 children with otitis media, aged 3-144 months who participated in various clinical trials, were analyzed. Data obtained from this period were compared against historical data collected between 1992 and 1997. RESULTS: Five hundred sixteen (52%) children had a baseline middle ear fluid pathogen isolated. The most common pathogens were Streptococcus pneumoniae 252 (49%), Haemophilus influenzae 190 (37%), S. pyogenes 38 (7%), and Moraxella catarrhalis 36 (7%). The overall proportion of H. influenzae (24-37%; P = 0.01) and the production of beta lactamase producing H. influenzae (2.6-7%; P = 0.02) increased from 1992-1997 to 1999-2004. There was a nonstatistically significant trend for a higher frequency of S. pneumoniae and H. influenzae isolates detected during the rainy season than during the dry season: S. pneumoniae 58% versus 42% but not significant (P = 0.1) and H. influenzae 68% versus 32% (P = 0.06), respectively. During the rainy season, penicillin-nonsusceptible S. pneumoniae was identified more frequently (38.5%) than during the dry season (18%) (P = 0.003; odds ratio: 2.94; 95% confidence interval: 1.4-6.45). Penicillin-nonsusceptible S. pneumoniae decreased from 46.5% (1999-2001) to 16% (2002-2003) and this was associated with a significant decline of a circulating 19F penicillin-resistant S. pneumoniae serotype (from 89% to 26%), respectively. CONCLUSIONS: S. pneumoniae and H. influenzae are the 2 most common pathogens producing otitis media in Costa Rican children. An increase in the number of H. influenzae and M. catarrhalis was observed in recent years. Penicillin-nonsusceptible S. pneumoniae isolates were more commonly observed during the rainy season, in which increased morbidity with respiratory pathogens is observed. PMID- 18162932 TI - Effects of the change from Stavudine to tenofovir in human immunodeficiency virus infected children treated with highly active antiretroviral therapy: studies on mitochondrial toxicity and thymic function. AB - BACKGROUND: Changing from drugs that have significant mitochondrial toxicity to less toxic compounds may be of benefit in human immunodeficiency virus (HIV) positive patients who receive highly active antiretroviral therapy. Few data on mitochondrial toxicity of antiviral drugs are available in HIV-positive children. METHODS: Eighteen HIV-positive children (median age, 10.9 years) receiving a stavudine-containing regimen were randomized to maintain stavudine (arm A) or change to tenofovir (arm B), while preserving the remaining drugs. Glucose, lipidic, and viro-immunologic factors were assessed at months 0, 1, 3, 6, 12, and 18. Thymic output and mtDNA content were measured in peripheral blood mononuclear cells at 0 and 6 months, mtDNA in isolated CD4+ and CD8+ T cells after 18 months. RESULTS: From baseline to month 6, arms A and B showed similar thymic output and mtDNA. After 18 months, a significant decrease in plasma HDL was observed in arm B, along with a small increase in blood glucose; mtDNA showed no difference. In the 2 arms other factors did not show significant differences from the baseline and from the previous values at 18 months. CONCLUSIONS: Changing from stavudine to tenofovir was well-tolerated, and viro-immunologic success was maintained. PMID- 18162933 TI - Early diagnosis of human immunodeficiency virus in infants using polymerase chain reaction on dried blood spots in Botswana's national program for prevention of mother-to-child transmission. AB - BACKGROUND: Botswana has high antenatal human immunodeficiency virus (HIV) prevalence (33.4%). The public health system provides free services for prevention of mother to child transmission of HIV (PMTCT) and antiretroviral therapy, which can reduce vertical HIV transmission from 35% to <5%. Infant HIV diagnosis is challenging in resource-limited settings, and HIV prevalence among HIV-exposed infants in Botswana is unknown. Dried blood spot (DBS) polymerase chain reaction (PCR) provides a feasible method to assess PMTCT programs and identify HIV-infected children. METHODS: We trained staff in 15 clinics and a hospital to obtain DBS on HIV-exposed infants age 6 weeks to 17 months receiving routine care. Samples were sent to the national HIV reference laboratory. Roche Amplicor 1.5 DNA PCR testing was performed. RESULTS: Between June-December 2005, 1931 HIV-exposed infants age 6 weeks to 17 months were tested for HIV, of whom 136 (7.0%) were HIV infected. Among infants 8 weeks between maternal seroconversion and first treatment [hazard ratio, 2.54; 95% confidence interval (CI), 1.14-5.65], female gender (hazard ratio, 2.02; 95% CI, 1.01-4.1), and cerebral calcifications at birth (hazard ratio, 4.3; 95% CI, 1.9-10). There was no correlation between gestational age at the time of maternal infection and risk for retinochoroiditis. CONCLUSIONS: A delay of >8 weeks between maternal seroconversion and the beginning of treatment, female gender, and especially cerebral calcifications are risk factors for retinochoroiditis during the first 2 years of life in infants treated for congenital toxoplasmosis. PMID- 18162935 TI - Vaccine Protection of Bangladeshi infants and young children against cholera: implications for vaccine deployment and person-to-person transmission. AB - BACKGROUND: Killed oral cholera vaccines are internationally licensed for older children and adults, but not for infants and young children. We investigated whether mass immunization of older children and adults can confer herd protection to children too young to be vaccinated. METHODS: We analyzed the first year of surveillance of an individually randomized, placebo-controlled trial of killed oral cholera vaccines in 89,596 older Bangladeshi children and adult women. Vaccine herd protection of children less than 2 years of age, who were too young to participate in the trial, was evaluated by determining whether the incidence of cholera during the first year of follow-up of this age group was lower in residential clusters with higher levels of vaccine coverage than in clusters with lower levels of vaccine coverage. RESULTS: Vaccine coverage of the targeted population ranged from 4% to 65% in different clusters. The incidence (cases per 1000) of cholera among children less than 2 years of age ranged from 18.9 in clusters in the lowest quintile of vaccine coverage to 8.6 in clusters in the highest quintile (P = 0.004 for the inverse association between vaccine coverage and risk of cholera) Vaccine coverage of adult women (relative risk of cholera = 0.95 for each percent increase in vaccine coverage; 95% confidence interval: 0.92 0.99; P < 0.01), but not of older children, was independently associated with a lower risk of cholera in children less than 2 years of age. CONCLUSIONS: Vaccination of older age groups was associated with protection of children too young to be vaccinated. The pronounced herd protection of young children associated with vaccination of adult women suggests that adult women may play a prominent role in the transmission of cholera to young children in this setting. PMID- 18162936 TI - RANTES promoter gene polymorphisms and susceptibility to severe respiratory syncytial virus-induced bronchiolitis. AB - BACKGROUND: Severe respiratory syncytial virus (RSV) infection is characterized by enhanced chemokine activity. Several studies have linked increased regulated on activation, normal T cell expressed and secreted (RANTES) expression with severe RSV disease. Three single nucleotide polymorphisms, -28C/G, -403G/A, and In1.1T/C in the RANTES gene, have been correlated with the gene's transcriptional activity. In the present study, we explored the possible correlation of the genetic variability of the RANTES gene with the clinical manifestation of RSV disease. METHODS: DNA samples were obtained from 106 children hospitalized for RSV bronchiolitis, in a 2-year period. One hundred twenty sex-matched healthy adults, without a history of severe lower respiratory tract infections, formed the control group. RESULTS: No association was established between -28C/G polymorphism and RSV-induced bronchiolitis, mainly because of its extreme rarity in the studied population. No statistically significant differences were observed in cases and controls regarding genotype and allele frequencies of each of the In.1.1T/C and -403G/A polymorphisms. By contrast, the -28C/C-403G/AIn1.1T/T combined genotype was significantly more common in cases than in controls. CONCLUSIONS: Our results indicate an association between a common genotype with severe RSV infection. This observation supports the previously reported results indicating RANTES as an important mediator of RSV infection. PMID- 18162937 TI - The use of dengue nonstructural protein 1 antigen for the early diagnosis during the febrile stage in patients with dengue infection. AB - BACKGROUND: To evaluate the use of dengue nonstructural protein 1 (NS1) antigen for the early diagnosis during the febrile stage in patients with dengue infection. METHODS: A total of 445 sera obtained from 165 patients [dengue fever (DF): 42, dengue hemorrhagic fever (DHF) grade I: 50, II: 63, III and IV: 10] and 8 other febrile illnesses 5-15 years of age, were assayed for the NS1 antigen, dengue-specific Ig M and Ig G antibodies. RESULTS: The positive rates of NS1 antigen among patients with either DF or DHF was 100% (7 of 7) on day 2, 92.3% (12 of 13) on day 3, 76.9% (40 of 52) on day 4, 56.5% (61 of 108) on day 5 of fever; and declined to 43.1% (59 of 137) on day 6 with defervescence and 29.8% (25 of 84) on day 7 (1 day after defervescence). The positive rates of patients with DF were higher than those with DHF but no statistically significant difference was found. However, patients with primary DHF infection had significantly higher positive rates than those with secondary DHF infection. The positive rates of Ig M antibodies were in reverse proportion to those of NS1 antigen. The additional Ig M antibody determination increased the positive rates to 90.4% (47 of 52) on day 4, 83.3% (90 of 108) on day 5 of fever; 95.6% (131 of 137) on day 6 with defervescence, and 88.1% (74 of 84) on day 7. CONCLUSIONS: Dengue NS1 antigen testing is suggested as a helpful tool for the early diagnosis of dengue infection after the onset of fever. The additional Ig M antibody determination increased the diagnostic rates. PMID- 18162938 TI - Direct detection of the multidrug resistance genome of Haemophilus influenzae in cerebrospinal fluid of children: implications for treatment of meningitis. AB - BACKGROUND: Multidrug resistance (MDR), specifically to ampicillin and chloramphenicol, has complicated the treatment of Haemophilus influenzae type b (Hib) meningitis. This is worsened by use of prior antibiotics, which limits identification of the causative agent by culture and increases reliance on antigen detection. OBJECTIVE: We aimed to develop a PCR assay for detecting the family of Haemophilus integrating and conjugative elements (ICEs) represented by ICEHin1056 among antibiotic resistant Hib, and then apply this directly to CSF to diagnose Hib meningitis and predict organism susceptibility, irrespective of culture results. STUDY DESIGN: Primers specific for orf 51 of ICEHin1056 were designed and multiplexed with Bex primers, specific for H. influenzae, and tested on culture positive and negative cases. RESULTS: Of 73 Hib isolates, orf 51 PCR amplicons, predicting the presence of ICEs, were found in all 33 MDR isolates while only in 1 of 33 sensitive strains. The remaining 7 ampicillin susceptible, chloramphenicol and tetracycline resistant strains did not produce a PCR product to orf 51. PCR amplification from CSF specimens of these culture positive cases produced identical results with 100% and 97% positive and negative predictive values, respectively. Multiplex PCR to detect Bex and orf 51 identified another 16 MDR Hib cases among 81 culture-negative CSF samples. CONCLUSIONS: Direct PCR for orf 51 in CSF identified resistance pattern of 51% more Hib strains than culture alone (110 versus 73). The ability to detect MDR, in culture negative Hib meningitis cases has significant implications for better directing antibiotic treatment of meningitis cases and thus for preventing disability and death. PMID- 18162939 TI - The clinical and molecular epidemiology of community- and healthcare-acquired rotavirus gastroenteritis. AB - BACKGROUND: Rotavirus is the most common etiologic agent of healthcare-acquired diarrhea in pediatric patients. There has been little published information on healthcare-acquired rotavirus infection. METHODS: This was a retrospective cohort study of children hospitalized with rotavirus gastroenteritis at our institution between December 1999 and May 2004. Patients with community- and healthcare acquired rotavirus gastroenteritis were compared with regards to age, time of infection, patient unit, and viral subtype as determined by reverse transcription polymerase chain reaction sequencing. RESULTS: Five hundred seventy-seven children were hospitalized with rotavirus gastroenteritis during the study period. One hundred twenty-one (21%) of these infections were healthcare acquired. The incidence of healthcare-acquired infection was 4.2 cases per 10,000 patient-days. With the exception of 1 outbreak on an isolated patient unit, community- and healthcare-acquired disease affected similar patient populations, had the same temporal distribution, and were caused by viruses with similar subtypes. However, there was a significant difference between the geographic distribution of community- and healthcare-acquired disease within the hospital (P < 0.001). The majority (83%) of community-acquired cases were admitted to general medicine-surgery units, but only 53% of the healthcare-acquired cases occurred on these units (P = 0.005). The remaining healthcare-acquired infections occurred on units that rarely admitted patients with community-acquired disease. CONCLUSIONS: Healthcare-acquired rotavirus gastroenteritis seems to be caused by repeated introduction of community strains into the hospital setting. Heightened attention to infection control practices and rapid rotavirus identification is necessary on all units, especially those that infrequently admit children with rotavirus gastroenteritis, to prevent the spread of healthcare-acquired disease. PMID- 18162940 TI - The descriptive epidemiology of Streptococcus pneumoniae and Haemophilus influenzae nasopharyngeal carriage in children and adults in Kilifi district, Kenya. AB - BACKGROUND: Transmission and nasopharyngeal colonization are necessary steps en route to invasive pneumococcal or Haemophilus influenzae disease but their patterns vary geographically. In East Africa we do not know how these pathogens are transmitted between population subgroups nor which serotypes circulate commonly. METHODS: We did 2 cross-sectional nasopharyngeal swab surveys selecting subjects randomly from a population register to estimate prevalence and risk factors for carriage in 2004. H. influenzae type b vaccine was introduced in 2001. RESULTS: Of 450 individuals sampled in the dry season, 414 were resampled during the rainy season. Among subjects 0-4, 5-9, and 10-85 years old pneumococcal carriage prevalence was 57%, 41%, and 6.4%, respectively. H. influenzae prevalence was 26%, 24%, and 3.0%, respectively. Prevalence of H. influenzae type b in children <5 years was 1.7%. Significant risk factors for pneumococcal carriage were rainy season (odds ratio [OR]: 1.65), coryza (OR: 2.29), and coculture of noncapsulate H. influenzae (OR: 7.46). Coryza was also a risk factor for H. influenzae carriage (OR: 1.90). Of 128 H. influenzae isolates, 113 were noncapsulate. Among 279 isolates of Streptococcus pneumoniae, 40 serotypes were represented and the distribution of serotypes varied significantly with age; 7-valent vaccine-types, vaccine-related types, and nonvaccine types comprised 47%, 19%, and 34% of strains from children aged <5 years. Among older persons they comprised 25%, 28%, and 47%, respectively (P = 0.005). CONCLUSIONS: The study shows that pneumococcal carriage is common up to 9 years of age and that the majority of serotypes carried at all ages are not covered specifically by the 7-valent pneumococcal conjugate vaccine. PMID- 18162941 TI - A toddler with shivering spells, leukocytosis, and thrombocytopenia. PMID- 18162942 TI - The treatment of community-acquired methicillin-resistant Staphylococcus aureus infections. PMID- 18162943 TI - Passive antibody prophylaxis for RSV. PMID- 18162944 TI - Randomized, controlled trial efficacy of pneumococcal conjugate vaccine against otitis media among Navajo and White Mountain Apache infants. AB - We report the phase III trial efficacy of 7-valent pneumococcal conjugate vaccine against clinical and culture proven otitis media (OM) among Navajo and White Mountain Apache infants. Efficacy was -0.4% (95% CI: -19.4 to 15.6) for clinically-diagnosed OM, 5.1% (95% CI: -51.5 to 40.6) for severe OM, and 64% (95% CI: -34% to 90%) for vaccine serotype pneumococcal OM suggesting that this vaccine is efficacious for pneumococcal OM in this high risk population. PMID- 18162945 TI - Real-time polymerase chain reaction detection of Bordetella pertussis DNA in acellular pertussis vaccines. AB - Using 2 real-time polymerase chain reaction (PCR) assays for Bordetella pertussis, 2 of 5 acellular pertussis vaccines were found to contain B. pertussis DNA. Because residual DNA in vaccines can cause environmental contamination, the administration of acellular pertussis vaccines to patients should be physically separated from the collection of patients' specimens for testing of B. pertussis DNA by real-time PCR. PMID- 18162946 TI - Foscarnet salvage therapy for acyclovir-resistant varicella zoster: report of a novel thymidine kinase mutation and review of the literature. AB - The authors describe an acyclovir-resistant varicella zoster virus infection in a pediatric patient after hematopoietic stem cell transplant, the use of foscarnet as salvage therapy, and review the literature to clarify the pediatric experience with foscarnet in this setting. A novel thymidine kinase mutation is described, along with a new phenotypic assay for characterizing acyclovir resistance in varicella zoster virus. PMID- 18162947 TI - Liposomal amphotericin B associated with severe hyperphosphatemia. AB - We report 4 patients who developed hyperphosphatemia while receiving liposomal amphotericin B to treat an invasive fungal infection. Resolution of the hyperphosphatemia occurred after transition to amphotericin B lipid complex. This phenomenon may occur more commonly in patients with mild to moderate renal insufficiency. PMID- 18162948 TI - Cutaneous leishmaniasis treated with azithromycin in a child. AB - This report describes the case of a 10-year-old boy with cutaneous leishmaniasis presumed to be caused by Leishmania major and successfully treated with oral azithromycin. Clinical studies using azithromycin for the treatment of cutaneous leishmaniasis are reviewed. PMID- 18162949 TI - Transmission of Mycobacterium marinum from fish to a very young child. AB - Mycobacterium marinum causes tuberculosis in fish and can cause skin infections in humans who swim in contaminated water or who have direct contact with infected fish. We report the case study of an 18-month-old girl with M. marinum abscesses, who acquired the infection through indirect contact with a contaminated bucket. Appropriate cleaning of aquarium equipment is very important, especially with young children in the household. PMID- 18162950 TI - Hemosiderin-laden macrophages in the cerebrospinal fluid of a neonate after traumatic lumbar puncture. AB - Macrophages in cerebrospinal fluid are described as indicators of pathology. We present findings from the lumbar puncture of a child without neurologic disease. Cerebrospinal fluid obtained after an initial, traumatic lumbar puncture attempt included a high proportion of macrophages, some containing erythrocyte fragments and hemosiderin. This suggests that although macrophages may indicate pathology, they can also accumulate after traumatic lumbar puncture. PMID- 18162951 TI - Brain abscess secondary to dental braces. AB - We report a case of Haemophilus aphrophilus brain abscess presenting 6 weeks after application of dental braces in an adolescent patient with a formerly undetected patent foramen ovale. Neither brain abscess nor invasive H. aphrophilus infection has previously been associated with dental braces. Application or tightening of dental braces may cause bacteremia and invasive disease from oral commensals. PMID- 18162952 TI - The conundrum of prior bacille calmette-guerin vaccination in children of temporary residents in the United States. AB - Bacille Calmette-Guerin (BCG) is one of the most widely used vaccines throughout the world. Children of temporary residents in the United States frequently undergo tuberculin skin testing as part of their health maintenance visits. Management of those children with a positive skin test can lead to doctor-parent disagreements, because of differences in tuberculosis (TB) policies between the United States and other countries that routinely administer BCG vaccine. Two British specialists compare their approach with that of the United States. They also discuss the potential for specific diagnosis of latent TB infection with interferon-based TB diagnostic blood testing, to distinguish positive skin tests caused solely by BCG vaccination. PMID- 18162953 TI - Adolescent jump case in Japan associated with influenza but not oseltamivir. PMID- 18162954 TI - Neonatal echovirus 30 infection associated with severe hepatitis in twin neonates. PMID- 18162955 TI - Parotitis and kawasaki disease in a child with Noonan syndrome. PMID- 18162963 TI - RACs, POA, and the unstoppable evolution of healthcare. PMID- 18162964 TI - "Americans with Disabilities Act" as a case management "resource". PMID- 18162965 TI - Understanding the Medicare "Extended Care Benefit" a.k.a. the 3-midnight rule. AB - PURPOSE: The purpose of this article is to discuss the overall features of the "Extended Care Benefit," also known as the 3-midnight rule for post-acute skilled nursing facilities (SNF) coverage. The goal is to support case managers in the effort to gain a better understanding of how the benefit applies to the Medicare beneficiaries with whom they are working. OBJECTIVES: (1) Give a brief history of the purpose of the "extended care benefit," (2) describe the circumstances under which a patient may be eligible for coverage of skilled nursing or rehabilitation services following a hospitalization, and (3) apply new knowledge to practice by incorporating case management tips into practice. PRIMARY PRACTICE SETTING(S): The primary setting for case managers who need to know this information is in acute care and in postacute care facilities that admit patients for extended care services. FINDINGS/CONCLUSIONS: The eligibility of Medicare beneficiaries for this benefit may be broader than is commonly known by case managers. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: 1. Case managers should not "assume" that the patient does not meet criteria for this benefit. For example: Even if the patient is not ready for rehabilitation services at the time of discharge, a case manager can advocate for deferred admission when the patient is ready. 2. Case managers should include in the patient's admission assessment, the history of previous hospitalizations, particularly in the past 30 days. The patient may have met the 3-midnight requirement prior to admission. 3. Hospital-based case managers must work closely with intake staff, case managers, or liaison staff from the SNF level of care to do a comprehensive assessment of the patient's continuing care needs. 4. Delivering the message to Medicare beneficiaries and their families that the patient does not meet criteria for "extended care services" is a very difficult task and should be done in collaboration with other team members, particularly social workers. 5. With 43% of all admissions of patients being in the "short-stay" category, early intervention is critical to determine the possibility of the patient's postacute needs. It is the short-stay patient who does not meet medical eligibility for 3 days who needs more attention early in the stay. 6. Keeping a patient in the hospital, who does not meet medical necessity standards, even for 1 day with the purpose of meeting the 3-day rule may be considered to be "fraudulent," and is a serious matter. Acute care setting is intended to provide acute care services and not to be used to help "needy" individuals meet criteria for extended care benefits. PMID- 18162967 TI - Predictive modeling & outcomes. AB - PURPOSE/OBJECTIVES: The intent of this article is to explain predictive modeling a statistical tool-as it applies to the practice of case management. While actuaries and financial experts focus on the statistical relevance of predictive risk scores, case managers will benefit from knowing what these scores mean and how interpreting and applying them into meaningful action can lead to improved patient outcomes. PRIMARY PRACTICE SETTING(S): Predictive modeling can be used by physician practice groups, managed care organizations, worksite wellness programs, and any organization desiring to identify the most actionable population for targeted outreach, education, and management. FINDINGS/CONCLUSIONS: Predictive modeling is a technological tool that functions as an electronic claims canvasser searching for predefined variables of interest. This tool is used to identify high-cost diagnoses that, in turn, provide a risk score indicative of the likelihood to utilize more healthcare resources and dollars than persons of the same age and gender. By targeting specific diagnoses or conditions, clinicians can define precise patient interventions such as appointment reminders, weight checks, and dietary compliance; assess the intended results of prescribed medications, or simply to provide education and support. The validation of predictive modeling's true value lies in the thorough evaluation of the outcomes of these interventions. The success of predictive modeling can be demonstrated only by a combination of specific data and evidence based intervention leading to improved models of healthcare delivery and improved patient outcomes. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Using predictive models, case managers will be able to target the most actionable patients who will benefit from targeted outreach and education. Case managers will gain an understanding of how a numerical value can lead to the development of a comprehensive collaborative care plan with the patient and other members of the interdisciplinary team to not only improve the patient's overall health status but to engage the patient in his or her own care, empowering the patient to take responsibility for his or her own health status. PMID- 18162969 TI - Nursing care management at a shelter-based clinic: an innovative model for care. AB - PURPOSE/OBJECTIVES: (1) Describe the nurse care manager role at a shelter-based clinic for women and children who have experienced domestic violence (DV). (2) Develop an innovative model for nursing care management in this unique setting and discuss implications for its implementation in other shelter-based clinics. (3) Provide historical benchmarking data on the use of health-related community resources and common barriers to obtaining these services among women and children following a nurse care management intervention. PRIMARY PRACTICE SETTING: A clinic-based, nurse care management model was developed and implemented at a not-for-profit emergency residential shelter for women and children who have experienced DV. However, DV victims do not just appear in the community: nurse case managers encounter DV in every healthcare setting from hospitals to physician offices/clinics. FINDINGS/CONCLUSIONS: A shelter-based clinic is a safe, confidential, and easily accessible place for women and children victimized by DV to receive immediate care and ongoing assistance for health-related needs. The nurse care manager, who practices in this setting and utilizes motivational interviewing techniques, has a unique opportunity to empower and support women to make informed decisions regarding the health and well-being of themselves and their children. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Provision of nursing care management services in a shelter-based clinic is an innovative approach to improving outcomes and reducing the burden of suffering among women and children who leave abusive situations. The nurse care manager role and, specifically, the model of care described in this project, which includes motivational interviewing techniques, may be generalized to other shelter-type settings to promote sustained, healthy lifestyle practices for victims of DV. Benchmarks developed in this study serve as an initial approach to evaluating outcomes when applying this nursing care management model in practice. Future studies on the effectiveness of this approach among victims of DV are warranted. PMID- 18162970 TI - The Medicare post-acute care payment reform initiative: impact and opportunity for case management. AB - PURPOSE/OBJECTIVES: The Deficit Reduction Act of 2005 mandated that a standardized patient assessment occurs for all Medicare recipients at hospital discharge (regardless of whether they are transferred to a post-acute care [PAC] setting) and during stays in PAC settings to support PAC payment reform. The purpose of this article is to describe the current progress on this initiative and the impact and opportunities it presents for case managers. PRIMARY PRACTICE SETTINGS: The mandate applies to acute care hospitals, skilled nursing and inpatient rehabilitation facilities (IRFs), home health (HHA), and long-term care hospitals. FINDINGS/CONCLUSIONS: The PAC payment reform demonstration project is under way. The same standardized assessment tool will be utilized to measure the patient's health and functional progress throughout the entire episode of care. A Web application for providers to submit the assessment data to Center for Medicare and Medicaid Services is also under development. A final report must be submitted in Congress in 2011. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: 1. An assessment of a patient's discharge health and functional status will be available to the healthcare team to determine appropriate placement into a PAC site. 2. A patient's health and functional progress will be monitored throughout an episode of care. 3. Case managers have long known that paying attention to patients' needs across the care continuum can impact the outcomes of the care provided. 4. You can impact this initiative by discussing the opportunity to participate in the effort with other members of your organization and/or providing information on patient characteristics important to the discharge/transfer decision to the study team. PMID- 18162972 TI - The heartbeat of case management. PMID- 18162974 TI - Strategies for effective occupational health case management. PMID- 18162975 TI - "Monsters, inc."-not the movie, but large medical claims! PMID- 18162976 TI - Patient advocacy, within the law. PMID- 18162978 TI - ABO antigen expression in graft tissue: is titration against donor erythrocytes relevant? AB - ABO-incompatible living donor renal transplantation has become an accepted treatment for end-stage renal disease. Two main factors appear to be important when crossing the ABO barrier, the donor organ A/B antigen expression and the amount of recipient anti-A/B antibody. Antigen expression depends on the ABO blood group and subgroup and may vary in different tissues and cells. The amount of recipient anti-A/B antibody, determined by titration, is very variable. One major drawback with titration is the lack of conformity between different laboratories, making comparisons difficult. For clinical use, the anti-A/B antibody titration technique has to be simple, rapid, and cheap, in addition to being accurate. Although there is a need for more standardized procedures for determination of ABO antibodies, existing techniques are sufficient in the clinical care of patients. To illustrate the variation in susceptibility of different graft tissues to ABO antibodies, in this paper we describe a case of an ABO-incompatible combined liver and kidney transplantation. PMID- 18162979 TI - Interinstitutional variation in the measurement of anti-A/B antibodies: the Japanese ABO-Incompatible Transplantation Committee survey. AB - Exact measurement of antiblood groups A and B antibody (Ab) titers is critical for successful ABO-incompatible living kidney transplantation (ABO-ILKT). However, no standard method for quantitative determination has yet been established. In most institutions performing ABO-ILKT, serial-doubling dilution of serum using a tube technique (TT) is a standard method for determining anti ABO Ab titers, but there are very large interinstitutional variations in the titration of anti-ABO Abs using the TT. The Japanese ABO-Incompatible Transplantation Committee has conducted a national survey to measure anti-ABO Ab titers among more than 30 Japanese institutions. Interinstitutional differences between maximum and minimum values were as large as 32-fold in immunoglobulin M (IgM) and 256-fold in immunoglobulin G (IgG). There seem to be many detailed technical differences in the TT among the institutions. Consequently, in the present study we tried to find the most reliable technique for measuring anti-ABO Ab titers. We compared four different techniques, the TT, BioVue Column Agglutination Technology, DiaMed-ID Micro Typing System, and flow cytometry (FCM). Theoretically, FCM seems to be the most reliable technique because it does not require a great number of agents, each of which may affect the results. The FCM technique yielded a very consistent outcome in repeated measurements and it is one of the most objective techniques because measured data are automatically analyzed by a computer system. In our experience, FCM shows very good reproducibility and seems to be the best technique for titration since it can be performed without use of agents. PMID- 18162980 TI - Comparing the tube and gel techniques for ABO antibody titration, as performed in three European centers. AB - Data from 60 consecutive ABO-incompatible kidney transplantations performed in Stockholm, Sweden; Freiburg, Germany; and Uppsala, Sweden, revealed significant variation in preoperative A/B antibody levels, with median titers of 1:32, 1:128, and 1:8, respectively. We wanted to investigate whether these differences were method-related. The same samples from 21 healthy blood donors were analyzed in the three centers using current local methods. Results confirmed method-related differences, with higher A/B titers in Freiburg and lower titers in Uppsala compared with Stockholm. Results for the same sample differed by a median of three (range 0 to 6) titer steps. When the same number of samples were analyzed in the three centers using the same gel method and the same test erythrocytes, results differed by a median of one titer step (range 0 to 4) for the same sample. In conclusion, gel hemagglutination technique significantly decreases intercenter variation compared with tube technique. PMID- 18162981 TI - The European experience. AB - A new protocol for ABO-incompatible kidney transplantation was introduced in 2001. In this protocol, antigen-specific immunoadsorption to remove existing AB antibodies is used in addition to rituximab to prevent rebound of antibodies and conventional immunosuppression (tacrolimus, mycophenolate mofetil, and prednisolone). This protocol has successively been implemented in many European centers, primarily in Sweden and Germany but also in the UK, Switzerland, the Netherlands, Norway, Denmark, Greece and Spain, and almost 200 ABO-incompatible transplantations have now been performed. In a recent 3-center pooled analysis of 60 consecutive ABO-incompatible living-donor transplantations, there were no graft losses that could be related to the ABO incompatibility and, when compared with ABO-compatible transplantations, no difference in graft or patient survival was found. PMID- 18162982 TI - Flow cytometric measurement of ABO antibodies in ABO-incompatible living donor kidney transplantation. AB - Due to different detection methods, a comparison of anti-A/B antibody (Ab) levels among transplantation centers after living donor ABO-incompatible kidney transplantation is problematic. In the present study, anti-A/B Ab levels were determined prior to, and after, blood group A-to-O kidney transplantation using a recently established semiquantitative flow cytometry-based method, ABO fluorescence-activated cell sorting (ABO-FACS), and compared with standard agglutination titers and indirect antiglobulin testing. Pretransplant agglutination titers were reduced from 1:64 to 1:4, by a total of 14 Glycosorb A column immunoadsorptions (IADSs). Compared with the agglutination titers, antidonor immunoglobulin (Ig) M ABO-FACS mean fluorescence intensity ratios (MFIRs) decreased faster and remained low. No difference was observed using donor type or third-party A red blood cells (RBCs) for the ABO-FACS. Glycosorb A columns were not specific, also reducing anti-B and antiporcine IgM levels, which was confirmed by detecting anti-A/B and antiporcine Abs in the column eluates. In conclusion, analysis of pre- and posttransplant Abs from ABO-incompatible kidney transplant recipients by ABO-FACS allows a better understanding of Ab kinetics, which may improve the design of future IADS protocols. PMID- 18162983 TI - Quantification of blood group A and B antibodies by flow cytometry using beads carrying A or B trisaccharides. AB - In the clinical management of patients receiving blood group ABO-incompatible organ allografts, it is of importance to determine the levels of blood group A and B antibodies before and after transplant. Currently used methods, which are mostly based on hemagglutination, are inexact and are associated with large intercenter variations. Here, we describe preliminary data from our efforts to establish a flow cytometry-based assay for the semiquantification of blood group A and B antibodies using beads carrying synthetic A or B trisaccharides. In agreement with previous investigations, blood group O individuals had greater levels of anti-A immunoglobulin G (IgG) than B individuals, whereas the levels of anti-A immunoglobulin M (IgM) were similar in sera from blood group O and B individuals. PMID- 18162984 TI - Present techniques for antibody removal. AB - Renal transplantation into a patient with a positive cytotoxic cross-match or with an incompatible blood group inevitably results in acute humoral rejection, unless the HLA or anti-A/B antibodies have been removed before transplantation. Although there are several procedures to remove HLA and anti-A/B antibodies, plasmapheresis and immunoadsorption are the most commonly used. In this report, presently available techniques for antibody removal are briefly reviewed. PMID- 18162985 TI - Double-filtration plasmapheresis. AB - It is well known that anti-ABO antibodies in the recipient's serum cause hyperacute rejection in ABO-incompatible kidney transplantation. To perform successful ABO-incompatible kidney transplantation, temporary elimination of the anti-ABO antibodies from the recipient's serum is mandatory. Several methods of removing anti-ABO antibodies have been reported: plasmapheresis or immunoadsorption is widely employed in ABO-incompatible kidney transplantation. Two plasmapheresis methods are available, namely, regular plasma exchange by centrifugation or a plasma separator, and double-filtration plasmapheresis (DFPP). DFPP was designed to selectively remove the immunoglobulin fraction from the serum and, as a result, to minimize the volume of substitution fluid required. In this procedure, we usually use 0.5-1.0 L of an 8% albumin solution as the replacement fluid. This is equivalent to 2.5-5.0 L of fresh plasma used in regular plasma exchange. For pretransplant preconditioning, we usually perform DFPP. Three immunosuppressive drugs, such as tacrolimus, mycophenolate mofetil, and methylprednisolone, are administered 7 days before renal transplantation. To remove anti-A and/or anti-B antibodies, the recipients receive 3 or 4 DFPP sessions before the transplantation until the anti-ABO antibody titers have decreased to < or =1:32. A low dose of rituximab (total dosage 200 mg) is given 7 days before renal transplantation and basiliximab is administered at the time of renal transplantation. Between 2005 and 2006, 39 patients were enrolled in this protocol, and only 1 patient could not undergo renal transplantation because their anti-ABO antibody titer was not reduced to the acceptance criterion of ABO incompatible living-related kidney transplantation (ABO-ILKT), which is 32x dilution. The desensitization success rate using our pretransplant conditioning regimen was 97%. DFPP effectively and safely eliminated anti-ABO antibodies from ABO-incompatible kidney transplantation recipients and contributed to successful ABO-ILKT. PMID- 18162986 TI - Pharmacodynamics of rituximab in kidney transplantation. AB - The B-cell depleting anti-CD20 antibody rituximab has become a therapeutic alternative in renal transplantation. However, understanding of the pharmacodynamics is limited. We have therefore studied the effect of single-dose rituximab, in combination with conventional triple immunosuppressive therapy, on the B-cell population in peripheral blood as well as in tissues, in kidney transplant recipients. Forty-nine kidney recipients received single-dose rituximab. The prevalence of B cells was assessed in peripheral blood, kidney transplant tissue, and in lymph nodes. In 88%, complete depletion of B cells in peripheral blood was observed and, 15 months after treatment, B cells were still undetectable in the majority of patients. In kidney tissue, B cells were also completely eliminated. In contrast, the B cells were not eliminated in lymph nodes, although a reduction was observed. In conclusion, single-dose rituximab in kidney transplant recipients evokes a long-term elimination of B-cells in peripheral blood as well as within the kidney transplant. PMID- 18162987 TI - Preapheresis immunosuppressive induction: necessary or harmful? AB - In ABO-incompatible kidney transplantation, only a few studies have addressed the necessity, duration, and content of immunosuppressive induction therapy. At our center (Karolinska Institute, Stockholm, Sweden), using a preconditioning regimen consisting of 13 days of tacrolimus, mycophenolate mofetil, and prednisolone, we have investigated both short- and long-term renal allograft function (up to 28 days and 1 year after transplantation, respectively) and correlated them to tacrolimus 12-hr trough levels. In summary, during the first 28 days after transplantation, renal allograft function in the ABO-incompatible group was impaired when compared with that observed in the ABO-compatible group. One possible explanation for this finding is the prolonged pretransplantation exposure to tacrolimus in the ABO-incompatible group, resulting in tacrolimus associated renal toxicity, which slows the reduction in plasma creatinine. In fact, the day before, and also immediately after, the transplantation (for the first 3-4 postoperative days), the tacrolimus 12-hr trough levels in the ABO incompatible group were greater than in the ABO-compatible group. Possibly, a shorter pretreatment period with tacrolimus or a reduced target tacrolimus trough level could eliminate this difference in postoperative renal allograft function. However, 1 year after transplantation, kidney allograft function in the two study groups was similar. PMID- 18162988 TI - Japanese experience of ABO-incompatible living kidney transplantation. AB - Since 1989, when we performed the first ABO-incompatible living-related kidney transplantation (ABO-ILKT) in Japan, many Japanese institutions have started their own ABO-ILKT programs. By the end of 2005, 851 ABO-ILKTs had been performed in Japan at 82 institutions. In the present study, we review the surveillance data of the Japanese ABO-Incompatible Transplantation Committee and our own, recent experience of ABO-ILKT. One-, 3-, 5-, and 10-year patient survival has been 95%, 92%, 90%, and 85%, respectively, whereas 1-, 3-, 5-, and 10-year graft survival has been 89%, 85%, 79%, and 61%, respectively. Between 1989 and 1999, a triplicate immunosuppressive regimen consisting of tacrolimus or cyclosporine A plus azathioprine or mizoribine plus methylprednisolone was administered at most institutions. Between 2000 and 2004, tacrolimus, mycophenolate mofetil, and methylprednisolone were used at most of the institutions. Splenectomy was performed in most recipients between 1989 and 2004. Recently, many institutions started to use anti-CD20 antibody (rituximab) as an alternative to splenectomy. In most cases, ABO-ILKT recipients underwent 3 or 4 sessions of plasmapheresis or double-filtration plasmapheresis before transplantation. A greater incidence of acute rejection was observed during the cyclosporine A era, but the incidence of rejection was markedly reduced in the tacrolimus era. Anti-CD20 antibody induction markedly reduced the incidence of antibody-mediated rejection and greatly improved the results. In conclusion, there were significant differences in graft survival and the incidence of rejection before and after the introduction of tacrolimus/mycophenolate mofetil. In addition, rituximab as an alternative to splenectomy is definitely an effective regimen for successful ABO ILKT. PMID- 18162989 TI - Preemptive postoperative antigen-specific immunoadsorption in ABO-incompatible kidney transplantation: necessary or not? AB - Several standard protocols for ABO-incompatible kidney transplantation use scheduled preemptive antigen-specific immunoadsorption during the postoperative period. Our center has developed a different approach. Our patients undergo antigen-specific immunoadsorption postoperatively only if their isoagglutinine titers (immunoglobulin G anti-A/B) exceed 1:8 in the first postoperative week and 1:16 in the second postoperative week. Using this strategy, 22 ABO-incompatible kidney transplantations have been performed at our center since 2004. Only 32% of these patients (7 of 22) needed to undergo postoperative immunoadsorption (mean 4.1 immunoadsorption sessions per patient). The renal outcome in patients receiving postoperative immunoadsorption treatment versus the outcome in patients without postoperative immunoadsorption remained equal at a mean follow-up of 17 months. We identified a shorter pretransplant time on dialysis, a blood type constellation of donor A1/recipient O, and high initial starting titers as predictors for the need for postoperative immunoadsorption treatment. A more detailed version of this study, with modified tables and figures, has been accepted for publication in Nephrology Dialysis Transplantation. PMID- 18162990 TI - Long-term results of ABO-incompatible kidney transplantation with antigen specific immunoadsorption and rituximab. AB - ABO-incompatible (ABOi) kidney transplantation has gained a renewed interest during the past years. In 2001, a protocol for ABOi kidney transplantation based on antigen-specific immunoadsorption and rituximab was introduced at our center. In this study long-term graft function using this protocol was assessed. All ABOi kidney recipients with >1-year follow-up (n=15) were compared with all ABO compatible (ABOc) living donor kidney recipients maintained on the same basic immunosuppression (n=27). Patient and graft survival as well as rejections and calculated glomerular filtration rate were analyzed. Mean follow-up was 3 years. There was no significant difference in patient and graft survival nor in rejection episodes. Mean glomerular filtration rate (79-83 ml/min) was equivalent at 1, 2, and 3 years in both groups. We conclude that ABOi kidney transplantation using antigen-specific immunoadsorption and rituximab is equivalent to standard ABOc living donor kidney transplantation. ABOi transplantation following this protocol does not have a negative impact on graft function long-term. PMID- 18162991 TI - Can ABO-incompatible organ transplantation pave the way for clinical xenotransplantation? AB - Because of the apparent mechanistic similarities between antibody-mediated rejection of ABO-incompatible organ allografts and vascularized xenografts, there is hope that strategies to enable transplantation across the ABO barrier may also be effective in curbing xenograft rejection. This paper discusses the molecular similarities and differences between an ABO-incompatible allograft and a porcine xenograft in terms of their interactions with the immune system. PMID- 18162992 TI - ABO-incompatible transplantation: less may be more. AB - Many have reported success with ABO-incompatible kidney transplantation using B cell ablative therapies such as anti-CD20 and splenectomy. However, splenectomy and anti-CD20 is associated with an increased risk of infection. We show how ABO incompatible kidney transplants can be accomplished with a low risk of antibody mediated rejection and graft loss using plasmapheresis preconditioning, low-dose intravenous immunoglobulin, and standard maintenance immunosuppression. The mean follow up for our cohort of 53 patients is 2 years. The mean creatinine clearance at 1 and 3 years is 58 mL/min and 63 mL/min, predicting excellent long-term function. Only long-term follow up of these patients will render definitive answers, however, these data demonstrate that ABO-incompatible kidney transplantation increases the donor pool by providing live donor kidneys that function promptly with minimal risk of early loss. This can be accomplished with a modest, brief escalation of immunosuppression and at a lower cost to the health care system than maintaining the patient on dialysis. PMID- 18162994 TI - Giant cell arteritis. AB - PURPOSE OF REVIEW: Giant cell arteritis is an important cause of blindness in patients over 60 years of age. We attempt to identify the important clinical aspects in the diagnosis and management of this disorder, so that treatment may be instituted expeditiously to limit visual loss. RECENT FINDINGS: In addition, we identify current areas of standards in the investigation and treatment of patients with suspected giant cell arteritis. SUMMARY: We review some newer treatment possibilities, based on recently proposed immunologic models of giant cell arteritis, that may lead to newer treatment modalities. PMID- 18162995 TI - Neuro-ophthalmic complications of endoscopic sinus surgery. AB - PURPOSE OF REVIEW: This review aims to provide an overview of powered endoscopic sinus surgery and raise awareness of the possible neuro-ophthalmic complications associated with the procedure. RECENT FINDINGS: The neuro-ophthalmic complications of endoscopic sinus surgery can be categorized into five anatomic sites: eye, orbit, optic nerve, extraocular muscle, and lacrimal drainage system. Double vision may be caused by various ocular motility abnormalities. Visual loss may be the result of orbital hemorrhage or direct optic nerve trauma. SUMMARY: A wide range of neuro-ophthalmic complications may occur during endoscopic sinus surgery. The use of powered cutting instruments has been a recent advance in endoscopic sinus surgery, but it has also been implicated in rapid, irreversible, and devasting complications. Multiple preoperative and intraoperative factors appear to be involved in the adverse events associated with endoscopic sinus surgery. Ophthalmologists may be asked to evaluate and manage ocular damage in patients who have undergone endoscopic sinus surgery, and in some cases emergent intervention may be required in an attempt to avoid permanent visual loss. PMID- 18162996 TI - Expanding the spectrum of neuromyelitis optica: friend or foe? AB - PURPOSE OF REVIEW: Neuromyelitis optica is a severe inflammatory disease with selective involvement of the optic nerve and spinal cord and relative sparing of the brain. This paper will review the recent literature on neuromyelitis optica with special reference to the recently proposed changes in the diagnostic criteria and the broadening clinical spectrum of the disease. RECENT FINDINGS: The revised proposed diagnostic criteria of neuromyelitis optica allow incorporation of the status of the recently discovered autoantibody, NMO-IgG, and the presence of brain abnormalities shown by magnetic resonance imaging. NMO-IgG has been found to have good sensitivity and specificity in distinguishing neuromyelitis optica from multiple sclerosis. About 60% of neuromyelitis optica patients can develop such brain abnormalities not typical of multiple sclerosis. The visual prognosis and ambulatory outcome of neuromyelitis optica is poor, and early, aggressive treatment may be warranted to prevent relapses resulting in severe disability. SUMMARY: The clinical spectrum of neuromyelitis optica has expanded since the original description of the disease. Clinicians need to be aware of the recent developments in the diagnostic approach to neuromyelitis optica. PMID- 18162997 TI - Paraneoplastic syndromes in neuro-ophthalmology. AB - PURPOSE OF REVIEW: The aim of this review is to discuss and highlight the recent advances in our understanding of paraneoplastic syndromes in neuro-ophthalmology and their significance. RECENT FINDINGS: The pathophysiologic mechanism in neuro ophthalmic paraneoplastic syndromes involves an immune response triggered by aberrant expression of onconeuronal antigens that cross-react with antigens in the visual system. Recently, 18-fluoro-deoxy-glucose/positron emission tomography with computed tomography scanning has emerged as a useful modality in diagnosing occult tumors responsible for paraneoplastic syndromes. Paraneoplastic optic neuropathy has been recently associated with the anti-CV2/CRMP-5 antibody. The use of serologic analysis of recombinantly expressed clones (SEREX) has led to the identification of new antigens associated with melanoma-associated retinopathy, such as visual arrestin, rhodopsin, titin, and mitofilin. Calcium channel blockers and alemtuzumab have been found to improve visual function in cancer-associated retinopathy. Rituximab has been found to be effective in childhood opsoclonus-myoclonus syndrome. SUMMARY: A high index of suspicion is needed to diagnose neuro-ophthalmic paraneoplastic syndromes. There have been recent advances in our understanding of the pathophysiology and treatment of these disorders. This will facilitate early treatment of causative occult tumors and improves the prognosis. PMID- 18162998 TI - Sarcoidosis. AB - PURPOSE OF REVIEW: Research conducted in recent years has added to our current understanding of the epidemiologic, immune, and genetic factors involved in the pathogenesis of sarcoidosis. RECENT FINDINGS: Exposure to an environmental or occupational antigen in a genetically susceptible individual is thought to trigger an immunologic response. The cells and cytokines that lead to granuloma formation have been an area of active study. Certain human leukocyte antigen genes appear to play roles in susceptibility and disease phenotype, and two genome scans have identified candidate genes. A genetic susceptibility has been suggested for ocular involvement as well. High-resolution computed tomography has been invaluable in selected patients, and new radionucleotide techniques are currently being developed. The tumor necrosis factor alpha antagonist infliximab has been used successfully in cases of refractory sarcoidosis, whereas the use of etanercept in the treatment of sarcoidosis has been disappointing, and may actually cause the disease. SUMMARY: Recent advances in our understanding of the immunologic events in sarcoidosis may lead to developments in treatments that would further decrease systemic and ocular morbidity. PMID- 18162999 TI - Seronegative spondyloarthropathies and the eye. AB - PURPOSE OF REVIEW: Ocular inflammation is a common and important manifestation of disease in patients with seronegative spondyloarthropathy. RECENT FINDINGS: Anterior uveitis is among the most common manifestations of eye disease in patients with seronegative spondyloarthropathies. Additionally, a strong correlation exists among such affected patients and the presence of the HLA-B27 gene. Steroid treatment is often effective for uveitis associated with spondyloarthropathies. Patients with ocular inflammation that is chronic in nature or uncontrolled by steroid treatment have shown good response to immunosuppressive agents known to be effective for articular inflammation, such as methotrexate. Recent studies have shown the possible benefits of tumor necrosis factor-alpha inhibitors, such as infliximab and etanercept. SUMMARY: Uveitis, a leading cause of blindness, is among the most common eye manifestations in patients affected by seronegative spondyloarthropathies. Evidence suggests both genetic and environmental factors in its pathogenesis. Immunomodulator treatments, including the recently popular biologics, may be effective in the control of chronic uveitis in patients with spondyloarthropathy. Ocular surgery is often needed in these patients, and appropriate cautions are required in order to achieve successful vision rehabilitation. PMID- 18163000 TI - Biologics in the treatment of uveitis. AB - PURPOSE OF REVIEW: This review summarizes the current evidence for biologic therapies in the treatment of uveitis. The review emphasizes published research in this field since 2005. RECENT FINDINGS: The anti-tumour necrosis factor-alpha infliximab and adalimumab have demonstrated significant efficacy in controlling uveitis associated with seronegative spondyloarthropathies and juvenile idiopathic arthritis; however, etanercept has failed to show a similar treatment effect in uveitis associated with these conditions. The majority of reports of biologic therapies in posterior uveitis have been uncontrolled trials, or retrospective studies, of uveitis resistant to immunosuppression. Encouragingly, successful control of such refractory intraocular inflammation has been consistently reported with infliximab and interferon alpha, particularly Behcet's disease-associated uveitis. A limited number of reports of anti-interleukin therapies, daclizumab and anakinra, have supported a role for these therapies in some types of uveitis. SUMMARY: Biologic therapies have increased the treatment options for sight-threatening uveitis. Despite experimental rationale, the lack of evidence from randomized controlled studies limits our understanding of when to commence therapy, which agent to choose and how long to continue treatment. Additionally, the high cost and potential side effects of all biologic agents have limited their current use to uveitis refractory to immunosuppression. PMID- 18163001 TI - Inflammatory cystoid macular edema. AB - PURPOSE OF REVIEW: The aim of this article is to update our current understanding and management of inflammatory cystoid macular edema. RECENT FINDINGS: Cystoid macular edema is a common cause of visual loss in uveitis, which occurs predominantly in older patients with chronic uveitis forms and might be heralded by subclinical changes on optic coherence tomography. Cystoid macular edema is emerging as a major cause of visual loss in HIV-infected patients with immune recovery uveitis. Elevated levels of proinflammatory cytokines and vascular endothelial growth factor were found in all types of cystoid macular edema. Treatment with anti-inflammatory and anti-vascular endothelial growth factor drugs is widely applied for all forms of cystoid macular edema and usually has a beneficial, but temporary effect. So far, there are no clear guidelines for the treatment of subclinical cystoid macular edema in uveitis. The effect of vitrectomy in inflammatory cystoid macular edema is not yet clear and might become more important in the future. Recent advances in management include intravitreal drug delivery systems of cystoid macular edema-modifying drugs. SUMMARY: This review summarizes current thoughts on inflammatory cystoid macular edema focusing on the new, clinically relevant findings. Upcoming data on aqueous constituents in cystoid macular edema and imaging with the new generation of optic coherence tomography offer the hope that a better treatment strategy will soon be established. PMID- 18163002 TI - Tuberculosis. AB - PURPOSE OF REVIEW: The purpose of this report is to present an update on the manifestations and management of ocular tuberculosis. RECENT FINDINGS: Tuberculosis affects one-third of the world's population. The incidence of tuberculosis has increased with the increase in the HIV infected population. Following a resurgence of the disease in the US, the incidence has recently declined. Patients may develop scleritis that can be focal, nodular or diffuse with or without keratitis. Anterior granulomatous uveitis may occur. The posterior segment reveals vitritis, choroiditis, and can mimic serpiginous choroiditis and other entities. Patients who are immunosuppressed or HIV infected may develop active mycobacterial disease in the eye leading to rapid destruction of the ocular structures. The diagnosis of ocular tuberculosis is made by isolation of Mycobacterium tuberculosis on Lowestein-Jensen medium or by PCR. The diagnosis is supported by the clinical findings, imaging techniques including optical coherence tomography, fluorescein angiography, indocyanine green and ultrasonography. Tuberculin skin test helps to confirm the diagnosis. SUMMARY: Ocular tuberculosis may occur in the absence of pulmonary disease. Patients present with a spectrum of clinical signs. The disease may mimic several clinical entities. Early diagnosis and prompt treatment of ocular tuberculosis may prevent ocular morbidity and blindness. PMID- 18163003 TI - Anti-vascular endothelial growth factor therapy for ocular neovascular disease. AB - PURPOSE OF REVIEW: Recent research has shown that vascular endothelial growth factor (VEGF) is responsible for many ocular pathologies involving neovascularization. Over the past several years several new agents targeting VEGF have become commercially available for intraocular use. These agents have revolutionized the care of neovascular age related macular degeneration and have great potential for other blinding conditions such as diabetic retinopathy, retinopathy of prematurity, and neovascular glaucoma. RECENT FINDINGS: The VEGF Inhibition Study in Ocular Neovascularization (VISION) trial first showed that an anti-VEGF agent (pegaptanib) was able to prevent vision loss in neovascular age related macular degeneration. The Minimally Classic/Occult Trial of Anti-VEGF Antibody Ranibizumab in the Treatment of Neovascular AMD (MARINA) and Anti-VEGF Antibody for the Treatment of Predominantly Classic Choroidal Neovascularization in AMD (ANCHOR) trials showed that ranibizumab prevented moderate vision loss in neovascular age related macular degeneration and for the first time that a substantial proportion of patients regained vision. Smaller case series have shown that bevacizumab can regress retinal, iris and disc neovascularization. Ongoing trials are investigating the utility of anti-VEGF therapy in retinopathy of prematurity, diabetic retinopathy, and neovascular glaucoma. SUMMARY: Newer anti-VEGF therapies have shown unprecedented efficacy in treating age related macular degeneration with many patients experiencing improvement in vision. Ongoing trials will help guide their use in age related macular degeneration and expand their indications to many other blinding diseases. PMID- 18163004 TI - Ocular surface reconstruction: recent advances and future outlook. AB - PURPOSE OF REVIEW: Ocular surface disorder underlies a diverse group of prevalent diseases in the United States, caused by biological aging, autoimmune conditions, trauma, or iatrogenic factors. Left untreated, these conditions can progress to vision loss or destruction of the globe itself. This review discusses the most recent and relevant clinical and experimental advances in the treatment options for ocular surface disorders. RECENT FINDINGS: Current literature suggests that recent progress in tissue bioengineering, and molecular and cellular biology research presents many potential interventional therapies for ocular surface diseases. Depending on the pathogenesis of each condition, treatment options include bioengineered amniotic membrane graft, limbal stem cell transplantation, conjunctival and extraocular tissue transplantation, multiagent immunosuppressant therapy, and bioartificial devices such as lacrimal gland microdevices and keratoprostheses, or tissue adhesives. SUMMARY: Much progress has been made in the fields of microbiology, stem-cell research, tissue engineering, and bioartificial devices for the treatment of the heterogeneous group of ocular surface disorders. Intensive efforts are underway to ensure the adaptation and accessibility of these therapeutic options to the general population. PMID- 18163005 TI - Bibliography. Current world literature. Neuro-ophthalmology. PMID- 18163008 TI - Aortic valve stenosis and coronary artery disease: pathophysiological and clinical links. AB - Aortic valve stenosis (AVS), including a range of disorder severities, from mild leaflet thickening without valve obstruction, 'aortic sclerosis', to severe calcific aortic stenosis, is a progressive, active process of valve modification, mediating by chronic inflammation (similar to atherosclerosis for cardiovascular risk factors) and biological features. AVS is the expression of early tissue damage due to endothelial damage and oxidative, inflammatory processes, and appears as a surrogate marker for cardiovascular events associated with coronary artery disease (CAD). AVS progression correlates with coronary artery risk factors, such as hypertension, age and cholesterol, and a quantitative evaluation of valve and coronary calcium score comprises a useful marker for cardiovascular prognosis. The low concordance of AVS with CAD appears to be due to other genetic or metabolic factors more specific for calcification processes. Moreover, both pathologies appear to be included within atherosclerotic disease and may be the object of the same clinical therapy and prevention. PMID- 18163009 TI - Is there a link between atrial fibrillation and certain bacterial infections? AB - The pathogenesis of atrial fibrillation (AF), the most common cardiac dysrhythmia, remains unknown. However, many recent studies point to an association between AF and inflammation because of a demonstrable significant correlation between the dysrhythmia and various biomarkers of inflammation. For example, C-reactive protein (CRP), a sensitive biomarker of systemic inflammation, has been reported to be significantly higher in patients with AF compared with a control group with no history of atrial dysrhythmias. Histological anomalies in the atria of patients with AF have also been observed. These anomalies may have an inflammatory basis, although it is not known if the structural changes within the atria of patients with AF are a cause or consequence of the dysrhythmia. Given the suggested involvement of inflammation with this dysrhythmia, an initiating factor for inflammation has been sought. Chronic bacterial infection is the most likely event to initiate and maintain an inflammatory process. Recently, bacteria infections have been hypothesized to be involved in the pathogenesis of AF, and Helicobacter pylori and Chlamydia pneumoniae are two bacteria that have aroused interest. Here, we give a brief overview of AF and then specifically explore the recent evidence that suggests that AF may be caused by bacterial infection(s) in certain patients. PMID- 18163010 TI - New echocardiographic technologies in the clinical management of hypertensive heart disease. AB - Doppler echocardiography is a fundamental instrument to understand heart damage during essential arterial hypertension. Left ventricular (LV) hypertrophy may also be conveniently studied in its morphological and functional aspects by ultrasound application. Echocardiography can also provide important morphological and functional information in hypertensive patients for therapeutic management and prognostic stratification. In recent years, echocardiography has been enriched by very refined techniques that are capable of studying the physiopathological intramyocardial phenomena: (i) tissue Doppler (which studies intramyocardial velocities and time intervals and allows the analysis of strain and strain rate); (ii) integrated backscatter (which analyzes variations of myocardial reflectivity in decibels); (iii) transthoracic Doppler derived coronary flow reserve (which quantifies the vasodilator response of coronary velocities to a hyperaemic stimulation); (iv) myocardial echo-contrast echocardiography (which studies the kinetics of ultrasound contrast microbubbles at the intramyocardial level); and (v) real-time three-dimensional (3-D) echocardiography [which allows a more precise evaluation of left ventricular (LV) volumes and LV mass]. These new methodological approaches have recently been used in the hypertensive clinical setting to provide a deeper knowledge of the complex physiopathological and histopathological mechanisms underlying the modifications induced by arterial hypertension at the myocardial tissue level (myocytes, collagen, microcirculation). This review shows the advancement of high-tech ultrasound applied to hypertensive heart disease, pointing out limitations and incremental potentialities in comparison with conventional echocardiography. PMID- 18163011 TI - The effects of simvastatin and levothyroxine on intima-media thickness of the carotid artery in female normolipemic patients with subclinical hypothyroidism: a prospective, randomized-controlled study. AB - BACKGROUND: Patients with subclinical hypothyroidism (sHT) have an increased risk of developing atherosclerosis, probably related to dyslipidemia. We compared the effects of levothyroxine (L-T4) versus simvastatin treatment on carotid intima media thickness (IMT) and lipid profiles in patients with sHT. METHODS: Forty female patients with newly-diagnosed sHT and 20 age- and-sex matched euthyroid female controls were enrolled. Patients were randomized to receive L-T4 (n = 20) or simvastatin (n = 20) for 8 months. Carotid IMT and the serum concentration of cholesterol were measured at baseline and at the end of the treatment. RESULTS: Compared with controls, sHT patients had higher IMT values (0.66 +/- 0.16 mm versus 0.54 +/- 0.10 mm, P < 0.001). A significant reduction in serum levels of total and low-density lipoprotein (LDL)-cholesterol, triglycerides and mean IMT was observed in patients randomized to the simvastatin group compared to their baseline values. No correlation was demonstrated between the changes in total or LDL-cholesterol concentration and IMT in the simvastatin group. L-T4 replacement resulted in a significant reduction in IMT but serum lipid levels did not change significantly. CONCLUSION: Our results demonstrate that simvastatin significantly reduces IMT in addition to the significant improvement in serum lipids in female patients with sHT. This reduction of IMT was independent of the decrease in serum cholesterol during simvastatin treatment. Although L-T4 substitution therapy also decreases IMT, it does not appear to significantly improve lipid levels. PMID- 18163012 TI - Intravenous contrast echocardiography after myocardial infarction: relationship among residual myocardial perfusion, contractile reserve and long-term remodelling. AB - OBJECTIVE: Previous studies have shown the potential role played by intracoronary myocardial contrast echocardiography (MCE) in predicting long-term remodelling and function after myocardial infarction (MI). Scanty data, however, are available on the role of intravenous MCE in this regard. The aim of this study was to assess the role of residual myocardial blood volume in the asynergic region in modulating ventricular volume changes over time post-MI. METHODS: Thirty-two consecutive patients with anterior MI were studied predischarge using low-dose dobutamine echocardiography (Dob) and intravenous triggered MCE. Videointensity plots were generated from the apical approach and fitted exponentially. Volumes were assessed at baseline, during Dob and at 8 months. RESULTS: Baseline volumes, which appeared related to the extent of the asynergic region (P < 0.01) but showed no relation with videointensity in that area, did not change at follow-up, although Dob had elicited significant contractile reserve. However, videointensity in the asynergic region showed a significant interaction (P = 0.044) with the change in diastolic volume over time, with patients with the highest videointensity reverting remodelling (n = 11, from 69 +/- 16 to 65 +/- 16 ml/m) as compared with the remaining population (n = 21, from 68 +/- 16 to 73 +/- 21 ml/m). This was not seen when Dob-derived parameters were used. Multivariate analysis ranked videointensity second (P = 0.066), after baseline stroke volume (P = 0.005), in predicting changes in volumes over time. CONCLUSIONS: Unlike inotropic reserve, residual myocardial blood volume in the dysfunctioning muscle, as assessed by predischarge quantitative intravenous MCE, has the potential to modulate remodelling in patients who suffered an anterior MI. PMID- 18163013 TI - B-type natriuretic peptide as a marker of asymptomatic re-stenosis after coronary implantation of a paclitaxel-eluting stent. AB - BACKGROUND: The aim of the present study was the detection of asymptomatic coronary re-stenosis after percutaneous coronary intervention (PCI). METHODS: We studied 26 subjects who had been recently implanted with a paclitaxel-eluting coronary stent by both a conventional exercise test and the determination of plasma B-type natriuretic peptide (BNP) levels. RESULTS: At control coronary angiography, nine months after initial PCI, six patients had re-stenosis and 20 were re-stenosis free. We found that re-stenosis was best predicted by the combination of a basal plasma BNP level > or = 50 pg/ml and a positive or uncertain conventional exercise test (positive likelihood ratio of the combination = 10). The best predictor of absence of re-stenosis was a low (< 50 pg/ml) plasma BNP level (negative likelihood ratio = 0.26). CONCLUSIONS: Accordingly, basal BNP level testing can be recommended in the follow-up evaluation of coronary patients after PCI, to improve both the detection and the exclusion of asymptomatic re-stenosis. PMID- 18163014 TI - Aminothiol redox alterations in patients with chronic heart failure of ischaemic or non-ischaemic origin. AB - OBJECTIVE: Oxidative stress plays a role in the progression of chronic heart failure (CHF), but whether and how ischaemic heart disease (IHD) or non-IHD aetiology may account for differential redox alterations is currently unclear. We assessed the relation between thiol redox state and lipid peroxidation, as a marker of oxidative stress, in patients with CHF of ischaemic or non-ischaemic origin. METHODS: Blood reduced glutathione, plasma total and reduced cysteine, cysteinylglycine, homocysteine, glutathione, plasma alpha-tocopherol, ascorbic acid, and free malondialdehyde were assessed in 43 CHF heart transplant candidates (24 IHD and 19 non-IHD) and 30 controls matched for age, gender and number of atherosclerotic risk factors. RESULTS: Reduced cysteine was increased in CHF patients compared with controls. The highest levels were found in IHD versus non-IHD patients versus controls. Malondialdehyde levels were significantly higher in IHD patients than in controls, whereas antioxidant vitamins did not differ among the three groups. CONCLUSIONS: Specific abnormalities in the thiol pattern are associated with heart failure aetiology in CHF patients. Our findings point to the possible role of reduced cysteine in the progression of chronic IHD to heart failure status, as an additional pro-oxidant stimulus for worsening oxidative stress. PMID- 18163015 TI - Late enhancement detected by cardiac magnetic resonance imaging in acute myocarditis mimicking acute myocardial infarction: location patterns and lack of correlation with systolic function. AB - OBJECTIVE: Clinical recognition of acute myocarditis is difficult. Late enhancement (LE) detected by magnetic resonance imaging appears to be a valuable tool for evaluating and monitoring inflammatory myocardial diseases. However, patterns of myocardial enhancement and its correlation with systolic function are not yet well defined. The aim of this study was to assess the extent and location of LE during the acute phase of active myocarditis mimicking acute myocardial infarction and at follow-up, in order to correlate LE with systolic function and to monitor disease progression. METHODS: Magnetic resonance imaging was performed in 13 patients with clinical symptoms and signs of acute myocarditis. RESULTS: All patients showed LE, which was located in the lateral free wall in 11 (85%) patients, at the basal level in eight (61%), at the mid level in nine (69%), and at the distal level in 10 (77%). Moreover, six (46%) patients showed LE in the posterior and inferior wall (P = 0.099 versus lateral wall), four (31%) patients in the septum and inferior wall (P = 0.017 versus lateral wall) and three (23%) patients in the anterior wall (P = 0.006 versus lateral wall). Left ventricular ejection fraction was 0.50 +/- 10 and it was severely reduced only in one patient (0.20). Conversely, in the remaining patients, left ventricular ejection fraction was preserved (> or =0.45) (P < 0.001). Multifocal LE was detected in eight (61%) patients, whereas pericardial effusion was observed only in two (15%) patients (P = 0.002). Twelve patients showed ST-segment elevation and chest pain, as it occurs in acute myocardial infarction, whereas one patient showed non-ST-segment elevation with dyspnoea, as it occurs in non-ST-elevation myocardial infarction. The mean follow-up duration was 4 +/- 12 months, and follow-up was complete for all but one patient who died. LE extent decreased in 11 (92%) patients, whereas new areas of LE were detected in one (8%) patient (P < 0.001). CONCLUSIONS: Contrast enhancement is a frequent finding in the clinical setting of suspected myocarditis and is associated with active and/or persistent myocardial inflammation. Myocarditis detected by magnetic resonance imaging predominantly occurs in the lateral free wall and very rarely affects systolic function. PMID- 18163016 TI - Post-cardioversion transesophageal echocardiography (POSTEC) strategy with the use of enoxaparin for brief anticoagulation in atrial fibrillation patients: the multicenter POSTEC trial (a pilot study). AB - OBJECTIVES: In patients with atrial fibrillation (AF), we sought to evaluate the feasibility and safety of a new transesophageal echocardiography (TEE)-guided strategy, aimed at selecting, 7 days post-cardioversion, those patients who are at low risk (i.e. who can terminate anticoagulation after a second TEE) and those at high risk (i.e. who have to continue it). METHODS: We enrolled 206 patients with non-valvular AF into a randomized, multicenter clinical trial. Group A patients underwent a TEE-guided cardioversion with heparin and at least 4 weeks of oral anticoagulation therapy (OAT) after cardioversion. Group B patients received enoxaparin and underwent a TEE-guided cardioversion. After 7 days, a second TEE was carried out. In the absence of TEE thromboembolic risk factors and left atrial appendage (LAA) dysfunction anticoagulation was discontinued. RESULTS: In group A, 88 out of 102 patients underwent TEE and cardioversion was efficacious in 77 of 78. In group B, 100 out of 104 patients underwent TEE and cardioversion was efficacious in 80 of 87 patients; 55 patients underwent the second TEE and enoxaparin was stopped in 50 without LAA dysfunction. In group A, one transient ischemic attack and one sudden cardiac death occurred. In group B, one patient with complex aortic plaques suffered a stroke during enoxaparin. There was a minor hemorrhage in groups A and B, and a severe hemorrhage in a patient during OAT because of persistent atrial stunning. Hospitalization length and duration of anticoagulation were significantly shorter in group B. CONCLUSIONS: The pre/post-cardioversion TEE strategy with enoxaparin in AF may constitute a feasible and safe approach in selecting patients at low thromboembolic risk who can benefit from precocious termination of anticoagulation (7 days after cardioversion). It may be also useful to identify those patients in whom a life-lasting anticoagulation could be beneficial. A larger trial to confirm these findings is under way. PMID- 18163017 TI - Contrast-induced nephropathy and mortality: possible crucial role of asymmetric dimethylarginine. PMID- 18163018 TI - Rationale and design of the COPPS trial: a randomised, placebo-controlled, multicentre study on the use of colchicine for the primary prevention of postpericardiotomy syndrome. AB - RATIONALE: Colchicine seems to be well tolerated and effective in the treatment and prevention of pericarditis. A preliminary clinical trial has shown that colchicine may be considered not only for the treatment of postpericardiotomy syndrome (PPS), but also for its primary prevention. STUDY DESIGN: The COPPS study is a multicentre, double-blind, randomised trial. On the third postoperative day, 360 patients, 180 in each treatment arm, will be randomised to receive placebo or colchicine (1.0 mg twice daily for the first day followed by a maintenance dose of 0.5 mg twice daily for 1 month in patients > or =70 kg, and halved doses for patients <70 kg or intolerant to the highest dose). The primary efficacy endpoint is the incidence of PPS at 12 months. Secondary endpoints are disease-related hospitalisation, cardiac tamponade, constrictive pericarditis, and relapses at 18 months. Additional analysis will include the time to PPS. IMPLICATIONS: The COPPS trial will evaluate the use of colchicine for the primary prevention of PPS. This study will also provide important information on the frequency, clinical presentation, and prognosis of this syndrome in clinical practice. PMID- 18163019 TI - Endovascular treatment of painless aortic ulcer detected by transesophageal echocardiography as an incidental finding. AB - Penetrating aortic ulcer (PAU) is defined as an ulcerating atherosclerotic lesion that disrupts the aortic wall layers, penetrating the internal elastic lamina. The natural history of the disease and the preferred treatment are still debated. We report a case of painless PAU in a 78-year-old man detected by transesophageal echocardiography at the level of proximal descending aorta as an incidental finding. The patient underwent endovascular stent-graft placement without any complications at early and late follow-up. PMID- 18163020 TI - A new variant of Tako-tsubo cardiomyopathy: transient mid-ventricular ballooning. AB - Stress cardiomyopathy is a reversible left ventricular dysfunction precipitated by emotional stress. Affected patients are generally women, whose symptoms are similar to myocardial infarction with reversible apical dyskinesis associated with hypercontractile basal segments and no evidence for hemodynamically significant coronary arterial stenoses by angiography. We report the case of an 82-year-old woman who presented with acute onset of chest pain after emotional stress and with reversible left ventricular dysfunction consisting of akinesis of the midventricular segments and hyperkinesis of the basal and apical segments. PMID- 18163021 TI - Coronary artery spasm as a cause of ST elevation and inappropriate implantable cardioverter defibrillator intervention. AB - Coronary artery spasm can cause both brady- and tachyarrhythmia, through induction of AV block (usually linked to coronary spasm of the right coronary artery) or ventricular tachycardia/fibrillation linked to extensive myocardial ischemia. The electrocardiographic aspect of coronary artery spasm is an ST segment elevation. We describe the case of patient implanted with an implantable cardioverter defibrillator (ICD) for unexplained syncope which, during coronary artery spasm, received an inappropriate device firing due to ST segment elevation, leading to a double count of the QRS by the ICD. PMID- 18163022 TI - Radiofrequency circumferential ablation limited to the left atrium showing fibrillatory activity in a patient with paroxysmal atrial fibrillation. AB - We report a case of a tailored approach to atrial fibrillation based on the substrate. In a patient with paroxysmal atrial fibrillation, electroanatomic mapping showed that atrial fibrillation was limited to a large area around the ostia of right pulmonary veins. The patient was successfully treated by catheter anatomic isolation of this area from the remaining part of the left atrium. PMID- 18163023 TI - Successful surgical treatment of a giant coronary aneurysm communicating with the right atrium. AB - Aneurysms of the coronary arteries are rare events. In most cases, such abnormalities are clinically silent and, when they cause symptoms, there are dyspnea and/or angina. Our report describes a case presenting with dyspnea; the admission electrocardiogram demonstrated largely spread ischemia in the lower lateral leads and myocardial necrosis in the inferior wall. Selective coronary angiography disclosed severe three-vessel disease; aortic angiography revealed a large vascular neoformation, partly thrombosed, with multilobes and a wide communication with the ascending aorta; the oxygen run showed a slight, non significant 'jump' at the level of the right atrium, confirming a possible left to-right shunt. On open chest surgery, these alterations were confirmed. The principal aneurysm (6-7 cm) originating in right auricle gave rise to a localized pseudoaneurysm above the superior vena cava; the other three aneurysmal structures were situated along the course of the right coronary artery. After 2 years of clinical and echocardiographic control, the patient is symptom-free under conventional medical treatment. PMID- 18163024 TI - Successful catheter ablation of a manifest left anterior accessory pathway. AB - Left anterior accessory pathways are considered to be rare. The junction between the mitral and aortic valves consists of a continuous fibrous tissue. Nevertheless, muscular pathways connecting the left atrial and ventricular myocardium through the fibrous ring between these valves have been observed very rarely. Few reports about successful ablation of left anterior accessory pathways are available. We describe the case of a young patient with episodes of recurrent palpitations due to atrioventricular tachycardia through a manifest left anterior accessory pathway. Complete elimination of the accessory pathway was achieved by means of an ablative procedure via the transaortic retrograde approach. PMID- 18163025 TI - Apical hypertrophic cardiomyopathy in encephalomyopathy. AB - Apical hypertrophic cardiomyopathy (AHC) is associated with neurological abnormalities such as transient ischemic attack, stroke, limb-girdle muscular dystrophy, or eosinophilic myositis in single cases. The association of AHC and metabolic myopathy has not been reported. In an 84-year-old woman with long standing gait disturbance, dementia, Parkinson syndrome, ptosis, ophthalmoparesis, tetraparesis, polyneuropathy, lactacidosis, polyarthralgia, dorsalgia, and osteoporosis, cardiac examination for long-standing anginal chest pain and palpitations, revealed supraventricular and monomorphic ventricular ectopic beats, hypertrophic signs, ST-depression and negative T waves on electrocardiogram (ECG), diastolic dysfunction with impaired relaxation, and AHC on transthoracic echocardiography. AHC was confirmed by cardiac magnetic resonance imaging, which additionally showed a small left ventricular apical aneurysm with a wall-thickness of only 3 mm. The patient was suspected to additionally have a multisystem disease, most likely due to impaired oxidative metabolism. This case shows that AHC may take a mild course and be associated with a number of extracardiac abnormalities. PMID- 18163026 TI - A large pulmonary artery pseudoaneurysm in a patient with Behcet's disease. PMID- 18163027 TI - Myocardial late gadolinium enhancement in specific cardiomyopathies by cardiovascular magnetic resonance: a preliminary experience. AB - Late gadolinium enhancement cardiovascular magnetic resonance (CMR) can visualize myocardial interstitial abnormalities. The aim of this study was to assess whether regions of abnormal myocardium can also be visualized by late enhancement gadolinium CMR in the specific cardiomyopathies. A retrospective review of all referrals for gadolinium CMR with specific cardiomyopathy over 20 months. Nine patients with different specific cardiomyopathies were identified. Late enhancement was demonstrated in all patients, with a mean signal intensity of 390 +/- 220% compared with normal regions. The distribution pattern of late enhancement was unlike the subendocardial late enhancement related to coronary territories found in myocardial infarction. The affected areas included papillary muscles (sarcoid), the mid-myocardium (Anderson-Fabry disease, glycogen storage disease, myocarditis, Becker muscular dystrophy) and the global sub-endocardium (systemic sclerosis, Loeffler's endocarditis, amyloid, Churg-Strauss). Focal myocardial late gadolinium enhancement is found in the specific cardiomyopathies, and the pattern is distinct from that seen in infarction. Further systematic studies are warranted to assess whether the pattern and extent of late enhancement may aid diagnosis and prognostic assessment. PMID- 18163028 TI - Giant coronary sinus in patient with double superior vena cava demonstrated by multislice computed tomography. PMID- 18163029 TI - Management of complex femoral artery occlusion with a re-entry catheter. PMID- 18163030 TI - The imaging of huge pulmonary artery aneurysm associated with Taussig-Bing anomaly. PMID- 18163031 TI - Post-processing using multislice computed tomography coronary angiography improves image interpretability in patients with fast heart rates and heart-rate variations. PMID- 18163033 TI - Reducing the risks of infiltration and extravasation. PMID- 18163034 TI - The ventral striatum as an interface between the limbic and motor systems. PMID- 18163035 TI - Williams syndrome: a genetic deletion disorder presenting clues to the biology of sociability and clinical challenges of hypersociability. AB - Williams syndrome is a neurodevelopmental disorder that results from the deletion of approximately 25-30 genes spanning about 1.5 megabases in the q11.23 region of chromosome 7. Patients with this syndrome present with a combination of a distinctive elfin-like facial appearance; growth retardation; mild mental retardation; an inconsistent cognitive profile that includes visuospatial impairments with good facial discrimination and relatively preserved expressive language skills; and cardiovascular abnormalities. In addition, a striking behavioral feature of the syndrome is the high sociability and empathy that these patients show for others. The study of patients with "partial" deletions of the chromosome band 7q11.23, mutated genes in this region and knockout mice with deletions of specific genes in the homologous G1-G2 region of mouse chromosome 5 are clarifying some genotype/phenotype relationships. Furthermore, genes located in this region that are prominently expressed have been implicated in brain development and function. The neuropsychological profile of patients with Williams syndrome is heterogeneous, highlights important dissociations between cognitive functions and suggests that the behavioral dimensions of sociability, empathy, engageability, and talkativeness may be independent of, or not easily explained by, the cognitive deficits. Williams syndrome has enormous heuristic value because its pathological feature of heightened "sociability" can be a "deficit" symptom of major complex neuropsychiatric disorders, such as schizophrenia and autism. Data consistent with a core inability of patients with Williams syndrome to inhibit social approach suggest that this disorder may afford an opportunity to study the biological basis of the "drive" toward socialization. From a research perspective, the syndrome lends itself to neurobiological studies of sociability as a dimension that varies independently of cognition (or at least many separable cognitive processes). Importantly, from a clinical perspective, the syndrome challenges us to administer strategic psychosocial interventions that take advantage of the opportunities that "pathological" sociability provide, while avoiding its threats. An illustrative example of an effective strategically planned psychosocial intervention for a patient with Williams syndrome is briefly presented. PMID- 18163036 TI - Somnambulism induced by quetiapine: two case reports and a review of the literature. AB - Somnambulism, a previously unreported side effect of quetiapine, is described in two cases. Both cases involved individuals who had no prior or family history of somnambulism and had attention-deficit/hyperactivity disorder. The possible significance of this will also be discussed. Somnambulism is a common parasomnia that reflects an impairment in the normal mechanisms of arousal from sleep in which motor behaviors are activated without full consciousness. Motor behaviors are initiated during deep non-rapid eye movement or slow-wave sleep (stages 3-4), and may be limited to relatively simple manifestations, such as sitting up, fumbling with objects or bedclothes, or mumbling. PMID- 18163037 TI - Comorbidity with axis I anxiety disorders in remitted psychotic patients 1 year after hospitalization. AB - INTRODUCTION: Comorbid anxiety disorders are frequently encountered in psychoses and mainly assessed during the hospitalization. METHODS: Comorbidity was investigated in 98 patients with schizophrenia, schizoaffective, or bipolar disorder, previously hospitalized for psychotic symptoms. Assessments, including Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Brief Psychiatric Rating Scale, and Clinical Global Impressions Scale, were performed during hospitalization (t0) and subsequently in a phase of remission (t1). Comorbidity was assessed at t1 only. RESULTS: One or more comorbid anxiety diagnoses were made in 46 (46.9%) patients. Of these, 15 (32.6%) received multiple anxiety diagnoses, while 31 (67.4%) single anxiety diagnoses. Schizophrenic patients had a rate of social anxiety disorder (SAD) higher (P<.05) than the others. Patients assessed with panic disorder or with obsessive-compulsive disorder at t1 showed significantly greater severity of illness at t0; patients with SAD demonstrated greater severity at t1. No significant differences in the rates of individual anxiety disorders were found in patients treated with typical or atypical antipsychotics or with both. CONCLUSION: Anxiety disorders, particularly obsessive-compulsive disorder, panic disorder and SAD, seem to be frequently comorbid in remitted psychotic patients; SAD would be more prevalent in schizophrenia and might negatively impact the course of the illness. PMID- 18163038 TI - Unexpected reduction in migraine and psychogenic headaches following rTMS treatment for major depression: a report of two cases. AB - Our objective is to report a coincident reduction in headache pain in patients treated with repetitive transcranial magnetic stimulation (rTMS) for major depressive disorder (MDD). Two patients with a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnosis of MDD, non-responsive to prior antidepressant treatment who were enrolled in a sham-controlled, double-blind study of rTMS for MDD. After the study, it was revealed that both were in the active-treatment arm. Both patients suffered from near daily headaches and kept logs of headache frequency and severity before, during, and after the study. Headache pain was significantly reduced under double-blind conditions with rTMS treatment, but returned to baseline following cessation of rTMS treatment. Ultimately, when receiving rTMS post-study as a maintenance intervention for MDD (approximately 2 rTMS sessions/week), the positive effects on headache amelioration were sustained. Headache pain is frequently comorbid with mood disorders and has been reported as the most common side effect with rTMS. In these subjects, rTMS was, in fact, associated with relief of depressive symptoms and preexisting headache pain. This indicates that rTMS may be beneficial for both disorders in some patients. PMID- 18163039 TI - Bipolar depression: best practices for the outpatient. AB - Although certain aspects of bipolar disorder are well understood, there is a need for more information concerning management of acute bipolar depression, the effect of comorbid conditions, and long-term management of bipolar disorder. The outpatient presenting with bipolar disorder often presents with many of the key problems related to the long-term course of the disorder, including misdiagnosis and treatment non-adherence. Depressive symptoms are also prevalent during the course of bipolar disorder, with studies finding that depression can cause a low grade "darkness" that longitudinally affects outpatients with bipolar disorder. These variable and persistent depressive symptoms may cause severe functional impairment and increased suicidality. Pharmacologic treatment of bipolar disorder typically includes anti-manic and mood-stabilizing medication. Although some studies find antidepressants have some positive effect, researchers have found that antidepressants, including selective serotonin reuptake inhibitors, when used as monotherapy or in conjunction with mood stabilizers, have little benefit for the treatment of bipolar disorder and may increase the likelihood of a switch into mania, hypomania, or mixed episodes. For long-term outpatient treatment, lamotrigine and lithium are proven to be highly effective. However, clinicians should also stress psychosocial treatment approaches, such as cognitive behavioral therapy, as a principle of chronic disease management for long-term outpatients. Data on pharmacotherapy and psychosocial treatments are emerging, and clinicians should integrate these two treatment options into the standard of care. This expert roundtable supplement focuses on the treatment and management of the bipolar outpatient at risk for a depressive relapse as well as patients experiencing both acute and long-term symptoms of the disorder. Two case studies are presented to elucidate the best practices for the varying clinical states of bipolar disorder. PMID- 18163042 TI - The impact, identification, and management of attention-deficit/hyperactivity disorder in adults. Introduction. PMID- 18163043 TI - [Thoracoskopy in diagnosis and treatment of lung injuries]. AB - The thoracoscopy was used at 150 patients with penetrating wounds of thorax. Severity of injury according to ISS scale was 9.02+/-1.1 scores, RTS criteria - 7.84+/-0.0 scores, and TRISS - 99.8+/-0.1%. Lung injuries were diagnosed at 71 (47.3%) patients. Features of various wounds of lung and methods of closure have been analyzed. Bleeding, air intake and intrapulmonary hematoma were the indications for hermetic closure of wound canal. Coagulation of lung wound was performed at 24 (33.8%) patients, closure with endosurgical techniques - 30 (42.2%); the wounds have not been closured at 8 (11.3%) patients. Conversion to thoracotomy was necessary at 5 (7%) patients with deep wounds of lung. The thoracoscopic surgical procedure has been completely performed at 58 (81.7%) patients. The better postoperative results were achieved after endosurgical closure of lung wounds. Computed tomography is perspective method of preoperative diagnosis of lung injury severity. PMID- 18163044 TI - [Median resection of pancreas]. AB - Experience of 16 median resections of pancreas due to serous or mucinous benign cystic tumor (13 patients) or small afunctional neuroendocrine tumor (3 patients) is analyzed. In all the cases the pathologic process was located at the zone of pancreas isthmus. The long-term results were followed-up at 13 patients from 2 to 14 years (mean followed-up - 8 ys). There were no recurrences of tumor. It is concluded that median resection of pancreas is effective and safe surgical operation for the patients with localization of benign tumor at the zone of pancreatic istmus. PMID- 18163045 TI - [Combined operations for prophylaxis of gastroesophageal bleedings at the patients with portal hypertension]. AB - Results of portocaval bypass (1st group), suture ligation of gastric and esophageal varicose veins (2nd group), and combined operations (3rd group) performed in early postoperative period at 759 patients with liver cirrhosis and extrahepatic portal hypertension are analyzed. Long-term results were studied at 549 patients. Early recurrence of bleeding was occurred at 41 (12.9%) patients of 1st group, at 17 (8.5%) of 2nd, and at 2 (4%) patients of 3rd group. Lethality due to bleeding recurrence at 1st group was 5.1% among the patients with liver cirrhosis and 1.9% among ones with extrahepatic portal hypertension. Recurrence of bleeding at 2nd group was seen at 52.8% patients with liver cirrhosis, and at 36.4% - with extrahepatic portal hypertension. Lethality due to bleeding recurrence in this group was 7%. The best results were achieved in 3rd group: the recurrence of bleeding was diagnosed at 2 (7.7%) cases, there was 1 lethal outcome. It is concluded that combined surgical procedures at the patients with portal hypertension permit to reduce the rate of early recurrences of bleeding in 2.5 times. It allow to perform widely the partial portocaval anastomoses at the patients with liver cirrhosis and the bypass procedures at extrahepatic portal hypertension. PMID- 18163046 TI - [Selective proximal vagotomy in combination of transverse gastroduodenoanastomosis for the treatment of ulcer pyloroduodenal stenosis]. AB - Results of proximal vagotomy in combination of transverse gastroduodenoanastomosis (GDA) at 36 patients with sub- and decompensated ulcer pyloroduodenal stenosis are analyzed. The principles of surgical method choice depending on stenosis localization and motor-evacuatory function of stomach are discussed. Obtained results are compared with ones after selective proximal vagotomy with longitudinal GDA by Jaboule and pyloroplasty by Finney. Clinical, radiological, endoscopic and morphological data demonstrate the advantages of transverse GDA compared with longitudinal GDA and pyloroplasty. PMID- 18163047 TI - [Treatment tactics of patients with pyloroduodenal stenoses]. AB - Overall 172 patients aged 29 to 87 years with stenosis of stomach pyloric part and duodenum were studied. The complications of gastric or duodenal ulcer (119), cancer of stomach (49) and pancreas (4) were the cause of stenosis. All the patients were divided into 3 groups: 1st group consisted of 43 patients who has undergone urgent surgery due to stenosis with bleedings and perforations without additional protein-energetic treatment before operation; 2nd group - 65 patients who were prepared preoperatively with complete parenteral nutrition; 3rd group - enteral tube feeding with balanced mixtures before surgery (64 patients). The best results were achieved in the patients with preoperative enteral tube feeding; it allowed to reduce lethality from 25.5 and 16.3% at 1st and 2nd group to 0% ant 3rd group. PMID- 18163048 TI - [Plastic closure of large ulcers of duodenum posterior wall complicated with bleeding]. AB - New method of organ-saving surgery for bleeding duodenal ulcer is described. Advantages of this method have been confirmed in experimental studies on 10 mongrel dogs and by successful clinical application at 3 patients with large bleeding ulcers of duodenum posterior wall. PMID- 18163049 TI - [Early diagnosis of hospital infection based on monitoring of wound microflora at purulent surgery unit]. AB - Microflora of purulent surgical wounds of patients of purulent surgery unit was studied, multiresistant strains were analyzed particularly. Data of 1994 and 1999 were compared with ones of 2004 bearing in mind the administration of new antibiotics (cefalosporins and phthorquinolones) since 2000. Etiology and causes of nasocomial infections, principles of early diagnosis are analyzed in detail. PMID- 18163050 TI - [Long-term results of surgical treatment of thymomas at the patients with generalized myasthenia]. AB - Results of diagnosis and treatment of 150 patients with thymoma and generalized myasthenia are analyzed. The results of examination were registered before surgery, and also in 1, 3, 5 and 10 years after one. It is demonstrated that thymoma is potentially malignant tumor with risk of recurrence and requires long follow-up of patient after operation. Morphologic type and stage of tumor, severity of myasthenia are the main predictors in the prognosis of surgical treatment. Thymomectomy with excision of fatty tissue of anterior mediastinum is absolutely indicated for this severe category of patients, and it decreases the risk of tumor progression. Mortality of patients depends not only on tumor growth or recurrence but also on concomitant myasthenic disorders. PMID- 18163051 TI - [Long-term results of surgical treatment of patients with IV stage of chronic lower limb ischemia]. AB - Results of surgical treatment of 210 patients with IV stage of atherosclerotic chronic lower limb ischemia are analyzed. Direct revascularization of lower limb permits to increase lifetime of patients and to improve quality of life. All the patients require permanent systematic complex therapy after surgery regardless of its variant. PMID- 18163052 TI - [Plastic closure of wound defects in complex treatment of hand purulent diseases]. AB - Results of different techniques of early plastic closure of wound defects at 457 patients with inflammatory and posttraumatic purulent and necrotic processes of fingers and hand are analyzed. Overall 12 methods of plastic closure including combination of them were performed depending on localization, size and condition of wound. Good esthetic and functional short-term results were achieved at 434 (94.9%) patients, long-term results - at 441 (96.5%). The complications were seen at 23 (5%) cases. PMID- 18163053 TI - [Treatment of amebiasis surgical complications at HIV-patients]. AB - Experience of treatment of 8 HIV-patients with amebiasis is analyzed. Five patients with provisional diagnosis "acute appendicitis" underwent diagnostic laparoscopy, sanation and drainage of abdominal cavity. Ameboma with acute intestinal obstruction was diagnosed at 2 patients; they underwent extraperitonization of inflammatory tumor. One patient with provisional diagnosis "peritonitis" had large purulent-necrotic total process in colon; subtotal resection of colon has been performed. Extraintestinal complications of amebiasis have been also diagnosed at 6 patients in short-term postoperative period. Mini invasive methods of surgical treatment are more preferable for HIV-patients. This approach in combination with traditional antibacterial therapy permits to reduce the rate of postoperative complications and recurrences. PMID- 18163054 TI - [Parodontal microhemodynamics status in patients with vascular dystonias]. AB - The state of peridontal microhemodynamics was investigated in 80 patients with an initial arterial hypotension (IAH) and 70 patients with an arterial hypertension (AH) of different age-groups (6, 15, 20-24, 40-45 years) according to the laser dopplerography method. A decrease of a bloodflow intensity in all agt-groups with IAH and in senior group with AH was estimated. Impairments of the regional hemodynamics were accompanied by the high rate of stomatological morbidity. PMID- 18163055 TI - [Periapical tissues microvessels reactivity study after pulp extirpation and on different stages of endodontic treatment]. AB - Reaction of the microvessels in the gingival periapical tissues on pulp extirpation was followed by hyperemia development in microcirculatory bed, its expression increased in proportion as parodontal inflammation intensified and further strengthened after root canal filling. Microcirculation normalization in the gingival periapical tissues after endodontic treatment of chronic pulpitis and pulp extirpation as part of endodontic treatment was evident in 1 month in intact parodontium and in 6 months in case of its inflammation. PMID- 18163056 TI - [Erythrocytic membrane status (as cell model) in cases of severe cranium-face trauma: possibilities for correction]. AB - Acute posttraumatic period of severe maxillofacial trauma in case of hyperbaric oxygenation therapy is followed by expressed activation of lipide-peroxide oxidation (LPO) in erythrocytic membrane on the background of significant reduction of the substrate for fermentative antioxidation system (AOS). Imbalance in the system LPO:AOS is followed by expressed structural reorganization in erythrocytic membrane and build-up extraerythrocytic haemoglobin in plasma. Klonidine promotes LPO activity reduction and prevents depletion of AOS fermentative capacity in membrane that is followed by normalization of structural functional interrelations in membrane and reduction of the level of extraerythrocytic haemoglobin in plasma. PMID- 18163057 TI - [Saliva microcrystallization: new methodical approaches]. AB - Results of the studies of saliva microcrystallization were summarized. It was shown that the use of new methods in silico let successfully divide saliva microcrystals in norma and in cases of different diseases of gastrointestinal tract, sugar diabetes of both types and prostate tumors. PMID- 18163058 TI - [Influence of correcting therapy course on salivary components of patients with fluorosis]. AB - Fluorosis, caused by long-term intake of high levels of fluoride, is characterized by clinical manifestations in bones and teeth. However, detrimental effects of high fluoride intake are also observed in soft tissues. Saliva, as a biological liquid of the human organism, may be a reflection of the metabolic state. The concentration of calcium (Ca2+), phosphate (PO(3)(4)-), chloride-ions (CL-) and activity of alkaline phosphatase were determined in saliva of 46 patients with fluorosis before and after antioxidant therapy. An imbalance of salivary components of the patients with fluorosis has been corrected by the therapy course partially. PMID- 18163059 TI - [Validation of Oral Health Impact Profile (OHIP) quality of life questionnaire in Russian patients with evidence of chronic generalized periodontitis]. AB - As part of research project translation and validation of Oral Health Impact Profile (OHIP) quality of life questionnaire has been administered. 25 patients have been interviewed (8 men and 17 women), the average age was 40+/-8 years, with evidence of chronic generalized periodontitis. The clinical picture of the disease development in those patients was common. All patients perceived questions of the pilot questionnaire version correctly. The relations between the quality of life and the sex of patients, social status and other factors have been found statistically significant. PMID- 18163060 TI - [Use of NO-therapy in correction nose deformations in patients with thick porous skin]. AB - If the patient has thick porous skin in the region of end part of nose the result of the operation satisfies neither patient nor surgeon. During recent 2 years the authors used NO-therapy (apparatus Plazon) both in preparation thick porous skin for the operation and in postoperative period for prevention crude scars development and their treatment. NO-therapy was conducted to 22 patients pre- and postoperatively, to 26 patients operated 1,5-2 years ago (who had some compactness in the region of end part of nose) and to 5 patients with crude scar deformations. NO-therapy promoted quicker reduction of oedema and liquidation postoperative scars had formed in cases of endonasal incisions. PMID- 18163061 TI - [Comparative evolution surgical accesses to temporo-mandibular joint]. AB - In experiment on 30 corpses of adult people criteria of an operational wound (depth of a wound, a corner of operational action, an axis of operational action, a corner of an inclination of operational action) were studied at preauricularis, intrauricularis, intrauriculo-temporalis and posterior mandibullaris access to temporo-mandibular joint (TMJ). New surgical intrauriculo-temporalis access to the joint is substantrated. On the basis of the analysis of 289 operations at 268 patients the indications to a choice of surgical access were developed at various diseases and damages of TMJ. PMID- 18163062 TI - [Concentration of proinflammatory and antiinflammatory cytokines in blood and wound of patients with odontogenic phlegmons]. AB - Levels of IL-1beta, IL-6, TNFalpha, IL-10 and TGF-beta in blood and wound of 56 patients with odontogenic phlegmons in dynamic of infection process were expected. In accordance with known facts production of proinflammatory cytokines (IL-1beta, IL-6, TNFalpha in blood in acute phase of the inflammation process was much more intensive than in normal conditions. Increased proinflammatory cytokines generation led to significant raising of IL-10 concentration, but elevation of its level in blood was only at the 10-14 the days of therapy. The dynamic of these cytokines concentrations in wound were similar. On other side the negative role in regeneration of wound plays TGFbeta. Minimal concentration of this cytokine in blood was found at 1-3 from treatment beginning. On the background of the intensive therapy the level of TGFbeta raised fastly and at 10 14 days was much more than in control conditions. Wound level of TGFbeta was more than in normal conditions from the third day. These changes may be one of the reason of hyper proliferation of keratinocytes, that led rough scars. PMID- 18163063 TI - [Spiral computer tomography use on the treatment stages of patients with defects and deformations of face bones and soft tissues]. AB - There are discussed possibilities of spiral computer tomography use in cases of defects and deformations of face bones and soft tissues elimination. PMID- 18163064 TI - [Clinical peculiarities of sialolithiasis course and method of treatment choice in the period of the disease exacerbation]. AB - There were examined and treated 181 patients with sialolithiasis in the period of the disease exacerbation. About 30% of the patients were directed to the clinic with mistaken diagnoses. Out of diagnostic techniques in the period of exacerbation there were employed external and bimanual palpation of the inflamed gland region. The most diagnostic value in the period of sialolithiasis exacerbation had X-ray and computer sialotomography (it allowed to diagnose salivary calculi in 98% of the cases). In the period of sialadenitis exacerbation sialotomography is not recommended for use. Ultrasound diagnostics let to reveal concretions of big sizes. Treatment of patients in the acute period of sialolithiasis included salivary gland duct system sanation with antiseptics use and external blockades. PMID- 18163065 TI - [Dynamic magnetotherapy use in comprehensive treatment phlegmons of maxillofacial region and mandible fractures]. AB - 145 patients (60 with phlegmons of submandibular and submental regions, 85--with fractures of mandible) were observed and treated with the use of moving pulse magnetic field (MPMF) produced by special apparatus (AMO-ATOS-E, <>, Saratov-city), 60 patients with the same pathology were treated by traditional physical methods and served as control. Use of MPMF led to quicker patient recovery: quicker reduction (if compared with traditional physical methods of treatment) of oedema and soft tissue inflammatory infiltration, quicker relief from inflammatory reaction (according to data of cytokinin profile), improvement of tissue blood supply in the region of fractures in patients with mandible fractures. As the result--we had shortening treatment terms of such patients and complication number reduction. PMID- 18163066 TI - [Comparative analysis of the results of patient prosthetic treatment by removable arch and elastic constructions in cases of end mandibular defects based on microcirculation study of prosthetic bed tissues]. AB - Study was performed of the microcirculation in gingival tissues of prosthetic bed in patients with end mandibular defects treated by removable arch and elastic constructions in order to increase prosthetic treatment efficacy. It was established that most favourable stimulating effect upon trophics was achieved in patients who had received removable plate constructions made according to the technology of thermoinjection system produced by the company Valplast on nylon base. PMID- 18163067 TI - [3-D analysis obtained in mandible movements in children 7-15 years old with different types of occlusion]. AB - 60 children with different types occlusion--normal, distal, mesial and transverse between the ages 7-15 years were selected. Using unique computer software programme and 3D digitizer MicroScribe-G2 (Company Immersion, USA) on the articulator with mounted casts, persuaded measuring the scores in different movements of mandible habitual occlusion, maximum forward movement and maximum lateral movements. Scores were calculated and results interpreted. PMID- 18163068 TI - [Situation analysis for community regional preventing dental caries programme planning with water fluoridation use]. AB - As the results of clarification of some technical parameters of community water supply systems, examination of 780 12-year-old children and F-concentration determination in 111 water samples in 13 cities (with population 50, 000 and more) of 7 regions of Russian Federation there were determined economical and technical conditions for planning and implementation regional communal dental caries preventing programmes with the use of water fluoridation. The received data in future let us make mapping of these regions for planning and choice substantiation of dental caries preventive programmes with endogenic use of fluoride. PMID- 18163069 TI - [Glomus tumors of the middle ear in children]. AB - Glomus tumor (GT) is a rare tumor of the middle ear in children. In spite of cytologically benign nature of this tumor, its course can be characterized as malignant because of GT infiltrative growth, destruction of the adjacent bone tissue, possible intracranial involvement. Because of scare symptoms, early GT diagnosis is problematic. At admission to hospital principal complaints of the children are poor hearing, discharge from the ear. These often underlie misdiagnosis -- acute otitis media. GT of the middle ear are detected with application of computed tomography which is able to show changes in the soft tissue and bone destruction, and angiography. Combination of the above two methods allows assessment of location and advance of the pathological process. GT treatment planning depends on the examination data and can include radiotherapy, endovascular occlusion of the vessels carrying blood to the tumor, surgical treatment and combination of the methods. Modern approaches to GT diagnosis and treatment are described. A case of GT of the temporal bone with involvement of the middle cranial fossa in a 4-year-old child is reported. PMID- 18163070 TI - [Infantile cerebral paralysis: modern approach to diagnosis and correction of hearing problems]. AB - The article describes clinical forms of infantile cerebral paralysis, provides the results of examination of the acoustic function in infants with introduction of modern techniques (otoacoustic emission, acoustic impedance technique, acoustic evolved potentials). PMID- 18163071 TI - [Effects of morphological changes in the middle ear mucosa on reparative processes in the neotympanic membrane in reconstructive sanative operation in patients with chronic otitis media]. AB - A complex clinicomorphological examination was performed in 39 patients with chronic otitis media (OM) who had undergone reconstructive sanative surgery. The analysis of the results of the above examination provided information on efficacy of the original method of semiquantitative assessment of local cell reactivity and qualitative composition of tissue elements (content of lymphocytes, plasmocytes, fibroblasts; the degree of development of the capillaries, fibrosis, stasis, edema, inflammation activity by neutrophil count). A retrospective analysis of tympanic membrane graft retention depending on morphological intraoperative condition of the middle ear mucosa and retrotympanic compartments enables prognosis of tympanoplasty outcome in chronic OM. PMID- 18163072 TI - [Changes in rigidity of the internal ear in body tilt]. AB - We tried to select information concerning changes in acoustic rigidity of the internal ear in 90% body tilt from tympanogram. In such conditions there is no correlation between a 10-fold rise of perilymph pressure and tympanogram parameters. This contradiction suggests that rigidity of tympanogram is a total of rigidity of the ear canal part, middle and internal ear; that inert rigidity of the middle ear in body tilt masks a weak dynamic component of the internal ear. Determination and isolation of the ear canal rigidity from tympanogram (compensation) is a standard procedure. For compensation of the dominating static component of the middle ear we used an original technique -- deduction of the numbered tympanograms. We estimated that deduction of the initial position tympanogram from subsequent tympanograms yields the dynamic component. The procedure of deduction for the tilt angles +75, 60, 45, 30, 15, O, -15 and -30 produced transformed tympanograms (TT) the rigidity of which was 7-10 times less that peak rigidity of standard tympanograms. Peak rigidity of TT of 5 volunteers (n=8) changed as the function of a tilt angle and significantly differed in different positions. TT rigidity may be formed by peripheral cochlear membranes. PMID- 18163073 TI - [Objective methods of fitting speech processors of cochlear implants Combi-40/40+ and Tempo+: impedance technique]. AB - Impedance measurement as an objective method of hearing examination is widely used in cochlear implantation. Stapedial reflex is measured intraoperatively and during the procedure of cochlear implant fitting. It was found that reflex threshold levels and most comfortable levels vary significally in one patient and between all patients. Thus, reflexometry is a convenient method of fitting speech processors of cochlear implants in children but the subjective method of fitting is more effective. PMID- 18163074 TI - [State of the larynx in children with congenital cleft lip and palate]. AB - Organic dysphonia was diagnosed in 27.4% of 215 children with congenital cleft lip and palate aged from 1.5 to 15 years. The above malformation was associated with such organic laryngeal diseases as edema, hypertrophy, nodules of the vocal cords and chronic laryngitis incidence of which directly correlates with duration and severity of the defect, age of the patient and time of reestablishment of the palatopharyngeal ring. After plastic repair of the oronasal defect, with age, laryngeal pathology gradually relieves. PMID- 18163075 TI - [Clinicocytological assessment of nasal mucosa and buccal epithelium in coal miners]. AB - The aim of the study was cytomorphological examination of the cells of nasal mucosa and buccal epithelium in coal miners. Two groups of miners were examined: non-exposed and exposed to industrial dust. It was found that coal dust inhalation provokes dystrophic processes in the upper airways, impairs intracellular regeneration resulting in atrophic changes in nasal mucosa epithelium. Morphometry (rhinocytogram) with estimation of cell composition in the upper airways of miners evaluates a cleaning function and respiratory system defense in long-term exposure of the miners to occupational dust hazards. PMID- 18163076 TI - [Nasal respiratory function in acute trauma as shown by anterior active rhinomanometry]. AB - Anterior active rhinomanometry (AARM) was used for objective investigation of nasal respiration in patients with nasal bone fractures. Twenty one patients with nasal septum deformation have undergone reconstructive septorhinoplasty in acute trauma, 33 patients -- standard nasal bone reposition. AARM proved its efficacy as a simple and convenient method of nasal obstruction assessment in acute trauma and can be successfully applied for determination of therapeutic policy and control of surgical intervention. Most patients with nasal bone fracture had preoperative low total volume flow and elevated nasal resistance. Nasal septum curvature and deformation of external nose structures are of importance in development of the pathology. One-stage reconstruction of the nasal septum and pyramid in acute trauma brings about significantly better long-term functional results compared to conventional closed nasal bone reposition. PMID- 18163077 TI - [Use of biomaterial alloplant and homeostatic material surgicel in reconstructive laryngeal and tracheal surgery]. AB - Posttraumatic stenosis of the larynx, cervical and upper choracic parts of the trachea was treated surgically using biomaterial (cartilage) alloplant and hemostatic material sergisel with an antibacterial effect. Cartilage implantation into the lateral tracheal walls was made in 10 patients, creation of the lateral and anterial walls of the larynx and trachea was made in 4 patients. Short before the cartilage placing in the tissues it was packed with material sergisel. Further adequate dilatation prosthetic reconstruction of the trachea for 3-4 months with application of silicon T-tube allowed complete decanulation of 7 patients (50%) -- they have undergone plastic surgery of the anterior tracheal wall. The rest 7 patients are at the stage of dilatation of the tracheal passage with a good response to therapy. Thus, a positive treatment result of alloplant and sergisel (implant and hemostatic materials, respectively) allows us to recommend these materials for reconstructive surgery of hollow organs of the neck. PMID- 18163078 TI - [Potentialities of conservative therapy of vocal disorders in children]. AB - The article concerns problems of classification and treatment of various vocal problems in children, presents treatment outcomes in patients with functional and organic dysphonia using complex homeopathic drugs. PMID- 18163079 TI - [A method of reconstructive surgery in persistent nasal scoliosis]. AB - The author proposes a novel method of reconstructive surgery in persistent nasal scoliosis. The device for conduction of the proposed technique has been developed. From 1976 to 2006 this surgical method was used in 627 patients aged 14-65 years. Of them, 43 patients had clinical manifestations of acute or chronic sinusitis. The reconstructive operation was performed after arrest of sinus inflammation. One-stage functional operations on the paranasal sinuses were made in 27 patients with sinusitis. The author emphasizes low traumatism, blood loss, duration of the proposed operation as well as the absence of serious complications and positive functional and cosmetic results. The method can be easily mastered by otorhinolaryngologists with experience in rhinological operations. PMID- 18163080 TI - [A septorhinoplastic technique for correction of nasal septum deformation in children]. AB - The authors propose a septorhinoplastic technique for correction of nasal septum deformation in children consisting in creation of a right-angled fragment connected in the upper part with "maternal" tissue due to an uninterrupted cut of the cartilage. The technique was used in surgical treatment of 112 children aged 8-17 years. Most frequent indication for surgical intervention was a defect of nasal respiration. Early and late after the operation the nasal septum was flat without signs of flotation in 103 (92%) patients. Parameters of rhinopneumotaxometry and mucociliary clearance improved significantly. The conclusion is that such correction of the septal defects is sparing and improves function of the operated organ. PMID- 18163081 TI - [Stepwise therapy of allergic rhinosinusitis]. AB - After examination of 240 patients with allergic rhinosinusitis (ARS) of whom 60 had edematose ARS and 60 edematose ARS associated with bronchial asthma (BA) treated unsuccessfully with topic glucocorticosteroids the patients received intranasal diprospan by an original scheme. Sixty patients with an initial stage of polypous ARS and associated ENT pathology and 60 ARS patients with hypertrophic mucosa of the middle and/or lower nasal cavities and unfavourable variants of intranasal anatomy were operated (functional endonasal interventions). A retrospective analysis of the efficacy of the therapeutic diagnostic algorithm of treatment policy showed that 92% patients benefited from the above treatment. PMID- 18163082 TI - [Efficacy of CO2 laser in the treatment of patients with chronic tonsillitis]. AB - Efficacy of surgical treatment of patients with chronic tonsillitis using CO2 laser was evaluated by complaints, recurrence rate, local cellular immunity, clinical examination. Follow-up of 50 patients with chronic tonsillitis after ablation of the palatine tonsils by means of CO2 laser demonstrated that palatine tonsils ablation with CO2 laser is an effective method of chronic tonsillitis treatment as a pertistent positive effect was achieved after this operation: the absence of long-term recurrences and complaints, normalization of cytological picture. PMID- 18163083 TI - [Current features of a mastoiditis course]. AB - A clinical course of mastoiditis according to recent literature data is characterized, therapeutic policy is discussed. The results of mastoidectomies made in the ENT clinic of the Moscow State Medical University in 2004-2007 are reviewed retrospectively. PMID- 18163084 TI - [Combined endolaryngeal videoendoscopic surgery and photodynamic treatment of patients with recurrent laryngeal and tracheal papillomatosis]. AB - Two-stage combined treatment of chronic recurrent papillomatosis of the larynx and trachea has been designed and tested in P.A. Herzen Research Cancer Institute. Stage I of the treatment consisted in endolaryngeal videoendoscopic surgery with Nd:YAG-laser destruction, argon-plasma coagulation and electroresection of the papilloma; stage II - postoperative photodynamic therapy (PDT) to prevent recurrence. In 1995-2007 the treatment was given to 32 patients aged 10-66 years with recurrent papillomatosis of the airways with the disease history 5-58 years. In two thirds of the patients papillomatosis involved several parts of the larynx and trachea. Squamous cell papilloma was accompanied with dysplasia of the first-second degree in 10 (31%) patients, dysplasia of the third degree - in 4 (12,5%), cancer in situ - in 3 (9,4%) patients. Human papilloma virus (HPV) was detected by hybridization in situ in 96%. The course of the treatment resulted in a complete regression (CR) of papilloma in 25 of 32 (78%), partial regression in 7 patients. The recurrence-free interval averaged 32 months (maximal 7 years) in 14 of 25 patients with CR. HPV was eradicated in a group of patients with persistent clinical remission. A 6 to 19 month follow-up recorded papilloma recurrence in 11 patients. The recurrence-free period increased 2,5 fold. In patients with dysplasia of degree I-III and cancer in situ (n=17) CR of dysplasia and preinvasive cancer foci was achieved in 15 (88%) patients. PMID- 18163085 TI - [Rhinological aspects of endoscopic endonasal removal of hypophysial adenomas]. AB - The authors analyse rhinological aspects of endoscopic endonasal transsphenoidal adenomectomy (EETA): a nasal operation, intra- and postoperative liquorrhea, liquorrhea-associated meningitis, postoperative nasal hemorrhage, local changes of nasal cavity anatomic structures; provide practical recommendations on management of the conditions many of which are life-threatening. PMID- 18163086 TI - [Sentinel lymph nodes in squamous cell cancer of the head and neck]. AB - Twenty five patients with squamous cell oral cancer and without clinical evidence of regional cervical metastases have undergone 99mTc lymphoscintigraphy of cervical lymph nodes to localize the sentinel node 24 hours before surgery. Intraoperative localization of the sentinel node with gamma-probe (the probe was removed afterwords) was followed by operation on the primary focus and radical neck fat dissection. The removed tissues were examined histologically. The sentinel nodes were detected in 22 (88%) patients. A total number of the sentinel lymph nodes was 44. Six patients had metastases in the sentinel nodes, in 3 of them metastases were detected in the removed fat. The results of the study show that the technique is informative in localization of the sentinel lymph node. PMID- 18163087 TI - [Endoscopic laser surgery in combined nasopharyngeal and middle ear pathology in children with conductive hypoacusis]. AB - Endoscopic laser surgery on lymphoid structures of the nasopharynx near the pharyngeal opening of the auditory tube was made with Lazon-10P laser in 67 children aged 3 to 14 years with documented exudative otitis media (EOM). All the children had conductive hypoacusis. Children who had tympanogram of type B and in whom otoscopy detected exudate behind the tympanic membrane were subjected to one stage laser tympanostomy in the anteroinferior quadrant of the tympanic membrane followed by transtympanic drug introduction into the tympanic cavity. Tympanostoma closed spontaneously 3 weeks later. Normal hearing recovered in all the children. Diagnostic endoscopy of the nasopharynx can reveal causes underlying dysfunction of the auditory tube. The results demonstrate high efficacy of the methods allowing elimination of mechanical occlusion of the auditory tube in the region of its pharyngeal opening with one-stage intervention on the middle ear. PMID- 18163088 TI - [New potentials in assessment of the acoustic analyzer in occupational selection and certification of drivers]. AB - As there is a need now in modern, informative and objective methods of diagnosis of early changes in the acoustic analyzer of drivers exposed to intensive occupational noise. To develop such methods, we studied parameters of evoked otoacoustic emission with reference to age of the drivers and duration of occupational exposure to noise. We established characteristics of delayed evoked otoacoustic emission and evoked distortion product otoacoustic emission. These parameters can be used in occupational selection and certification of transport drivers as markers of initial signs of a negative noise effect on the hearing organ. PMID- 18163089 TI - [Characteristics of a negative effect of aviation noise on hearing organ of aircraft maintenance personnel]. AB - Otolaryngological examination was carried out of 80 engineers and technicians engaged in maintenance of aircrafts in the airports. They are continuously exposed to occupational aviation noise and therefore are at risk for chronic neurosensory hypoacusis. Pure tone audiogram registers in them a rise in hearing thresholds throughout the whole band of frequencies studied. This means that aviation maintenance staff is exposed to noise containing highly intensive infrasound and high-frequency components. The detected pathology directly correlated with duration of exposure to noise. Development of neurosensory hypoacusis in aviation maintenance specialists starts earlier than in other specialists exposed to noise. PMID- 18163090 TI - [Modern views on diagnosis and therapeutic policy in non-purulent pathology of the internal ear]. AB - The authors review pathogenesis, clinical characteristics, modern pharmacological, physical and surgical treatment of patients with non-purulent diseases of the internal ear: Menier's disease, cochleovestibulopathy in vertebro basilar insufficiency, benign paroxysmal postural vertigo, various forms of neurosensory hypoacusis. PMID- 18163091 TI - [Transmeatal controlled barohydrodynamic diagnosis of otitis media with perforation of the tympanic membrane]. AB - The article presents a novel method of topical diagnosis and treatment of acute and chronic otitis media purulenta with perforation in the tympanic membrane; describes indications and contraindications for the method application, its technique with use of two devices: otobarohydroscope and transmeatal catheter. The treatment was performed in a group of patients with acute otitis media purulenta at the stage of tympanic membrane perforation complicated with exudative serous mastoiditis. Transmeatal controlled barohydrodynamic therapy proved its high efficacy in topical diagnosis and therapy of patients with the above condition. PMID- 18163092 TI - [Efficacy of implants in stapedoplasty]. AB - Stapedoplasty efficacy was compared in use of one of the following implants: Shea, Schuknecht and K-piston. Two stapedoplasty techniques were also compared: stapedectomy and stapedotomy. The analysis of 279 cases of tympanic and mixed otosclerosis shows that the greatest number of complications (neurosensory hypoacusis, deafness, postoperative tympanic granulomas) occurred in use of Shea prosthesis (200 implantations) both in stapedectomy and stapedotomy. Schuknecht (n=56) and K-piston (n=23) implants caused no complications. Postoperative audiological examination showed better functional results in patients treated with stapedotomy (98%) versus stapedectomy (92%). PMID- 18163093 TI - [General oxygen supply in males with rhonchopathy]. AB - Supply, transport and release of oxygen in tissues were studied in 100 patients with rhonchopathy. Blood was obtained from the lobe of the ear and was examined on the gas analyzer (348 model). The results of the study show that respiratory hypoxemia is characteristic for rhonchopathy, hypoxemia is compensated with gemic factors, nocturnal hypoxemia leads to hypoxia in 7% patients with rhonchopathy. PMID- 18163094 TI - [Usage of platelet-enriched plasma as hemostatic and analgenic medication in tonsillectomy]. AB - The rate of posttonsillectomy hemorrhage, pain severity were assessed in the study group (80 tonsillar niches) and in the control group (110 tonsillar niches). Treatment of postoperative tonsillar niches with platelet-enriched plasma (PEP) was performed in the study group and was not performed in the controls. No hemorrhage was registered in the study group and 9 cases of hemorrhage occurred in the control group. Swallowing test and analysis of the visual-analogue scales demonstrated that pain after tonsillectomy was less severe after local use of PEP. Thus, usage of PEP in the region of the postoperative wound is effective in prevention of hemorrhage and pharyngeal pain attenuation after tonsillectomy. PMID- 18163095 TI - [Enforcing osseointegration of dental implantates spray-coated by bioceramics with the help of hyaluronic acid and hydroxyapatite gel in experimental conditions]. AB - Influence of HAP-gel (2 g of 2% solution of hyaluronic acid mixed with 0,5 g of hydroxyapatite and 0,1 ml of colloidal silver) upon osseointegration in case of delayed introduction of titanium implantates in dog jaw. By scanning electron microscopy it was shown that solely use either of HAP-gel or of ceramic spraying increased direct contact area between bone and implantates in the 6 and 9 months time period. Combination of spray-coated ceramic with HAP-gel was effective in 3 months after implantation, when solely the HAP-gel or the ceramic spraying were little effective. In the following terms of experiment (6 and 9 months) significant differences between groups 3 and 4 (implantate with ceramic spraying but without HAP-gel in the alveolus and implantate with ceramic spraying and with HAP-gel in the alveolus) were not found. The area of implantate integration with jaw bone (cortical part of it was excluded) was equal to 80% and was maximal for the given conditions of the experiment. PMID- 18163096 TI - [Experimental substantiation of hormone replacing therapy by estrogens in treatment and prevention of inflammatory infectious diseases of maxillo-facial region]. AB - Estrogen play the important role in normal current of inflammatory process. At age oppression of synthesis or at surgical menopause in an organism it is formed estrogen-deficiency syndrome shown in particular, in salivary glands change, loss of a jaw bones, disseminated current of inflammatory processes. For study of hormone replacing estrogen's therapy role in experiment, we determine the influence of transdermal estrogen-consisting systems on structure and function of large salivary glands, density of a bone jaws and current of infectious inflammatory process in laboratory animals. For hormone-replacing estrogen therapy we used an equivalent doze of estrogen-consisting plaster with a doze of estradiol 0.32 mg per day. The plaster pasted once a week on the ear. Criteria of efficiency were: calculation of bone destruction areas, pathology study of glandulars parotid and densitometries. The received experimental data testify to efficiency of application hormone-replacing estrogen's therapy in inflammatory diseases of maxillofacial area proceeding on a background of estrogen-deficiency. PMID- 18163097 TI - [Study of biomimetic apatite formation on dentine surface]. AB - The aim of this study was investigation of the opportunity of biomimetic growth of apatite on a dentine surface at various methods of its processing. Artificial blood human plasma--simulated body fluid (SBF)--was used as a source of ions. According to the scanning electronic microscope (SEM) and confocal laser scanning microscope (CLSM) and energy dispersive X-ray analysis (EDX) they have revealed the growth of crystals of calcium--deficient apatite. The structure of the formed apatite layer differed depending on a kind of processing of a dentine surface. After acid etching the globules of the apatite are located chaotically and the part of a dentine surface remains free. The removal of the collagen fibres by NaClO promotes uniform, controllable growth of crystals, forming a monolithic layer. The hypermineralisated areas of a tooth create the best conditions for growth. The transformation hydroxyl apatite in a superficial dentine layer can be caused by increasing of pressure at preparing with diamond bur. The growth of crystals of brushite in the smear layer is possible at a storage in water at 37 degrees capital ES, Cyrillic. The crystals have lamellar form and are well integrated in a dentin surface. PMID- 18163098 TI - [Study of the chronic apical periodontitis influence upon the patient's organism status]. AB - During clinical-immunological study of the patients with chronic apical periodontitis (CAP) some differences were found between the following categories- CAP involving not more than 2 teeth and plural CAP. There were found the changes of both general immunity indices and its local components. Chronic inflammation course in periodontal tissues led to imbalance between cellular and humoral local immune reactions. It was shown that the presence of protracted multiple infectious focuses in periodontal tissues led to organism sensitizing and secondary immune deficiency. PMID- 18163099 TI - [Changes of gustatory perception in elderly patients with type II sugar diabetes. Surgical section]. AB - In 73 elderly patients with type II sugar diabetes there were found changes of gustatory perception of sweet that were most pronounced on primary stage of the disease and that can be considered as manifestation of diabetic neuropathy of oral cavity organs and tissues. PMID- 18163100 TI - [Experience in making up for the defects of mandible with titanium reconstructive plates]. AB - A brief review of the adoption and the use of reconstructive titanium plates (RTP) to make up for the defects of the mandible is presented in the article. 33 patients aged from 19 to 63 were observed. The defects of various areas of the mandible from 3 to 15 cm were made up with RTP. In 10 patients there were combinations with other materials. The periods of observation were up to 8 years. Absence of complications during the whole period of observations is stated. Secondary osteogenesis induction has been noted as a result of RTP interaction with the bone and the periosteum. Attention is paid to the necessity to observe technical requirements of RTP application. It allows to use the plates in making up for the defects of mandible (including vast defects) in the course of long (not less than 8 years) periods. Respectivity of the study of conditions and factors influencing optimum reparative regeneration of bone tissue surrounding RTP is conformed. PMID- 18163101 TI - [Peculiarities of the use of surgical preauricular approach to TMJ in cases of traumas]. AB - Preauricular approach to the temporomandibular joint is the ideal approach in patients with high mandibular condyle fractures and for most arthroplastic procedures. The article presents indications and our modifications of this surgical procedures. Clinical example of surgical treatment of the perforation of the meniscus by preauricular approach is also presented in the article. PMID- 18163102 TI - [Oxidative stress in victims with heavy combined cranium-facial trauma]. AB - For acute posttraumatic period of heavy combined cranium-facial trauma (CFT) considerable activation of peroxide lipids oxidation in the liquor is typical beginning from the 1st day of posttraumatic period on the background of speedy and drastic depletion of fermentative and low-molecular antioxidant liquor system (in spite of introduction of antioxidants particularly a-tocopherol acetate). Non adequate functioning of the system of antioxidant defense on the background of free radical activity splash can be considered as breakdown of the process of adaptive reaction forming. It leads to weighting the course of posttraumatic period of heavy combined cranium-facial trauma and its outcome as a whole. PMID- 18163103 TI - [Experience of the osteoplastic material Osteoplast-K use for surgical interventions on parodontal tissues]. AB - Information of the composition and properties of osteoplastic material based on sulphated glycosammoglykans (Osteoplast-K) is given, results of the material use in cases of surgical interventions on parodontium are presented. PMID- 18163104 TI - [Peculiarities of visual perception of dentition and smile aesthetic parameters]. AB - As the result of the studies it was determined in which limits the dentition central line displacement from the face middle line and the change of smile line tilt angle become noticeable for visual perception. And also how much visual perception of the dentition aesthetic parameters were differed in doctors with different experience, dental technicians and patients. PMID- 18163105 TI - [Morphological description of distraction regenerates in children with congenital and acquired defects and facial skeleton bones hypoplasia]. AB - Results of the morphological study of regenerates received after distraction upper and lower jaws in children were described. It was established that maternal bone and regenerate (retention during 3 months) in children of different age groups varied according to the stage of their structures differentiation; regenerate bone tissue was always less mature than maternal bone irrespective of its localization; lag in differentiation of new growth regenerate bone structures in the group of patients with congenital pathology if compared with the same indicator in the group of children with acquired pathology. PMID- 18163106 TI - [Clinical value, diagnosis and treatment of thyroid differentiated microcarcinoma]. AB - Results of surgical treatment of 240 patients with thyroid differentiated microcarcinoma (less 1 cm in diameter) are analyzed. Papillary and follicular tumor was diagnosed at 212 (88.3%) and 28 (11.7%) respectively. Microcarcinoma had no clinical symptoms in majority cases. In some cases microcarcinoma had aggressive clinical course: at 10.8% patients the tumor was extended beyond the bounds of thyroid capsule (T3), at 2.1% patients the primary symptoms of disease were metastases into cervical nodes. Long-term results of surgical treatment were followed-up until 25 years after surgery, 5-year survival rate and recurrence free period were 99.5 and 99% respectively. Recurrence of tumor occurred at 2 (0.8%) patients with initial III and IVA stages of disease despite previous radical operation, one of them died in 4 years after surgery. Microcarcinoma is regarded as early stage of thyroid gland cancer. Ultrasonic examination and fine needle biopsy permit to diagnose timely this malignant tumor. PMID- 18163107 TI - [Diagnosis and treatment of lung injuries]. AB - Overall 1218 patients with lung injuries were treated during 11 years. The rate of deep wounds and injuries of central zone and root of lung was 24.9%. Endoscopic and x-ray methods (especially computed tomography) were used for diagnosis. Surgical tactics depended on type of injury. Resection of lung and pneumonectomy were performed at 10.2% patients, closure of lung wound--at 41.3%. Surgical treatment of lung wound with dissection and revision of wound canal was performed at 42.6% patients with deep injuries. Classified surgical tactics permits to reduce the rate of postoperative pulmonary complications from 62 to 11.6%, and lethality--from 11.2 to 3.1%. PMID- 18163108 TI - [Endoscopic examination for diagnosis of cancer of major duodenal papilla]. AB - Endoscopic procedures were used for diagnosis and treatment of cancer of major duodenal papilla (MDP) at 31 patients. Gastroduodenoscopy (with visual assessment) and endoscopic retrograde cholangiopancreatography were performed at all the patients. Morphological study of tissue sampling permitted to confirm preliminary visual diagnosis only in half of patients. Detection of x-ray symptoms and bile passage disorders at retrograde cholangiopancreatography were high informative. All the patients underwent endoscopic papillosphincterotomy which was effective at 26 (83.9%) patients. PMID- 18163109 TI - [Treatment of bile ducts injuries after laparoscopic cholecystectomy]. AB - Features of bile ducts injuries after laparoscopic cholecystectomy were analyzed at 54 patients. High level of lesions (H. Bismuth III-IV type) was revealed at 80% cases. Along with mechanical (transsection, excision, clipping) thermal injury of the bile ducts is usual at laparoscopic cholecystectomy. The injury was diagnosed during the operation at 21 (37.7%) cases, in 2-30 days after operation- at 30 (56.6%), and in 3 months--2 years--at 3 (5.7%) patients. Surgical tactics depended on level and terms of injury, state of bile duct wall and complications. Biliodigestive anastomosis with Roux techniques and transhepatic drainages was the main surgical method (76% cases). Peritonitis and other severe complications required two-stage treatment. Overall 40 (74.1%) patients followed-up more 3 years; there were no cases of stricture recurrence. PMID- 18163110 TI - [The ways for reduction of complication rate after laparoscopic cholecystectomy at acute cholecystitis]. AB - Results of 1831 laparoscopic cholecystectomies (LCE) are analyzed. Symptoms of obstructive jaundice were at 170 (9.3%) patients. All these patients underwent endoscopic papillosphincterotomy before LCE for removal of bile hypertension. Conversion to open surgery was necessary in 44 (2.4%) cases. Intraoperative injuries of extrahepatic bile ducts occurred at 5 (0.27%), bleeding--at 10 (0.6%) patients. Postoperative complications were seen at 36 (2.0%) patients including intraabdominal bleeding (4), drainage bile less (8), subhepatic abscess (4), epigastric wound infection (16). There were 2 (0.1%) lethal outcomes. Accurate surgical technique and timely conversion to open surgery prevent complications of LCE. PMID- 18163111 TI - [Diagnosis of hepatic failure at the patients with abdominal sepsis]. AB - Results of prospective study of 398 patients with abdominal sepsis are analyzed. It is demonstrated that acute hepatic failure occurs at 45% of these patients and it is direct cause of death at 28% of them. Blood level of triglycerides and alkaline phosphatase are reliable markers of hepatic dysfunction at abdominal sepsis. Concentrations of these markers are associated with severity of patients and systemic inflammation syndrome, and also with the number of performed laparotomies. PMID- 18163112 TI - [Extended D3 lymphadenectomy in surgical treatment of stomach cancer]. AB - Results of 178 operations with extended D3 lymphadenectomy (study group) performed at the patients with stomach cancer are analyzed. Control groups consisted of 175 patients undergone operations with D2 lymphadenectomy, and 266 patients undergone surgery of typical extension. Study and control groups were similar according to prognostic factors. Indications for extended lymphadenectomy are formulated. It was demonstrated that short-term results in both group were equal. Safety of extended D3 lymphadenectomies wasalso confirmed. Extended D3 lymphadenectomies permitted to increase the respectability on 6.7%. PMID- 18163113 TI - [Association of surgical treatment and oxygen active forms with fertility at patients with varicocele]. AB - Study group consisted of 115 patients with varicocele undergone laparoscopic clipping of left seminal vein, control group--121 patients after Ivanisevich's operation. Chemiluminescence of seminal fluid with detection of oxygen active forms were studied at all the patients before surgery, in 14 and 72 days after it. Volume of ejaculate, concentration, mobility and morphology of spermatozoa were also examined. It is demonstrated that level of oxygen active forms may be regarded as criteria of efficacy of surgical treatment. Laparoscopic clipping of left seminal vein demonstrates better results (less postoperative complications and recurrences) compared with Ivanisevich's operation. PMID- 18163114 TI - [Combination of minimally invasive technologies in the surgery of nephrolithiasis]. AB - Overall 62 patients coral calculi underwent percutaneous nephrolithotripsy and distant shock-wave lithotripsy (DSWL). Percutaneous nephrolithotripsy as monotherapy was effective at 47 (75.8%) patients. At 15 (24.2%) patients the residual calculi were fragmented with DSWL. Obtained positive results permit to recommend percutaneous nephrolithotripsy both as monotherapy and in combination with DSWL for the treatment of patients with coral nephrolithiasis. PMID- 18163115 TI - [Autohemotransfusion in multifield hospital]. AB - Autohemotransfusions were used at 120 patients before surgery and in first day after it. Control group of patients was treated with donor blood components. Level of hemoglobin and hematocrit was controlled in both groups. It was demonstrated that these parameters returned to normal level on 5-7 days earlier at the patients of study group (with autohemotransfusions) than at ones of control group. PMID- 18163116 TI - [The hand-assisted laparoscopic resections of colon's left parts]. AB - Authors have an experience of 88 hand-assisted laparoscopic procedures. Results of 32 hand-assisted resections of colon's left parts are analyzed. The special original sealing device was used for making of pneumoperitoneum during laparoscopic procedures. The study group consisted of 19 female and 13 male patients. Mean age was 60.0 +/- 9.8 years (42-76 ys). Overweight was seen at 23 (72%) patients, 12 of them had obesity of I-III stage. Previous abdominal operations have been performed at 11 (34.4%) patients. Mean time of surgery was 181 +/- 53 min, operative bleeding--92 +/- 65 ml, no intraoperative complications occurred. Length of minilaparotomy was 7.3 +/- 0.8 cm. Poltoperative complications were seen at 4 (12.5%) cases, there were no lethal outcomes. Mean hospital stay was 9.8 +/- 3.5 bed-days. PMID- 18163117 TI - [Prediction of surgical outcome for thoracoabdominal aortic aneurysm]. AB - Prediction model for lethal outcome after operation for thoracoabdominal aortic aneurysm (TAAA) has been constructed based on assessment of preoperative risk factors. The main risk factors of lethal outcome at TAAA repair are: Crawford's operation (OR 12.25), deep hypothermia and circulatory arrest (OR 10.86), renal failure (OR 6.8), coronary heart disease (OR 2.49), chronic non-specific lung diseases (OR 2.29), age >50 years (OR 2.18), TAAA of II type by Crawford (OR 2.12). The prognostic system permits to access individual risk with high accuracy including at the patients with combination of different risk factors. PMID- 18163118 TI - [Late repeated operations after correction of heart isolated septal defects]. AB - Overall 812 patients with heart isolated septal defects have been operated. Repeated operations were performed at 23 patients. The follow-up ranged 6 months to 15 years. The causes of repeated surgeries and structure of complications are analyzed in details. PMID- 18163119 TI - [Reconstructive surgery of prolonged and multifocal stenoses of trachea]. AB - Overall 553 patients aged 9 to 77 years with cicatrical stenoses of trachea were treated. The causes of stenoses were tracheostomy (345 patients), intubation of trachea (155), trauma (29), unknown (24). Surgical methods of treatment were used at 448 (81.0%) patients, endoscopic methods--at 105 (19.0%); overall 1184 operations have been performed. Transplantation of donor thyrotracheal complex has been performed at 1 patient with subtotal cicatrical stenosis of trachea. Combination of staged reconstructive and plastic operations with endoscopic procedures permits to achieve good functional results. Treatment of these patients should be performed at special medical centers which have all the necessary technologies. PMID- 18163120 TI - [Rational prophylaxis and treatment of infectious complications in kidney transplantation]. AB - Clinical and bacteriological assessment of purulent complications in kidney transplantation has been carried out. The main infectious agents were S. epidermalis, E. coli, E. faecalis. Based on these data the optimal antibiotic therapy has been used that permitted to reduce the rate of infectious complications from 41 to 25.3%. PMID- 18163121 TI - [Current trends in surgical treatment of peritonitis]. AB - Results of treatment using laparoscopic operations, traditional laparotomy with passive abdominal drainage and laparostomy at 1835 patients with peritonitis are analyzed. It is demonstrated that choice of surgical policy depends on ethiology, generalization of peritonitis, abdominal microphlora, degree of endogenous intoxication and organs failure, prognosis of disease. Lethality after laparoscopic operations was 1.8%, after traditional laparotomy--3.4%, after laparostomy--47.7%. Lethality at local peritonitis was 1.3%, at generalized- 13.8%, at postoperative--22%. General lethality was 7.4%. Experimental studies on 55 mongrel dogs demonstrated the advantage of mechanical suture at peritonitis that prevents insufficiency of anastomoses. PMID- 18163122 TI - [Treatment of duodenal fistulas]. AB - Successful treatment of non-formed duodenal fistulas using original obturative aspiration device is reported. This simple device is described in details. Overall 122 patients with external non-formed intestinal fistulas were treated, 11 (9.1%) of them had duodenal fistulas. Traditional methods of treatment were used at 5 of 11 patients; original method was applied at 6 patients. Four of five patients treated with traditional methods died at postoperative period due to various complications. All the 6 patients treated with original method cured completely. PMID- 18163125 TI - Copy-number analysis goes more than skin deep. PMID- 18163126 TI - Genomic rearrangements in the spotlight. PMID- 18163127 TI - BRCA1--sowing the seeds crooked in the furrow. AB - Breast tumors with deficiency in DNA double-strand break repair might be expected to show aneuploidy. A new study shows that microdeletions in PTEN, resulting in complete loss of PTEN protein, are signature lesions in these cancers, particularly those arising in BRCA1-mutation carriers. PMID- 18163128 TI - Basal bodies, kinocilia and planar cell polarity. PMID- 18163131 TI - The emerging landscape of breast cancer susceptibility. AB - The genetic basis of inherited predisposition to breast cancer has been assiduously investigated for the past two decades and has been the subject of several recent discoveries. Three reasonably well-defined classes of breast cancer susceptibility alleles with different levels of risk and prevalence in the population have become apparent: rare high-penetrance alleles, rare moderate penetrance alleles and common low-penetrance alleles. The contribution of each component to breast cancer predisposition is still to be fully explored, as are the phenotypic characteristics of the cancers associated with them, the ways in which they interact, much of their biology and their clinical utility. These recent advances herald a new chapter in the exploration of susceptibility to breast cancer and are likely to provide insights relevant to other common, heterogeneous diseases. PMID- 18163132 TI - Spermatoprotective activity of the leaf extract of Psidium guajava Linn. AB - BACKGROUND: The leaves of Psidium guajava Linn. (guava) contain several natural antioxidants. We therefore designed the present study to evaluate the effect of ethanol extract of guava leaves on gossypol-associated sperm toxicity in Wistar rats. METHODS: Animal groups 1, 2, and 3 (n=6 each) were treated orally with crude cottonseed oil to provide 14 mg/kg/d of free gossypol for 53 d. Besides, groups 1 and 2 rats were supplemented orally with 250 mg/kg/d and 500 mg/kg/d respectively of guava leaf extract (GLE) for the same period. Group 4 animals (control, n=6) received normal saline. RESULTS: No significant difference (P>0.05) occurred in the sperm count of group 1 rats compared to control. In animal group 2, significant increase (P<0.05) in sperm count occurred, as opposed to group 3 animals, where this parameter decreased significantly (P<0.05). Besides, mean values of 78 %, 82 %, 30 %, and 65 % respectively were obtained for sperm motility in animal groups 1, 2, 3, and 4. CONCLUSION: Our findings suggest that ethanol extract of guava leaves possesses beneficial effect on gossypol associated sperm toxicity, and may therefore enhance male fertility, possibly owing to its rich constituents of natural antioxidants. PMID- 18163133 TI - Congenital malformations in singleton and twin births in rural Nigeria. AB - BACKGROUND: The presence of a congenital malformation at birth is a cause of anxiety at an otherwise joyous occasion. Congenital malformations are a significant contributor to perinatal mortality. STUDY DESIGN: A retrospective study of external congenital abnormalities in singleton and twin births in rural eastern Nigeria over a 20 year period. RESULT: The incidence of congenital defects for all deliveries was 110.8/10,000 births. Of 1453 twins and 32206 singleton births, there were 58 and 315 congenital abnormalities, with incidence of 97.8/10,000 births and 399.2/10,000 births respectively. Twins were significantly (x(2) =115.22; p< 0.0000) more likely to have a congenital malformation than singletons (RR 4.08, 95% CI 3.10 - 5.7). The pattern of defects was similar for singleton and twin births and the leading system affected was the musculo-skeletal system, distantly followed by the central nervous system. For both groups the commonest malformation was ulnar polydactyly, followed by hydrocephaly and ankyloglossia. Surprisingly no conjoined twins were recorded and there were only 7 cases of congenital umbilical hernia, abnormalities previously considered to be very common in Nigerians and Africans respectively. CONCLUSION: Twins are about four times more likely to have congenital malformations than singletons. The overall prevalence of congenital malformations recorded is comparatively low. There is need for more detailed routine documentation of all birth defects including post-mortem report and the conduct of prospective population-based epidemiological surveys of birth defects in Nigeria. PMID- 18163134 TI - Triplet pregnancy as seen in the Jos University Teaching Hospital. AB - BACKGROUND: Triplet pregnancies are high-risk pregnancies and fortunately the prevalence is low. They are associated with high perinatal morbidity and mortality. DESIGN: All the records of the 16 triplet pregnancies managed in Jos University Teaching Hospital, North Central Nigeria, were retrieved and analysed. RESULTS: There were a total of 41,159 deliveries and 16 triplet pregnancies giving a prevalence of 1 triplet pregnancy in 2,572 deliveries. Over 60% of the mothers were above the age of 24 years, and about 94% were multipara. The female to male sex ratio was 1:0.85, that is, 1 female in 0.85 males. The pre-term delivery rate was 68.7% of the total triplet pregnancies. Booked patients had better foetal and maternal outcome compared with unbooked patients. Most (81.3%) of the women were allowed vaginal delivery. There was no significant difference between the average Apgar scores of babies delivered by caesarean section and vaginal delivery. All the patients had postpartum haemorrhage (a blood loss of 500 ml and above), but none of them required blood transfusion. CONCLUSION: Triplet pregnancies are not managed often in Jos University Teaching Hospital probably because they do not occur commonly on the Jos Plateau, Nigeria. PMID- 18163135 TI - Comparison of clinical, microscopic and rapid diagnostic test methods in the diagnosis of Plasmodium falciparum malaria in Enugu, Nigeria. AB - OBJECTIVE: To compare the use of clinical, microscopic and immunochromato-graphic methods in the diagnosis of Plasmodium falciparum malaria. METHODS: Children with history of fever were consecutively recruited. Information on personal characteristics, and clinical features were obtained with the parental consent before enrolment. Malaria parasite density was done with quantitative microscopy, as well as specie identification in both local and reference laboratories. Rapid diagnostic test (RDT) kits were also used for parasite detection. RESULTS: Eighty three children had clinical diagnosis of malaria, while the remaining 6 had other diagnoses. On microscopy 89 and 26 patients had parasitaemia by the local and reference laboratories respectively. With the RDT, 15 patients had parasitaemia with corresponding minimum densities of 1200 and 716 parasites per ul from the local and reference laboratories. The sensitivity and specificity of the RDT to the reference laboratory were 42.31% and 93.65% respectively, while concordance was 57.69% for RDT, 30.49% for clinical diagnosis (all symptoms), 43.75% for clinical diagnosis (fever alone), and 29.21% for local laboratory. CONCLUSION: There was a significant deficiency in malaria diagnosis using any of the methods alone. For reliable diagnosis of malaria a combination of the methods of diagnosis is recommended. Degradation of the RDTs might have contributed to its low sensitivity. Training and better quality assurance should be established for the medical and laboratory workers in order to improve malaria diagnosis. PMID- 18163136 TI - Intestinal parasitism in Magama Gumau rural village and Jos township in north central Nigeria. AB - OBJECTIVE: To determine the prevalence of intestinal parasitosis in one rural village and one urban centre in North Central Nigeria. METHODS: A total of 111 single stool specimens from all the volunteered rural dwellers and 93 specimens from randomly selected urban dwellers were examined using Formol-ether and modified Ziehl-Neelsen techniques; during the months of June and July 2005. A questionnaire was completed for each subject and the nutritional status of the adults was assessed using the anthropometric measurements (weight and height for age and Biomass index). RESULTS: The results suggest very high prevalence rates of intestinal parasitosis of 72.1% and 69.9% for the rural and urban populations respectively. All the age groups were infected. The males in the rural area had a prevalence of 69.2% as against 74.6% in females (P>0.05); while in the urban area, the males were more significantly infected (77.4%) compared with the females with 66.1% (P< 0.05). Those with normal BMI (18.5-24.9 kg/m2) had the highest prevalence of 79.3% and 72.4% for the rural and urban populations respectively. The prevalence of the parasites in the rural and urban populations respectively were: Entamoeba coli (16.2% and 9.7%); E. histolytica (18.9% and 18.3%); E. hartmani (1.8% ad 0.0%); Endolimax nana (16.2% and 18.3%); Iodamoeba butschlii (0.0% and 1.1%); Giardia lamblia (7.2% and 4.3%); Schistosoma mansoni (9.9% and 0.0%); Strongyloides stercoralis (0.9% and 0.0%); Hookworm (4.5% and 5.4%); Ascaris lumbricoides (1.8% and 0.0%); Enterobius vermicularis (0.0% and 1.1%); Cryptosporidium parvum (29.7% and 19.4%); and Enterocytozoon bieneusi/Encephalitozoon intestinalis (39.6% and 47.3%). Polyparasitism was recorded in 48.6% of the rural subjects and 36.6% of the urban subjects. CONCLUSION: The study has shown a very high prevalence of intestinal parasitosis in both the rural and urban populations and that C. parvum and E. bieneusi/E. intestinalis are harboured by apparently healthy individuals. PMID- 18163137 TI - Patients' perception of quality of emergency care in a Nigerian teaching hospital: The influence of patient-provider interactions. AB - BACKGROUND: Attention is rarely given to patients' opinions regarding the quality of care they received, which is an important feedback to healthcare providers, planners and policy makers. AIM: To assess how patients who survived life threatening/emergency conditions percieved the quality of care they received. METHOD: This prospective study was carried out among adult patients who had received emergency care at the Accident & Emergency (A & E) unit of Olabisi Onabanjo University Teaching Hospital (OOUTH), Sagamu, between March and December 2004 using a semi-structured questionnaire. RESULTS: There were 1129 respondents, 81 males and 48 females. Their mean age was 35.3 years. 62% were treated for surgical, and 37.2% for medical emergencies. The mean duration of stay at A & E was 2.4 days. Although 91% of the respondents regarded available equipments as very adequate, 38.8% perceived the overall quality of care as sub-optimal. Many of the patients were displeased with their interactions with care providers. They longed for urgent improvement in waiting time, speed of issuing drugs, imterpersonal relationship with health workers and attending to emotional distress of emergency victims. They also wished to have free treatment during emergencies CONCLUSION: Majority of the patients who received care in A & E of this tertiary hospital perceived the quality of care received as satisfactory. However, a substantial proportion regarded the quality of care as sub-optimal. Although most thought equipments were adequate, many of them expressed displeasure with their interactions with care providers. To improve patient's satisfaction with emergency care, greater emphasis needs to be placed on enhancing the interpersonal relationships between health workers and patients than is currently done. PMID- 18163138 TI - Pattern of the cases seen in the accident and emergency department in a Nigerian Tertiary Hospital over a period of twelve months. AB - INTRODUCTION: There is a dearth of information on emergency medical services in Nigeria. This study was conducted to determine the age, sex distribution and the pattern of patient presentation in the Accident and Emergency Department of a Nigeria teaching hospital. METHODS: A retrospective study of all cases seen at the accident and emergency department of the University College Hospital in 2003 was carried out. The information extracted from the records includes age, sex, and diagnosis, department to which the patient was referred, the month of presentation and the outcome within the first twenty four hours of presentation. RESULTS: A total of 4674 patients attended the casualty, with a male: female ratio of 1.2:1. The third decade was the peak age distribution. There was a predominance of surgical cases (61%). In the treatment outcome, 52.1% were referred to other departments while there were ten (0.2%) mortalities. Trauma related cases constituted 45.1%. Road traffic accidents were the commonest cause of trauma. CONCLUSION: The largest proportion of patient were in the active third decade of life. Trauma is the commonest cause of presentation in the Accident and Emergency Department. A significant proportion of patients do not require admission. The doctor in the Accident and Emergency Department must be skilled in basic trauma care. PMID- 18163139 TI - Pattern of abnormal liver enzymes in HIV patients presenting at a Nigerian Tertiary Hospital. AB - AIM: To demonstrate the pattern of abnormal liver enzymes in HIV patients presenting at the University of Port Harcourt Teaching Hospital, Port Harcourt. BACKGROUND: Abnormalities of liver function tests (LFT) have been shown to be common in HIV/AIDS in developed countries. Studies have shown that these abnormalities may be due to direct inflammation induced by the HIV virus on the liver cell. It may also be due to gall bladder disease and infection with bacterial, viral or other opportunistic agents. PATIENTS AND METHODS: We reviewed the clinical notes and biochemistry results of HIV positive patients presenting at the HIV clinic from January 2003 to October 2003. Efforts were made to elicit symptoms suggestive of hepatobiliary disease. Their hepatitis B surface antigen status and their hepatitis C status were obtained. Attempts were made to determine the pattern of liver injury in each patient seen. RESULTS: A total of one hundred and twenty -nine (129) cases were recruited. The mean age was 35.6 +/ 9.0 years with a male to female ratio of 1.1 : 1. One hundred and thirteen patients (87.6%) had abnormalities of their LFT'S. Sixteen of these (14.5%) were classified as cholestatic liver injury while ninety-four (85.5%) were classified as having hepatocellular injury. Three patients (2.3%) were HbSAg positive while one (0.8%) was hepatitis C antibody positive. None of the patients was on anti retroviral drugs at the time of recruitment. CONCLUSION: We therefore conclude that abnormalities of liver enzymes are common in patients with HIV in this environment. It is therefore important to characterise the nature of this abnormality and to institute appropriate management. However further studies are required in this field of HIV related liver disease in our locality. PMID- 18163140 TI - Problems of management of primary congenital glaucoma in Benin City, Nigeria. AB - OBJECTIVES: To determine the problems encountered in the management of primary congenital glaucoma in developing countries. METHODS: A prospective study of new patients with a diagnosis of primary congenital glaucoma attending the eye clinic of the University of Benin Teaching hospital, Benin City, Nigeria over a 5 year period, from July 2001 to June 2006. The age, sex, presenting complaints and duration were recorded. They were examined by the aid of a pen torch, hand-held slit lamp and direct ophthalmoscope. Intraocular pressure and corneal diameter were measured under anaesthesia. The patients were followed up till June 2006 and the problems encountered in management were recorded. RESULTS: Eight children comprising 5 males (62.5%) and 3 females (37.5%) with primary congenital glaucoma were seen. All the patients (100%) presented with buphthalmos and 7 (87.5%) had corneal opacity. Four (50%) accepted surgery but only 2 (25%) eventually had surgery. Maximum follow-up was 7 months. CONCLUSION: Presentation with features of advanced disease, poor acceptability of surgery, poor follow-up and low surgical rate are problems associated with the management of congenital glaucoma in Benin City, Nigeria. PMID- 18163141 TI - Early childhood caries: experience in Nigerian children at Lagos. AB - Objective of the paper was to assess the prevalence of Early Childhood Caries among Nigerian children aged 6 months to 5 years, describe the pattern and relate it to the socioeconomic status of their parents/guardians. METHODS: It was a descriptive study carried out in three randomly selected local government areas in Lagos State. Questionnaires designed to provide information on the socio economic status of the parents were completed by one of the authors. All the children were examined using sterile instruments for the presence of early childhood caries. RESULTS: The prevalence of early childhood caries amongst the children was 10.5%, among them 4.8% had severe early childhood caries. Early childhood caries was observed in children as early as 13 months. The primary incisors were most affected. Only 17 (4.4%) had visited the dentist. CONCLUSION: There was a tendency towards higher caries level in children of higher socio economic status. Preschool children had great demand for restorative care. PMID- 18163142 TI - The views of some selected Nigerians about mental disorders. AB - AIM: To determine the views of sampled subjects about mental disorders and their treatment recommendations of such disorders. METHODS: This is a cross sectional descriptive study. The subjects were asked to list the causes of mental disorders they know, explain what they think the role of evil spirits in mental disorders is, if any, and to name the treatment options available for these disorders. Medical students who participated in the survey were also re-administered the study questionnaire following their completion of four week's rotation in Mental Health. RESULTS: Medical students (30), nurses (20), medical records officers (20) and 64 others including 10 religious ministers participated in the study. Thirty-eight- (28.4%) respondents thought that evil spirits cause mental disorders, and 88 (65.7%) would recommend prayer houses for the treatment of mental disorders. Medical students, even after their 4 weeks of rotation in mental health, did not differ in their beliefs from the rest of the groups. CONCLUSION: The knowledge of most interviewees about mental disorders is based on religious beliefs without scientific foundation. Adequate training in psychiatry for medical students, interaction of psychiatrists with their non-psychiatrist counterparts, public enlightenment about mental disorders may be necessary to raise the level of awareness/general knowledge of, and reduce superstitious beliefs about psychiatric disorders. PMID- 18163143 TI - Factors influencing non-compliance with anti-hypertensive drug therapy in Nigerians. AB - BACKGROUND: Non-compliance with antihypertensive drug therapy can have a negative impact on cardiovascular outcome. The objective of this study was to determine compliance rate with anti-hypertensive therapy and factors affecting compliance among patients attending a Nigerian tertiary hospital. METHODS: A cross sectional study was conducted on 225 black hypertensive patients attending a tertiary clinic in Lagos, Nigeria. Demographic data, current anti-hypertensive therapy, compliance with prescribed therapy, factors affecting compliance and BP were documented via an investigator administered questionnaire and open-ended interviews. RESULTS: In this study, 77(34.2%) of the hypertensive patients were non-compliant. Blood pressure control was significantly better among compliant patients (45.9%) than in non-compliant patients (27.3%) (x(2) = 7.35 p=0.007). Gender, age, number of drugs used, educational level and presence of co morbidities did not affect compliance. The major reasons for non-compliance were miscellaneous factors (60%) related to both patient's attitudes and beliefs (reflecting ignorance),and consultation failure on the part of clinicians. Lack of finances and side effects of medications accounted for 23.8% and 16.2% of non compliances respectively. CONCLUSION: Although lack of finances is the single most self reported reason, miscellaneous factors related to patients' attitudes and belief contribute frequently. Incorporating patient education and counselling in routine follow-up may improve compliance, BP control, and ultimately impact positively on cardiovascular outcome. PMID- 18163144 TI - The use of thromboembolic prophylaxis by surgeons: A multicentre Nigerian study. AB - BACKGROUND: Despite the well known contributions of deep vein thrombosis and pulmonary embolism to perioperative deaths, these major causes of morbidity and mortality appear not to be given adequate attention by surgeons in our environment. OBJECTIVE: To study the practice of thromboembolic prophylaxis among surgeons in some sub-specialties in three Nigerian tertiary institutions. MATERIALS AND METHODS: A structured questionnaire was used to collect information from consultants in the surgical sub-specialties on the use of thromboembolic prophylaxis in their practice. RESULTS: Fifty nine adequately completed questionnaires were returned. Of this, only 28 (47.5%) used prophylaxis routinely in major surgeries. Use was most frequent in orthopaedics and least in ophthalmology and otorhinolaringology. Subcutaneous heparin was the most commonly employed agent and the most important indication for prophylaxis was a previous history of thromboembolism. CONCLUSION: The results obtained suggest low use of prophylaxis. Evidenced-based guidelines are available on antithrombotic and thrombolytic therapy with the benefits outweighing risks, burdens and costs. Therefore greater attention should be paid to antithrombotic measures by all the surgical sub-specialties and patients stratified according to risk. PMID- 18163145 TI - Perception of Nigerian internal medicine residents on the diagnosis and management of heart failure. AB - AIM: To evaluate the perception of internal medicine residents on diagnosis and management of heart failure in Nigeria. METHOD: A modified version of the Euro-F study questionnaire was administered to internal medicine residents (IMRs) from Nigeria who were attending the pre part I Fellowship examination of the Faculty of Internal Medicine of the National Postgraduate Medical College. Responses were compared with data from the Nigerian primary physicians (PCP) survey and the Euro HF study. RESULTS: The commonest symptoms used by the IMRs in heart failure diagnosis were exertional dyspnoea 68.85%(42), pedal oedema 63.93%(39) and orthopnoea 50.82%(31), while the signs included elevated jugular venous pressure 50.82%(31), basal crepitations 47.50%(29) and a gallop heart rhythm 39.34%(24). Categorisation using the Framingham criteria for diagnosis showed that 52% of the IMRs knew three or more major signs of heart failure. The IMRs use of investigations in diagnosing heart failure was fair to good and was significantly higher than results from the Euro-F study and the primary care physicians (PCPs) in Lagos study; echocardiogram, (p=0.007and <0.00001); electrocardiogram, (p= 0.0002 and p=0.001); chest x-ray (p=0.05 and 0.5) respectively. However the mean proportion of IMRs that would rely on investigation to make a diagnosis was significantly lower than in the Euro-F study (p=0.0001) and in the PCPs (p = 0.02). Although the mean proportion of the IMRs patients using ACE-inhibitors was significantly higher than in the Euro-F survey (p<0.001) and the PCPs (p <0.00001), majority (93.94%) of the IMRs were using less than half the trial doses for treatment. On the other hand, the knowledge of survival benefits with the use of b-blockers was very poor in the all the groups, p>0.05. CONCLUSION: A substantial knowledge gap still exists among the IMRs as regards the diagnosis and management of heart failure and this need to be addressed by the trainers. PMID- 18163146 TI - Mass casualty incidents and disasters in Nigeria: The need for better management strategies. AB - AIMS AND OBJECTIVES: The aim of this article is to discuss principles involved in disaster management, disasters in Nigeria, examine the current level of preparedness in the country and make recommendations for improvement. METHODS: An overview of various disaster events in the country coupled with review of the literature. RESULT: Fatality figures for disaster in Nigeria are high. CONCLUSION: There is need for a strong political will from government at all levels regarding disaster management in order to mitigate its occurrence and impact. PMID- 18163147 TI - A three-year review of mortality patterns in the medical wards of Aminu Kano Teaching Hospital, Kano, Nigeria. AB - BACKGROUND: The pattern of morbidity and mortality reflects the burden of disease in a particular community. This pattern shows geographic variations between communities and countries. The knowledge of the pattern of diseases and their contribution to mortality in a given country is very important in evaluating its health care delivery system. Such knowledge is important for health planning and for improving the healthcare services in that particular nation. We set out to study the mortality pattern in our medical wards. MATERIALS AND METHODS: This is a retrospective study that reviewed the causes of death in the medical wards of Aminu Kano Teaching Hospital (AKTH) Kano between January 2001 and December 2003 (three years). The data were obtained from the admission and discharge/death register, patients' case records, the weekly as well as quarterly mortality reviews. The SPSS version 10 was used to analyse the data. RESULTS: A total of 3369 patients were admitted over the 3 year study period. Of these 2518 (74.7%) were discharged or referred and 851 patients died, giving an overall mortality rate of 25.3%. The male to female admission ratio was 1.6:1. Majority of deaths, 714 (83.9%) occurred after 24 hours of admission. The most important causes of death were infectious diseases other than HIV/AIDS (17.9%), cerebrovascular disease (17%), HIV/AIDS (13.6%), chronic renal failure (12.5%) and diseases of the circulatory system (11.9%). CONCLUSION: Mortality in the medical wards reflects the emerging trend of mixed disease spectrum burden comprising communicable and non communicable diseases. Public health education, raising the socio-economic status of our people as well as improving the standards of our health care facilities and personnel would prevent a large proportion of deaths from medical wards. PMID- 18163148 TI - Severe gastrointestinal haemorrhage due to primary intestinal Kaposi's sarcoma - a case report. AB - Kaposi's sarcoma (KS) was previously a relatively rare disease. With the advent of HIV/AIDS pandemic however, AIDS-related KS has been on the increase and so has interest in the disease. Ninety per cent of patients with KS present with skin lesions. While the gastrointestinal tract is a fairly common site of metastatic KS, primary gastrointestinal KS is uncommon. The presentation of gastrointestinal KS with severe gastrointestinal bleeding is rarer still. In this report, we present a 56yr old HIV-negative patient who presented with severe gastrointestinal bleeding without any skin lesions. Multiple haemorrhagic polypoidal lesions were found on the walls of the jejunum and ileum as well as the liver at exploratory laparotomy and these were found to be KS on histopathologic examination. We also discuss the diagnostic and therapeutic challenges we had with this rare cause of severe GI bleeding. PMID- 18163149 TI - Endometrial carcinoma following treatment for breast carcinoma in a Nigerian female. A case report and review of the literature. AB - Tamoxifen is the primary hormonal therapy for breast cancer as well as its chemoprevention. It is used in the management of breast cancer because of its anti oestrogenic effect. It is however an agonist on the endometrium with undesirable effects of endometrial proliferation with the risk of endometrial hyperplasia and carcinoma. Several authors have reported cases of endometrial hyperplasia and carcinoma following tamoxifen therapy for breast carcinoma. No such report has been made from Nigeria . This paper presents the first case of endometrial carcinoma following tamoxifen therapy for breast carcinoma in a 52 year old Nigerian female. It also highlights the recommended guidelines for the follow up of women with breast cancer on tamoxifen therapy. PMID- 18163150 TI - Acute renal impairment following use of intravenous contrast agent: A case report and reminder for increased awareness of the problem. AB - OBJECTIVE: Contrast-induced nephropathy is one of the adverse reactions to the use of radiocontrast media. Its incidence can be reduced if preventive measures are instituted. This report presents a case of acute renal impairment following use of radiocontrast media. PATIENT, METHOD AND RESULTS: We report a case of an elderly hypertensive diabetic who developed acute renal impairment within 24 hours of undergoing an intravenous urography. He was not a known asthmatic and had no previous history of allergy. His renal function improved subsequently with hydration, diuretics and strict input/output monitoring. He was discharged after 23 days in hospital and renal function returned to pre-morbid state 3 months after discharge. CONCLUSION: The report is a reminder of the possibility of nephropathy associated with the use of intravascular contrast media. There is need for extra care to be taken to identify risk factors in patients that are to undergo intravascular contrast studies and adequate attention paid to these. PMID- 18163151 TI - Non-cirrhotic portal vein thrombosis, an uncommon cause of portal hypertension - a case report. AB - Most cases of portal hypertension follow chronic liver disease (cirrhosis) while non cirrhotic causes are occasionally seen. A case of portal hypertension secondary to non-cirrhotic portal vein thrombosis is reported. The patient was managed at the Aminu Kano Teaching Hospital Kano, Nigeria in the year 2006. She presented with recurrent massive upper gastrointestinal bleeding and was resuscitated, followed by clinical, radiological and endoscopic evaluation. She had massive splenomegaly and grade four oesophageal varices with evidence of recent bleed. There were, however, no other stigmata of chronic liver disease or portal hypertension. Abdominal computed tomography scan confirmed portal vein thrombosis. Splenectomy and ligation of short gastric veins was performed and the patient has not had any repeat episode of haematemesis or malaena six months post surgery. PMID- 18163152 TI - Simple Reaching Is Not So Simple: Association Between Hand Use and Grip Preferences in Captive Chimpanzees. AB - We assessed the relationship between grip preference and hand use in chimpanzees in 2 experiments. In experiment 1, we evaluated consistency in hand use and grip preference across 4 food types. The chimpanzees showed population-level right handedness and there are significant positive associations for both hand and grip use across food types. In experiment 2, we assessed validity of hand use in relation to grip preference in 2 colonies of chimpanzees via the same methodology. Differences in hand preferences between colonies were associated with variation in the observed grip preferences. There was no evidence of rearing effects on handedness in either colony. We discuss the overall results in the context of the evolution of handedness in relation to increasing motor demands as manifest in variation on grasping behavior. PMID- 18163153 TI - Masked Cross-Modal Repetition Priming: An Event-Related Potential Investigation. AB - We report three experiments that combine the masked priming paradigm with the recording of event-related potentials in order to examine the time-course of cross-modal interactions during word recognition. Visually presented masked primes preceded either visually or auditorily presented targets that were or were not the same word as the prime. Experiment 1 used the lexical decision task, and in Experiments 2 and 3 participants monitored target words for animal names. The results show a strong modulation of the N400 and an earlier ERP component (N250 ms) in within-modality (visual-visual) repetition priming, and a much weaker and later N400-like effect (400-700 ms) in the cross-modal (visual-auditory) condition with prime exposures of 50 ms (Experiments 1 & 2). With a prime duration of 67 ms (Experiment 3), cross-modal ERP priming effects arose earlier during the traditional N400 epoch (300-500 ms) and were also larger overall than at the shorter prime duration. PMID- 18163154 TI - Fluctuation and synchronization of gait intervals and gait force profiles distinguish stages of Parkinson's disease. AB - We study the effects of Parkinson's disease (PD) on the long-term fluctuation and phase synchronization properties of gait timing (series of interstride intervals) as well as gait force profiles (series characterizing the morphological changes between the steps). We find that the fluctuations in the gait timing are significantly larger for PD patients and early PD patients, who were not treated yet with medication, compared to age-matched healthy controls. Simultaneously, the long-term correlations and the phase synchronization of right and left leg are significantly reduced in both types of PD patients. Surprisingly, long-term correlations of the gait force profiles are relatively weak for treated PD patients and healthy controls, while they are significantly larger for early PD patients. The results support the idea that timing and morphology of recordings obtained from a complex system can contain complementary information. PMID- 18163155 TI - The Role of Recognition Memory in Anaphor Identification. AB - In studies of anaphor comprehension, the capacity for recognizing a noun in a sentence decreases following the resolution of a repeated-noun anaphor (Gernsbacher, 1989). In studies of recognition memory, the capacity for recognizing a noun in a scrambled sentence decreases following the recognition that another noun has occurred before in the scrambled sentence (Dopkins & Ngo, 2002). The results of the present study suggest that these two phenomena reflect the same recognition memory process. The results suggest further that this is not because participants in studies of anaphor comprehension ignore the discourse properties of the stimulus materials and treat them as lists of words upon which memory tests are to be given. These results suggest that recognition processes play a role in anaphor comprehension and that such processes are in part the means by which repeated-noun anaphors are identified as such. PMID- 18163156 TI - Light induced EPR spectra of reaction centers from Rhodobacter sphaeroides at 80K: Evidence for reduction of Q(B) by B-branch electron transfer in native reaction centers. AB - Photosynthetic reaction centers (RCs) from Rhodobacter sphaeroides capture solar energy by electron transfer from primary donor, D, to quinone acceptor, Q(B,) through the active A-branch of electron acceptors, but not the inactive B-branch. The light induced EPR spectrum from native RCs that had Fe(2+) replaced by Zn(2+) was investigated at cryogenic temperature (80K, 35 GHz). In addition to the light induced signal due to formation of D(+*)Q(A) (-*) observed previously, a small fraction (~5%) of the signal displayed very different characteristics: (1) The signal was absent in RCs in which the Q(B) was displaced by the inhibitor stigmatellin. (2) Its decay time (tau=6 s) was the same as observed for D(+*)Q(B) (-*) in mutant RCs lacking Q(A,) which is significantly slower than for D(+*)Q(A) (-*) (tau=30 ms). (3) Its EPR spectrum was identical to that of D(+*)Q(B) (-*). (4) The quantum efficiency for forming the major component of the signal was the same as that found for mutant RCs lacking Q(A) (Phi =0.2%) and was temperature independent. These results are explained by direct photochemical reduction of Q(B)via B-branch electron transfer in a small fraction of native RCs. PMID- 18163157 TI - Spatial Modelling Using a New Class of Nonstationary Covariance Functions. AB - We introduce a new class of nonstationary covariance functions for spatial modelling. Nonstationary covariance functions allow the model to adapt to spatial surfaces whose variability changes with location. The class includes a nonstationary version of the Matern stationary covariance, in which the differentiability of the spatial surface is controlled by a parameter, freeing one from fixing the differentiability in advance. The class allows one to knit together local covariance parameters into a valid global nonstationary covariance, regardless of how the local covariance structure is estimated. We employ this new nonstationary covariance in a fully Bayesian model in which the unknown spatial process has a Gaussian process (GP) prior distribution with a nonstationary covariance function from the class. We model the nonstationary structure in a computationally efficient way that creates nearly stationary local behavior and for which stationarity is a special case. We also suggest non Bayesian approaches to nonstationary kriging.To assess the method, we use real climate data to compare the Bayesian nonstationary GP model with a Bayesian stationary GP model, various standard spatial smoothing approaches, and nonstationary models that can adapt to function heterogeneity. The GP models outperform the competitors, but while the nonstationary GP gives qualitatively more sensible results, it shows little advantage over the stationary GP on held out data, illustrating the difficulty in fitting complicated spatial data. PMID- 18163158 TI - Protocol for emergency EPR dosimetry in fingernails. AB - There is an increased need for after-the-fact dosimetry because of the high risk of radiation exposures due to terrorism or accidents. In case of such an event, a method is needed to make measurements of dose in a large number of individuals rapidly and with sufficient accuracy to facilitate effective medical triage. Dosimetry based on EPR measurements of fingernails potentially could be an effective tool for this purpose. This paper presents the first operational protocols for EPR fingernail dosimetry, including guidelines for collection and storage of samples, parameters for EPR measurements, and the method of dose assessment. In a blinded test of this protocol application was carried out on nails freshly sampled and irradiated to 4 and 20 Gy; this protocol gave dose estimates with an error of less than 30%. PMID- 18163159 TI - EPR dosimetry in chemically treated fingernails. AB - By using EPR measurements of radiation-induced radicals it is possible to utilize human fingernails to estimate radiation dose after-the-fact. One of the potentially limiting factors in this approach is the presence of artifacts due to mechanically induced EPR signals (MIS) caused by mechanical stress during the collection and preparation of the samples and the so-called background (non radiation) signal (BKS). The MIS and BKS have spectral parameters (shape, g factor and linewidth) that overlap with the radiation-induced signal (RIS) and therefore, if not taken into account properly, could result in a considerable overestimation of the dose. We have investigated the use of different treatments of fingernails with chemical reagents to reduce the MIS and BKS. The most promising chemical treatment (20 min with 0.1 M dithiothreitol aqueous solution) reduced the contribution of MIS and BKS to the total intensity of EPR signal of irradiated fingernails by a factor of 10. This makes it potentially feasible to measure doses as low as 1 Gy almost immediately after irradiation. However, the chemical treatment reduces the intensity of the RIS and modifies dose dependence. This can be compensated by use of an appropriate calibration curve for assessment of dose. On the basis of obtained results it appears feasible to develop a field deployable protocol that could use EPR measurements of samples of fingernails to assist in the triage of individuals with potential exposure to clinically significant doses of radiation. PMID- 18163160 TI - Comparison of the validity of anthropometric and bioelectric impedance equations to assess body composition in adolescent girls. AB - OBJECTIVE: The purpose of this study was to examine the validity of two anthropometric and four bioelectric impedance (BIA) equations to estimate body composition from dual-energy x-ray absorptiometry (DXA) in adolescent girls of various ethnicities. The rationale for this study was to develop a prediction equation for percent body fat in a multi-ethnic, representative sample of sixth to eighth grade girls. DESIGN: One-hundred and sixty-six girls (51 African American, 45 non-Black Hispanic, 55 non-Hispanic Caucasian, 15 multi-ethnic) participated. Estimates of percent fat and fat-free mass (FFM) from six published BIA and anthropometric equations and the equation developed from this study were compared to body composition determined from DXA. An RJL Systems analyzer was used to measure BIA. Anthropometry included body weight, height, and triceps and calf skinfolds. RESULTS: Average (+/- SD) age, size and body composition was as follows: age, 12.1+/-1.2 yrs, body mass 52.7+/-15.9 kg, height, 154.6+/-8.1 cm; DXA percent fat, 27.9+/-10.4; fat mass (FM), 15.6+/-10.2 kg; and fat free mass (FFM) 35.7+/-6.8 kg. No ethnic differences were found in the relationships between estimated and DXA measured body composition, with the exception of the skinfold equation. The six equations explained on average 82% of the variance in percent fat, 94% of the variance in fat mass, and 88% in fat free mass. Bland Altman analysis indicated that none of the equations performed satisfactorily in our sample. CONCLUSIONS: The BIA and anthropometric equations were significantly related to DXA body composition parameters, however none met the criteria for cross-validation. PMID- 18163161 TI - Bayesian Smoothing with Gaussian Processes Using Fourier Basis Functions in the spectralGP Package. AB - The spectral representation of stationary Gaussian processes via the Fourier basis provides a computationally efficient specification of spatial surfaces and nonparametric regression functions for use in various statistical models. I describe the representation in detail and introduce the spectralGP package in R for computations. Because of the large number of basis coefficients, some form of shrinkage is necessary; I focus on a natural Bayesian approach via a particular parameterized prior structure that approximates stationary Gaussian processes on a regular grid. I review several models from the literature for data that do not lie on a grid, suggest a simple model modification, and provide example code demonstrating MCMC sampling using the spectralGP package. I describe reasons that mixing can be slow in certain situations and provide some suggestions for MCMC techniques to improve mixing, also with example code, and some general recommendations grounded in experience. PMID- 18163163 TI - Biomimetic modeling of oxidative drug metabolism : Strategies, advantages and limitations. AB - The prediction of drug metabolism is an important task in drug development. Besides well-established in vitro and in vivo methods using biological matrices, several biomimetic models have been developed. This review summarizes three different nonenzymatic strategies, including metalloporphyrins as surrogates of the active centre of cytochrome P450, Fenton's reagent, and the electrochemical oxidation of drug compounds. Although none of the systems can simulate the whole range of cytochrome P450-catalyzed reactions adequately, the biomimetic models show some advantages over standard in vitro methods. For example, metalloporpyhrin catalysts allow the synthesis of certain metabolites in sufficient amounts and with sufficient purities to permit characterization and further pharmacological and toxicological tests. The electrochemical generation of metabolites coupled on-line to liquid chromatography/mass spectrometry is a promising tool for studying reactive metabolites and can be applied in automated high-throughput screening approaches. In this paper, detailed comparisons with cytochrome P450 catalysis are drawn, advantages and disadvantages of the respective methods are revealed, and possible applications are discussed. PMID- 18163162 TI - Critical role of hypoxia and A2A adenosine receptors in liver tissue-protecting physiological anti-inflammatory pathway. AB - Whole body exposure of wild type control littermates and A2A adenosine receptor (A2AR) gene deleted mice to low oxygen containing inspired gas mixture allowed the investigation of the mechanism that controls inflammatory liver damage and protects the liver using a mouse model of T cell-mediated viral and autoimmune hepatitis. We tested the hypothesis that the inflammatory tissue damage associated hypoxia and extracellular adenosine --> A2AR signaling plays an important role in the physiological anti-inflammatory mechanism that limits liver damage during fulminant hepatitis. After induction of T cell-mediated hepatitis, mice were kept in modular chambers either under normoxic (21% oxygen) or hypoxic (10% oxygen) conditions for 8 h. It was shown that the whole body exposure to hypoxic atmosphere caused tissue hypoxia in healthy animals as evidenced by a decrease in the arterial blood oxygen tension and increase of the plasma adenosine concentration (P < 0.05). This "hypoxic" treatment resulted in significantly reduced hepatocellular damage and attenuated levels of serum cytokines in mice with acute liver inflammation. The anti-inflammatory effects of hypoxia were not observed in the absence of A2AR in studies of A2AR gene deficient mice or when A2AR have been pharmacologically antagonized with synthetic antagonist. The presented data demonstrate that total body hypoxia triggered pathway provides protection in acute hepatitis and that hypoxia (upstream) and A2AR (downstream) function in the same immunosuppressive and liver tissue-protecting pathway. PMID- 18163164 TI - A novel approach for monitoring of cyanobacterial toxins: development and evaluation of the passive sampler for microcystins. AB - We have investigated the ability of an integrative sampler for polar organic chemicals to sequestrate a group of common and highly hazardous cyanobacterial toxins-microcystins. In a pilot experiment, commercially available passive samplers were shown to effectively accumulate microcystins after 7 days' exposure in the field. To find the most efficient configuration for sequestration of microcystins, four different porous membranes (polycarbonate, polyester, polyethersulfone and nylon) and two sorbents (Oasis HLB and Bondesil-LMS) were evaluated in the laboratory experiments, where samplers of different configuration were exposed to microcystins (microcystin-RR and microcystin-LR) for 14 days under steady conditions. We observed differences in sampling rates and amounts of accumulated microcystins depending on the sampler configurations. The samplers constructed with the polycarbonate membrane and Oasis HLB sorbent (2.75 mg/cm2) provided the highest sampling rates (0.022 L/day for microcystin-RR and 0.017 L/day for microcystin-LR). To the best of our knowledge, the present study is the first reporting application of passive samplers for microcystins, and our results demonstrate the suitability of this tool for monitoring cyanotoxins in water. PMID- 18163165 TI - The effect of folic acid supplementation on the pharmacokinetics and pharmacodynamics of oral methotrexate during the remission-induction period of treatment for moderate-to-severe plaque psoriasis. AB - OBJECTIVE: We assessed the effect of folic acid (FA) on the pharmacokinetics and pharmacodynamics of low-dose oral methotrexate (MTX) during the remission induction phase of psoriasis treatment. METHODS: In a 32-week, open-label, two way cross-over study, patients (n=20, seven men, aged 35-70 years) with moderate to-severe plaque psoriasis were randomly assigned to receive MTX plus FA (20 mg/week) for 16 weeks followed by MTX monotherapy (three doses of MTX separated by 12-h intervals once a week) for an additional 16 weeks (treatment arm A, n=10) or to receive the opposite sequence of treatments (arm B, n=10). Dosing of MTX was individualised with the help of pre-study evaluation of plasma MTX pharmacokinetics. The Psoriasis Area and Severity Index (PASI), biochemistry and haematology tests and erythrocyte concentration of MTX polyglutamates (MTXPG) were evaluated throughout the study. RESULTS: In arms A and B, the mean (range) concentrations of MTXPG (nmol/L) were comparable [week 16: 96.2 (32.0-157) vs. 111 (73.7-175), P=0.32; week 32: 103 (55.8-173) vs. 83.6 (27.4-129), P=0.24]. After 16 weeks, the mean+/-SEM PASI decreased from 20.1+/-2.1 to 8.8+/-1.3 in arm A, while a greater reduction from 27.2+/-2.1 to 5.1+/-1.0 occurred in arm B (P<0.001). Positive correlations were found between the percent improvement in PASI at week 16 and the ratios of the concentration of MTXPG to plasma folate (rho=0.59, P=0.008) or RBC folate concentration (rho=0.56, P=0.013). Due to an accelerated decline in PASI in arm A and a trend to its worsening in arm B after crossing over of treatments, the mean absolute PASI scores in both arms were comparable at week 32. CONCLUSION: The antipsoriatic effect of MTX during the remission-induction phase of treatment is influenced by folate status and may be significantly less if combined treatment with FA is used, irrespective of pre treatment folate levels. The individual tailoring of MTX dosing needs further attention because the mean percent PASI improvement from baseline was 83% and the inter-patient variability in response was low after 16 weeks of monotherapy with MTX. PMID- 18163166 TI - A lifelike patient simulator for teaching robotic colorectal surgery: how to acquire skills for robotic rectal dissection. AB - BACKGROUND: The use of robotic systems for colorectal resections is well documented, but robotic surgery is not yet established as a substitute for all laparoscopic colorectal procedures. The features of the new-generation robotic system seem to be well suited for proper mesorectal excision, with the identification and preservation of autonomic pelvic nerves. Proper training in the use of robotic skills is essential. METHODS: This report describes the creation of a pelvic model that can be used to teach the complex skills needed for successful completion of robotic rectal dissection. The model was designed to be cost effective, portable, and reusable in multiple teaching programs. Both the setup and size of the trainer were designed to be the same as those for a real patient and to allow for proper simulation of port placement in a true robotic rectal dissection. The operative field was molded directly onto a replica of a human skeleton, and the materials that make up the trainer closely replicate the consistency of a real patient. RESULTS: To date, no adequate artificial pelvic models have been available for rectal dissection. Cadaveric models are expensive, and virtual reality trainers, although offering an attractive alternative for some procedures, currently are not available for complex robotic tasks such as rectal dissection. One major advantage of this trainer is that it allows for the surgeon to develop proficiency in both the areas of robotic setup and console without the assistance of a second surgeon. CONCLUSIONS: The trainer described in this report provides an accurate simulation of true robotic rectal dissection. Its portability makes it easy to use at various hospitals. As robotic surgery becomes more common, this training tool has the potential to help surgeons quickly build the skills necessary for the successful use of robotic surgery in the area of rectal dissection. PMID- 18163167 TI - Transoral access for endoscopic thyroid resection. AB - BACKGROUND: Endoscopic neck surgery is requested by an increasing number of patients. The access trauma of the axillary, breast, and chest approaches is greater than with open or video-assisted surgery. The authors tested the feasibility of the sublingual transoral access, which they believe is the most promising minimally invasive endoscopic access to the thyroid gland from outside the neck region. METHODS: The sublingual transoral access was first evaluated in two fresh human cadavers. An experimental investigation then was performed using a porcine model. A total of 10 endoscopic transoral thyroidectomies were performed in 10 pigs using a modified axilloscope with an obturator, ultrasonic scissors, and a neuromonitoring system to identify the recurrent laryngeal nerve. RESULTS: A complete transoral thyroid resection was achieved with both the human cadavers and all the living pigs. Despite the complexity of the anatomic region, the transoral procedure was astonishingly easy to perform. In the animal study, the time from the introduction of the obturator just above the larynx to its removal was 59 s. The average overall operation time was 50 min. The neuromonitoring system permitted the regular function of the recurrent laryngeal nerves on both sides to be proved after removal of the thyroid gland. The pigs were observed for another 2 h after the operation. No complications occurred during the operation or afterward. CONCLUSIONS: Endoscopic transoral thyroid resection is possible. It proved to be a safe procedure in living pigs and astonishingly easy to perform. The results may be helpful for thyroid resections in humans using a similar access, as suggested by the thyroidectomies in human cadavers preceding this study. PMID- 18163168 TI - Systematic review of the role of thoracoscopic splanchnicectomy in palliating the pain of patients with chronic pancreatitis. AB - BACKGROUND: The management of opiate-dependent intractable abdominal pain caused by chronic pancreatitis remains challenging. The published series on the role, safety, feasibility, and efficacy of thoracoscopic splanchnicectomy are reviewed. METHODS: The MEDLINE, EMBASE, and PREMEDLINE databases were searched, and relevant English language publications were systematically retrieved. Data were pooled by two independent reviewers. RESULTS: Between 1994 and 2006, 302 patients were featured in 16 reports. The reports described 202 procedures as bilateral and 100 as unilateral. These procedures were associated with rates of 16.6% for morbidity, 1.3% for conversion to thoracotomy, 1.3% for reoperation to manage complications, and 0% for mortality. The mean postoperative hospital stay was 2.7 days. The mean success rate was 90% up to 6 months of follow-up evaluation, 75% at >6 to 15 months of follow-up evaluation, and 49% at >15 months to 5.7 years of follow-up evaluation. Further intervention for pain relief was required for 12.9% of the patients. CONCLUSION: Splanchnicectomy reduces pain and improves quality of life for patients with chronic pancreatitis. Patient selection determines success rates, but the early good results achieved decline with time elapsed after thoracoscopic splanchnicectomy. PMID- 18163169 TI - Laparoscopic adrenalectomies: A nationwide single-surgeon experience. AB - BACKGROUND: Laparoscopic adrenalectomy (LA) was first performed in Iceland in 1997. Since then, all procedures for presumed benign lesions of the adrenals have been performed laparoscopically in a single center. Compared with conventional adrenalectomy, LA appears to achieve superior results in terms of recovery, hospital stay, and morbidity. This study aimed to evaluate the results of LA in Iceland. METHODS: The hospital records of all patients who underwent LA in Iceland from 1997 through 2005 were reviewed. The preoperative diagnosis was documented, as well as the pathologic diagnosis, operative details, complications, and length of hospital stay. RESULTS: In 49 operations, 53 adrenal glands were removed from 48 patients (37 women and 11 men). The mean patient age was 53.6 years (range, 24.4-78.8 years). The left adrenal was removed from 29 patients, the right adrenal from 14 patients, and both adrenals from 5 patients. The most common indications and diagnoses included 17 nonsecreting tumors (12 adenomas, 3 hyperplasias, 1 complex adrenal cyst, and 1 hemangioma), 12 aldosteronomas (10 aldosteronomas and 2 nodular hyperplasias), and 10 pheochromocytomas (9 confirmed, 1 adrenal hyperplasia). Other indications and diagnoses were less common. The mean operative time was 168 min (range, 87-370 min) for unilateral operations and 412 min (range, 345-480 min) for bilateral operations. The mean blood loss was 117 ml (range, 0-650 ml) for unilateral operations and 200 ml (range, 0-350 ml) for bilateral operations. The complications were mild pancreatitis (n = 1), urinary tract infection (n = 1), atelectasis (n = 1), mild congestive heart failure (n = 2), and transient corneal abrasion (n = 1). No conversion to open procedure was needed. The mean tumor size was 3.5 cm (range, 1.5-6.2 cm), and the mean postoperative hospital stay was 2.6 days (range, 1-6 days). CONCLUSION: The results of laparoscopic adrenalectomies in Iceland for benign lesions of the adrenals are comparable with published results from large referral centers. PMID- 18163170 TI - A consensus document on robotic surgery. PMID- 18163171 TI - Tumor necrosis rate adjusted by tumor volume change is a better predictor of survival of localized osteosarcoma patients. AB - BACKGROUND: We assessed whether new parameter that considers both tumor volume change and necrosis rate predicts metastasis-free survival of localized osteosarcoma patients. We also evaluated relationship between tumor volume change and necrosis rate or metastasis-free survival. METHODS: We retrospectively reviewed 151 patients with stage II osteosarcoma who were treated with surgery and neoadjuvant chemotherapy. The tumor volume change was measured and calculated based on pre- and postchemotherapy magnetic resonance images. The mean metastasis free interval was 83.1 months. We calculated adjusted tumor necrosis rate as following formula: 100 - (100 - necrosis rate) x postchemotherapy/prechemotherapy tumor volume. Survival and logistic regression analyses were used to evaluate the correlation among size parameters, tumor necrosis rate and survival. RESULTS: The 5-year metastasis-free survival rate of 151 patients was 71.4% (95% CI, 67.7 75.1%). American Joint Committee on Cancer (AJCC) stage IIB (RR 2.27; 95% CI, 1.11-4.62; P = 0.025) and poor adjusted tumor necrosis rate (RR 2.02; 95% CI, 1.05-3.89; P = 0.035) independently correlated with metastasis-free survival period. Further, tumor volume change independently correlated with necrosis rate. Decreased tumor volume could predict good response, with sensitivity of 80.2%, specificity of 68.6%, and positive predictive value (PPV) of 74.7%. Increased or stable tumor volume could predict poor response, with sensitivity of 68.6%, specificity of 80.2%, and PPV of 75.0 %. CONCLUSION: The necrosis rate adjusted by the tumor volume change is an independent prognostic factor in osteosarcoma. This adjusted tumor necrosis rate may serve as a basis for risk-adapted therapy in combination with other prognostic factors. PMID- 18163172 TI - Staging of invasive breast carcinoma patients with T4anyNM0 and T1-3N3M0. PMID- 18163173 TI - Neoadjuvant chemoradiation and local excision for T2-3 rectal cancer. AB - BACKGROUND: Local excision (LE) of T1 low-risk (G1-2/L0/V0) rectal cancer is an established approach with local recurrence (LR) rates of approximately 5%, whereas LE of > or = T2 high-risk tumors or inadequate resections (R1/RX/R < or = 1 mm) showed high recurrence rates. Because of the favorable results after neoadjuvant chemoradiotherapy (nCRT) and radical surgery of disease that completely responds (CR) with almost absent LR even of T3-4 tumors, an extension of the indication for LE is controversially discussed, and therefore, we assessed this therapeutic option. METHODS: Including our own data, seven studies about LE after nCRT of cT2-3 tumors (n = 237) were analyzed after a PubMed search for cT categories, tumor height, nCRT regimens, schedule and technique of surgery, complications, freedom of stoma, response rates (ypT0-3), length of follow-up, LR, and metastases. RESULTS: Subgroups that we formed (retrospective vs. prospective/retractor vs. transanal endoscopic microsurgery) showed differences in the distribution of cT categories. However, neither the studies we considered nor our own patients showed LR in CR (ypT0). In addition, patients with ypT1 tumor consistently showed low LR rates of 2% (range, 0%-6%), whereas in ypT2 findings, less favorable LR rates of 6% to 20% were observed, and disease that did not respond to therapy (ypT3) displayed LR rates in up to 42%. CONCLUSIONS: Despite of a highly selected patient collective, an extended indication for LE of cT2-3 rectal cancer after nCRT may be considered. The strongest prognostic factors were a CR (ypT0) or responses on submucosa level (ypT1). These first results will have to be confirmed in a prospective trial with an appropriate sample size to ensure high statistical power. PMID- 18163174 TI - Factors associated with negative margins of lumpectomy specimen: potential use in selecting patients for intraoperative radiotherapy. AB - BACKGROUND: Lumpectomy followed by radiation is standard treatment for early breast cancer. Recently, the use of partial breast intraoperative radiation (IORT) has been developed, and patients selected for IORT should not have positive margins. This study's purpose was to identify factors predicting negative margins after lumpectomy. METHODS: Patient age, preoperative investigations, surgery, final pathology, and margin status were examined using a prospective database between 1999 and 2005. Univariate and multivariate logistic regression analysis were performed to identify patient and tumor factors predicting an increased rate of negative margins. The results were used to generate a patient selection algorithm. RESULTS: The rate of positive margins at first resection was 17% in 730 lumpectomies (708 patients). Multivariate analysis revealed that older age (P = .0006), smaller tumor size (P < .0025), type of surgery (OR = 3.4 for ultrasound vs mammogram-guided wire localization, P = .003), and having a core needle biopsy (CNB) with preoperative cancer diagnosis (P < .0001) were predictive for having a negative margin. Patients older than age 50 with a preoperative CNB showing invasive cancer less that 3 cm that can be localized under ultrasound had a negative margin rate of 98% (n = 178). These patients would be ideal for consideration of IORT. CONCLUSIONS: Negative margin rates after lumpectomy are predicted by age, tumor size, preoperative investigations, and localization technique. These variables can be used to select patients for IORT with a 2.2% chance of positive margins. PMID- 18163175 TI - Focus on cyclo(His-Pro): history and perspectives as antioxidant peptide. AB - Cyclo(His-Pro) is an endogenous cyclic dipeptide structurally related to tyreotropin-releasing hormone that was originally discovered in brain. In the central nervous system it has been described to exert multiple biological activities, which seem to be related to a presynaptic dopaminergic mechanism and include among the others a leptin-like function. It can be found in several body fluids and in the gastrointestinal tract where it has been suggested to act as a gut peptide with influence on the entero-insular axis. The oral administration of cyclo(His-Pro) and zinc was described to improve with a synergistic mechanism the glycaemic control in diabetes. The most intriguing function of this cyclic dipeptide is related with its neuroprotective role that was first reported in traumatic injuries of the spinal cord, and then confirmed in other models of experimental injuries of the nervous system. The mechanism that lies behind the neuroprotective activity of cyclo(His-Pro) remain poorly understood. Recent in vitro studies on rat pheochromocytoma PC12 cells have shown that it is a protective factor against stress stimuli and there is early pre-clinical evidence strongly suggesting that it enhances the expression of small heat shock proteins and antioxidant protection at the cellular level. Future research is underway to better characterize the possible use of this cyclic dipeptide in the therapy of neurodegenerative and metabolic disorders. PMID- 18163176 TI - Serum phenylalanine in patients post trauma and with sepsis correlate to neopterin concentrations. AB - Increased blood concentrations of phenylalanine in patients with trauma and sepsis are common but unexplained. We examined the potential relationship between serum concentrations of phenylalanine and the immune activation marker neopterin in 84 specimens of 18 patients (14 males and 4 females) post-trauma during 12-14 days of follow up. Compared to healthy controls, average phenylalanine and neopterin concentrations were elevated in patients, and there existed a positive correlation between concentrations of the two analytes (r (s) = 0.375, p < 0.001). No such association existed between neopterin and tyrosine concentrations (r (s) = -0.018), but neopterin concentrations correlated to the phenylalanine to tyrosine ratio (r (s) = 0.328, p = 0.001). Increased phenylalanine implies insufficient conversion by phenylalanine (4)-hydroxylase (PAH). Oxidative stress due to immune activation and inflammation may destroy cofactor 5,6,7,8 tetrahydrobiopterin and impair PAH activity. This assumption is further supported by the correlation found between higher neopterin concentrations and higher phenylalanine to tyrosine ratio, which estimates efficacy of PAH. PMID- 18163177 TI - X-ray sequence ambiguities of Sclerotium rolfsii lectin resolved by mass spectrometry. AB - X-ray crystallography, although a powerful technique for determining the three dimensional structure of proteins, poses inherent problems in assigning the primary structure in residues Asp/Asn and Glu/Gln since these cannot be distinguished decisively in the electron density maps. In our recently published X-ray crystal structure of the Sclerotium rolfsii lectin (SRL) at 1.1 A resolution, amino acid sequence was initially deduced from the electron density map and residues Asp/Asn and Glu/Gln were assigned by considering their hydrogen bonding potential within their structural neighborhood. Attempts to verify the sequence by Edman sequencing were not successful as the N terminus of the protein was blocked. Mass spectrometry was applied to verify and resolve the ambiguities in the SRL X-ray crystal structure deduced sequence. From the Matrix assisted laser desorption/ionization time-of-flight-mass spectrometry (MALDI TOF-MS) and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI MS/MS) analysis of tryptic and chymotryptic peptides of SRL, we could confirm and correct the sequence at five locations with respect to Asp/Asn and Glu/Gln. Analysis data also confirmed the positions of Leu/Ile, Gln/Lys residues and the sequence covering 118 of the total 141 residues accounting to 83.68% of the earlier deduced sequence of SRL. PMID- 18163178 TI - The effect of taurine on mesenteric blood flow and organ injury in sepsis. AB - Endotoxin decreases mesenteric blood flow and inflicts organ injury via free radicals. We investigated whether taurine, an endogenous antioxidant and vasodilator, could attenuate the deleterious effects of endotoxin in a mouse model of sepsis. Swiss albino mice were allocated into four groups and treated either with taurine (150 mg/kg, i.p. at 0(th), 8(th), 16(th) h) or its solvent sterile saline (NaCl 0.9%, w/v) while E. coli endotoxin (20 mg/kg, i.p.) or its solvent saline were also given at 8(th) h. At 24(th) h the animals were anaesthetized and the mesenteric blood flow was measured by using perivascular ultrasonic Doppler-flowmeter. The animals were then exsanguinated, the spleen, liver, and kidneys were isolated for histopathological examination. Thiobarbituric acid-reacting substances (TBARS), glutathione, and myeloperoxidase activity were determined in the liver samples. Endotoxin significantly decreased the mesenteric blood flow and glutathione levels in liver while TBARS and myeloperoxidase activity were increased. However, taurine did not block the deleterious effects of endotoxin nor it did attenuate the histopathological injury. Therefore, we concluded that endotoxin-induced organ injury via free radicals is resistant to blockade by taurine. PMID- 18163179 TI - Chemotactic tripeptides incorporating at position 2 alpha-aminoacid residues with unsaturated side chains. AB - New N-For-Met-Leu-Phe-OMe (fMLF-OMe) analogues incorporating three different gamma-delta-didehydro-alpha-aminoacid residues (namely: Alg = (S)-Allylglycine; Dag = Diallylglycine; Cpg = 1-Aminocyclopent-3-ene-1-carboxylic acid) replacing the native (S)-Leucine have been synthesized and their activity towards human neutrophils has been evaluated in comparison with that shown by the reference tripeptide fMLF-OMe. Chemotaxis, lysozyme release and superoxide anion production have been measured. (1)H NMR titration experiments and NOESY spectrum of the Cpg containing model 10 have been discussed in order to ascertain the preferred solution conformations. A fully extended (C(5)) conformation at position 2 and a folded conformation with two consecutive gamma-turns (C(7) structure) have been proposed for the Dag and Cpg containing tripeptides, respectively. PMID- 18163180 TI - Post exercise carbohydrate-protein supplementation: phosphorylation of muscle proteins involved in glycogen synthesis and protein translation. AB - The enzymes Akt, mTOR, p70(S6K), rpS6, GSK3, and glycogen synthase interact in the control of protein and/or glycogen synthesis in skeletal muscle, and each has been found to respond to exercise and nutrient supplementation. In the present study, we tested the hypothesis that nutrient supplementation post exercise, in the form of a carbohydrate-protein (CHO-PRO) supplement, would alter the phosphorylation state of these enzymes in a manner that should increase muscle protein and glycogen synthesis above that produced by exercise alone. After a 45 min cycling session followed by sprints and again 15 min later, the subjects (n = 8) ingested 400 ml of a CHO-PRO drink (7.8% dextrose and 1.8% protein electrolyte) or a placebo drink, as assigned using a randomized, counter-balanced design with repeated measures. Biopsies of the vastus lateralis were taken before exercise and at 45 min of recovery. At 45 min after supplementation, CHO-PRO treatment yielded greater phosphorylation of Akt (65%), mTOR (86%), rpS6 (85 fold), and GSK3alpha/beta (57%) than pre-exercise levels (p < 0.05). Although p70(S6k) showed an exercise response after 45 min, there were no differences between treatments. Glycogen synthase (GS) phosphorylation was significantly reduced 45 min after exercise for both treatments, but the reduction in phosphorylation was greatest during the CHO-PRO treatment (3-fold decrease; p < 0.05), indicating greater activation of GS following supplementation. No difference between treatments was detected prior to exercise for any of the enzymes. These results suggest that a post exercise CHO-PRO supplement alters the phosporylation levels of the enzymes tested in a manner that should accelerate muscle glycogen synthesis and protein initiation during recovery from cycling exercise. PMID- 18163181 TI - Protein expression of BACE1, BACE2 and APP in Down syndrome brains. AB - Down syndrome (DS) is the most common human chromosomal abnormality caused by an extra copy of chromosome 21. The phenotype of DS is thought to result from overexpression of a gene or genes located on the triplicated chromosome or chromosome region. Several reports have shown that the neuropathology of DS comprises developmental abnormalities and Alzheimer-like lesions such as senile plaques. A key component of senile plaques is amyloid beta-peptide which is generated from the amyloid precursor protein (APP) by sequential action of beta secretases (BACE1 and BACE2) and gamma-secretase. While BACE1 maps to chromosome 11, APP and BACE2 are located on chromosome 21. To challenge the gene dosage effect and gain insight into the expressional relation between beta-secretases and APP in DS brain, we evaluated protein expression levels of BACE1, BACE2 and APP in fetal and adult DS brain compared to controls. In fetal brain, protein expression levels of BACE2 and APP were comparable between DS and controls. BACE1 was increased, but did not reach statistical significance. In adult brain, BACE1 and BACE2 were comparable between DS and controls, but APP was significantly increased. We conclude that APP overexpression seems to be absent during the development of DS brain up to 18-19 weeks of gestational age. However, its overexpression in adult DS brain could lead to disturbance of normal function of APP contributing to neurodegeneration. Comparable expression of BACE1 and BACE2 speaks against the hypothesis that increased beta-secretase results in (or even underlies) increased production of amyloidogenic A beta fragments. Furthermore, current data indicate that the DS phenotype cannot be fully explained by simple gene dosage effect. PMID- 18163182 TI - AAIndexLoc: predicting subcellular localization of proteins based on a new representation of sequences using amino acid indices. AB - Identifying a protein's subcellular localization is an important step to understand its function. However, the involved experimental work is usually laborious, time consuming and costly. Computational prediction hence becomes valuable to reduce the inefficiency. Here we provide a method to predict protein subcellular localization by using amino acid composition and physicochemical properties. The method concatenates the information extracted from a protein's N terminal, middle and full sequence. Each part is represented by amino acid composition, weighted amino acid composition, five-level grouping composition and five-level dipeptide composition. We divided our dataset into training and testing set. The training set is used to determine the best performing amino acid index by using five-fold cross validation, whereas the testing set acts as the independent dataset to evaluate the performance of our model. With the novel representation method, we achieve an accuracy of approximately 75% on independent dataset. We conclude that this new representation indeed performs well and is able to extract the protein sequence information. We have developed a web server for predicting protein subcellular localization. The web server is available at http://aaindexloc.bii.a-star.edu.sg . PMID- 18163183 TI - Cloning and functional characterization of the HRASLS2 gene. AB - The HRAS-like suppressor 2 (HRASLS2) gene belongs to the H-REV107 gene family involved in the regulation of cell growth and differentiation. HRASLS2 is expressed at high levels in normal tissues of the small intestine, kidney, and trachea. We cloned HRASLS2 cDNA from human SW480 colon cancer cells. Most wild type, and some N- and C-terminal truncated HRASLS2 (HRASLS2DeltaNDeltaC) were expressed as a granular pattern located at perinuclear region in HtTA cervical cancer cells, while truncation at the C-terminus only (HRASLS2DeltaC) resulted in a diffuse pattern. Wild-type HRASLS2 significantly suppressed colony formation of HeLa and HCT116 cells. HRASLS2DeltaNDeltaC significantly inhibited colony formation of HCT116 cells, but HRASLS2DeltaC did not affect cell growth. HRASLS2 suppressed the RAS-GTP levels and total RAS protein by 44% and 25%, respectively in HtTA cells; however, the suppression was not observed in truncated HRASLS2 variants. In conclusion, the HRASLS2 protein suppressed growth and RAS activities of cancer cells, and the C-terminal hydrophobic domain appeared to be indispensable for both activities. PMID- 18163184 TI - Intracerebroventricular injection of L-arginine induces sedative and hypnotic effects under an acute stress in neonatal chicks. AB - L-arginine participates in many important and diverse biochemical reactions associated with the normal physiology of the organism. In the present study, we investigated the effect of central administration of L-arginine on the stress response and its mechanism in neonatal chicks. Intracerebroventricular (i.c.v.) injection of L-arginine clearly attenuated the stress response in a dose dependent manner, and induced sleep-like behavior during 10 min. To clarify the mechanism by which L-arginine induces sedative and hypnotic effects in chicks, we investigated the effects of nitric oxide (NO) synthase (NOS) inhibitors on L arginine-induced sedative and hypnotic effects, and as well as the effects of a NO donor. L-Arginine-induced (1.9 micromol) sedative and hypnotic effects were attenuated by i.c.v. co-injection with a non-selective NOS inhibitor N(G)-nitro-L arginine methyl ester HCl (400 nmol). In addition, the effects of L-arginine were slightly attenuated by the inactive isomer of the NOS inhibitor N(G)-nitro-D arginine methyl ester HCl (400 nmol). The i.c.v. injection of 3 morpholinosylnomine hydrochloride, a spontaneous NO donor, had little effect on postures. The i.c.v. injection of L-arginine had no effect on NOx concentration at various brain sites. These results suggested that the contribution of NO generation via NOS may be low in the sedative and hypnotic actions of L-arginine. Therefore, L-arginine and/or its metabolites, excluding NO, may be necessary for these actions. PMID- 18163185 TI - Proton magnetic resonance spectroscopy predicts proliferative activity in diffuse low-grade gliomas. AB - The aim of the study was to investigate the ability of (1)HMRS to reflect proliferative activity of diffuse low-grade gliomas (WHO grade II). Between November 2002 and March 2007, a prospective study was performed on consecutive patients with suspected supratentorial hemispheric diffuse low-grade tumors. All the patients underwent MR examination using uniform procedures, and then surgical resection or biopsy within 2 weeks of the MR examination. Proliferative activity of the tumors was assessed by Ki-67 immunochemistry (Mb-1) on paraffin embedded tumor sections. Spectroscopic data was compared with Ki-67 labeling index and other histologic data such as histological subtype, cellular atypia, cellular density using univariate and multivariate analysis. 82 of 97 consecutive patients had histologically confirmed WHO grade 2 gliomas. Ki-67 proliferation index (PI) was correlated with specific spectral patterns: (1) low PI (<4%) was associated with increased Cho/Cr and absence of both free lipids or lactates; (2) intermediate PI (4-8%) was associated with resonance of lactates; and (3) high PI (>8%) was characterized by a resonance of free lipids. On multivariate analysis, resonance of lactates and resonance of free lipids appeared as independent predictors of intermediate PI (P < 0.001) and high PI (P < 0.001), respectively; moreover, free lipids resonance was correlated with cellular atypia (P < 0.05). This study suggests that (1)HMRS is a reliable tool to evaluate the proliferation activity of WHO grade 2 glioma and to identify potentially more aggressive clinical behavior. PMID- 18163186 TI - Molecular docking studies on tetrahydroimidazo-[4,5,1-jk][1,4]-benzodiazepinone (TIBO) derivatives as HIV-1 NNRT inhibitors. AB - At present, chemotherapy seems to be the main weapon in the arsenal of remedies for the ongoing crusade against AIDS. The mode of binding of the TIBO family of inhibitors has been of interest because these compounds do not fit the two-hinged ring model as generally observed in the NNRTIs. Flexible docking simulations were performed with a series of 53 TIBO derivatives as NNRTIs. Binding preferences as well as the structural and energetic factors associated with them were studied. A good correlation (r(2)=0.849, q(2)=0.843) was observed between the biological activity and binding affinity of the compounds which suggest that the identified binding conformations of these inhibitors are reliable. Further screening of PubChem database yielded novel scaffolds. Our studies suggest that modifications to the TIBO group of inhibitors might enhance their binding efficacy and hence, potentially, their therapeutic utility. PMID- 18163187 TI - Functional characterization of the HOXB13 promoter region. AB - Homeobox (HOX) genes are crucial regulators of cell growth and differentiation. These genes initiate and control gene expression cascades that drive development. More recently, the absent or aberrant expression of HOX genes has been implicated in cancer development. Despite the observance of these expression changes, the regulation of the HOX genes in adult tissues and how these genes become deregulated in cancerous tissues still needs much investigation. We characterized the promoter region of the HOXB13 gene. A 3 kb region upstream of the HOXB13 gene, which included the 5'UTR, increased reporter gene expression in LNCaP cells by approximately 99 fold over the promoterless control construct. A highly conserved 179 base pair fragment containing only the 5'UTR of the HOXB13 gene constituted a minimal promoter in the LNCaP cell line. Strong promoter activity was seen in the presence or absence of testosterone, although testosterone exposure did decrease expression in LNCaP cells by 50%. In an androgen insensitive cell line Du145, no sensitivity to testosterone was detected and a consistent low basal level of expression was observed. Since HOXB13 expression is highly tissue specific, we investigated the ability of the promoter to drive expression in tissues other than prostate. We observed highest expression in LNCaP cells with low levels of expression in lung, retinoblastoma, and colon cancer cells and higher expression in MCF7 breast cancer cells. PMID- 18163188 TI - The use of the Camberwell Assessment of Need in treatment: what unmet needs can be met? AB - BACKGROUND: A useful way of operationalising treatment effects in routine outcome assessment data may be to assess the rate at which unmet needs at time point t change to met needs at time point t + 1. METHODS: Data were obtained from the local Cumulative Needs for Care Register (CNCR), a cumulative data set of needs (Camberwell Assessment of Need), psychopathology, well being and functioning of psychiatric patients living both inside and outside the hospital, in a circumscribed geographical area. RESULTS: In the group of relatively new patients, the number of met needs (sum score) increased over time. Higher unmet needs sum score predicted higher met needs at time point t + 1. Unmet needs in the areas of accommodation, household skills, self-care, safety to others (in new patients only), alcohol, drugs, money and benefits were associated with met needs on these items at time point t + 1, but there was no such association for occupation/daytime activities, psychotic symptoms, psychological distress and self-harm. CONCLUSION: Treatment outcomes in psychiatric practice can be usefully tracked and quantified using the rate of change from unmet to met needs. Needs in the area of the ability to live independently may represent outcomes that are more sensitive to treatment effects than needs in the realm of psychopathology and daytime activities. PMID- 18163189 TI - Larvicidal activity of some Euphorbiaceae plant extracts against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). AB - Larvicidal activity of ethyl acetate, butanol, and petroleum ether extracts of five species of Euphorbiaceae plants, Jatropha curcas, Pedilanthus tithymaloides, Phyllanthus amarus, Euphorbia hirta, and Euphorbia tirucalli, were tested against the early fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus (Say). The larval mortality was observed after 24 h of exposure. All extracts showed low larvicidal effects; however, the highest larval mortality was found in petroleum ether extract. The LC50 value of petroleum ether extracts of J. curcas, P. tithymaloides, P. amarus, E. hirta, and E. tirucalli were 8.79, 55.26, 90.92, 272.36, and 4.25 ppm, respectively, against A. aegypti and 11.34, 76.61, 113.40, 424.94, and 5.52 ppm, respectively, against C quinquefasciatus. Of the various ratios tested, the petroleum ether extracts of J. curcas and E. tirucalli were observed to be more efficient than the other plant extracts. It is, therefore, suggested that E. tirucalli can be applied as an ideal potential larvicide against A. aegypti and C. quinquefasciatus. This is an ideal ecofriendly approach for the control of the dengue vector, A. aegypti, and the lymphatic filariasis vector, C. quinquefasciatus. PMID- 18163190 TI - Applicability of the use of charcoal for the evaluation of intestinal motility in a murine model of Trypanosoma cruzi infection. AB - Chagas disease, caused by the protozoan Trypanosoma cruzi, remains a serious public health problem in Latin America. In relation to digestive problems, 4.5% of patients show mega syndromes (megacolon) in the chronic phase. In this article, we evaluated intestinal motility at the acute phase of T. cruzi infection through charcoal ingestion in adult mice. After infection, Swiss mice were administered an aqueous suspension of charcoal in water by gavage. Decrease in intestinal motility was determined by increased time of appearance of charcoal in the feces. The uninfected group showed a mean time of charcoal elimination of 109.0 +/- 14.6 min throughout the assay. On the other hand, infected mice presented a significant increase in charcoal defecation time during infection. At 15 days postinfection, infected mice showed a significant increase in charcoal defecation time, 310.2 +/- 67.4 min when compared to the uninfected group, which presented 97.8 +/- 31.8 min, indicating that the T. cruzi infection interferes with intestinal motility. Our results demonstrate that the use of charcoal is an ethical and efficient procedure to evaluate the intestinal motility in the murine model of T. cruzi infection. PMID- 18163191 TI - The impact of expressions of treatment efficacy and out-of-pocket expenses on patient and physician interest in osteoporosis treatment: implications for pay for-performance programs. AB - BACKGROUND: Clinical practice guidelines (CPGs) are increasingly used as the basis for pay-for-performance (P4P) programs. It is unclear how support for guidelines varies when treatment efficacy is expressed in varying mathematically equivalent ways. OBJECTIVES: To assess: (1) how patient and provider compliance with osteoporosis CPGs varies when pharmacotherapy efficacy is presented as relative risk reduction (RRR) versus absolute risk reduction (ARR) and (2) the impact of increasing out-of-pocket drug expenditures on acceptance of guideline concordant therapy. DESIGN: Cross-sectional survey of patients and physicians. SUBJECTS AND SETTING: Female patients age >50 years and providers drawn from academic and community outpatient clinics. MEASUREMENTS: Patient and provider acceptance of pharmacotherapy when treatment efficacy (reduction in hip fractures) was expressed alternatively in relative terms (35% RRR) versus absolute terms (1% ARR); acceptance of pharmacotherapy as patient drug copayment increased from 0% to 100% of the total drug costs. RESULTS: Compliance with CPGs fell significantly when the expression of treatment benefit was switched from RRR to ARR for both patients (86% vs 57% compliance; P < .001) and physicians (97% vs 56% compliance; P < .001). Increasing drug copayment from 0% to 10% of total drug cost decreased patient compliance with CPGs from 80% to 57% (P < .001) but did not impact physician compliance. With increasing levels of copay, both patient and provider interest in treatment decreased. LIMITATIONS: Respondents may not have fully understood the risks and benefits associated with osteoporosis and its treatment. CONCLUSION: Patient and provider interest in CPG-recommended treatment for osteoporosis is reduced when treatment benefit is expressed as ARR rather than RRR. In addition, minimal increases in drug copayment significantly decreased patient, but not provider, interest in osteoporosis treatment. Designers of P4P programs should consider details including expressions of treatment benefit and patients' out-of-pocket costs when developing measures to assess quality-of-care. PMID- 18163192 TI - Need for multivitamin use in the postoperative period of gastric bypass. AB - BACKGROUND: Based on the reduced gastric volume and the malabsorption produced by Roux-en-Y gastric bypass (RYGBP) and diet therapy, it is essential in the postoperative period to obtain and maintain an adequate nutritional state, with the aim of preventing malnutrition and seeking a healthy life. It is observed that patients have difficulty in understanding the new food choices that must considered, as they have eating habits that are very divergent from those currently proposed. There is often the need for vitamin and mineral replacement after laboratory tests. METHODS: This study calculated and evaluated the 24-h eating records of 210 patients, collected in the course of nutritional visits in follow-ups of the first, third, sixth, ninth, 12th, 18th, and 24th months postoperative. RESULTS: It was possible to observe an increase in the consumption of nutrients in the course of the study period, but it was not regular and significant for all the nutrients. Also, it is noted that the minimal requirements for vitamin A, vitamin C, calcium, iron and B-complex vitamins (except for cyanocobalamin and riboflavin) were not attained. The nutrients in which satisfactory results were obtained were total proteins of high biological value: cyanocobalamin and riboflavin. CONCLUSION: This study demonstrated the concern for nutrient supplementation in the postoperative period of RYGBP. Thus, the routine use of multivitamins is deemed necessary after the first month postoperatively, with its maintenance preferably for the rest of the patient's life, without abandoning periodic clinical and laboratory follow-up. PMID- 18163193 TI - Jejunal obstruction caused by migrated gastric band. AB - We describe the rare complication of an eroded gastric band in the gastric cavity that passed through the pylorus and obstructed the proximal jejunum at the point allowed by the length of the connecting tube. At surgery, in addition to the expected finding, multiple necrotic pressure ulcerations in the jejunal wall were found in step ladder locations. Anyone who has adopted laparoscopic gastric banding as the modality for surgical treatment of morbid obesity should be familiar with this rare but potentially dangerous complication. PMID- 18163194 TI - Correlations of laparoscopy with histology and laboratory studies on liver diseases in bariatric patients. AB - BACKGROUND: Non-alcoholic fatty liver disease is prevalent in obese patients. Liver biopsy remains the best diagnostic tool for confirmation. We evaluated the effectiveness of macroscopic parameters of laparoscopy in diagnosis of liver diseases. Moreover, correlations of laparoscopy with histology and laboratory data were also studied. METHODS: From December 2004 to April 2006, 126 morbidly obese patients submitted to laparoscopic bariatric surgery at the En-Chu-Kong Hospital were prospectively studied. RESULTS: There were correlations of histologic steatosis with liver surface fat spot density, liver margin shape, and liver size. Histologic inflammation was related to liver color, vascularity beneath hepatic capsule, liver margin shape, liver size, and liver surface nodularity. Histologic fibrosis had relations to liver color, liver surface nodularity, liver size, varices of ligamentum teres. Spleen size was related to liver surface nodularity and spleen congestion. Relationships of laboratory data with laparoscopic findings included: aspartate transaminase (AST) level with liver size, alanine transaminase (ALT) level with liver color and liver size, albumin level with liver margin shape and liver surface fibrosis and liver size, total protein level with liver size, alkaline phosphatase (ALP) level with liver surface fibrosis, blood glucose level with liver surface nodularity and spleen size, C-peptide with liver size. Besides, there were relations of gamma-GT level with liver color, liver margin shape, liver and spleen size. CONCLUSION: Besides histology and laboratory studies, laparoscopic inspection of the abdominal cavity provides important and additional information, which contributed to the final diagnosis of chronic liver diseases and detection of possible pathology in patients. PMID- 18163196 TI - Eptifibatide does not suppress the increase of inflammatory markers in patients with non-ST-segment elevation acute coronary syndrome. AB - BACKGROUND: Platelets are involved in inflammatory reactions which play an important role in the development of atherosclerosis and its acute complications. The objective of this study was to test the ability of glycoprotein (GP) IIb-IIIa antagonist eptifibatide to suppress the increase of inflammatory markers in non ST-segment elevation acute coronary syndrome (ACS). METHODS: Twenty-five patients with unstable angina and non-ST-segment elevation myocardial infarction received eptifibatide on admission (two 180 microg/kg boluses followed by infusion at 2.0 and 1.3 microg/kg/min for 24 and 48 h, respectively) and 25 were treated without GP IIb-IIIa antagonists. Plasma von Willebrand factor (vWF) and soluble P selectin were determined at baseline, 48 h and 2 weeks after onset of ACS, and were also measured in a group of healthy volunteers. Serum C-reactive protein (CRP), tumor necrosis factor alpha (TNFalpha), and interleukin 6 (IL6) were measured at baseline, 48 h, 2 weeks and 6 months. RESULTS: P-selectin was increased at baseline and vWF at baseline, 48 h and 2 weeks in comparison with healthy donors. CRP, TNFalpha, but not IL6 were increased at baseline, 48 h and 2 weeks in comparison with their levels at 6 months. Maximal values of CRP, TNFalpha and vWF were detected at 48 h. At any time point eptifibatide failed to decrease the levels of all tested markers. CONCLUSION: Eptifibatide does not suppress elevated levels of inflammatory markers in patients with non-ST-segment elevation ACS. PMID- 18163197 TI - Fluorimetric study of the pro-oxidant activity of EUK8 in the presence of hydrogen peroxide. AB - The catalase mimetic complex Mn(III)-salen chloride (EUK8) was found to be pro oxidant under low hydrogen peroxide concentrations. The increase in the fluorescence rate of the probe 1,2,3-dihydrorhodamine (DHR) in solution, as well as the carbonyl content of human serum albumin were found to be maximum at H2O2:EUK8 molar ratios ranging from 0 to 2, supporting previous findings regarding the mechanism of EUK8 catalase activity and the formation of highly oxidative Mn(V)-O2- species. This pro-oxidant effect is precluded by the presence of glutathione. Cytotoxicity to HeLa cells, as probed by increased rate of oxidation of intracellular DHR, was not observed. Our findings suggest that the combination of H2O2 and EUK8 at specific molar ratios, in the absence of reductants/antioxidants, induces the oxidation of organic molecules. It is shown that the fluorimetric determination of pro-oxidant activity of metal complexes is more sensitive than the colorimetric quantification of protein carbonyl content. The implications of our findings with respect to the somewhat confusing results arising from in vivo studies of EUK8 and other Mn(III) anti-oxidant metal complexes are discussed. PMID- 18163198 TI - Spectroscopic and electrochemical characterization of cytochrome c encapsulated in a bio sol-gel matrix. AB - Sol-gel technique represents a remarkably versatile method for protein encapsulation. To enhance sol-gel biocompatibility, systems envisaging the presence of calcium and phosphates in the sol-gel composition were recently prepared and investigated. Unfortunately, the low pH at which solutions were prepared (pH < 2.5) dramatically limited their application to proteins, because the acidic environment induces protein denaturation. In this paper we apply a new protocol based on the introduction of calcium nitrate to the inorganic phase, with formation of a binary bioactive system. In this case protein encapsulation results versatile and secure, being achieved at a pH close to neutrality (pH 6.0); also, the presence of calcium is expected to enhance system biocompatibility. To determine the properties of the salt-doped sol-gel and the influence exerted on entrapped biosystems, the structural and functional properties of embedded cytochrome c have been investigated. Data obtained indicate that the salt-doped sol-gel induces no significant change in the structure and the redox properties of the embedded protein; also, the matrix increases protein stability. Interestingly, the presence of calcium nitrate appears determinant for refolding of the acid-denatured protein. This is of interest in the perspective of future applications in biosensoristic area. PMID- 18163199 TI - Effects of tensile stress on the alpha1 nicotinic acetylcholine receptor expression in maxillofacial skeletal myocytes. AB - This study was to investigate the alterations of the alpha1 nicotinic acetylcholine receptor (nAChR) levels under tensile stress stimulation in maxillofacial skeletal myocytes. The skeletal muscle satellite cells from two to three days post-natal BALB/C mice's maxillofacial muscle were collected for primary cell culture. The second passage cells in the loaded groups were subjected to cyclic tensile stress (0.5 Hz, 2000 micro strain) produced by a four point bending system for 2, 4, 6, 8, or 12 h. In the control groups, cells were cultured on similar plates and kept in the same incubator without mechanical stress loading. The examination of nAChR alpha1 receptor expression was performed by receptor binding of [125I] a-bungarotoxin. The nAChR alpha1 mRNA transcript level was analyzed by semi-quantitative RT-PCR. The result showed that the nAChR alpha1 receptor expression was elevated significantly in stress-stimulated group (P < 0.05). An increase of nAChR alpha1 in mRNA transcript level was also observed in stress groups as compared with controls (P < 0.05). It is concluded that nAChR was a possible molecular mechanism which might play an important role in mechanotransduction of tensile stress loading on maxillofacial skeletal myocytes. PMID- 18163200 TI - Thr11 phosphorylated H3 is associated with centromere DNA during mitosis in MCF-7 cells. AB - In this study, indirect immunofluorescence labeling was used to examine the cellular dynamic distribution of Thr11 phosphorylated H3 at mitosis in MCF-7 cells. The Thr11 phosphorylation was observed beginning at prophase at centromeres. Upon progression of mitosis, fluorescence signal was enhanced in the central region of the metaphase plate and maintained till anaphase at centromeres. During telophase, the fluorescent signal of Thr11 phosphorylated H3 disappears from centromeres, but the signal appears again at the midbody during cytokinesis, which suggests that the modified histones may take part in the formation of the midbody and play a crucial role in cytokinesis. Chromatin immunoprecipitation (ChIP) was used to confirm that Thr11 phosphorylated H3 is specifically associated with centromere DNA at prophase to metaphase, which is coincident with the results observed by immunofluorescence. In conclusion, there was a precise spatial and temporal correlation between H3 phosphorylation of Thr11 and stages of chromatin condensation. The timing of Thr11 phosphorylation and dephosphorylation in mitosis were similar to that reported for Ser10 phosphorylation of H3. The Thr11 phosphorylated H3 localized at centromeres during mitosis, which was different from the Ser10 phosphorylated H3 localized at telomere regions and Thr3 phosphorylated H3 localized along the chromosome arms. The results suggest that the Thr11 phosphorylation of histone H3 may play a specific role which was different from Ser10 and Thr3 phosphorylation in mitosis. PMID- 18163201 TI - Mesosome formation is accompanied by hydrogen peroxide accumulation in bacteria during the rifampicin effect. AB - Ultrastructural alteration and hydrogen peroxide localization were examined in Xanthomonas campestris pv. phaseoli during rifampicin effect using transmission electron microscopy. Bacterial cells were treated with rifampicin and then were examined by electron microscopy to observe the changes of ultrastructure or hydrogen peroxide accumulation in living cells that took place before lysis. Intriguingly, rifampicin treatment led to presence of an additional location of hydrogen peroxide accumulation within the cells. There was an association between the frequency and size of the additional location of hydrogen peroxide accumulation and the concentration of rifampicin. Furthermore, an additional ultrastructure, mesosomes, was also present in cells during rifampicin effect. The frequency and size of mesosome increased with the increasing concentration of rifampicin. Result of multiple linear regression showed that the size of mesosome plays as a key factor in the quantity of excess hydrogen peroxide accumulation in cells during rifampicin effect. Linear correlation was confirmed between quantity of excess hydrogen peroxide accumulation and the size of mesosome in cells during rifampicin effect. This finding intensely indicated that mesosomes are just the additional location of hydrogen peroxide accumulation in cells under cellular injury caused by rifampicin treatment. The mesosome formation is always accompanied by excess hydrogen peroxide accumulation in X. campestris pv. phaseoli during rifampicin effect. PMID- 18163202 TI - Myosin light chain kinase inhibition: correction of increased intestinal epithelial permeability in vitro. AB - PURPOSE: To examine whether myosin light chain kinase (MLCK) inhibitors can reduce intestinal epithelial permeability increases in vitro. MATERIALS AND METHODS: Isolated rat, mouse and human colonic tissue mucosae and Caco-2 monolayers were exposed to cytochalasin D (cD) and sodium caprate (C10), in the absence and presence of the MLCK inhibitors, ML-9 and D PIK. Transepithelial electrical resistance (TEER) and Papp of [14C]-mannitol or FITC-dextran 4000 (FD 4) were measured. Western blots were used to measure MLC phosphorylation. RESULTS: Increases in Papp of [14C]-mannitol and decreases in TEER were induced by tight junction openers. These changes were attenuated by ML-9. D-PIK offset the FD-4 Papp increase induced by C10 in Caco-2 only, while ML-9 and PIK inhibited MLC directly, cD induced constriction of peri-junctional actin in Caco 2 monolayers, but this was prevented by ML-9. Although mannitol fluxes across colonic mucosae from dextran-sulphate (DSS)-treated mice were higher than control, they were not ameliorated by either ML-9 or PIK in vitro. CONCLUSIONS: ML-9 inhibits paracellular permeability increases in several intestinal epithelial models. D-PIK reduced stimulated paracellular fluxes in Caco-2 monolayers, but not in tissue. Pre-established increases were not modified by two MLCK inhibitors in a mouse model of IBD. PMID- 18163203 TI - Trust and memory: organizational strategies, institutional conditions and trust negotiations in specialty clinics for Alzheimer's disease. AB - Clinicians aim to establish trust during medical encounters because, without it, health consumers may not seek medical care, consider their diagnoses legitimate, or adhere to treatment regimens. This paper examines the identification and treatment of memory loss within two specialty clinics to understand how cultural dynamics, such as organizational ethos and work practices, influence the social fabric of cognitive evaluations. Ethnographic data suggest important historical and cultural differences in the approaches to Alzheimer's disease (AD). Organizational routines, however, support a common goal, that of moving individuals from "potential patients" to patients, and ultimately research subjects, through establishing trust. Although the processes through which trust is potentially achieved, or the social conditions of trust, were similar at the sites, the object of trust was different. Whereas one clinic encouraged trust in collective medical expertise, the other focused on trust in specific clinicians. These conditions affect the clinical consequences of trust, particularly how and when the diagnosis is delivered, use of the AD label and other terminology, and the level of standardization. The individual consequences include perceptions of patients and depictions of the prognosis. Whether cognitive impairment is viewed as a scientific puzzle to be solved or is seen as a chronic illness significantly shapes the organizational processes of clinical evaluation. Alzheimer's disease, as a cultural object, is a particularly salient exemplar of the clinical negotiation of ambiguous diagnostic categorizations and the unpredictable patient in daily biomedical practice. PMID- 18163205 TI - Partial phase synchronization of neural populations due to random Poisson inputs. AB - We show that populations of identical uncoupled neurons exhibit partial phase synchronization when stimulated with independent, random unidirectional current spikes with interspike time intervals drawn from a Poisson distribution. We characterize this partial synchronization using the phase distribution of the population, and consider analytical approximations and numerical simulations of phase-reduced models and the corresponding conductance-based models of typical Type I (Hindmarsh-Rose) and Type II (Hodgkin-Huxley) neurons, showing quantitatively how the extent of the partial phase synchronization depends on the magnitude and mean interspike frequency of the stimulus. Furthermore, we present several simple examples that disprove the notion that phase synchrony must be strongly related to spike synchrony. Instead, the importance of partial phase synchrony is shown to lie in its influence on the response of the population to stimulation, which we illustrate using first spike time histograms. PMID- 18163206 TI - Brief report: enhanced picture naming in autism. AB - Language and communication deficits are key diagnostic criteria for autism. However, not all aspects of language are equally affected. Here we present evidence of enhanced performance of a critical aspect of language-word processing in children with autism. The results have implications for explanatory theories of autism and language, and for the development of therapeutic approaches. PMID- 18163207 TI - Fluctuating asymmetry and sexual orientation in men and women. AB - The present study measured handedness and bodily fluctuating asymmetry (FA), two markers of developmental instability, in 89 heterosexual and 78 gay men and women. Asymmetry in ear breadth, ear length, ankle breadth, second digit length, fourth digit length, and two composite indices were calculated for each participant and a modified Edinburgh Inventory was used to assess handedness. Results showed that, for men, there was a significant positive correlation between three measures of FA and sexual orientation scores (SOS). As ear breadth FA and two composite FA scores increased so did SOS on a modified Kinsey scale (where 0 indicated exclusive heterosexuality and 6 indicated exclusive homosexuality). No significant relationships were found between SOS and FA for women. Similarly, sexual orientation was not related to handedness for either sex. The theoretical implications of the present results are discussed. PMID- 18163208 TI - Mating systems of Psychotria tenuinervis (Rubiaceae): distance from anthropogenic and natural edges of Atlantic forest fragment. AB - The aim of this study was to determine and compare the mating systems among Psychotria tenuinervis populations at anthropogenic edges, natural edges, and the forest interior using allozyme electrophoresis of naturally pollinated progeny arrays. P. tenuinervis showed low outcrossing rates, varying from 37% to 50% of the mating attributable to outcrossing and 50% to 63% attributable to self fertilization, in the three habitats. The forest interior had the highest outcrossing rate (t(m) = 0.50 and t(s) = 0.43) among the three habitats. However, there were no differences in either multilocus or single-locus rates among the three habitats, indicating that the contribution of biparental inbreeding to the apparent selfing rate in these populations was very low. The multilocus (t(m)) and single-locus (t(s)) outcrossing rates for the P. tenuinervis in the sample plots within each habitat showed great heterogeneity. In conclusion, edge creation seems not to influence its mating systems. Additionally, although P. tenuinervis is a distylous species, the population's inbreeding can be attributed almost entirely to self-fertilization. PMID- 18163209 TI - Association of interleukin-6, interleukin-12, and interleukin-10 gene polymorphisms with essential hypertension in Tatars from Russia. AB - Essential hypertension is a common disease with fatal clinical complications. Epidemiological and family studies have confirmed the role of genetic predisposition in its development. Hypertensive patients have been shown to have an altered profile of pro- and anti-inflammatory cytokines. The aim of our investigation was to reveal the association of interleukin-6, interleukin-12, and interleukin-10 gene polymorphisms with essential hypertension and its clinical complications in a Tatar ethnic group from Bashkortostan, Russia. The study involved 362 hypertensive patients and 244 healthy subjects from this Tatar ethnic group (Bashkortostan, Russia). DNA was isolated from whole venous blood using phenol-chloroform extraction by the standard method. IL6 -572 G/C, IL12B 1159 C/A, and IL10 -627 C/A genotypes were typed using polymerase chain reaction followed by restriction enzyme digestion. We found that the IL10 -627 *C/*C genotype was associated with decreased risk of hypertension (OR = 0.64, P = 0.035). IL6 genotypes and allele distribution did not differ significantly between subjects with and without hypertension, but the IL6 -572 *G/*G genotype frequency was found to be significantly higher among those patients who had stroke, compared with normotensive control subjects (P = 0.036). Carriers of the IL12B 1159 *A/*A genotype had a lower risk of stroke (OR = 0.38, P = 0.028). Our study has shown the association between IL10 -627 C/A polymorphism and essential hypertension in the Tatar ethnic group from Bashkortostan, Russia. The IL10 627*C/*C genotype was found to be protective against hypertension. We also demonstrated that hypertensive patients with the IL12B *A/*A and IL6 *G/*G genotypes had increased risk of stroke. Our results suggest a role for cytokines in cardiovascular disease development in the Tatar ethnic group, but further investigation is needed. PMID- 18163210 TI - Growth-inhibitory effect of adiponectin via adiponectin receptor 1 on human breast cancer cells through inhibition of S-phase entry without inducing apoptosis. AB - Adiponectin is one of the most important adipocytokines secreted from adipose tissue. In addition to its effects on glucose and fatty acid metabolism, it has been reported that adiponectin has a direct growth-inhibitory effect on breast cancer cells. However, it still remains to be established how adiponectin affects cell cycle and apoptosis and whether or not its inhibitory effect is mediated through adiponectin receptors. Here, we demonstrated that adiponectin treatment resulted in a significant dose-dependent growth inhibition of both MDA-MB-231 and T47D cells. In both cell lines, the G0/G1 population significantly increased after adiponectin treatment, but apoptosis was not induced. High expression of mRNA and protein of adiponectin receptor 1 was observed, but expression of adiponectin receptor 2 was very low in both cell lines. Treatment with small interference RNA against adiponectin receptor 1 significantly reduced the growth inhibition induced by adiponectin in both cell lines. Taken together, adiponectin decreases breast cancer cell proliferation by inhibiting the entry into S-phase without inducing apoptosis, and this inhibitory effect is mediated through adiponectin receptor 1. PMID- 18163211 TI - Medical therapy: options and uses. AB - Since the initial use of medical treatment for acromegaly, several advances have been made in the understanding of the pathophysiology of growth hormone producing tumors, resulting in the development of multiple medical options and novel treatments. Currently there are three major classes of medication available for the treatment of acromegaly: somatostatin receptor ligands, growth hormone receptor antagonists, and dopamine agonists. Somatostatin receptor ligands are the treatment of choice for acromegaly due to their effectiveness in controlling growth hormone excess in approximately 60% of patients and their beneficial effects on tumor volume. Clinical trials have demonstrated efficacy of pegvisomant in up to 97% of patients, but long term data and safety have yet to be established. Dopamine agonists are inexpensive, but their use is hampered by their lack of efficacy compared to other medications. Medical therapy has an established role as adjuvant therapy after non-curative surgery, as well as primary therapy for selected patients unsuitable for surgical resection. Medical treatment to control growth hormone hypersecretion is often needed after radiation therapy until the effects are evident. Preliminary data suggest a potential role for medical treatment prior to surgical resection, surgical debulking to improve medical efficacy, and combination therapy with multiple medications from the three classes. More studies are required, however, to validate the utility of these approaches in treating acromegaly. With the available therapies, disease control can be achieved in nearly all patients with acromegaly. PMID- 18163212 TI - Medical consequences of acromegaly: what are the effects of biochemical control? AB - This chapter discusses the effects of biochemical control of acromegaly on cardiovascular diseases, metabolic complications, respiratory abnormalities, malignancies and bone alterations. Acromegaly is associated with increased morbidity and mortality for cardiovascular and respiratory complications, whereas neoplasms seem to be a minor cause of increased risk of death. Other associated diseases are osteoarthritis, carpal tunnel syndrome, fatigue, visual abnormalities and reproductive disorders. Acromegaly results in premature death because of prolonged elevation of GH an IGF-I levels, and a strong biochemical control improves well-being and restores life expectancy to normal. The main goals of medical treatment of acromegaly include normalization of biochemical markers of disease activity, improvement in signs and symptoms of the disease, removal or reduction of tumor mass and preservation of pituitary function. PMID- 18163213 TI - Management of acromegaly: is there a role for primary medical therapy? AB - Acromegaly is a chronic, debilitating disease caused by chronic growth hormone (GH) hypersecretion which results in chronic medical comorbidities, poor quality of life and high mortality rates. Successful treatment can improve clinical signs and symptoms and normalize mortality rates. Over 95% of acromegaly is caused by a somatotroph adenoma of the pituitary, and the first-line treatment is generally transsphenoidal surgery, which can be curative in 50-60% of patients. Nonetheless, high rates of persistent acromegaly following surgery and the limited efficacy of radiation therapy necessitate chronic medical treatment for many patients. Somatostatin analogues have become the preferred first-line medical therapy for many practitioners, as they achieve better biochemical and direct tumor control than the dopamine agonists, and long-acting preparations make once monthly administration possible. Cabergoline, a dopamine agonist, offers a lower-cost option and may be effective in patients with a pituitary tumor that co-secretes GH and prolactin. Pegvisomant is a GH receptor antagonist that produces exceptional biochemical response rates but lacks any direct effects on the tumor, which may limit its effectiveness as life-long monotherapy. Combinations of these three drug classes have not been rigorously studied, and preliminary trials do not suggest improved clinical outcomes. While medical treatment options for acromegaly have significantly improved over the last 30 years, limitations remain, and a multi-specialty team approach is necessary for the effective long-term management of patients with acromegaly. PMID- 18163214 TI - Tripling of methamphetamine/amphetamine use among homeless and marginally housed persons, 1996-2003. AB - Methamphetamine/amphetamine (MA)-related morbidity and mortality has been increasing in the United States. MA use is associated with high-risk sexual behavior and syringe-sharing practices. Homeless and marginalized housed persons (H/M) have high rates of substance use and mental health disorders. Little is known about trends of MA use among the H/M. The objective of this study was to quantify increases in MA use among H/M in San Francisco and to determine which demographic and behavioral subgroups have experienced the greatest increases in MA use. We conducted serial cross-sectional population-based studies in three waves: 1996-1997, 1999-2000, and 2003 and studied 2,348 H/M recruited at shelters and lunch lines. The main outcome was self-reported current (30-day) MA use. We found a tripling of current MA use among H/M persons from 1996 to 2003, with a sevenfold increase in smoked MA use. MA use doubled to tripled in most demographic and behavioral subgroups, whereas it quadrupled in those under age 35, and there was a fivefold increase among HIV-infected persons. The increase in MA use among H/M places a vulnerable population at additional increased risk for HIV infection and MA-use related morbidity and mortality. Among HIV-infected H/M, the increase in MA use has important public health implications for the development and secondary transmission of drug-resistant HIV caused by synergistic neurocognitive decline, poor adherence to HIV medications, and increased sexual risk behavior. Clinicians caring for H/M persons should inquire about MA use, refer interested MA users to MA dependence treatment programs and provide targeted HIV sexual risk reduction counseling. For HIV-infected H/M MA users, clinicians should closely monitor adherence to HIV or other chronic medications, to avoid unnecessary morbidity and mortality. Further research is needed to elucidate the most effective prevention and treatment for MA use and dependence among the H/M. PMID- 18163215 TI - Tuning the HNN experiment: generation of serine-threonine check points. AB - We describe here the tunability of the HNN experiment to obtain certain residue specific peak patterns in the spectra of ((15)N, (13)C) labeled proteins. This is achieved by tuning a band-selective 180 degrees pulse on the carbon channel in the pulse sequence, whereby one can tamper with the C(alpha)-C(beta) coupling evolutions for the different residues. Specifically, we generate distinctive peak patterns for serine and threonine and their neighbors in the different planes of the three dimensional spectrum. These provide useful anchor points during sequential assignment of backbone resonances. The performance of this experiment, referred to as HNN-ST here, is demonstrated using two proteins, one properly folded and the other completely denatured. With the availability of high field spectrometers, techniques such as TROSY, and ever increasing sensitivities in the probes, this experiment with its large number of check points has a great potential for rapid and unambiguous backbone resonance assignment in large proteins. PMID- 18163216 TI - Apolipoprotein E gene is related to mortality only in normal weight individuals: the Rotterdam Study. AB - OBJECTIVE: To investigate the relationship between the apolipoprotein E (APOE) gene and the risk of mortality in normal weight, overweight and obese individuals. METHODS AND RESULTS: In a population-based study of 7,983 individuals aged 55 years and older, we compared the risks of all-cause and coronary heart disease (CHD) mortality by APOE genotype, both overall and in subgroups defined by body mass index (BMI). We found significant evidence for interaction between APOE and BMI in relation to total cholesterol (p=0.04) and HDL cholesterol (p<0.001). Overall, APOE*2 carriers showed a decreased risk of all-cause mortality. Analyses within BMI strata showed a beneficial effect of APOE*2 only in normal weight persons (adjusted hazard ratio (HR) 0.7[95% CI 0.5 0.9]). APOE*2 was not associated with a lower risk of all-cause mortality in overweight or obese persons. The effect of APOE*2 in normal weight individuals tended to be due to the risk of CHD mortality (adjusted HR 0.5 [95% CI 0.2-1.2]). CONCLUSION: The APOE*2 allele confers a lower risk of all-cause mortality only to normal weight individuals. PMID- 18163217 TI - Atmospheric polycyclic aromatic hydrocarbons (PAHs) in the urban air of Delhi during 2003. AB - Atmospheric concentrations of polycyclic aromatic hydrocarbons (PAHs) in Delhi were evaluated to study particulate PAHs profiles during the different seasons of 2003. Samples of urban suspended particulate matter were collected during January 2003 to December 2003 at three locations (Okhla, Dhaulakuan and Daryaganj), using a high volume sampler provided with glass fiber filters. Samples were analyzed using the gas chromatography technique. The annual average concentrations of total PAHs were found as 1,049.3 ng/m(3) at Okhla, 1,344.37 ng/m(3) at Daryaganj, and 1,117.14 ng/m(3) at Dhaulakuan. The seasonal average concentrations were found to be maximum in winter and minimum during the monsoon season. Principal Component Analysis (PCA) of the data was also carried out and the results indicate that diesel and gasoline driven vehicles are the principal sources of PAHs at all the three sites under investigation. Other sources might come from stationary combustion sources such as cooking fuel combustion and industrial emission. PMID- 18163218 TI - Spatio-temporal variations of organic carbon and chlorophyll degradation products in the surficial sediments of Izmir Bay (Aegean Sea/Turkey). AB - The aim of this research is to determine the effects of Izmir Big Channel Waste Water Treatment Project on the sediment quality of Izmir Bay. Wastewater treatment improves the water quality. However, sediment does not respond to this treatment as fast as water column. Monitoring of bottom water and sediment quality is necessary for identification of the recovery of the whole ecosystem. For this purpose, bottom water and sediment samples were collected from three stations which are located in the middle and inner parts of the Izmir Bay on a monthly basis between January 2003 and December 2003. Values measured at stations ranged between; 0.54-12.82 microg/L for chlorophyll-a, 0.09-9.32 microg/L for phaeopigment, 0.05-1.91 mg/L for particulate organic carbon in bottom waters, 11.88-100.29 microg/g for chlorophyll degradation products and 1.12-5.39% for organic carbon in sediment samples. In conclusion, it was found that grazing activity explained carbon variations in sediment at station 2, but at station 1 and station 3 carbon variations in sediment were not related to autochthonous biological processes. PMID- 18163219 TI - Selenium and vitamin E increases polymorphonuclear cell phagocytosis and antioxidant levels during acute mastitis in riverine buffaloes. AB - Antioxidant, antiinflammatory and phagocytic activities were studied in milk polymorphonuclear cells (PMNs) isolated from healthy buffaloes (group I) and during clinical mastitis with the treatment of Enrofloxacin alone (group II) and combined treatment with Enrofloxacin and Vitamin E plus selenium (group III). On days 0,3, 8 and 15 the milk Somatic cell count (SCC) were significantly higher in mastitic milk than in milk obtained from healthy buffaloes. In group II SCC decreased significantly on day 3 and day 8, however in group III reduction in SCC was observed on day 3, day 8 and day 15 (P < 0.05). The antiinflammatory activity was evaluated by determining nitrite plus nitrate (NOx) production in the milk PMNs before treatment and on day 8. NOx activity was significantly higher in mastitic milk than from healthy controls, both before and after treatment (P < 0.05). In group II and group III the activity decreased significantly on day 8 (P < 0.05). The Glutathione peroxidase (GSH-Px) activity was estimated in the milk polymorphonuclear cell (PMNs) supernatant. GSH-Px activity was significantly lower in mastitic buffaloes than in healthy controls, both before and after treatment (P < 0.05). In group II levels did not change in response to treatment, whereas in group III levels had increased significantly on day 8 (P < 0.05). The phagocytic activity (PA) (percentage of neutrophil that had phagocytosed 1-6 bacteria) and phagocytic index (PI) (average number of bacteria/ leukocytes counted in 100 cells) of the milk PMNs was significantly lower in mastitic buffaloes (P < 0.05). In group II the PA and PI did not change in response to treatment, whereas in group III both the parameters had increased significantly on day 8 (P < 0.05). The results of the present experiment indicated enhancement of antioxidative and cellular defense and reduction of somatic cell count in the mastitic animals treated with Enrofloxacin and Vitamin E plus Selenium as compared to the Enrofloxacin treatment alone. Hence Vitamin E plus selenium therapy may be added along with the antibiotics for effective amelioration of intramammary infection in buffaloes. PMID- 18163220 TI - Molecular cloning and characterisation of a pattern recognition protein, lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) from Chinese shrimp Fenneropenaeus chinensis. AB - A pattern recognition protein (PRP), lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) cDNA was cloned from the haemocyte of Chinese shrimp Fenneropenaeus chinensis by the techniques of homology cloning and RACE. Analysis of nucleotide sequence revealed that the full-length cDNA of 1,275 bp has an open reading frame of 1,098 bp encoding a protein of 366 amino acids including a 17 amino acid signal peptide. Sequence comparison of the deduced amino acid sequence of F. chinensis LGBP showed a high identity of 94%, 90%, 87%, 72% and 63% with Penaeus monodon BGBP, Litopenaeus stylirostris LGBP, Marsupenaeu japonicus BGBP, Homarus gammarus BGBP and Pacifastacus leniusculus LGBP, respectively. The calculated molecular mass of the mature protein is 39,857 Da with a deduced pI of 4.39. Two putative integrin binding motifs, RGD (Arg-Gly-Asp) and a potential recognition motif for beta-1,3-linkage of polysaccharides were observed in LGBP sequence. RT-PCR analysis showed that LGBP gene expresses in haemocyte and hepatopancreas only, but not in other tissues. Capillary electrophoresis RT-PCR method was used to quantify the variation of mRNA transcription level during artificial infection with heat-killed Vibrio anguillarum and Staphylococcus aureusin. A significant enhancement of LGBP transcription was appeared at 6 h post-injection in response to bacterial infection. These results have provided useful information to understand the function of LGBP in shrimp. PMID- 18163221 TI - Determinants of objective and subjective quality of life in first-time-admission schizophrenic patients in Poland: a longitudinal study. AB - OBJECTIVES: This study involved a naturalistic, prospective project assessing objective and subjective dimensions of quality of life (QoL) and the range of its putative determinants in first-episode schizophrenia. METHODS: Seventy-four patients, mean age 24.7 years, participated in the study. The patients were assessed 1 month following the first hospitalisation (Time 1), 12 months later (Time 2), and 4-6 years after Time 1 (Time 3). The Social Functioning Scale and World Health Organisation Quality of Life-BREF (WHOQOL-BREF) were applied to evaluate objective and subjective QoL, respectively. The Positive and Negative Syndrome Scale was used to examine psychopathological status, and the Global Assessment Scale was applied to evaluate social adjustment prior to hospitalisation. RESULTS: The objective and subjective QoL was relatively low at Time 1. Over time, the objective QoL improved in three domains, decreased in one domain, and remained stable in three domains. The subjective QoL did not change. The most important predictors of both dimensions of QoL were psychopathology and duration of untreated psychosis. CONCLUSIONS: Because psychopathology seems to have the greatest impact on the QoL, there is a need to develop community psychosocial treatment to reduce these symptoms and to support patients in the early phase of the disease. PMID- 18163222 TI - Allergens in atopic dermatitis. AB - Allergens play an essential role in atopic dermatitis, either intrinsic or extrinsic. They provoke cutaneous inflammation via IgE-dependent and cell mediated immune reactions. Food allergens have a well-known contribution to disease activity of atopic dermatitis, especially in infants and young children. However, the importance of inhaled allergens is still under investigation. For clinical implication, identification of individualized allergens is an ideal strategy for better control of atopic dermatitis and avoidance of atopic march. The aim of this article is to discuss the common allergens in atopic dermatitis (AD), the specificity and sensitivity of laboratory tests for allergens, and the clinical effect of various preventions. PMID- 18163223 TI - Role of bacterial pathogens in atopic dermatitis. AB - The skin of atopic dermatitis (AD) patients exhibits a striking susceptibility to colonization and infection with Staphylococcus aureus. This review summarizes our understanding about the role of S. aureus in AD. Indeed, S. aureus colonization is both a cause and a consequence of allergic skin inflammation. The mechanisms that allergic skin inflammation of AD promotes the increase of S. aureus colonization include skin barrier dysfunction, increased synthesis of the extracellular matrix adhesins for S. aureus, and defective innate immune responses due to decreased production of endogenous antimicrobial peptides. On the other hand, the exotoxins secreted by S. aureus are superantigens. Staphylococcal superantigens (SsAgs) may penetrate the skin barrier and contribute to the persistence and exacerbation of allergic skin inflammation in AD through the stimulation of massive T cells, the role of allergens, direct stimulation of antigen-presenting cells and keratinocytes, the expansion of skin homing cutaneous lymphocyte-associated antigen-positive T cells, and the augmentation of allergen-induced skin inflammation. SsAgs also induce corticosteroid resistance. In therapeutic interventions, anti-inflammatory therapy alone is very effective in reducing S. aureus colonization on the skin, but antibiotic treatment alone is unable to improve the allergic skin inflammation of AD. Therefore, we recommend the combination therapy of anti inflammatory drugs and antibiotics in the AD patients with secondary bacterial infection, exacerbated AD, or poorly controlled AD. However, when AD is well controlled by anti-inflammatory drugs alone, we do not recommend the antibiotic therapy. PMID- 18163224 TI - The genetics of atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a complex genetic disorder influenced by environmental factors. The mode of inheritance and genes involved are not clear. RESULTS: This report here is focusing on the current progress in searching the disease-susceptibility genes of AD via both the linkage studies and candidate gene approaches. Genome-wide linkage studies have identified multiple susceptibility loci on 3q and 17q. Candidate region linkage studies identify other susceptibility loci on 5q23-33, 11q13, and 13q12-14. At least 28 candidate genes have to date been verified in association studies, but only association with genes of interleukin (IL)-4, IL-13, IL-4RA, mast cell chymase, and serine protease inhibitor, kazal-type 5 have been replicated in more than two different studies. More halpotype tests and family-based association studies may help to shed more light for the candidate gene approach. CONCLUSION: Determining the candidate susceptibility genes for AD is not only helping understanding the pathophysiology but also affecting the response to therapy, which is important in pharmacogenetics. The effect of environmental trigger may also have to be considered to elucidate the real face of the disease. PMID- 18163225 TI - Cellular aspects of atopic dermatitis. AB - Atopic dermatitis (AD) is a highly pruritic, chronic, and relapsing inflammatory skin disease. Recent interest in AD has been sparked by reports of its increasing prevalence and its contribution to increasing health care costs. A precise understanding of immunologic mechanisms is crucial for the development of effective treatment strategies for AD. Various studies reveal that AD has a multifactorial cause with the activation of complex immunologic and inflammatory pathways. This review will discuss cellular-mediated immunological pathomechanisms of AD. Emphasis will be given to the role played by T cells, antigen-presenting cells, eosinophils, and keratinocytes. We also examine the immunological effect of superantigens on various inflammatory cells including T regulatory cells. PMID- 18163226 TI - Emerging treatment of atopic dermatitis. AB - Atopic dermatitis is a chronically relapsing eczematous disease of the skin. A wide range of therapeutic regimens has been used for atopic dermatitis. A better understanding of its pathogenesis will also lead to the development of novel approaches to treating this disease. This article reviews the recent advances in allergen-specific sublingual immunotherapy and therapy with antileukotriene drugs, probiotics, mycophenolate mofetil, leflunomide, and intermittent fluticasone propionate ointment, which the authors expect will be clinically useful therapies in the near future. PMID- 18163228 TI - A novel subcellular collagen organization process visualized by total internal reflection fluorescence microscopy. AB - The alpha1beta1 and alpha2beta1 integrins belong to a family of cell-surface molecules involved in structural contacts and signal-transduction events across the cell membrane. Employing two-dimensional substrates coated with fluorescently labeled type I collagen, we have discovered a novel subcellular matrix remodeling event that is particular to cells that express the fibrillar collagen receptor alpha2beta1. Cells expressing alpha1beta1 also perform this collagen organization process, but less proficiently. This work will provide a basis for subsequent studies of cell-mediated collagen fibril assembly. PMID- 18163227 TI - Treatment of eczema. AB - Eczema is a chronic inflammatory skin disease that has reached nearly epidemic proportions in childhood. Moreover, it is a difficult disease to control and, with its onset in childhood, is often the first manifestation of atopy. The clinical features of eczema include itchy red skin accompanied by dryness and lichenification. In the past, treatment options consisted primarily of avoidance of soap and water. These options have considerably improved with both nonpharmacologic and pharmacologic approaches. However, eczema is still a treatment challenge. Part of the problem in developing new treatment options has been the relative failure in translating basic science information into clinical application. It is hoped that the newer biologics will help bridge this gap and lead to greater success rates. PMID- 18163229 TI - Influence of systematically varied nano-scale topography on cell morphology and adhesion. AB - The types of cell-matrix adhesions and the signals they transduce strongly affect the cell-phenotype. We hypothesized that cells sense and respond to the three dimensionality of their environment, which could be modulated by nano-structures on silicon surfaces. Human foreskin fibroblasts were cultured on nano-structures with different patterns (nano-post and nano-grate) and heights for 3 days. The presence of integrin alpha(5), beta(1), beta(3), paxillin and phosphorylated FAK (pFAK) were detected by western blot and immunofluorescence. Integrin beta(3) exhibited stronger signals on nano-grates. pFAK and paxillin were observed as small dot-like patterns on the cell-periphery on nano-posts and as elongated and aligned patterns on nano-grates. Collectively, our observations highlighted the presence of focal (integrin beta(1), beta(3), pFAK, paxillin), fibrillar (integrin alpha(5), beta(1)) and 3-D matrix (integrin alpha(5), beta(1), paxillin) adhesions on nano-structures. The presented nano-structures offer interesting opportunities to study the interaction of cells with topographical features comparable to the size of extracellular matrix components. PMID- 18163230 TI - Early response of endothelial cells to flow is mediated by VE-cadherin. AB - Endothelial cells are known to respond to flow onset by increasing actin turnover rate. Current models assume that an increase in the actin turnover rate should result in a rise in cell crawling speed. Here we report that confluent endothelial monolayer shows an unexpected behavior: cell crawling speed decreases by approximately 40% within the first 30 min of flow onset. A drop in crawling speed has not been observed in either subconfluent endothelial cells or in VE cadherin-deficient cells. We found that flow onset caused an increase in the number of VE-cadherin-GFP molecules in the junctions and elicited changes in the cytoskeleton-associated fractions of alpha, beta -catenins and VE-cadherin. Flow application also increased the strength of interactions of endothelial cells with surfaces coated with recombinant VE-cadherin. These observations suggest that endothelial cell junctional proteins respond to flow transiently by increasing the strength of intercellular attachments early after flow onset and support the view on the active role of intercellular adhesions in mechanotransduction. PMID- 18163231 TI - Akt phosphorylates Connexin43 on Ser373, a "mode-1" binding site for 14-3-3. AB - Connexin43 (Cx43) is a membrane-spanning protein that forms channels that bridge the gap between adjacent cells and this allows for the intercellular exchange of information. Cx43 is regulated by phosphorylation and by interacting proteins. "Mode-1" interaction with 14-3-3 requires phosphorylation of Ser373 on Cx43 (Park et al. 2006). Akt phosphorylates and targets a number of proteins to interactions with 14-3-3. Here we demonstrate that Akt phosphorylates Cx43 on Ser373 and Ser369; antibodies recognizing Akt-phosphorylated sites or phospho-Ser "mode-1" 14-3-3-binding sites recognize a protein from EGF-treated cells that migrates as Cx43, and GST-14-3-3 binds to Cx43 phosphorylated endogenously in EGF-treated cells. Confocal microscopy supports the co-localization of Cx43 with Akt and with 14-3-3 at the outer edges of gap junctional plaques. These data suggest that Akt could target Cx43 to an interaction with 14-3-3 that may play a role in the forward trafficking of Cx43 multimers and/or their incorporation into existing gap junctional plaques. PMID- 18163232 TI - The murine CD99-related molecule CD99-like 2 (CD99L2) is an adhesion molecule involved in the inflammatory response. AB - CD99, a glycoprotein found on the surfaces of leukocytes and concentrated at the borders of endothelial cells, plays a major role in the migration of leukocytes across endothelial cells into sites of inflammation, and has other roles in thymocyte development. The human and mouse genomes encode only two proteins related to CD99. One of these, XGA, is a red blood cell surface antigen. The function of the other, CD99-like 2 (CD99L2), is not known. We cloned mouse CD99L2 and used CD99L2 isolated from transfected cells to raise specific antibodies. Similar to human CD99, CD99L2 was expressed at the borders between transfected cells as well as on mouse leukocytes and vascular endothelial cells in situ. Transfection of L cell fibroblasts with CD99L2 imparted to them the ability to adhere to each other in a divalent cation-dependent, homophilic manner. Anti CD99L2 antibody blocked influx of neutrophils and monocytes into a site of inflammation in vivo. PMID- 18163233 TI - Repeated simulated ischemia and protection against gap junctional uncoupling. AB - Ischemic preconditioning increases the heart's tolerance to a subsequent longer ischemic period. The aim of this study was to investigate the effect of early and delayed preconditioning on gap junction communication, connexin abundance, and phosphorylation in cultured neonatal rat cardiac myocytes. Prolonged ischemia followed 5 minutes after preconditioning in the early protocol, whereas 20 hours separated preconditioning and prolonged ischemia in the delayed preconditioning protocol. Gap junctional intercellular communication (GJIC) was assessed by Lucifer yellow dye transfer. An initial reduction in communication in response to sublethal ischemia was observed. This may be one mechanism whereby neighboring cells are protected from damaging substances produced during the first phase of subsequent regional ischemia in early preconditioning protocols. With respect to delayed preconditioning, the transient decrease in GJIC disappeared prior to prolonged ischemia, indicating that other mechanisms are responsible for delayed protection. Both early and delayed preconditioning preserved intercellular coupling after prolonged ischemia and this correlated with presence of less connexin43 dephosphorylation assessed by immunoblot. PMID- 18163234 TI - 2006 Annual Report of the American Association of Poison Control Centers' National Poison Data System (NPDS). AB - BACKGROUND: The American Association of Poison Control Centers (AAPCC; http://www.aapcc.org ) maintains the National Poison Data System (NPDS). Today, 60 of the nation's 61 US poison centers upload case data automatically. Most upload every 1- 60 minutes (median 11 minutes) to NPDS creating a real-time national exposure database and surveillance system. METHODOLOGY: We analyzed the case data tabulating specific indices from NPDS. The methodology was similar to that of previous years. Where changes were introduced, the differences are identified. Fatalities were reviewed by a team of 27 medical and clinical toxicologists and assigned to 1 of 6 categories according to Relative Contribution to Fatality (RCF). RESULTS: Over 4 million calls were captured by NPDS in 2006: 2,403,539 human exposure calls, 1,488,993 information requests, and 128,353 nonhuman exposure calls Substances involved most frequently in all human exposures were analgesics. The most common exposures in children less than age 6 were cosmetics/personal care products. NPDS documented 1,229 human fatalities. CONCLUSIONS: Poisoning continues to be a significant cause of morbidity and mortality in the US. NPDS represents a valuable national resource to collect and monitor US poisoning exposure cases. It offers one of the few real-time surveillance systems in existence, provides useful data and is a model for public health surveillance. PMID- 18163236 TI - Understanding the limitations of retrospective analyses of poison center data. PMID- 18163235 TI - Atypical antipsychotic medication poisoning: an evidence-based consensus guideline for out-of-hospital management. AB - The objective of this guideline is to assist poison center personnel in the appropriate out-of-hospital triage and out-of-hospital management of patients with suspected acute ingestions of atypical antipsychotic medications by 1) describing the process by which an ingestion of an atypical antipsychotic medication might be evaluated, 2) identifying the key decision elements in managing cases of atypical antipsychotic medication ingestion, 3) providing clear and practical recommendations that reflect the current state of knowledge, and 4) identifying needs for research. This guideline applies to ingestion of atypical antipsychotic medications alone. Co-ingestion of additional substances could require different referral and management recommendations depending on the combined toxicities of the substances. This guideline is based on an assessment of current scientific and clinical information. The expert consensus panel recognizes that specific patient care decisions might be at variance with this guideline and are the prerogative of the patient and the health professionals providing care, considering all of the circumstances involved. This guideline does not substitute for clinical judgment. The grade of recommendation is in parentheses. 1) Patients with stated or suspected self-harm or the recipient of a potentially malicious administration of an atypical antipsychotic medication should be referred to an emergency department immediately. This activity should be guided by local poison center procedures. In general, this should occur regardless of the dose reported (Grade D). 2) Patients without evidence of self harm should have further evaluation, including determination of the precise dose ingested, presence of signs or symptoms of toxicity, history of other medical conditions, and the presence of co-ingestants (Grade C). 3) Asymptomatic patients without evidence of attempted self-harm are unlikely to develop symptoms if the interval between the ingestion and the call is greater than 6 hours. These patients do not need referral and should receive follow-up based on local poison center protocols (Grade C). 4) All patients less than 12 years of age who are naive to atypical antipsychotic medications and are experiencing no more than mild drowsiness (lightly sedated and can be aroused with speaking voice or light touch) can be observed at home unless they have ingested more than four times the initial adult dose for the implicated antipsychotic medication or a dose that is equal to or more than the lowest reported acute dose that resulted in at least moderate toxicity, whichever dose is smaller (i.e., aripiprazole 15 mg, clozapine 50 mg, olanzapine 10 mg, quetiapine 100 mg, risperidone 1 mg, ziprasidone 80 mg) (Grade D). 5) All patients 12 years of age or older who are naive to atypical antipsychotic medications and are experiencing no more than mild drowsiness can be observed at home unless they have ingested more than five times the initial adult dose for the implicated antipsychotic medication (i.e., aripiprazole 50 mg, clozapine 62.5 mg, olanzapine 25 mg, quetiapine 125 mg, risperidone 5 mg, ziprasidone 100 mg) (Grade D). 6) Patients who use atypical antipsychotic medications on a chronic basis can be observed at home unless they have acutely ingested more than 5 times their current single dose (not daily dose) of the implicated antipsychotic medication (Grade C). 7) Patients who have ingested less than a threshold dose (see Recommendations 4-6) and are exhibiting no more than mild drowsiness can be observed at home with instructions to call the poison center if symptoms develop or worsen. If mild drowsiness is present at the time of the initial call, the poison center should make follow-up calls until at least 6 hours after ingestion. Consideration should be given to the time of day that home observation will take place. Observation during normal sleep hours might not be reliable. Depending on local poison center policy, patients could be referred to an emergency department if the observation would take place during normal sleeping hours of the patient or caretaker (Grade D). 8) Any patient already experiencing any signs or symptoms, other than mild drowsiness, thought to be related to atypical antipsychotic medication toxicity should be transported to an emergency department. Transportation via ambulance should be considered based on the condition of the patient and the length of time it will take the patient to arrive at the emergency department (Grade D). 9) Do not induce emesis (Grade D). 10) There are no specific data to suggest benefit from out-of-hospital administration of activated charcoal in patients exposed to atypical antipsychotic medications. Poison centers should follow local protocols and experience with the out-of-hospital use of activated charcoal in this context. Do not delay transportation in order to administer charcoal (Grade D). 11) For patients who merit evaluation in an emergency department, transportation via ambulance should be considered based on the condition of the patient and the length of time it will take the patient to arrive at the emergency department. Continuous cardiac monitoring should be implemented given reports of conduction disturbances associated with this class of medications. Provide usual supportive care en route to the hospital, including airway management and intravenous fluids for hypotension (Grade D). 12) Depending on the specific circumstances, follow-up calls should be made to determine outcome at appropriate intervals based on the clinical judgment of the poison center staff (Grade D). PMID- 18163237 TI - Unproven ingestion: an unrecognized bias in toxicological case series. AB - BACKGROUND: Case series of ingestion in preschool children may include patients without significant exposure if the substance is not measured. METHODS: In order to evaluate the unproven ingestion bias, we conducted, between January 2000 and June 2004, a retrospective analysis of a poison control center-based series of children <6 years old with a history of toxic methanol or ethylene glycol ingestion. RESULTS: Over the 54 month period, 115 children were referred to obtain a level. Of these, 102 children, aged 25 +/- 10 months, actually had a level analyzed. Only 21 patients had positive levels measured a median of 90 minutes post-ingestion. CONCLUSION: Our findings suggest that a significant fraction of purported cases were not confirmed. When a study aims at determining the toxicity of the substance, measurements of the xenobiotic should be required in any case series involving preschool aged children in order to decrease the unproven ingestion bias. PMID- 18163238 TI - Images in clinical toxicology. Abdominal pain and rectal bleeding after an enema. PMID- 18163239 TI - A provincial and territorial review of hepatitis A in men who have sex with men. PMID- 18163240 TI - The burden of varicella and zoster in British Columbia 1994-2003: baseline assessment prior to universal vaccination. PMID- 18163241 TI - Addendum. National Advisory Committee on Immunization (NACI) Statement on influenza vaccination for the 2007-2008 season. PMID- 18163242 TI - The process to establish and implement national goals and recommendations for vaccine preventable diseases in Canada under the National Immunization Strategy. PMID- 18163244 TI - Statement of retraction. PMID- 18163243 TI - ATP and adenosine trigger the interaction of plasma membrane IP3 receptors with protein kinase A in oviductal ciliated cells. AB - We have demonstrated that adenosine did not produce any change of intracellular free Ca2+ concentration ([Ca2+]i) in oviductal ciliated cells; however, it increased the ATP-induced Ca2+ influx through the activation of protein kinase A (PKA). Uncaging of IP3 and cAMP triggered a larger Ca2+ influx than did IP3 alone. Furthermore, the IP3 effect was abolished by Xestospongin C, an IP3 receptor blocker. Whole-cell recordings demonstrated the presence of an ATP induced Ca2+ current, and the addition of adenosine increased the peak of this current. This effect was not observed in the presence of H-89, a PKA inhibitor. Using excised macro-patches of plasma membrane, IP3 generated a current, which was higher in the presence of the catalytic PKA subunit and this current was blocked by Xestospongin C. We show here that activation of plasma membrane IP3 receptors directly triggers Ca2+ influx in response to ATP and that these receptors are modulated by adenosine-activated PKA. PMID- 18163245 TI - Repair of facial fractures in the Iraq war combat theater. PMID- 18163246 TI - Morphogenesis in bat wings: linking development, evolution and ecology. AB - The evolution of powered flight in mammals required specific developmental shifts from an ancestral limb morphology to one adapted for flight. Through studies of comparative morphogenesis, investigators have quantified points and rates of divergence providing important insights into how wings evolved in mammals. Herein I compare growth,development and skeletogenesis of forelimbs between bats and the more ancestral state provided by the rat (Rattus norvegicus)and quantify growth trajectories that illustrate morphological divergence both developmentally and evolutionarily. In addition, I discuss how wing shape is controlled during morphogenesis by applying multivariate analyses of wing bones and wing membranes and discuss how flight dynamics are stabilized during flight ontogeny. Further, I discuss the development of flight in bats in relation to the ontogenetic niche and how juveniles effect populational foraging patterns. In addition, I provide a hypothetical ontogenetic landscape model that predicts how and when selection is most intense during juvenile morphogenesis and test this model with data from a population of the little brown bat, Myotis lucifugus. PMID- 18163247 TI - Modulation of intracellular signaling pathways to induce apoptosis in prostate cancer cells. PMID- 18163248 TI - Not up to speed. Boards still don't grasp gravity of some issues: report. PMID- 18163249 TI - Survey says ... voters rank healthcare as their biggest issue. PMID- 18163250 TI - 'Alarming' VA problems. Questions continue over quality of vets' healthcare. PMID- 18163251 TI - Probe spotlights director. Roper on DaVita, NQF, Robert Wood Johnson boards. PMID- 18163252 TI - Donations in conflict. Medical groups get $1.5 million from devicemakers. PMID- 18163253 TI - Overseas patients. Foreign exchange. PMID- 18163254 TI - Grassley hears, but will he listen? Community-benefit draft needs work, execs say. PMID- 18163255 TI - HHS tries EHR bonus plan. But docs are less enthusiastic, calling it a 'tiny step'. PMID- 18163256 TI - Cigna's N.Y. pact may be model. PMID- 18163257 TI - Mixing business and academics? Researchers may need to disclose their affiliations. PMID- 18163258 TI - A critical role. Roundtable discussion. PMID- 18163259 TI - [What is wrong with the shoulder. Sequelae of a bad foul]. PMID- 18163260 TI - [Cryptorchism. Treatment should be finished by age 2]. PMID- 18163261 TI - [Why the change in life style is ineffective in heart patients. Lack of compliance is even rewarded]. PMID- 18163262 TI - [Family practice highlights of stomach, liver and intestinal reference points. Your arsenal against abdominal problems]. PMID- 18163263 TI - [General practice study reveals simplification of therapy for physician and patients. Patient satisfaction and better quality of life]. PMID- 18163264 TI - [Thorough clarification of national health insurance statistics. This prevents regress actions]. PMID- 18163265 TI - Primary care. Match of the day. AB - Preston North End's Deepdale redevelopment is a prime example of the new wave of partnerships between primary care trusts and sports clubs. Warrington Wolves rugby league club was a pioneer, with a 1.3m pound sterling PCT health centre at its ground since 2005. Financial issues include sports club business stability. Benefits include health promotion opportunities. PMID- 18163266 TI - Occupational health. Get the blues all over again. PMID- 18163267 TI - Patient involvement. Young clients can become the boss. PMID- 18163268 TI - Safety loophole for foreign workers. PMID- 18163269 TI - Scotland health spending 'flatlining' for three years. PMID- 18163271 TI - Recruitment advertising. The rules of attraction. PMID- 18163270 TI - Workforce. State of the union. PMID- 18163272 TI - Maternity services. A lot of bottle. PMID- 18163273 TI - Equality. Getting up to speed on race. PMID- 18163274 TI - Effectiveness of influenza vaccination. PMID- 18163275 TI - Effectiveness of influenza vaccination. PMID- 18163276 TI - Cystic duct biliary adenoma. PMID- 18163277 TI - Eugenic practice and genetic science in Scandinavia and Germany: some comments on Peter Weingart's comparison of Sweden and Germany. PMID- 18163278 TI - Putting the state in its place: the domestic education debate in New Zealand. PMID- 18163279 TI - Whose mothers? Generational difference, war, and the Nazi cult of motherhood. PMID- 18163280 TI - [Scouting and Jewish youth in Liechtenstein during the Nazi era]. PMID- 18163281 TI - [Physical culture, sports, and military preparedness: on the upswing in physical education and public health in Sweden during World War II]. PMID- 18163283 TI - [Racial selection and living space: two core principles in the blood-and-soil doctrine of Richard Walther Darre]. PMID- 18163282 TI - ["I'm not going to jail, am I?" A study of Lex Veneris, its creation, practice, and effect on the lives of individuals, 1919-45]. PMID- 18163284 TI - Interethnic unions and the regulation of sex in colonial Samoa, 1830-1945. PMID- 18163285 TI - [An invisible frontier? Hajnal's European marriage pattern in the Netherlands and Taiwan, 1830-1945]. PMID- 18163287 TI - [Evaluation of the DOTS program by the analysis of successful factors in therapy for tuberculosis patients in Osaka city]. PMID- 18163286 TI - [Swiss women who had become foreigners by marriage and their children during World War II: a search in the Vaud cantonal archives]. PMID- 18163288 TI - [Microscopic variation of normal structure of mucosal cells of the middle ear and its clinical significance]. PMID- 18163289 TI - [History and development of the epidemiology service in Serbia]. PMID- 18163290 TI - Spelling out the Act. PMID- 18163291 TI - [Composition and carbon storage of woody debris in moist evergreen broad-leaved forest and its secondary forests in Ailao Mountains of Yunnan Provinve]. AB - This paper studied the composition and carbon storage of woody debris in the primary moist evergreen broad-leaved forest and its main secondary forests (regenerated Lithocarpus forest, Populus bonatii forest, and Alnus nepalensis forest) in Ailao Moutains of Yunnan Province. The results showed that in the primary forest, the carbon storage of woody debris amounted to 36.56 t x hm(-2). Castanopsis wattii, Lithocarpus xylocarpus and L. chintungensis were the main contributors, and most of them were the logs with larger diameter and at intermediate stage of decay. The unique environment of richer precipitation, higher humidity and lower temperature in the study area, and the decay-resistance of hardwood were favorable to the accumulation of woody debris. The three secondary forests had a carbon storage of 1.2-5.0 t x hm(-2), which decreased in the order of regenerated Lithocaropus forest > P. bonatii forest > A. nepalensis forest, showing a tendency of increasing carbon storage with succession course. PMID- 18163292 TI - [Photosynthetic capacity and water use efficiency of tropical montane rainforest seedlings or saplings in Hainan Island]. AB - By using a Li-6400 portable photosynthesis system, this paper measured the maximal photosynthetic capacity (P(max)) and water use efficiency (WUE) of the seedlings or saplings of 150 species of tropical montane rainforest in Hainan Island. The results showed that among the test 150 species, Quercus bambusifolia (Fagaceae) had the highest P(max) (13.27 micromol x m(-2) x s(-1)) and Macaranga denticulate (Euphorbiaceae) had the highest WUE (5.99 mmol x mol(-1)), while Decaspermum cambodianum (Myrtaceae) had the lowest P(max) (1.35 micromol x m(-2) x s(-1)) and Sapium discolor (Euphorbiaceae) had the lowest WUE (2.4 mmol x mol( 1)). The average P(max) was the highest in Moraceae but the lowest in Myrtaceae, and the average WUE was the highest in Symplocaceae but the lowest in Moraceae. However, there were no significant differences (P > 0.05) among the families. No significant differences (P > 0.05) in P(max) and WUE were also observed among arbors and shrubs. The P(max) value increased significantly (P < 0.05) with the widening of families' geographical distribution range, in the order of narrow distribution families < medium distribution families < wide distribution families, while no significant differences were existed among their WUE (P > 0.05). The P(max) value also increased with the widening of species' geographical distribution range, in the order of endemic species in Hainan Island < tropical species < tropical-subtropical species < tropical-subtropical-temperate species, but WUE was opposite to P(max), namely, in the order of endemic species in Hainan Island > tropical-subtropical species > tropical species > tropical-subtropical temperate species. PMID- 18163293 TI - [Responses of litter decomposition and nutrient release to simulated nitrogen deposition in an evergreen broad-leaved forest in southwestern Sichuan]. AB - An in situ experiment was conducted in an evergreen broad-leaved forest in southwestern Sichuan to study the responses of litter decomposition and nutrient release to simulated nitrogen deposition. Four treatments were installed, i.e., null level (CK), low nitrogen level (LN, 50 kg N x hm(-2) x a(-1)), moderate nitrogen level (MN, 100 kg N x hm(-2) x a(-1)) and high nitrogen level (HN, 150 kg N x hm(-2) x a(-1)). The results showed that it would take 4.72-6.33 years to decompose 95% of litter mass, with the highest decomposition rate in CK and the lowest one in HN. After 365 days, the litter decomposition rate in N-amended treatments was lower than that in CK, but significant difference was only observed between HN and CK (P < 0.05). The remained C was higher, and the remained N and K were significantly higher in N-amended treatments than in CK (P < 0.05). The remained P was also higher in N-amended treatments than in CK, but significant difference was only observed between LN and CK (P < 0.05). Compared with CK, all N-amended treatments had a 3.9%-23.7% increase of litter C/N ratio. During litter decomposition, element N featured a pattern of accumulation at early stage and release later, while C, P and K released all the time. Nitrogen deposition inhibited both the nutrient release from the litter and the decomposition of its lignin and cellulose. The effects of nitrogen deposition on litter decomposition in the forest changed from positive to negative as time passed, and the negative effect could be strengthened with the increase of deposited nitrogen concentration. PMID- 18163294 TI - [Characteristics of Caragana microphylla sap flow and water consumption under different weather conditions on Horqin sandy land of northeast China]. AB - Employing heat balance Dynamax packaged sap flow measuring system and automatic weather recording system, the sap flow of artificial Caragana microphylla community on Horqin sandy land of northeast China was monitored consecutively in 2006, and the photosynthetically effective radiation, air temperature, relative humidity and wind velocity were measured synchronously. According to the manual records of weather conditions, four most representative weather conditions were gathered up to analyze the relationships of C. microphylla sap flow and its single branch water consumption with test meteorological factors. The results showed that under high air temperature and intense radiation on sunny days, the diurnal variation of C. microphylla sap flow appeared a broad peak curve, so as to adapt the circumstance of drought and water shortage via lower transpiration. The diurnal variations of sap flow and its dominant affecting factors differed with weather conditions, and photosynthetically effective radiation was always the dominant factor affecting the sap flow. The variation of the sap flow was the result of comprehensive effects of multi-meteorological factors, and the overall variation trend of water consumption of single branch was declined in the order of sunny days > cloudy days > windy days > rainy days, with the mean value being 459, 310, 281 and 193 mg x d(-1), respectively. PMID- 18163295 TI - [Amount and vertical distribution of macropores in forest soils in Changbai Mountains]. AB - By using tension infiltrometer, this paper studied the amount and vertical distribution of macropores with the radius of > or = 0.5 mm, 0.25-0.5 mm and 0. 1 0. 25 mm in dark brown forest soil, brown coniferous forest soil, mountain soddy forest soil, and mountain tundra soil in Changbai Mountains, and the effects of these macropores on soil saturated flux. The results showed that in dark brown forest soil, the amount of macropores decreased with increasing soil depth, being faster from surface soil down to 40 cm deep and slower beneath the depth of 40 cm. In brown coniferous forest soil and mountain tundra soil, the amount of macropores increased with increasing soil depth; while in mountain soddy forest soil, there was a little change in the amount of macropores with soil depth. Though the macropores only occupied a small proportion to the soil volume, they still played an important role in the transfer of soil saturated flux. PMID- 18163296 TI - [Estimation models of understory shrub biomass and their applications in red soil hilly region]. AB - With 16 familiar species of understory shrub at Qianyezhou ecological experimental station in red soil hilly region under Chinese Academy of Sciences as test objects, crown area (A(c)) and projected volume (V(c)) were used as the variables for building quadratic and power allometric equations, respectively, to estimate the biomass of individual populations, and mixed-model was used to estimate the biomass of the 16 species. The best-fit models were applied to estimate the biomass of understory shrub in different forest types. The results showed that the biomass of shrub layer varied significantly among different stand types. With species-specific models, the biomass in deciduous, secondary, and coniferous forests was estimated as 4 773, 3 175 and 733 kg x hm(-2), respectively; while with mixed model, the estimation result was a little lower, being 3 946, 2 772 and 840 kg x hm(-2), respectively. Under the conditions of species-specific models being not established, mixed model was more convenient and practical in estimating the biomass of understory shrub. PMID- 18163297 TI - [Chinese pine tree ring width chronology and its relations to climatic conditions in Qianshan Mountains]. AB - Taking Chinese pine in Qianshan Mountains as a sample, the tree ring width chronology including standard, residual, and Arstan chronologies was established. The results showed that the tree ring width of Chinese pine had a higher correlation with the temperature in May - July and in September - November, and significant positive correlations were observed between the tree ring width and the extreme minimum temperature in July and mean minimum temperature in September. The chronology had significant or very significant correlations with the extreme minimum temperature in December and next January, mean minimum temperature in January, annual precipitation, and the precipitation in April, May and last December. Chinese pine had stronger responses to the monthly/yearly water vapor pressure and relative humidity. The yearly and most monthly evaporation had negative effects on the growth, being most significant for the evaporation in April - July. The narrowed tree rings recorded by the chronology demonstrated the 30 times of extreme drought since 1 800. The growth of Chinese pine in Qianshan Mountains was also affected by the climate changes on global and hemisphere scales. There existed 11-, 23- and 50- year- common periodicity between the chronology and solar activity, and 10-, 20- and 45- year- common periodicity between the chronology and geomagnetic activity. PMID- 18163298 TI - [Edge effects of forest landscape in upper reaches of Minjiang River]. AB - By using GIS and RS techniques, the edge effects of forest landscape in the upper reaches of Minjiang River, Sichuan Province of China were studied on landscape scale. The results showed that there was a distinct boundary between forestland and farmland, with the vegetation changed significantly. The biomass at forestland edge was lower than that in forestland interior, with the edge effect width being 60 m, whereas the biomass in farmland exterior was higher than that in farmland interior, with the edge effect width being 60-90 m. Forestland grassland boundary was naturally formed and changed gradually. The biomass at forestland edge was lower than that in forestland interior, while that at grassland edge was higher than that in grassland interior, with the edge effect being 60 m for forestland and 45-75 m for grassland. The edge effect of forestland-shrub land boundary was similar to that of forestland-grassland boundary, with the edge effect width being 60 m for forestland and 45-75 m for shrub land. PMID- 18163299 TI - [Growth response of Broussonetia papyrifera seedlings to VA mycorrhizal fungi inoculation]. AB - In an experiment with single inoculation (SI) and co-inoculation (CI) of three VA mycorrhizal fungi, i. e., Glomus mosseea (GM), Glomus versiforme (GV) and Glomus diaphanum (GD), the growth response of Broussonetia papyrifera seedlings in limestone area was studied. The results showed that after 3 months of growth, the aboveground-, underground-, and total biomass were increased significantly by the inoculation, being 2.49-8.19 times as much as the control. Treatment CI had the highest biomass, but the leaf number had little difference with the control. In SI, GD inoculation had the greatest effect; and CI was more effective than SI. The ground diameter, height, and total leaf area in CI were 1.5, 2.2, and 6.0 times as much as those in CK, respectively, and the root/shoot ratio in CI was the highest (0.446). There existed an interactive selection between host plants and VA mycorrhizal fungi. PMID- 18163300 TI - [Effects of nitrogen supply on Parthenocissus tricuspidata seedling morphology and nitrogen allocation]. AB - With hydroponic culture, this paper studied the effects of different nitrogen (N) supply levels (0, 0.15, 0.3, 0.45, 0.6, 0.75 g N x L(-1)) on the seedling morphology and N allocation of Parthenocissus tricuspidata. The results showed that lower N supply retarded the development of P. tricuspidata seedling's branch; while higher N supply had significant effects in promoting the seedling's early branching and lowering the branching position on the main stem. Supplying higher amount of N increased the plant biomass and the N concentration in root, leaf and stem significantly. Compared with stem and root, leaf had the highest biomass, accounting for 50% of the total biomass, and the highest N accumulation, accounting for 60%-70% of the total accumulated N. PMID- 18163301 TI - [Ecophysiological responses of Festuca arundinacea to high temperature stress]. AB - The measurement of leaf relative water content (RWC), chlorophyll content, cell membrane lipid peroxidation, anti-oxidative system, and photosynthesis of two F. arundinacea cultivars (Barlexas and Crossfire II) and Cynodon dactylon under high temperature (38 degrees C / 30 degrees C, day/ night) showed that with the increasing time of exposure to high temperature, the leaf RWC, chlorophyll content, net photosynthetic rate (P(n)) and photochemical efficiency (F(v)/F(m)) of two F. arundinacea cultivars had a decreasing trend, and the average decrement was smaller for Barlexas than for Crossfire II. After exposed to high temperature for 9 days, the average activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in Barlexas leaves were 19.7%, 17.9% and 17.7% higher than those in Crossfire II leaves, and the P(n) of Barlexas and Crossfire II was decreased by 60.7% and 81.9%, respectively. Under high temperature, the F(v)/F(m) of Barlexas leaves was higher than that of Crossfire II leaves, which could be helpful to mitigate the damage of high temperature to the photosynthetic apparatus of Barlexas. No significant change was observed for each test physiological parameter of C. dactylon leaves with the increase of exposure time under high temperature. The adaptation ability to high temperature was in the order of C. dactylon > Barlexas > Crossfire II. PMID- 18163302 TI - [Appropriate soil nitrate N content for a winter wheat/summer maize rotation system in North China Plain]. AB - A field experiment with 12-level N fertilization in winter wheat growth season and zero-N in summer maize growth season was conducted to study the appropriate soil nitrate N content for a winter wheat/summer maize rotation system in North China Plain. The results showed that when the soil mineral N content before sowing was higher, a split application of 150 kg N x hm(-2) in winter wheat growth season could meet the N demand of both winter wheat and summer maize in the rotation system. The N use efficiency of winter wheat in different N treatments was only from 11% to 23%, while the residual N use efficiency of summer maize ranged from 30% to 52%. When the nitrate N content in top 90 cm soil layer before maize sowing was up to 82 and 151 kg x hm(-2), respectively, the N demand of summer maize at its ten-leaf stage and in its whole growth season could be met with no N application. However, when the nitrate N content in top 90 cm soil layer was less than 46 and 65 kg x hm(-2) at maize ten-leaf stage and after harvest, respectively, no N application could impact maize growth, and decrease its grain yield. With the maize yield and environmental protection considered comprehensively, the nitrate N content in top 90 cm soil layer should be maintained at an appropriate level from 65 to 151 kg x hm(-2) during maize growth season. PMID- 18163303 TI - [Effects of long-term fertilization on distribution of organic nitrogen components in soil aggregates in sub-humid agroecosystem]. AB - A 25-year long-term fertilization experiment was conducted on a Eun-Orthic Anthrosols in the sub-humid agroecosystem on Loess Plateau to study the effects of different fertilization on the distribution of organic nitrogen components in soil aggregates. The results indicated that under long-term fertilization, the distribution of hydrolyzed ammonia N and hydrolyzed unknown N (HUN) in soil aggregates was affected most significantly, followed by that of amino acid nitrogen, and of amino sugar nitrogen, suggesting that applying chemical and organic fertilizers in long-term could promote the combination of soil aggregates with hydrolyzed ammonia N and HUN, while amino sugar nitrogen was more stable during the processes of nitrogen cycling and transformation in soil. There were significant positive correlations of soil total hydrolyzed nitrogen with soil total organic carbon, total nitrogen, and fractal dimensions of aggregates, the correlation coefficient being 0.942, 0.981 and 0.910, respectively, illustrating that soil organic nitrogen components had significant effects on the formation and characteristics of soil aggregates. Correlation analysis, indicated that soil total N and organic C had greater effects on the organic nitrogen components in 1 2 mm and 0.25-1 mm soil aggregates. PMID- 18163304 TI - [K fertility and K deficit index of aquic brown soil under different fertilization systems]. AB - A 15-year fertilization experiment was conducted on an aquic brown soil to study the variations of its K fertility and the index of K deficit under different fertilization systems. The results indicated that no K application accelerated the depletion of soil K, representing a certain decrease of soil available and slow-release K. Applying K alone could not keep soil K in balance, still having a decrease of soil available and slow-release K. The application of recycled compost combined with appropriate amount of fertilizer K could compensate soil K expenditure to a certain degree, and keep the concentrations of soil available and slow-release K somewhat constant. The statistic analysis on the past years test results showed that the critical values of soil available K in corn- and soybean fields obtained by crossing method were all 73 mg x kg(-1). PMID- 18163305 TI - [Effects of temperature on organic carbon mineralization in paddy soils with different clay content]. AB - An incubation test with three kinds of paddy soil (sandy loam, clay loam, and silty clay soils) in subtropical region was conducted at 10, 15, 20, 25 and 30 degrees C to examine the response of the mineralization of soil organic carbon (SOC) to temperature change. The results showed that during the period of 160 d incubation, the accumulative mineralized amount of SOC in sandy loam, clay loam, and silty clay soils at 30 degrees C was 3.5, 5.2 and 4.7 times as much as that at 10 degrees C, respectively. The mineralization rate was lower and relatively stable at lower temperatures (< or = 20 C), but was higher at the beginning of incubation and decreased and became stable as the time prolonged at higher temperatures (> or = 25 degrees C). During incubation, the temperature coefficient (Q10) of SOC mineralization in test soils fluctuated, with an average Q10 in sandy loam, clay loam, and silty clay soils being 1.92, 2.37 and 2.32, respectively. There was a positive exponential correlation between SOC mineralization constant k and temperature (P < 0.01), and the response of SOC mineralization to temperature change was in the order of clay loam soil > silty clay soil > sandy loam soil. PMID- 18163306 TI - [Organic carbon decomposition rate in different soil types]. AB - With incubation experiment, this paper studied the decomposition rate of organic carbon in black soil, fluvo-aquic soil, and red soil. It was shown that these three soil types had significantly different decomposition rate constant of organic carbon (P < 0.05 ), with the corresponding value being 2.2 x 10(-4) x d( 1), 6.0 x 10(-4) x d(-1), and 3.4 x 10(-4) x d(-1), respectively. The decomposition rate constant had a significant correlation with soil pH, which was increased greatly when the soil pH was < 5.5 or > 8.0, and a significant negative correlation with the contents of soil clay and silt, indicating that soil clay and silt content was one of the dominant factors affecting the decomposition rate of organic carbon in these soil types. PMID- 18163307 TI - [Effects of spatial coupling of watering and fertilization on winter wheat photosynthetic characteristics and grain yield]. AB - A soil column experiment was conducted with a fertile Eum-Orthic Anthrosols to study the effects of spatial coupling of watering and nitrogen (N)- and phosphorus (P) fertilization on the photosynthetic characteristics and grain yield of winter wheat. The column consisted of three layers, each layer being 30 cm in thickness and with an underlying 2 cm layer of coarse sand to obstruct water- and nutrient exchange. The results showed that relative content of chlorophyll (SPAD), net photosynthetic rate (P(n)), and grain yield were significantly lower in the treatment of 0-30 cm soil drought and 30-90 cm soil wetness (D) than in that of 0-90 cm soil wetness (W). In these two watering treatments, a combined application of N and P got the highest SPAD, P(n) and grain yield, followed by applying P, and N. The SPAD, P(n) and grain yield were significantly higher when the fertilizer N was applied in 0-90 cm layer than in 0 30, 30-60 and 60-90 cm layers, respectively. No significant differences in SPAD, P(n) and grain yield were observed when fertilizer P was applied in 0-90 cm layer and in 0-30 cm layer. In treatment W, the SPAD, P(n) and grain yield were the highest when N and P were applied in 0-90 cm layer, had no significant differences with those when the N and P were applied in 0-30 cm layer, but were significantly higher than those when the fertilization was carried out in 30-60 and 60-90 cm layers. In treatment D, no significant difference in SPAD was observed when the N and P were applied in different layers, but P(n) and grain yield were significant higher when the N and P were applied in 0-90 cm than in 30 60 cm layer, and in 30-60 cm than in 60-90 cm layer. It was concluded that whether in treatment W or in treatment D, a combined application of N and P should be carried out in 0-30 cm soil layer. PMID- 18163308 TI - [Estimation of nitrogen concentration in cotton leaf based on canopy reflectance spectra]. AB - Through analyzing the relationships of nitrogen concentration in cotton leaf under different nitrogen supply levels with canopy multi-spectral reflectance and its derived ratio vegetation index (RVI, rholambda1/rholambda2), normalized difference vegetation index (NDVI, (rho(lambda1) - rho(lambda2))/(rho(lambda1)) + rho(lambda2)) and differential vegetation index (DVI, rho(lambda1) - rho(lambda2)), the sensitive wave bands and prediction functions of cotton leaf nitrogen concentration were worked out. The vegetation index composed of visible region (610, 660, 680 and 710 nm) and near infrared region (760, 810, 870, 950, 1 100 and 1 220 nm) had a higher correlation with the nitrogen concentration in cotton leaf, and the RVI composed of 950 nm and 710 nm could best predict the leaf nitrogen concentration. The validation with independent field experimental data indicated that RVI (950 nm and 710 nm) -based model was suitable for estimation of leaf nitrogen concentration of different cotton cultivars at their different growth stages. PMID- 18163309 TI - [Meteorological conditions affecting the quality of strong gluten- and medium gluten- wheat and climate division in Shandong Province]. AB - With stepwise and one-factor non-linear regression analyses, and based on the meteorological data during the period from anthesis to maturity at 31 experimental sites in Shandong Province, this paper studied the relationships of the kernel quality of 4 strong gluten- and 12 medium gluten wheat cultivars with main meteorological factors in 1999-2003. The results showed that for strong gluten- and medium gluten wheat cultivars, the optimum mean temperature for dough stability time was 20.0 degrees C and 20.5 degrees C, the optimum mean difference of diurnal temperature for protein content was 12.7 degrees C and 11.7 degrees C, the optimum precipitation for sedimentation volume and dough stability time was 48.6 mm and 52.1 mm, and 53.5 mm and 53.9 mm, and the optimum total sunshine for sedimentation volume and dough stability time was 297 h and 299 h, and 295 h and 298 h, respectively. The effects of single meteorological factor on the protein content, sedimentation volume, and dough stability time of wheat cultivars were not identical. Based on the comprehensive evaluation of weather condition, Shandong Province was divided into three production zones for the production of wheat, i.e., central and east Shandong adaptable for strong gluten- and medium gluten- wheat cultivars, northwest and southwest Shandong sub-adaptable for strong gluten- and medium gluten- wheat cultivars, south Shandong sub-adaptable for strong gluten wheat cultivars and adaptable for medium gluten wheat cultivars. PMID- 18163310 TI - [Microbial flora in Cerasus sachalinensis rhizosphere]. AB - By using selected culture media, the microbes in Cerasus sachalinensis rhizosphere were isolated, identified and classified, with their community structure and dynamic changes at different growth stages of C. sachalinensis studied. The bacteria isolated were belonged to 15 genera, among which, Bacillus, Pseudomonas and Flavobacterium were the dominant ones. Flavus and Albosporus were the two dominant genera in seven groups of Actinomyces, and Mucor, Aspergillus and Penicillium were the main genera of fungi. The microbial flora varied with C. sachalinensis growth stage, being the richest at defoliation stage and the least at budding stage. PMID- 18163311 TI - [Spatiotemporal pattern and its driving forces of urban growth in Shenyang City]. AB - By using time series Landsat TM satellite images and adopting GIS spatial analysis and landscape pattern analysis methods, this paper studied the spatiotemporal diversity of urban growth and the evolution of urban landscape pattern in Shenyang, and examined their driving forces. The results showed that in 1988-2004, the urban area in Shenyang increased persistently, and the growth intensity enhanced consistently, with the peaks occured in 2000-2004. The spatial differentiation of urban growth in the City was also distinct, with the southwest direction as the leading orientation, and the urban edges and different level economic development zones as the main growth areas. The urban landscape pattern became more and more complex, and the compactness index of urban development decreased. The evolution of urban landscape pattern was related to the characteristics of urban growth, which also showed spatiotemporal diversity. The urban growth and urban landscape pattern evolution in Shenyang were mainly attributed to the development of industrialization and the construction of different level economic development zones, the proper policies of local governments and the urban planning, as well as the development of traffic infrastructure. PMID- 18163312 TI - [A new category system of China nature reserves based on their attributes]. AB - Based on the numeric attributes of 114 typical nature reserves in China, a new category system was established by using cluster analysis method. Compared with the China national category system of nature reserves, the new category system was simpler and more applicable. The 4 categories in the new system matched well to the categories I , II , IV, and V in IUCN category system, respectively. The new category system, characterized by the consistency of category criteria and scale, could avoid the deviation caused by subjective category to some degree, and provide a quantitative standard for the classification of nature reserves in China. PMID- 18163313 TI - [Noise-reduction function and its affecting factors of urban plant communities in Shanghai]. AB - The factor analysis on the relationships between excess noise attenuation (decrement after noise propagating 30 m) and 8 structural characteristics of 19 urban plant communities in Shanghai showed that all the plant communities had notable effects on reducing noise, and the noise attenuation ability of the communities was significantly higher than that of lawn (P < 0.01). The plant communities could be divided into three groups base on their noise attenuation ability, i.e., those of > or = 10 dB(A), 6-10 dB(A), and < or = 6 dB(A). The main factors affecting the noise attenuation ability of the communities were leaf area index, average bifurcate height, average height, coverage, and average canopy diameter, and their correlation coefficients with noise attenuation were 0.343, 0.318, 0.285, 0.226 and 0.193, respectively. These five factors had a cumulative contribution rate of 65.47%, suggesting that they should be considered in stress when designing urban greenbelt for noise reduction. PMID- 18163314 TI - [Ecotourism carrying capacity of Hangzhou Xixi National Wetland Park in China]. AB - In this paper, an integrated estimation on the ecotourism carrying capacity of Hangzhou Xixi National Wetland Park in China was made from the aspects of ecological carrying capacity, spatial carrying capacity, facility carrying capacity, management carrying capacity, and psychological carrying capacity. The results indicated that the tourism carrying capacity of the Park was 4 145 - 6 450 persons per day. The rational distance between man and bird was first adopted to determine the ecotourism carrying capacity of wetland, which provided an effective solution both to fully ensure bird safety and to appropriately develop wetland tourism. The estimation of psychological carrying capacity based on tourist satisfaction degree reflected more objectively the extent the tourist demands satisfied at the planning, construction and management of tour places. Such an integrated estimation method based on the distance between man and bird and the tourist satisfaction degree could be of practical and instructive significances in the planning and management of wetland parks. PMID- 18163315 TI - [Distribution of phthalic acid esters in agricultural soils in typical regions of Guangdong Province]. AB - To understand the distribution characteristics of phthalic acid esters (PAEs) in agricultural soils in typical regions of Guangdong Province, surface soil (0-20 cm) samples were collected, and the concentrations of 6 PAEs were determined by gas chromatography. The results showed that the total concentration of the PAEs (sigma PAEs) ranged from not detected to 25.99 mg x kg(-1), and was < or = 1.00 mg x kg(-1) in 92.8% of the samples. The sigma PAEs differed with land use types and regions, and decreased in the sequences of paddy soil > banana soil > vegetable soil > sugarcane soil > orchard soil, and Dongguan > Shantou > Shunde > Zhanjiang > Zhongshan > Zhuhai > Huizhou. Among the test PAEs, DEHP had the highest detection rate (85.1%), DnBP had the highest concentration (not detected to 17.51 mg x kg(-1)). Comparing with the corresponding control limits of soil PAEs in USA, all test PAEs except DnOP were exceeded the limits to some extent, and DnBP, DMP and DEP exceeded seriously, indicating that the agricultural soils in the typical regions of Guangdong Province were contaminated by PAEs. PMID- 18163316 TI - [Characteristics of iodine uptake and accumulation by vegetables]. AB - With seaweed iodine and KI as exogenous iodine sources, a pot experiment was conducted to study the characteristics of iodine uptake and accumulation by pakchoi cabbage, celery, capsicum, and radish. The results showed that the iodine content in the edible parts of test vegetables increased with increasing amount of exogenous iodine, but the iodine accumulation rate differed with the kinds of vegetables, in the order of pakchoi > celery > radish > capsicum. The majority of iodine was accumulated in roots, with lesser amount transferred to shoots. The distribution of iodine in vegetables was commonly in the order of root > leaf > stem > fruit, but the iodine in radish is lower in its rhizome than in its shoot. Low concentrations (0-25 mg x kg(-1)) of exogenous iodine had little effects on the growth of vegetables, while high concentrations (> or = 50 mg x kg(-1)) of it had inhibitory effects, resulting in a decreased vegetable biomass. The sensitivity of test vegetables to the adverse effect of exogenous iodine was in the order of capsicum > pachoi > celery > radish. Compared with seaweed iodine, KI decreased the biomass of first cutting significantly (P < 0.05), but for the second cutting, little difference was observed between these two iodine sources. The uptake and accumulation of these two iodine sources by vegetables also differed with cuttings, i.e., the first cutting vegetables absorbed more KI, while the second cutting vegetables absorbed more seaweed iodine (P < 0.05), suggesting that seaweed iodine had a longer efficacy than KI. PMID- 18163317 TI - [Off-line control of runoff pollution by filtering ditch-pond system in urban tourist areas]. AB - An off-line filtering ditch-pond system for controlling storm runoff pollution in urban tourist areas was developed, which could retain the first flush effectively, resulting in the decrease of pollutant concentration and suspended solid average grain size, and the improvement of pollutant retention in runoff. This system could be an effective treatment system for storm runoff pollution, particularly for the scarcity of available land use in urban areas. In 2005, the yearly retention rates of TSS, COD, TN and TP were 86.4%, 85.5%, 83.9% and 82.9%, and during a storm event on June 26, the retention rates of runoff volume, TSS, COD, TN and TP were 67.9%, 97.0%, 89.2%, 94.9% and 96.2%, respectively. This system could also retain most of the suspended solids in runoff. PMID- 18163318 TI - [Parasitic functional response of Coccobius azumai Tachikawa (Hymenoptera: Aphelinidae) to Hemiberlesia pitysophila Takagi (Homoptera: Diaspididae)]. AB - With female Hemiberlesia pitysophila adult as the host, this paper studied the parasitic functional response of Coccobius azumai in laboratory. The results showed that the response of C. azumai fitted Holling Type II equation, and was affected by temperature and the densities of host and parasitoid. At the same temperature, the number of parasitized host increased with increasing host density. At 15 degrees C-25 degrees C, the parasitized number increased with temperature; while at 25 degrees C-35 degrees C, it was in adverse. There was a stronger intraspecific interference in the parasitic functional response of C. azumai, i.e., the number of parasitized host decreased with increasing C. azumai density. The relationship between searching efficiency (E) of C. azumai and its density (P) could be well simulated as E = 0.1659P(-0.5597) by Hassell (1969) model E = QP(-m) and as E = 0.1437T/ [1 + 0.2691 (P -1)] by Beddington (1975) model E = aT/[1 + bt(w) (P-1)]. PMID- 18163319 TI - [Effect of landscape pattern on mouse pest in Dongting Lake area]. AB - The relationship between landscape pattern and ecological process is one of the key issues in landscape ecology. Both the species protection and the bio-disaster prevention are closed linked with the landscape pattern of organism's habitat. Taking Dongting Lake area as a case, this paper studied the characteristics of landscape pattern in typical sampling plots, and analyzed the correlation of mouse capture rate in farmland and in house with the indices reflecting the characteristics of landscape pattern. The results showed that the mouse capture rate in farmland correlated significantly with the landscape aggregation, connection, and mosaic shape, while that in house had a significant correlation with the mosaic shape of construction land. Some suggestions on reducing mouse pest were put forward from the viewpoint of landscape pattern. PMID- 18163320 TI - [Regulation effects of arbuscular mycorrhizal fungi on the interactions between barnyard grass and upland rice under enhanced nitrogen supply]. AB - A pot experiment was conducted to study the regulation effects of arbuscular mycorrhizal fungi (AMF) on the interactions between Echinochloa crus-galli var. mitis L. and Oryza sativa L. under enhanced N supply (4.0 g N x m(-2) x a(-1)). The results showed that under monoculture condition, the AMF colonization on E. crus-galli increased but that on O. sativa decreased. In the treatments with and without AMF inoculation, upland rice biomass and its P and N uptake increased by 42.35% and 13.48%, 4.07% and 2.55%, and 30.35% and 62.09%, respectively, and barnyard grass biomass and its P and N uptake increased by 20.24% and 15.65%, 3.88% and 4.06%, and 15.10% and 30.35%, respectively. Under mixed cropping, the AMF colonization on E. crus-galli increased but that on O. sativa had little change. In the treatments with and without AMF inoculation, the biomass ratio of O. sativa to E. crus-galli decreased, but N uptake ratio changed a little. The P uptake ratio of O. sativa to E. crus-galli increased in treatment without AMF inoculation but decreased in treatment with AMF inoculation. It was suggested that AMF tended to enhance the competition of E. crus-galli to O. sativa under enhanced N supply. PMID- 18163321 TI - [Ovicidal activity of nine essential oils against Chrysomya megacephara in bacon and kipper]. AB - To exploit safe ovicides against the flies that attack bacon and kipper, the ovicidal activity of 9 plant essential oils was investigated by a modified egg dipping method. The results indicated that the essential oils from Cinnamomun cassia, Eugenia caryophyllata and Illicum verum had higher ovicidal activity, with their LC50 values being 0.428, 1.605 and 2.489 mg x ml(-1), respectively. Through GC/MS analysis, a total of 22 components were identified from C. cassia oil, among which, cinamaldehyde was the most predominant one, accounting for 92.33% of the total. The LC50 of synthesized cinamaldehyde (97.33%) was 0.281 mg x ml(-1), being a little higher than the LC50 of C. cassia oil, suggesting that cinamaldehyde was the main ovicidal component of C. cassia oil and could replace the latter as an ovicide against harmful flies. PMID- 18163322 TI - [Pelagic Mysidacea in East China Sea and its significance as the indicator of water mass]. AB - Based on the investigation data of four seasonal oceanographic censuses in the area 23 degrees 30' - 33 degrees N and 118 degrees 30' - 128 degrees E of East China Sea in 1997-2000, this paper studied the ecotypes and geographic and seasonal distribution of pelagic Mysidacea in this sea area, with its significance as the indicator of water mass approached. The results showed that among the species of pelagic Mysidacea, Acanthomysis latiscauda, Gastrosaccus pelagicus, Siriella sinensis and Hemisiriella pulchra were the coastal subtropical species with some features of warm temperate species, while Pseudanchialina pusilla, Promysis orientalis, Erythrops minuta and Anisomysis bipartoculata were the offshore subtropical species with some features of tropical species. Different from other zooplanktons, most of the pelagic Mysidacea species had lower optimal temperature and salinity and distributed more at near-shore in spring and winter, showing the features of warm temperate near shore species. The areas with high abundance of A. latiscauda, S. sinensis and H. pulchra were just the converging areas of the Changjiang dilute waters and warm current masses and of coastal waters and offshore high salinity waters. Erythrops minuta and Anisomysis bipartoculata could be used as the indicators of warm current, P. orientalis and Doxomysis quadrispinosa were the indicators of the Taiwan warm current, while the rest were not good enough to be the indicators of water masses. PMID- 18163323 TI - [Four hypotheses about the effects of soil nitrogen availability on fine root production and turnover]. AB - With global changes such as increasing temperature and enhanced N deposition, soil nitrogen (N) availability is predicted to increase substantially, and how fine root dynamics responds to the altered soil N has become one of the key questions in terrestrial ecology. As such, a number of hypotheses have been proposed to explain the relationship between increasing soil N availability and fine root production, mortality, and turnover. This article considered four major hypotheses: with increasing soil N availability, 1) both fine root production and turnover rate would increase, 2) both fine root production and turnover rate would decrease, 3) fine root production would decrease while fine root turnover rate would increase, and 4) fine root production would increase while fine root turnover rate would decrease. Current evidence suggests that the patterns depicted in hypothesis 1) and 2) could both occur in nature and may reflect characteristics of different species. Hypotheses 3) and 4) were thought to characterize only a transient stage of the responses of fine root dynamics to increasing N availability. To better understand the response of root dynamics to increasing soil N, future studies should consider: 1) the definition of fine roots and heterogeneity in fine root structure and function; 2) methods used in estimating fine root production and turnover rate; 3) changes of soil N availability both in space and time. More attention should also be paid to the influences of mycorrhizal infection on root dynamic responses to soil N availability. PMID- 18163324 TI - [Relationships of soil physical and microbial properties with nitrous oxide emission under effects of freezing-thawing cycles]. AB - Freezing-thawing cycles often occurs in the regions of mid-high latitude and high altitude. This process can affect soil physical and biological properties, e.g., soil water status, aggregate stability, and microbial biomass and community structure. Under its effects, the bio-indicators of soil microbes, i.e., the kinds and quantities of some specific amino sugars varied, and the course and intensity of soil nitrogen transformation changed, which resulted in an increase of nitrous oxide (N2O) production and emission, and made the soil be a major source of N2O emission. This paper summarized the research progress on the aspects mentioned above, with the further research directions on the theoretical problems of soil N2O production and emission under effects of freezing-thawing suggested. PMID- 18163325 TI - [Some ideas about ecological assets assessment]. AB - Ecological assets assessment is of great significance in building harmonious society and implementing scientific concept of development. Based on the achievements in ecological assets assessment research, the concept and scope of ecological assets were re-examined, and the contents of ecological assets assessment were summarized in this paper. It was considered that ecosystem quality, ecosystem pressure and ecological assets value should be the focus in assessing ecological assets. With the ecological assets assessment on forest ecosystem as an example, some referenced indicators and calculation models were given. By the end of the paper, some suggestions on the further study were presented. PMID- 18163326 TI - [Distribution characteristics of copper in soil and rape around Tongling mining area]. AB - Soil and rape samples around Tongling mining area were collected, and their copper (Cu) contents were investigated. The results showed that the upland soil developed on the slope deposit around the mining area as well as the paddy soil distributed in lower reaches was heavily polluted by Cu, while the fluvo-aquic soil further from the mining area was less contaminated. Though the Cu content in paddy soil and upland soil was nearly the same, its bioavailability was higher in paddy soil, due to the Cu pollution of irrigated water. There was a significant correlation between available and total Cu in these three types of soil. The activation rate of soil Cu (percentage of available Cu in total Cu) was 15.0% on average, which was positively correlated with soil total Cu and organic matter while negatively correlated with soil pH and Mn. The average Cu content in rape seed and stalk was 4.0 and 5.8 mg x kg(-1), respectively. The rape Cu content increased obviously with increasing soil available Cu content when the soil available Cu content was relatively low, but the Cu absorption and accumulation by rape decreased gradually when the soil available copper content was higher than 30 mg x kg(-1). PMID- 18163327 TI - [Effects of imidacloprid on arthropod community structure and its dynamics in alfalfa field]. AB - With species richness (S), individual number (N), Shannon's index (H') and evenness (E) as the indices of biodiversity, this paper studied the arthropod community structure and its seasonal dynamics in alfalfa field under effects of imidacloprid. The results showed that imidacloprid more affected the species number of natural enemies rather than that of insect pests, resulting in a significant increase of the dominance concentration of both total arthropod community and pest sub-community, but spraying the insecticide by the end of May had little effects on the dominance concentration of pollination bee sub community. The numbers of dominant pests such as aphid and thrip reduced significantly at the initial stage of spraying imidacloprid, increased rapidly after 7 days, and exceeded the control after 40 days. As for the natural enemy sub-community, its species number decreased greatly and its species diversity and evenness were lower at the initial stage of spraying imidacloprid, its species number increased gradually after 7 days and approached to the level of the control thereafter, while its individual number was lower than the control all the time. PMID- 18163328 TI - [Predation of micro-protozoa on bacteria in Taihu Lake]. AB - With dilution method, this paper studied the predation of different size micro protozoa on bacteria in Taihu Lake, and approached the effects of the predation on bacterial growth and of the water temperature on the predation. The results showed that in the water body of Taihu Lake, the predation rate of micro-protozoa with its size less than 32 microm was 5.07 d(-1), and the nano-size (less than 16 microm) protozoa contributed about 90.7%. The predation of nano-protozoa reduced the abundance of bacteria significantly. With the increase of water temperature, the predation rate of nano-protozoa and the growth rate of bacteria increased obviously. PMID- 18163329 TI - Activity of intracellular phospholipase A1 and A2 in Giardia lamblia. AB - Neither phospholipase A1 (PLA A1) nor phospholipase A2 (PLA A2), nor their respective genes, have been identified in Giardia lamblia, even though they are essential for lipid metabolism in this parasite. A method to identify, isolate, and characterize these enzymes is needed. The activities of PLA A1 and PLA A2 were analyzed in a total extract (TE) and in vesicular (P30) and soluble (S30) subcellular fractions of G. lamblia trophozoites; the effects of several chemical and physicochemical factors on their activities were investigated. The assays were performed using substrate labeled with 14C, and the mass of the 14C-product was quantified. PLA A1 and PLA A2 activity was present in the TE and the P30 and S30 fractions, and it was dependent on pH and the concentrations of protein and Ca2+. In all trophozoite preparations, PLA A1 and PLA A2 activities were inhibited by ethylenediaminetetraacetic acid and Rosenthal's inhibitor. These results suggest that G. lamblia possesses several PLA A1 and PLA A2 isoforms that may be soluble or associated with membranes. In addition to participating in G. lamblia phospholipid metabolism, PLA A1 and PLA A2 could play important roles in the cytopathogenicity of this parasite. PMID- 18163330 TI - Parasite community and mortality of overwintering young-of-the-year roach (Rutilus rutilus). AB - The composition and development of the parasite community of overwintering young of-the-year (YOY) roach (Rutilus rutilus) was studied in outdoor artificial ponds. Cumulative mortality of the roach from November until May was about 20%. The prevalences of the epizoic ciliates Trichodina sp. and Apiosoma sp., and of the monogeneans Dactylogyrus suecicus, Dactylogyrus nanus, and Gyrodactylus sp. in general tended to peak in the beginning of April at a water temperature of about 8 C. However, mean parasite intensity remained constant over this interval. Prevalence of metacercariae of the digeneans Diplostomum cf. spathaceum and Tylodelphys clavata remained constant with values around 60% and 10%, respectively. Larvae of the nematode Philometra obturans and the cestode Ligula intestinalis and females of the copepod Neoergasilus japonicus were found sporadically. A concept of 2 thresholds, a critical condition factor (attained by a temperature-induced energy deficiency), and a condition-dependent critical infection intensity, which determine winter mortality of YOY, is suggested. PMID- 18163331 TI - Environmental factors influencing community composition of gastropods and their trematode parasites in southern Ontario. AB - Agricultural activity and landscape features have previously been associated with diversity and prevalence of trematode species in amphibian second intermediate hosts. In this study, the density, diversity, and size of snail first intermediate hosts, and the diversity and prevalence of their trematode species, were assessed in 2 types of ponds, i.e., those adjacent to cornfields and those from the same region in southwestern Ontario that were adjacent to nonagricultural settings. Species of trematodes included, but were not restricted to, those that are known parasites of larval and adult frogs. We also assessed landscape factors likely to influence use by definitive hosts. Presence of the herbicide atrazine in ponds was measured to check that ponds adjacent to agriculture had potential to be affected by agricultural runoff. Both snail size and the proportion of snails releasing cercariae were greater in nonagricultural ponds, contrasting with a previous finding of lower trematode infection in tadpoles from nonagricultural ponds. Percentage of forest cover was associated with prevalence of certain trematode species, but not with estimates of combined prevalence. Absence of relations of trematode prevalence to measures of road density also contrasted with previous studies. We interpret our results in light of how agricultural activity might influence trematode viability, snail growth, and use by wildlife definitive hosts, independently of landscape factors. PMID- 18163332 TI - Habitat and host specificity of trematode metacercariae in fiddler crabs from mangrove habitats in Florida. AB - Fiddler crabs (Uca spp.) are common inhabitants of temperate and tropical coastal communities throughout the world, often occupying specific microenvironments within mangrove and salt marsh habitats. As second intermediate hosts for trematodes, we investigated patterns of host distribution and parasitism for 3 species of sympatric fiddler crabs in mangrove habitats adjacent to the Indian River Lagoon, Florida. Fiddler crab distribution varied among species, with Uca speciosa dominating the low and mid intertidal regions of mangrove banks. This species also exhibited higher prevalence and abundance of Probolocoryphe lanceolata metacercariae compared with Uca rapax, which is relatively more abundant in the high intertidal zone. We conducted a field experiment to test whether U. speciosa was more heavily parasitized by P. lanceolata as a result of its habitat distribution by raising U. speciosa and U. rapax under identical environmental conditions. After exposure to shedding cercariae under the same field conditions, all individuals of U. speciosa became parasitized by P. lanceolata, whereas no U. rapax were parasitized, suggesting that differences in parasitism were driven by host selection. PMID- 18163333 TI - Piscinoodinium, a fish-ectoparasitic dinoflagellate, is a member of the class dinophyceae, subclass gymnodiniphycidae: convergent evolution with Amyloodinium. AB - All dinoflagellates that infest the skin and gills of fish have traditionally been placed within the class Blastodiniphyceae. Their relatedness was primarily based upon a similar mode of attachment to the host, i.e., attachment disc with holdfasts. Results of recent molecular genetic analyses have transferred these parasites, including Amyloodinium, to the class Dinophyceae, subclass Peridiniphycidae. In our study, a small subunit rDNA gene from a parasitic dinoflagellate that has features diagnostic for species in the genus Piscinoodinium, i.e., typical trophont with attachment disc having rhizocysts, infesting the skin of freshwater tropical fish, places this organism within the dinophycean subclass Gymnodiniphycidae. This suggests a close relationship of Piscinoodinium spp. to dinoflagellates that include symbionts, e.g., species of Symbiodinium, and free-living algae, e.g., Gymnodinium spp. These molecular and morphological data suggest that evolution of this mode of fish ectoparasitism occurred independently in 2 distantly related groups of dinoflagellates, and they further suggest that the taxonomic status of parasites grouped as members of Piscinoodinium requires major revision. PMID- 18163334 TI - Structure of the pharynx in the adult nematode Anguillicoloides crassus (Nematoda: Rhabditida). AB - The structure of the pharynx of the adult female nematode Anguillicoloides crassus (Spirurina) has been studied for the first time using light and transmission electron microscopy. The cylindrical pharynx consists of a short anterior muscular corpus and an enlarged posterior glandular and muscular postcorpus. The main cellular components of the pharynx of A. crassus include the muscle cells, the marginal cells, the nerve cells, and 1 dorsal and 2 subventral glands. New observations for nematodes include: (1) the non-contractile regions of pharyngeal musculature in the corpus have specific appearance; (2) the ventrosublateral longitudinal nerve in the pharynx has an enlarged, enucleated anterior part, with a pronounced palmate projections; and (3) abundant lysosomelike membranous bodies consisting of myelinlike figures of varied size present in marginal cells and pharyngointestinal valve. The 2 subventral glands and, apparently, the single dorsal gland, have their openings at the same level, i.e., at the border between the corpus and postcorpus. The pharyngeal-intestinal valve joins the pharynx to the intestine. Knowledge of the ultrastructure of these complex characters may be useful in understanding of functional features, and for comparative morphology as well as evolutionary considerations within the Chromadorea. PMID- 18163335 TI - Low seroprevalence of Neospora caninum infection associated with the limousin breed in cow-calf herds in Andorra, Europe. AB - Neospora caninum seroprevalence and risk factors affecting seroprevalence in beef cattle in Andorra were investigated. Antibodies to N. caninum were evaluated by enzyme-linked immunosorbent assay performed on a yearly basis in 1,758 animals older than 6 mo, belonging to 26 herds. Mean seroprevalence of antibodies to N. caninum for the herds was 7.4 +/- 1.2% (130/1,758). Logistic regression analyses were performed on data from each animal, considering N. caninum seropositivity as the dependent variable, and herd, grazing area, year of sampling, repeat-test animal (animals sampled twice or more), sex, breed, age (animals <4 yr old or > or =5 yr old), and country of birth as possible risk factors. Based on the odds ratio, the prevalence of infection was 2.1 times higher (P < 0.01) in animals from the Ordino grazing area, 1.64 times higher in animals older than 5 yr (P < 0.01), and 6.7 times (1/0.15) lower in Limousin-mixed Limousin cattle (P < 0.002). The results suggest that the particular grazing location could promote the horizontal transmission of this parasite and that certain breeds are less susceptible to N. caninum infection than others. PMID- 18163336 TI - Neospora caninum and Toxoplasma gondii antibodies in dogs from Durango City, Mexico. AB - Toxoplasma gondii and Neospora caninum are structurally similar parasites, with many hosts in common. The prevalence of antibodies to T. gondii and N. caninum was determined in sera from dogs from Durango City, Mexico. Using a modified agglutination test, antibodies to T. gondii were found in 52 (51.5%) of the 101 dogs with titers of 1:25 in 27, 1:50 in 11, 1:100 in 5, 1:200 in 4, 1:400 in 2, 1:800 in 2, and 1:3,200 or higher in 1. Antibodies to N. caninum were determined by the indirect immunofluorescent antibody test (IFAT) and the Neospora sp. agglutination test (NAT). Two of the 101 dogs had N. caninum antibodies; these dogs did not have T. gondii antibodies, supporting the specificity of the tests used. The N. caninum antibody titers of the 2 dogs were: 1:400 by IFAT and 1:200 by NAT in 1, and 1:25 by NAT and IFAT in the other. Results indicate that these 2 structurally similar protozoans are antigenically different. PMID- 18163337 TI - T-helper-1 and T-helper-2 immune responses in mice infected with the intestinal fluke Neodiplostomum seoulense: their possible roles in worm expulsion and host fatality. AB - Neodiplostomum seoulense is highly pathogenic and lethal to experimental mice; most worms are expelled within 2 mo of acquisition. In this study, T-helper (Th) cell immune responses were studied in N. seoulense-infected BALB/c mice. Spleen and mesenteric lymph node (MLN) cells of infected mice proliferated in response to parasite antigens; CD4+ T cells proliferated more than CD8+ T cells. Antigen induced interferon (IFN)-gamma (a Th1 cytokine) secretion began to increase at day 7 postinfection (PI) in spleen and MLN cells, and this was maintained at day 28 PI, whereas interleukin (IL)-4 (a Th2 cytokine) secretion was somewhat lower. Similar results were observed for mRNA signals of IFN-gamma and IL-4. Antigen specific serum total immunoglobulin (Ig)G, IgG1, IgM, and IgA levels (Th2 induced) were elevated from days 7 to 14 to day 28 PI, and IgG2a (Th1-induced) was elevated at days 21 to 28 PI. Interestingly, the numbers of macrophages (Th1- or Th2-induced), which were found to kill N. seoulense worms in vitro, increased remarkably during days 14-28 PI in spleens and small intestines of infected mice. This study shows that mixed Th1 and Th2 responses occur during the course of N. seoulense infection in BALB/c mice. Heavy infiltrations of macrophages in the small intestine may participate in host damage and worm expulsion. PMID- 18163338 TI - Vaccination with microneme protein NcMIC4 increases mortality in mice inoculated with Neospora caninum. AB - NcMIC4 is a Neospora caninum microneme protein that has been isolated and purified on the basis of its unique lactose-binding properties. We have shown that this protein binds to galactosyl residues of lactose; antibodies directed against NcMIC4 inhibit host cell interactions in vitro, thus making it a vaccine candidate. Because of this feature, NcMIC4 was first purified on a larger scale in its native, functionally active form using lactose-agarose affinity chromatography. Second, NcMIC4 was expressed in Escherichia coli as a histidine tagged recombinant protein (recNcMIC4) and purified through Ni-affinity chromatography. Third, NcMIC4 cDNA was cloned into the mammalian pcDNA3.1 DNA vector and expression was confirmed upon transfection of Vero cells in vitro. For vaccination studies, we employed the murine cerebral infection model based on C57Bl/6 mice, employing experimental groups of 10 mice each. Two groups were injected intraperitoneally with purified native NcMIC4 and recNcMIC4, respectively, employing RIBI adjuvant. The third group was vaccinated intramuscularly with pcDNA-NcMIC4. Control groups included an infection control, an adjuvant control, and a pcDNA3.1 control group. Following 3 injections at 4-wk intervals, mice were challenged by i.p. inoculation of 2 x 10(6) N. caninum tachyzoites (Nc-1 isolate). During the course of parasite challenge (3 wk), mice from the 3 different test groups showed varying degrees of symptoms bearing a semblance to neosporosis, i.e., walking disorder, rounded back, apathy, and paralysis of the hind limbs. Control groups showed no symptoms at all. Most notably, vaccination with pcDNA-MIC4 proved antiprotective, with 60% of mice succumbing to infection within 3 wk, and all mice lacking a measurable anti NcMIC4 IgG response. NcMIC4 in its native form elicited a substantial humoral IgG1 immune response and a reduction in cerebral parasite load compared to the controls, but 20% of mice succumbed to infection. Vaccination with recNcMIC4 also resulted in 20% of mice dying; however, in this group, cerebral parasite load was similar to the controls, and recNcMIC4 vaccination elicited a mixed IgG1/IgG2 response. In conclusion, vaccines based on NcMIC4, especially pcDNA-NcMIC4, render mice more susceptible to cerebral disease upon challenge with N. caninum tachyzoites. PMID- 18163339 TI - Parasite communities of common dolphins (Delphinus delphis) from Patagonia: the relation with host distribution and diet and comparison with sympatric hosts. AB - We studied the helminths of 18 common dolphins, Delphinus delphis, from northern Patagonia. Parasites were found only in the gastrointestinal tract. Four species were in the stomach, the nematode Anisakis simplex, the acanthocephalan Corynosoma cetaceum, and the digeneans Braunina cordiformis and Pholeter gastrophilus, plus 1 digenean in the hepatopancreatic ducts, Oschmarinella rochebruni. Infection levels were low (0-155 parasites). Braunina cordiformis and C. cetaceum were the most prevalent species. Anisakis simplex was mainly concentrated in the forestomach, B. cordiformis in the main stomach, and C. cetaceum in the pyloric stomach. Component diversity was low and component evenness was intermediate. Infracommunity diversity was also low, and the mean evenness was higher than at the component community level. Low prevalence in common dolphins is consistent with parasite assemblages of other cetaceans off Patagonia. None of the parasites found is specific to this host species within the study area. We suggest that potential prey are probably important in parasite transmission in this and other dolphins from the Patagonian region. PMID- 18163340 TI - Isolates of Plasmodium inui adapted to Macaca mulatta monkeys and laboratory reared anopheline mosquitoes for experimental study. AB - Plasmodium inui is a parasite of macaques and other nonhuman primates in Asia that is studied as a model for the human malaria parasite P. malariae. Presented here are descriptions of the isolation, passage histories into Macaca mulatta monkeys, and infectivity to different Anopheles spp. mosquitoes of 18 different isolates of this parasite. PMID- 18163341 TI - Geographic and host range of the nematode Soboliphyme baturini across Beringia. AB - The nematode Soboliphyme baturini Petrov, 1930, was found to represent a single species with a relatively broad geographic range across Beringia and northwestern North America on the basis of the assessment of molecular sequence data for adult and juvenile parasites. Refuted are hypotheses suggesting that several cryptic species could be partitioned either among an array of mustelid definitive hosts or across the vast region that links North America and Eurasia. Host specificity for this species is examined on the basis of a comprehensive list for definitive hosts, derived from new field surveys and existing literature for S. baturini. Only 5 mustelids (Gulo gulo, Martes americana, M. caurina, M. zibellina, and Neovison vison) appear to have significant roles in the life history, persistence, and transmission of this nematode. Soboliphyme baturini readily switches among M. americana, M. caurina, Mustela erminea, or N. vison at any particular locality throughout its geographic range in North America, although Martes spp. could represent the source for nematodes in a broader array of mustelids. Molecular analyses (243 base pairs of mitochondrial gene nicotinamide dehydrogenase [ND4]) suggest that hypotheses for host specificity across an array of mustelid definitive hosts are not supported. The life cycle of S. baturini is explored through a review of diet literature for 2 marten species, M. americana and M. caurina, and other mustelids across the Holarctic. Shrews (Soricomorpha: Soricidae) comprise >8% of prey for these species of Martes, suggesting their putative role as paratenic hosts. Juvenile nematodes found in the diaphragms of soricids are genetically identical to adult S. baturini found in the stomachs of mustelids at the same locations in both Asia and North America, corroborating a role in transmission for species of Sorex. PMID- 18163342 TI - The sugar glider (Petaurus breviceps): a laboratory host for the nematode Parastrongyloides trichosuri. AB - Parastrongyloides trichosuri is a nematode parasite of the Australian brush tailed possums that can be propagated through many generations in vitro. This makes P. trichosuri uniquely suited for genetic investigations, including those involving transgenesis. However, an obstacle to its use as an experimental model has been the fact that its host is limited to Australia and New Zealand and that it cannot be exported because of its status as a protected species or agricultural pest, respectively. In previous studies, conventional laboratory animals such as rats, mice, rabbits, ferrets, and chickens have failed to support infections. In the present study, gerbils and short-tailed opossums proved similarly refractory to infection. In contrast, the sugar glider (Petaurus breviceps, family Petauridae) proved to be a good host for P. trichosuri. Patent infections resulted using as few as 6 infective larvae (L3i) and as many as 2,000 L3i. Large numbers of L3i (1,000-2,000) produced patent infections of much shorter duration than those seen when 100 L3i were initially given to the sugar glider. In one case, an infection initiated with 100 L3i was patent for over 1 yr. Parastrongyloides trichosuri is easily cryopreserved using a method developed for Strongyloides stercoralis. Thus, we have identified an experimental host for P. trichosuri that will make it possible to conduct research on this parasite in laboratories outside the endemic sites. PMID- 18163343 TI - Life cycle of Stephanoprora uruguayense (Digenea: Echinostomatidae) in Argentina. AB - The life cycle of Stephanoprora uruguayense Holcman et Olague, 1989, was experimentally resolved. In an artificial pond in the Zoological Garden in Buenos Aires City, Argentina, Heleobia parchappei (Hydrobiidae) was found to be releasing large-tailed cercariae with a prepharyngeal body, but lacking collar spines and corpuscles in the excretory system. Metacercariae, which encysted on the gills of naturally and experimentally infected Cnesterodon decemmaculatus (Poecilidae), developed collar spines and corpuscles in the excretory system in 7 days. Sexually mature adults were recovered from chicks and immature adults from mice fed metacercariae from C. decemmaculatus. Eggs shed in chick feces developed to miracidia within 10 days; sporocysts were found on the gills of snails. Stephanoprora uruguayense and S. denticulata from Europe are similar in adult morphology, but can be distinguished by morphological and behavioral features of larvae. Likewise, although S. denticulata and S. paradenticulata from Venezuela are similar to S. uruguayense in adult morphology, they differ considerably in larval morphology and intermediate hosts. PMID- 18163344 TI - Cichlidogyrus sclerosus (Monogenea: Ancyrocephalinae) and its host, the Nile tilapia (Oreochromis niloticus), as bioindicators of chemical pollution. AB - Experimental results showed that the gill monogenean Cichlidogyrus sclerosus and its host, the Nile tilapia Oreochromis niloticus, exhibited significant numerical and physiological responses after exposure to sediments polluted with polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls, and heavy metals in comparison with control fishes. After 15 days of exposure, C. sclerosus abundance significantly increased in treatments with low to fairly high sediment pollutant concentrations, but declined at high sediment pollutant concentrations. Hypertrophy and hyperplasia in secondary gill lamellae and the spleen melanomacrophage centers were significantly higher at extremely high sediment pollutant concentrations compared with the controls. Spleen lymphocyte and monocyte counts were significantly lower at extremely high sediment pollutant concentrations and were significantly correlated with high fluorescent aromatic compound concentrations measured as PAH exposure indicators. A multivariate redundancy analysis showed significant statistical association between sediment pollutant concentration, C. sclerosus abundance, and tilapia physiological variables. The polluted sediments negatively affected monogenean abundance and induced immunosuppression in hosts, consequently increasing histological damage in hosts and allowing persistent C. sclerosus infection. This study documents evidence suggesting that C. sclerosus and its host are indeed excellent models to test environmental quality in tropical freshwater ecosystems. PMID- 18163345 TI - Characterization of the antigen SO7 during development of Eimeria tenella. AB - The developmental expression of the antigen SO7, which has been previously shown to protect chickens against infection by several Eimeria species, was investigated. Using RT-PCR, mRNA for SO7 was found to be restricted primarily to unsporulated oocysts (0 hr). Western blot (WB) analysis with an antibody to recombinant SO7 (rbSO7) revealed expression of the protein from 6 to 72 hr (fully sporulated) of sporulation and in sporozoites (SZ). SO7 was absent in host derived second-stage merozoites (MZ) and was present in culture-derived first stage MZ but at a level of only 25% of that exhibited by SZ. During invasion of Madin-Darby bovine kidney (MDBK) cells by SZ in vitro, the level of SO7 within cells, as determined by WB analysis, remained relatively constant until 48 hr of development and then decreased by about 40% at the next time point (72 hr). The SO7 secreted into the culture media during in vitro development increased to a relative maximum at 48 hr and then decreased to about 20% of maximum at 72 hr. Immunostaining with anti-rbSO7 indicates that SO7 is highly concentrated in both refractile bodies (RB) of SZ, with some limited distribution in the apical complex. Anti-rbSO7 intensively stained the intracellular parasites and the first stage schizonts during in vitro development of E. tenella in MDBK cells. Upon release from the schizonts, the first-stage merozoites stained with 1 or 2 bright spots typically at each end. The results suggest that SO7 is closely associated with the SZ RB and is developmentally regulated but may not play a direct role in cellular invasion. PMID- 18163346 TI - Role of activated macrophages in Acanthamoeba keratitis. AB - The purpose of this study was to determine whether activating the conjunctival macrophages would affect the course of Acanthamoeba spp. keratitis in a Chinese hamster model of this disease. Chinese hamster spleen cells were stimulated with concanavalin A (Con A), and interferon gamma (IFN-gamma) -containing supernatants were collected 24 hr later. The IFN-gamma-containing supernatants were loaded into liposomes, which were fed to peritoneal macrophages in vitro. Macrophage activation was assessed by testing for production of nitric oxide (NO) with the use of Griess reagent. Conjunctival macrophages were activated in situ by subconjunctival injection of liposomes containing Con A-activated spleen cell culture supernatants. Control liposomes were loaded with phosphate-buffered saline (PBS). Macrophages exposed to supernatants from Con A-stimulated spleen cells produced 4-fold-higher amounts of NO than unstimulated macrophages. Activation of macrophages via subconjunctival injection of liposomes containing supernatants from Con A-stimulated spleen cell cultures resulted in rapid resolution of the corneal infection. Approximately 80% of animals treated with PBS-containing liposomes demonstrated evidence of corneal disease at day 14 compared to 10% incidence of infection in the Con A-treated group. Moreover, at all time points examined, the clinical appearance of the keratitis in animals treated with liposomes containing Con A supernatant was significantly reduced compared to the group treated with liposomes containing PBS (P < 0.05). Macrophages stimulated with IFN-gamma-containing supernatants killed significant numbers of the trophozoites in vitro (P < 0.05). Killing was inhibited by cytochalasin D, but not by L-N6-1-iminoethyl-L-lysine dihydrochloride (L-NIL), which is a selective inhibitor of inducible NO synthase (INOS). PMID- 18163347 TI - Substance P is associated with heart enlargement and apoptosis in murine dilated cardiomyopathy induced by Taenia crassiceps infection. AB - Dilated cardiomyopathy (degeneration of heart muscle and heart enlargement) is an important cause of heart failure among young adults. Dilated cardiomyopathy may be a complication during or after various viral, bacterial, or parasitic diseases. Substance P (SP) is a neurotransmitter that is involved in the pathogenesis of various diseases. To determine whether SP is associated with cardiac changes in murine cysticercosis, we compared heart-weight to body-weight ratio, cardiac pathology, cardiomyocyte size, and cardiac-apoptosis (TUNEL assay) in hearts from Taenia crassiceps-infected (wild-type vs. SP-knockout) mice. We noted that, as compared with control uninfected wild-type mice, elevated protein levels of SP and its receptor as studied by ELISA or immunohistochemistry, respectively, were elevated in the hearts of parasite-infected wild-type mice. The heart-weight to body-weight ratios were significantly higher in the parasite infected wild-type mice versus those of the infected SP-knockout mice. Furthermore, wild-type infected mice developed dilated cardiomyopathy with increased chamber size of both ventricles, decreased ventricular wall thickness, compensatory cardiomyocyte hypertrophy, and increased cardiac apoptosis. This cardiac pathology did not develop in mice lacking SP activity (i.e., in infected SP knockout mice) or in uninfected mice. These data indicate that SP is associated with cardiac changes in an animal model of parasitic dilated cardiomyopathy. PMID- 18163348 TI - A new enteric nematode of Camallanus (Nematoda: Camallanidae) from Jiyu Island of Amoy (Taiwan Strait). AB - A new enteric nematode, Camallanus jiyuensis n. sp., is described from the intestine of the shokihaze goby, Tridentiger barbatus (Gunther, 1861), collected from the beach of Jiyu Island, Amoy, southeast of China. The new species differs from its congeners mainly in the following combination of characters. The body is rather small in size, with 2 squarish, chitinized thickenings on the anterior border of the buccal capsule. The prongs of the trident are unequal. The nerve ring is near the posterior end of tridents. Cervical papillae are located in the middle of muscular esophagus. The excretory pore is located in the posterior third of muscular esophagus. Twelve pairs of pedunculate caudal papillae are present, of which 7 pairs are preanal and 5 are postanal. Spicules are similar, but subequal in length; the right and the left account for 9.8% and 8.9% of body length, respectively. The vulva does not protrude conspicuously from body surface and is located postequatorially. PMID- 18163349 TI - A new species of Dentiphilometra (Nematoda: Philometridae) from the musculature of the gray snapper Lutjanus griseus (osteichthyes) off the Caribbean coast of Mexico. AB - A new nematode, Dentiphilometra lutjani n. sp. (Philometridae), is described from gravid females (the male is unknown) collected from the body musculature of the marine perciform fish gray snapper, Lutjanus griseus (Lutjanidae), from the Bay of Chetumal and southern coast of Quintana Roo, off the Caribbean coast of Mexico. The new species differs from the only other congener, Dentiphilometra monopteri, from the swamp eel Monopterus albus in China, mainly in the body length of gravid female (15.40-53.21 mm), the shape of the posterior body end (not markedly narrowed, with low caudal projections), the esophageal gland (maximum width near its posterior end), and the length (344-483 microm) of larvae from the uterus; both species also differ in their host types (marine perciform fish vs. freshwater swamp eel) and geographical distribution (Mexico vs. China). PMID- 18163350 TI - A new species of Oochoristica (Eucestoda: Cyclophyllidea) parasite of Ctenosaura oaxacana (Reptilia: Iguanidae) from Mexico. AB - Oochoristica whitfieldi n. sp., parasitizing the intestine of the Oaxacan black iguana Ctenosaura oaxacana (Kohler and Hasbun, 2001), in Ruinas de Guiengola, Oaxaca state, Mexico, is described. The new species can be distinguished from all 4 congenera infecting Iguanidae in the neotropical realm in possessing a lower mean number of testes (122 in O. acapulcoensis Brooks, Perez-Ponce de Leon, and Garcia-Prieto, 1999; 62 in O. guanacastensis Brooks, Perez-Ponce de Leon, and Garcia-Prieto, 1999, and 95 in O. leonregagnonae Arizmendi-Espinosa, Garcia Prieto, and Guillen-Hernandez, 2005, vs. 35 in O. whitfieldi), and a wider scolex (0.450-0.600, 0.475-0.537, 0.5-0.8, vs. 0.25-0.26, respectively). Oochoristica iguanae Bursey and Goldberg, 1996 differs from the new Mexican species in having a longer strobila (60-110 mm vs. 14.4-33.7 mm, respectively), fewer ovarian sublobes (6 vs. 11-17), and a cirrus pouch that hardly reaches excretory canals (whereas in O. whitfieldi the cirrus pouch widely overpasses these canals). PMID- 18163351 TI - Emended description and redetermination of Sarwaria caballeroi n. comb. (nematoda: Ostertagiinae) with details of the synlophe and esophageal characters. AB - Among genera of the ostertagiine nematodes, structural attributes of Spiculopteragia caballeroi are consistent with criteria that diagnose the genus Sarwaria. Specifically, the following characters are compatible with referral to this genus: (1) species characterized by monomorphic males; (2) tapering lateral synlophe in the cervical zone; (3) minuscule, thornlike cervical papillae; (4) length of Ray "4" < Ray "5"; (5) relatively thick or robust Ray "4"; (6) a substantially reduced dorsal lobe and dorsal ray that are disposed or curved ventrally relative to Rays "8"; and (7) a broad, laterally inflated dorsal lobe. Consequently, we propose Sarwaria caballeroi n. comb., and we provide further validation for the genus Sarwaria. Additionally, we propose that the ostertagiines are represented by 12 valid genera, including those characterized by a bursal formula of 2-2-1 (Cervicaprastrongylus, Hyostrongylus, Mazamastrongylus, Spiculopteragia, and Teladorsagia in addition to Sarwaria), and those in which the lateral rays describe a 2-1-2 pattern (Camelostrongylus, Longistrongylus, Marshallagia, Orloffia, Ostertagia, and Pseudomarshallagia). PMID- 18163352 TI - Hydromermis osami n. sp. (Nematoda: Mermithidae) from chironomids eclosing from northern Minnesota waterways. AB - Hydromermis osami n. sp. (Nematoda: Mermithidae) is described from the hemocoel of 2 species of Rheotanytarsus chironomid imagos eclosing from Lake Ozawindib and Gulsvig Landing in northern Minnesota during August 2002 and June and July 2006. The species is distinguished from the other 26 described members of the genus by the terminal mouth; oval, opaque, thick walled amphids, wider than long; acute posterior end; single spicule not bifurcated proximally; well-developed uterine and vulval limbs of the S-shaped vagina; absence of bursal sleeve; absence of nutrient vesicles in the trophosome; and an esophagus length over 40% of body length. Members of the new species emerge from the hosts as adults. The other described Hydromermis species, number of specimens, location, and known hosts are tabulated. PMID- 18163353 TI - New and emended descriptions of gregarines from flour beetles (Tribolium spp. and Palorus subdepressus: Coleoptera, Tenebrionidae). AB - The following new gregarine taxa are described from larvae of flour beetles (Coleoptera: Tenebrionidae): Awrygregarina billmani, n. gen., n. sp., from Tribolium brevicornis; Gregarina cloptoni, n. sp., from Tribolium freemani; Gregarina confusa, n. sp., from Tribolilum confusum; and Gregarina palori, n. sp., from Palorus subdepressus. In addition, the description of Gregarina minuta Ishii, 1914, from Tribolium castaneum, is emended. Scanning electron micrograph studies of these species' oocysts reveal differences in surface architecture. The Gregarina species have oocysts with longitudinal ridges, visible with SEM, whereas Awrygregarina billmani oocysts have fine circumferential striations; surface architecture is the main feature distinguishing the 2 gregarine genera. Although parasites from adult beetles are not included in the descriptions, adults of all host species can be infected experimentally using oocysts from the new taxa. PMID- 18163354 TI - New species of Rhabdias (Nematoda: Rhabdiasidae) from Bufo occidentalis (Anura: Bufonidae) from Sierra Madre del Sur, Mexico. AB - Rhabdias kuzmini n. sp., a parasite of the lungs of Bufo occidentalis, is described and illustrated. This Mexican taxon differs from the related species in the genus by the possession of 4 lips (2 subdorsal and 2 subventral) and 2 lateral pseudolabia, corpus not inflated, a larger barrel-shaped buccal capsule, equatorial vulva, and the presence of a slightly swollen cuticle in the anterior and posterior ends of the body. This is the 16th species described in the Neotropical Realm and the first species of Rhabdias described from endemic anurans in Mexico. PMID- 18163355 TI - Two new species of Ascarophis (Nematoda: Cystidicolidae) in marine fishes from Chile. AB - In this study, we describe 2 new species of Ascarophis van Beneden, 1871 (Nematoda: Cystidicolidae), found in fishes from southern Chile. Ascarophis carvajali n. sp. was found in Austrolycus depressiceps and Patagonotothen cornucola, whereas Ascarophis draconi n. sp. was taken from Champsocephalus gunnari. These new Ascarophis species differ from other species in a combination of several morphometric and morphological characteristics. Although A. carvajali n. sp. was morphologically close to Ascarophis minuta, the new species has a larger ratio between glandular and muscular esophagus, filaments on both egg poles, and a shorter right spicule than A. minuta. Ascarophis draconi n. sp. was morphologically similar to Ascarophis adioryx and Ascarophisfiliformis. However, A. adioryx has eggs without filaments, a smaller ratio between glandular and muscular esophagus length, and a smaller ratio between left and right spicule lengths in contrast to A. draconi n. sp., whereas A. filiformis has a shorter glandular esophagus and left spicule length than A. draconi n. sp. Only 1 Ascarophis species has been recorded in a single fish from Chile (i.e., Ascarophis sebastodis in Sebastes capensis). Consequently, this study constitutes not only new species and records of Ascarophis in fishes from Chile, but also new records for the Pacific coast of South America. PMID- 18163356 TI - A new species of Hepatozoon (Apicomplexa: Adeleorina) from Python regius (Serpentes: Pythonidae) and its experimental transmission by a mosquito vector. AB - Hepatozoon ayorgbor n. sp. is described from specimens of Python regius imported from Ghana. Gametocytes were found in the peripheral blood of 43 of 55 snakes examined. Localization of gametocytes was mainly inside the erythrocytes; free gametocytes were found in 15 (34.9%) positive specimens. Infections of laboratory reared Culex quinquefasciatus feeding on infected snakes, as well as experimental infection of juvenile Python regius by ingestion of infected mosquitoes, were performed to complete the life cycle. Similarly, transmission to different snake species (Boa constrictor and Lamprophis fuliginosus) and lizards (Lepidodactylus lugubris) was performed to assess the host specificity. Isolates were compared with Hepatozoon species from sub-Saharan reptiles and described as a new species based on the morphology, phylogenetic analysis, and a complete life cycle. PMID- 18163357 TI - A new species of Subulura (Nematoda: Subuluroidea) from ground squirrels, Spermophilus spilosoma bennett, 1833, in New Mexico. AB - A description is presented of a new species of Subulura Molin, 1860, Subulura novomexicanus, collected from the spotted ground squirrel, Spermophilus spilosoma Bennett, 1833, in New Mexico. The males are 24 to 29 mm long, precloacal sucker 1.7-2.0 mm from posterior end, spicules subequal 0.83-1.0 long, and gubernaculum Y-shaped 0.21-0.39 mm long. Females are 34-39 mm long, vulva near middle of body 14.06-22.00 mm from anterior end, and eggs 0.04-0.05 mm long by 0.03-0.04 mm wide. The new species is distinguished from Subulura ungulatus Erickson, 1938 in being longer and having spicules that are distinctively different in size and form. It also differs from Subulura nevadense Babero, 1973 in being longer and having a larger egg size and smaller spicules. The new species is most similar to Subulura andersoni (Cobbold, 1876) (Thwaite, 1927); however, the males of this species are smaller and they have broad cervical alae, and the adults occur in squirrels of India. Reexamination of S. ungulatus showed 11 pairs of caudal papillae, including 4 pairs of preanals, 1 pair adanal and lateral, and 6 pairs of postanal papillae located at the end of the tail. Spermophilus richardsoni (Sabine, 1822) is a new host record for S. nevadense, and Montana is a new distributional record for this nematode. Eimeria callospermophili Henry, 1932, is a new record for S. spilosoma. PMID- 18163358 TI - Effect of alkyl-lysophospholipids on some aspects of the metabolism of Leishmania donovani. AB - Alkyl-lysophospholipids (ALPs), developed initially to be antitumor agents, have proved highly effective in the treatment of visceral leishmaniasis, a disease caused by the species making up the protozoan complex Leishmania donovani. Although their effectiveness is known, the mode of action against this parasite is not completely understood. In the present work, we have studied the effect of 3 derivatives, edelfosine, miltefosine, and ilmofosine. Using nuclear magnetic resonance spectroscopy ('H-NMR), we have examined the excreted catabolites from glucose metabolism in the promastigote forms treated with these compounds. The ALPs at concentrations of 19 and 38 microM inhibit the excretion of acetate, succinate, and pyruvate. The effect of edelfosine, miltefosine, and ilmofosine on the activity of the enzymes hexokinase, glycerolkinase 3-PD, phosphoglucose isomerase, superoxide dismutase, and phospholipase C were also examined. Glycerolkinase 3-PD and phosphoglucose isomerase are generally insensitive to the compounds, whereas hexokinase and superoxide dismutase are inhibited by miltefosine and ilmofosine. The ALPs exhibited an activated effect against the phospholipase C activity. Alkyl-lysophospholipids were shown to have a significant effect on several enzymes in important biochemical pathways indispensable for the survival of L. donovani promasigotes. PMID- 18163359 TI - Clonorchicidal properties of the synthetic trioxolane OZ78. AB - Because the synthetic trioxolane OZ78 is active against the liver fluke Fasciola hepatica, we were motivated to investigate the in vivo and in vitro activity against another liver fluke, namely Clonorchis sinensis. Rats infected with C. sinensis for 2 and 5 wk were treated orally with single doses of OZ78 (75, 150, or 300 mg/kg). Worm burden reductions were assessed against untreated control rats. Scanning electron microscopy (SEM) was used to observe adult C. sinensis after recovery from rats 1-3 days posttreatment with a single 300 mg/kg oral dose of OZ78 and after in vitro exposure to concentrations of 1, 10, and 100 microg/ml of OZ78. A single 300 mg/kg oral dose of OZ78 resulted in worm burden reductions of 78.5% and 98.5% against juvenile and adult C. sinensis, respectively. SEM observations revealed tegumental surface alterations, including blebbing and sloughing. OZ78 emerges as a new compound with a broad spectrum of activity against major foodborne trematode infections. PMID- 18163360 TI - Seroprevalence of Toxoplasma gondii antibodies in cats from Durango City, Mexico. AB - The prevalence of antibodies to Toxoplasma gondii was determined in sera from 105 domestic cats from Durango City, Mexico. Using a modified agglutination test, antibodies to this parasite were found in 21% of the 105 cats, with titers of 1:25 in 3 cats, 1:50 in 4 cats, 1:200 in 5 cats, 1:400 in 2 cats, 1:800 in 2 cats, 1:1,600 in 4 cats, and 1:3,200 or higher in 2 cats. Cats older than 1 yr had a significantly higher frequency of infection than that found in cats younger than 0.5 yr (41 vs. 13.2%, respectively; odds ratio = 4.55; 95% CI = 1.24-17.18; P = 0.01). Overall, the seroprevalence of T. gondii antibodies in cats in Durango, Mexico, is much lower compared with those reported in other countries. PMID- 18163361 TI - Prevalence of Neospora caninum and Toxoplasma gondii antibodies in wild ruminants from the countryside or captivity in the Czech Republic. AB - In the Czech Republic, sera from 720 wild ruminants were examined for antibodies to Neospora caninum by screening competitive-inhibition enzyme-linked immunosorbent assay and confirmed by indirect fluorescence antibody test (IFAT); the same sera were also examined for antibodies to Toxoplasma gondii by IFAT. Neospora caninum antibodies were found in 14% (11 positive/79 tested) roe deer (Capreolus capreolus), 14% (2/14) sika deer (Cervus nippon), 6% (24/ 377) red deer (Cervus elaphus), 1% (2/143) fallow deer (Dama dama), 3% (3/105) mouflon (Ovis musimon), and none of 2 reindeer (Rangifer tarandus). Toxoplasma gondii antibodies were found in 50% (7/14) sika deer, 45% (169/377) red deer, 24% (19/79) roe deer, 17% (24/143) fallow deer, 9% (9/105) mouflon, and 1 of 2 reindeer. In 42 samples of wild ruminants that tested positive for N. caninum antibodies, 28 (67% of the positive N. caninum samples) reacted solely to N. caninum. This is the first evidence of N. caninum infection in mouflon, the first N. caninum seroprevalence study in farmed red deer, and the first survey of N. caninum in wild ruminants from the Czech Republic. PMID- 18163362 TI - Survey of the metazoan ectoparasites of the European flounder Platichthys flesus (Linnaeus, 1758) along the north-central Portuguese coast. AB - A survey was undertaken to identify metazoan ectoparasite species on the European flounder, Platichthys flesus (Linnaeus, 1758), in 4 different locations off the north-central Portuguese coast. Parasites of 7 different taxa were found: Caligus diaphanus, Caligus sp., and Lepeophtheirus pectoralis (Copepoda: Caligidae); Acanthochondria cornuta (Copepoda: Chondracanthidae); Holobomolochus confusus (Copepoda: Bomolochidae); Nerocila orbignyi (Isopoda: Cymotholdae); and praniza larvae (Isopoda: Gnathiidae). Lernaeocera branchialis, a common European flounder parasite in the North and Baltic Seas, was not observed among the surveyed fish. Caligus diaphanus, Caligus sp., and Nerocila orbignyi are new host records. The high prevalence and intensity values recorded for L. pectoralis and A. cornuta suggest that both parasite species are common to the European flounder along the north-central Portuguese coast. In contrast, infection levels with respect to the other parasite taxa were, in most cases, comparatively lower, thereby indicating that they only occur occasionally among flounders in the surveyed area. PMID- 18163363 TI - Early migration of Sarcocystis neurona in ponies fed sporocysts. AB - Sarcocystis neurona is the most important cause of equine protozoal myeloencephalitis (EPM), a neurologic disease of the horse. In the present work, the kinetics of S. neurona invasion is determined in the equine model. Six ponies were orally inoculated with 250 x 10(6) S. neurona sporocysts via nasogastric intubation and killed on days 1, 2, 3, 5, 7, and 9 postinoculation (PI). At necropsy, tissue samples were examined for S. neurona infection. The parasite was isolated from the mesenteric lymph nodes at 1, 2, and 7 days PI; the liver at 2, 5, and 7 days PI; and the lungs at 5, 7, and 9 days PI by bioassays in interferon gamma gene knock out mice (KO) and from cell culture. Microscopic lesions consistent with an EPM infection were observed in brain and spinal cord of ponies killed 7 and 9 days PI. Results suggest that S. neurona disseminates quickly in tissue of naive ponies. PMID- 18163364 TI - A human case of Plagiorchis vespertilionis (Digenea: Plagiorchiidae) infection in the Republic of Korea. AB - Plagiorchis vespertilionis (Digenea: Plagiorchiidae) is generally considered a bat parasite, but here it is reported for the first time in a human. The patient was a 34-yr-old male who lived in a coastal village of Haenam-gun (county), Jeollanam-do (province), Republic of Korea. Only 1 worm, 2.6 mm long and 0.7 mm wide, was recovered after praziquantel treatment and purging with magnesium salts. The fluke was characterized by a large body size, a sucker ratio of 1:1, a straight cirrus organ, a short distance between the ventral sucker and ovary, well-developed vitellaria, a uterus with descending and ascending loops, and fully developed eggs with an average size of 32.5 x 17.5 microm. The patient had habitually eaten the raw flesh of snakehead mullet and gobies that had been caught near his village. The present case represents the first record of a human P. vespertilionis infection. PMID- 18163365 TI - Influence of a thermal discharge on parasites of a cold-water flatfish, Pleuronectes americanus, as a bioindicator of subtle environmental change. AB - A study was conducted to ascertain the influence of a thermal discharge on the health and parasites of a coastal cold-water flatfish, the winter flounder (Pleuronectes americanus), a species sensitive to environmental change. Flounder were sampled in spring 1998 and 1999 beneath the plume and at reference sites north and south up to 1 km from the discharge. Species diversity and estimates of abundance of macroscopic algae, invertebrates, and fish were also recorded. After capture by scuba divers, a comparison of condition factor, organ indices, blood values, histology, and parasites was made between groups of fish from the discharge and reference sites. Diversity and abundance of algae, invertebrates, and fish were considerably greater beneath the plume than at the reference sites. The thermal water had no apparent effect on flounder taken beneath the plume, but it affected both its ecto- and endoparasites. Prevalence and mean abundance of Cryptocotyle lingua metacercariae were significantly greater, whereas Trichodina jadranica and Gyrodactylus pleuronecti were less on the gills of fish sampled beneath the plume than at the reference sites. Four endoparasites, i.e., Ceratomyxa drepanopsettae, Steringophorus furciger, Macvicarius soleae, and Lecithaster gibbosus were significantly more abundant in the reference samples. These results suggest that environmental change affected transmission of the parasites of winter flounder exposed to the thermal effluent. PMID- 18163366 TI - Species of coccidia (Apicomplexa: Eimeriidae) infecting pikas from Alaska, U.S.A. and northeastern Siberia, Russia. AB - Eighty-eight fecal samples from 2 species of pika, Ochotona collaris and Ochotona hyperborea, collected in Alaska (N = 53) and Russia (N = 35), respectively, were examined for the presence of coccidia (Apicomplexa: Eimeriidae). Five oocyst morphotypes were observed. In O. collaris, we found Eitmeria calentinei, Eimeria cryptobarretti, and Eimeria klondikensis, whereas in O. hyperborea, we found Eimeria banffensis, E. calentinei, E. cryptobarretti, E. klondikensis, and Isospora marquardti. This study represents new geographic records for all 5 coccidia and new host records for E. cryptobarretti and I. marquardti. Only minor quantitative differences were seen between the sporulated oocysts we studied and those reported in their original descriptions. PMID- 18163367 TI - A Ribeiroia spp. (class: Trematoda)--specific PCR-based diagnostic. AB - Increased reporting of amphibian malformations in North America has been noted with concern in light of reports that amphibian numbers and species are declining worldwide. Ribeiroia ondatrae has been shown to cause a variety of types of malformations in amphibians. However, little is known about the prevalence of R. ondatrae in North America. To aid in conducting field studies of Ribeiroia spp., we have developed a polymerase chain reaction (PCR)-based diagnostic. Herein, we describe the development of an accurate, rapid, simple, and cost-effective diagnostic for detection of Ribeiroia spp. infection in snails (Planorbella trivolvis). Candidate oligonucleotide primers for PCR were designed via DNA sequence analyses of multiple ribosomal internal transcribed spacer-2 regions from Ribeiroia spp. and Echinostoma spp. Comparison of consensus sequences determined from both genera identified areas of sequence potentially unique to Ribeiroia spp. The PCR reliably produced a diagnostic 290-base pair (bp) product in the presence of a wide concentration range of snail or frog DNA. Sensitivity was examined with DNA extracted from single R. ondatrae cercaria. The single-tube PCR could routinely detect less than 1 cercariae equivalent, because DNA isolated from a single cercaria could be diluted at least 1:50 and still yield a positive result via gel electrophoresis. An even more sensitive nested PCR also was developed that routinely detected 100 fg of the 290-bp fragment. The assay did not detect furcocercous cercariae of certain Schistosomatidae, Echinostoma sp., or Sphaeridiotrema globulus nor adults of Clinostomum sp. or Cyathocotyle bushiensis. Field testing of 137 P. trivolvis identified 3 positives with no overt environmental cross-reactivity, and results concurred with microscopic examinations in all cases. PMID- 18163368 TI - Human neurocysticercosis: rightward hemisphere asymmetry in the cerebral distribution of a single cysticercus. AB - The distribution of single cysticerci between cerebral hemispheres was studied in 227 adult cases of calcified and vesicular neurocysticercosis (NC). A rightward lateralization of calcified cysticerci was significant only in women, whereas vesicular cysticerci were equally distributed in both hemispheres. Factors related with the differences in the inflammatory response and in the regional cerebral blood flow between genders could be involved. PMID- 18163369 TI - Ribeiroia ondatrae cercariae are consumed by aquatic invertebrate predators. AB - Trematodes amplify asexually in their snail intermediate hosts, resulting in the potential release of hundreds to thousands of free-living cercariae per day for the life of the snail. The high number of cercariae released into the environment undoubtedly increases the probability of transmission. Although many individual cercariae successfully infect another host in their life cycle, most fail. Factors that prevent successful transmission of cercariae are poorly understood. Microcrustaceans and fish have been observed to eat cercariae of some species, although the possibility that predation represents a significant source of mortality for cercariae has been largely unexplored. We tested the cercariophagic activity of several freshwater invertebrates on Ribeiroia ondatrae, a trematode that causes limb deformities in amphibians. Individuals of potential predators were placed into wells of multiwell plates with 10-15 cercariae, and numbers of cercariae remaining over time were recorded and compared with numbers in control wells that contained no predators. Of the species tested, Hydra sp., damselfly (Odonata, Coenagrionidae) larvae, dragonfly (Odonata, Libellulidae), larvae, and copepods (Cyclopoida) consumed cercariae. In some cases, 80-90% of the cercariae offered to damselfly and dragonfly larvae were consumed within 10 min. In most cases, predators continued to consume cercariae at the same average rates when offered cercariae together with individuals of an alternate prey item. Hydra sp. ate fewer cercariae in these trials. Our findings suggest the need for field and laboratory studies to further explore the effects of predators on transmission of R. ondatrae to amphibian larvae. In addition, the results suggest that conservation of the biodiversity and numbers of aquatic predators may limit adverse impacts of trematode infections in vertebrate hosts. PMID- 18163370 TI - Phenotypic variation in infectivity of Diplostomum spathaceum cercariae within a population. AB - The present study examined phenotypic variation in infectivity of Diplostomum spathaceum (Trematoda) cercariae within a natural population. Twelve infected Lymnaea stagnalis were collected from the field, and the infectivity of cercariae from individual snails was assessed under constant laboratory conditions. At a water temperature of 16.3 C, the mean infectivity of cercariae from the snails varied between 55.5% and 87.5%. Depending on the source of variation, this may have important ecological and evolutionary implications for both natural parasite populations and those occurring in aquaculture. PMID- 18163371 TI - Effect of climatic warming on the Pacific walrus, and potential modification of its helminth fauna. AB - The decreasing extent of sea-ice in the arctic basin as a consequence of climatic warming is modifying the behavior and diets of pagophilic pinnipeds, including the Pacific walrus, Odobenus rosmarus divergens Illiger, the species emphasized here. Mammals such as the walrus and bearded seal, Erignathus barbatus (Erxleben), cannot remain associated with the sea-ice, and continue to feed on their usual diet of benthic invertebrates inhabiting coastal waters to a depth of approximately 100 m, when the northwestward retreating ice reaches deep waters beyond the margins of the continental shelf. With reduction of their customary substrate (ice), the walrus has become more pelagic and preys more often on ringed seals, Phoca hispida Schreber. Dietary changes, with modifications of helminth faunas, may be induced by various factors. Increased consumption of mammals or their remains by walruses may lead to a higher prevalence of trichinellosis in them and to more frequent occurrence in indigenous peoples inhabiting the arctic coasts. To assess predicted effects on the composition of helminth fauna of the walrus, we recommend systematic surveys of their helminths as part of research on effects of climatic warming. PMID- 18163372 TI - Total synthesis and structure assignment of the anthrone C-glycoside cassialoin. PMID- 18163373 TI - Allylation of ketones with a ferrocene-based planar chiral Lewis acid. PMID- 18163374 TI - Highly efficient ruthenium(II) porphyrin catalyzed amidation of aldehydes. PMID- 18163375 TI - Two-dimensional tiling by rubrene molecules self-assembled in supramolecular pentagons, hexagons, and heptagons on a Au111 surface. PMID- 18163376 TI - Nanofibrous poly(lactic acid)/hydroxyapatite composite scaffolds for guided tissue regeneration. AB - The production of nanofibrous PLA/HA composite scaffolds is described. The morphological, mechanical, surface, and thermal properties of the composites were extensively investigated. The results show that the mixture of PLA and HA formed smooth nanofibers without lumps. The incorporation of HA increased the mechanical strength of the nanofibers and changed the morphology, increasing the mean fiber diameter and pore size. Surface and internal properties confirmed that HA was homogeneously distributed inside the nanofibers and oriented towards their surface. The nanofiber composites allowed the adhesion and proliferation of pre osteoblasts for up to 3 weeks. PMID- 18163377 TI - MMTCA recognition by molecular imprinting in interpenetrating polymer network hydrogels based on poly(acrylic acid) and poly(vinyl alcohol). AB - A novel IPN hydrogel designed to recognize MMTCA is prepared by applying the molecular-imprinting method. The IPN is characterized by FT-IR, DSC, and SEM. Langmuir analysis shows that an equal class of adsorption is formed in the hydrogel. The adsorption equilibrium constant and the maximum adsorption capacity are evaluated, and the effect of the pH on MMTCA adsorption is discussed. The selectivity of the imprinted polymer for MMTCA is studied in aqueous solutions of MMTCA/aspirin/riboflavin. The results suggest that the MMTCA-imprinted polymer shows superior selectivity for MMTCA as compared to riboflavin and aspirin. The reproducibility of the imprinted polymer to MMTCA is also studied. PMID- 18163378 TI - Distinct functions of H-Ras and K-Ras in proliferation and survival of primary hepatocytes due to selective activation of ERK and PI3K. AB - Ras proteins mediate signals both via extracellular signal-regulated kinase 1 and 2 (ERK), and phosphoinositide 3-kinase (PI3K). These signals are key events in cell protection and compensatory cell growth after exposure to cell damaging and pro-apoptotic stimuli, thus maintaining homeostasis. By transfection techniques, we found that both H-Ras and K-Ras were expressed and appeared functionally active in primary hepatocytes. We compared the ability of H-Ras and K-Ras homologues to preferentially activate one of the two pathways, thereby differentially controlling cell survival and growth. We found that ectopic expression of dominant negative (DN) H-RasN17, but not DN K-RasN17, efficiently inhibited both phosphorylation and translocation of ERK to the nuclear compartment, which are prerequisites for cell cycle progression. Furthermore, ectopic expression of constitutive active (CA) H-RasV12, but not CA K-RasV12, potentiated EGF-induced proliferation. We also found that expression of CA mutants of either H-Ras or K-Ras protected hepatocytes from transforming growth factor-beta1 (TGF-beta1)-induced apoptosis. However, H-Ras-induced survival was mediated by ERK/RSK as well as by PI3K, whereas K-Ras-induced survival was mediated by PI3K only. In conclusion, H-Ras and K-Ras had differential functions in proliferation and survival of primary hepatocytes. H-Ras was the major mediator of ERK-induced proliferation and survival, whereas H-Ras and K-Ras both mediated PI3K-induced survival. PMID- 18163379 TI - Decorin enhances the proliferation and differentiation of myogenic cells through suppressing myostatin activity. AB - Decorin, a small leucine-rich proteoglycan, plays an important role in the regulation of cell growth. Our recent study has shown that immobilized decorin in the collagen matrix sequesters myostatin into the extracellular matrix and prevents its inhibitory action to myoblast proliferation in vitro. However, it still remains unclear whether free decorin could affect the proliferation and differentiation of myogenic cells by regulating myostatin activity. In the present study, we generated stable clonal C2C12 myoblasts that were over expressing decorin, and showed that decorin over-expressing cells had an increased rate of proliferation as compared to control cells. Decorin over expressing cells formed multi-giant hypertrophic myotubes with an elongated morphology and larger size as compared to control cells, although the initiation of differentiation in decorin over-expressing cells was somewhat delayed as compared to control cells. Western blot analysis demonstrated that MyoD expression in decorin over-expressing cells was lower than that in control cells until 12 h after induction to differentiate. At 48-h differentiation, the expressions of MyoD, p21 and myogenin were dramatically increased in cells that over-expressed decorin. Furthermore, we revealed that over-expression of decorin suppressed the activity of myostatin endogenously synthesized in C2C12 myoblasts and attenuated the signaling of exogenous myostatin. Consistent with these results, knock-down of decorin impairs C2C12 myoblast growth by increasing the sensitivity to exogenous myostatin. These results clearly show that decorin enhances the proliferation and differentiation of C2C12 myoblasts through suppressing myostatin activity. PMID- 18163380 TI - Oxidative stress induces GLUT4 translocation by activation of PI3-K/Akt and dual AMPK kinase in cardiac myocytes. AB - In response to metabolic stress, GLUT4, the most abundant glucose transporter, translocates from intracellular vesicles to the plasma membrane. This appears to play an important role in protecting cardiac myocytes from ischemic injury. To investigate the precise mechanisms of GLUT4 translocation in cardiomyocytes, we have established a method for quantifying the relative proportion of sarcolemmal GLUT4 to total GLUT4 in these cells. Stimulation with H2O2 resulted in a concentration-dependent increase in GLUT4 translocation, which peaked at 15 min after stimulation. The dominant-negative form (DN) of AMP-activated protein kinase (AMPK) alpha2 inhibited the H2O2-induced translocation of GLUT4. We further examined the role of two known AMPK kinases (AMPKKs), calmodulin dependent protein kinase kinase (CaMKK)beta and LKB1. The DN of CaMKKbeta or LKB1 alone inhibited H2O2-induced GLUT4 translocation only partially compared to the inhibition produced by the DN of AMPKalpha2. However, the combination of DN-LKB1 and DN-CaMKKbeta inhibited translocation to an extent similar to with DN AMPKalpha2. Stimulation with H2O2 also activated Akt and the inhibition of PI3 K/Akt prevented GLUT4 translocation to the same extent as with AMPK inhibition. When the DN of AMPKalpha2 was applied with DN-PI3-K, there was a complete reduction in the GLUT4 membrane level similar to that seen at the 0 time-point. These results demonstrate that AMPK and PI3-K/Akt have an additive effect on oxidative stress-mediated GLUT4 translocation. PMID- 18163381 TI - Expression of fibroblast growth factor 10 and its receptor, fibroblast growth factor receptor 2B, in the bovine corpus luteum. AB - There is evidence that several fibroblast growth factors (FGFs) are involved in growth and development of the corpus luteum (CL), but many FGFs have not been investigated in this tissue, including FGF10. The objective of this study was to determine if FGF10 and its receptor (FGFR2B) are expressed in the CL. Bovine CL were collected from an abattoir and classed as corpus hemorrhagica (stage I), developing (stage II), developed (stage III), and regressed (stage IV) CL. Expression of FGF10 and FGFR2B mRNA was measured by reverse transcription polymerase chain reaction (RT-PCR). Both genes were expressed in bovine CL, and FGF10 expression did not differ between stages of CL development. FGF10 protein was localized to large and small luteal cells by immunohistochemistry. FGFR2B expression was approximately threefold higher in regressed compared to developing and developed CL (P < 0.05). To determine if FGF10 and FGFR2B expression is regulated during functional luteolysis, cattle were injected with PGF2alpha and CL collected at 0, 0.5, 2, 4, 12, 24, 48, and 64 hr thereafter (n = 5 CL/time point), and mRNA abundance was measured by real-time RT-PCR. FGF10 mRNA expression did not change during functional luteolysis, whereas FGFR2B mRNA abundance decreased significantly at 2, 4, and 12 hr after PGF2alpha, and returned to pretreatment levels for the period 24-64 hr post-PGF2alpha. These data suggest a potential role for FGFR2B signaling during structural luteolysis in bovine CL. PMID- 18163382 TI - Development and psychometric evaluation of the instrument: attitude toward patient advocacy. AB - Nurses' patient advocacy can influence patient outcomes. There is a lack of reliable and valid instruments on patient advocacy. The purpose of this study was to develop an instrument to measure nurses' attitudes toward patient advocacy. Bu and Jezewski's theory of patient advocacy and Fishbein and Ajzen's definition of attitude were used to guide the development and evaluation of the scale. Seven experts evaluated scale content validity. Two samples (N = 200; N = 2,500) were randomly selected from the Oncology Nursing Society membership to allow us to examine psychometric properties of the scale. The scale possessed satisfactory psychometric properties. PMID- 18163383 TI - Evidence for a heritable unidimensional symptom factor underlying obsessionality. AB - The division of obsessive-compulsive symptoms (OCS) into specific factors is now widely accepted. However, the utility of these categories for genetic studies remains unclear, as studies examining their heritability have been inconsistent. Less attention has been paid to the possibility that clinically significant obsessionality is primarily determined by a "core" group of OCS that crosses the boundaries between symptom subgroups. The aim of this study is to determine whether such a core group exists, and to compare its heritability to that of the more traditionally derived symptom factors. We examined the properties and heritability of obsessive-compulsive symptoms in college students, medical students, and obsessive-compulsive disorder (OCD) families using the Leyton Obsessional Inventory. In each of the three samples, we identified a core group of symptoms that comprised a single unique construct and accounted for over 90% of the variation of the four more traditional symptom factors. This core construct was highly correlated with OCD in our families and had a heritability estimate of 0.19 when OCD was not included as a covariate and 0.49 when OCD was included as a covariate. In contrast, the four symptom factors were not heritable. There appears to be an underlying unidimensional component to obsessionality, both in non-clinical and clinical samples. This component, which is heritable, accounts for the majority of the variation of the more traditionally derived symptom factors in our sample, and is composed of OCS that are not specific to any of the symptom subgroups. PMID- 18163384 TI - Examining pain-related distress in relation to pain intensity and psychological distress. AB - Despite frequent use of the term symptom distress in the pain literature, symptom distress is often confused with symptom intensity and psychological distress, contributing to inadequate assessment of symptoms and less than ideal symptom management. In this article we address these issues and propose a hybrid model, combining Price's interaction of pain sensation, pain unpleasantness, and secondary pain affect model with an information processing model. Recommendations on methods and techniques to reduce this confusion would assist healthcare professionals and researchers to better distinguish among these terms as they manage patient symptoms and design symptom management studies. Thus, the purpose of this article is to examine the terms symptom distress, symptom intensity, and psychological distress using pain as the example symptom. PMID- 18163385 TI - Association study between the serotonin 1A receptor (HTR1A) gene and neuroticism, major depression, and anxiety disorders. AB - The serotonin neurotransmitter system in general, and the serotonin 1A receptor in particular, has been broadly implicated in the pathophysiology of mood and anxiety disorders, although the results of genetic association studies have been mixed. In this study, we examined the serotonin 1A receptor gene, HTR1A, for its association with shared genetic risk across a range of anxiety and depression related phenotypes. Using multivariate structural equation modeling, we selected twin pairs from the population-based Virginia Adult Twin Study of Psychiatric and Substance Use Disorders scoring at the extremes of a latent genetic risk factor that underlies susceptibility to neuroticism, major depression, and several anxiety disorders. One member from each selected pair was entered into a 2-stage, case-control association study for the HTR1A gene. In the resulting sample of 589 cases and 539 controls, four SNPs spanning the HTR1A locus, including the C( 1019)G functional promoter polymorphism (rs6295), were screened in stage 1, the positive results of which were tested for replication in stage 2. While one marker met threshold significance criteria in stage 1, this association was not replicated in stage 2. Post-hoc analyses did not reveal association to any of the specific psychiatric phenotypes. Our data suggests that the HTR1A gene may not play a major role in the genetic susceptibility underlying depressive and anxiety related phenotypes. PMID- 18163386 TI - New functional single nucleotide polymorphism (Ala72Ser) in the COMT gene is associated with aggressive behavior in male schizophrenia. AB - A new functional single nucleotide polymorphism (SNP) Ala72Ser in the COMT gene was discovered recently. The purpose of our study is to examine the association between Ala72Ser and Val158Met functional polymorphisms in COMT gene and homicidal behavior in schizophrenia. DNA was genotyped for the Ala72Ser and Val158Met SNPs of the COMT gene in a sample of 93 schizophrenic patients who committed homicide (H-SCZ) and 100 schizophrenic patients who had never committed homicide (NH-SCZ). A statistically significant difference was found in genotype distribution and allele frequencies in SNP Ala72Ser of COMT gene between H-SCZ and NH-SCZ group. In haplotype analysis, the frequency of the combination of high high activity allele (Ala-Val) was lower in H-SCZ group than in NH-SCZ group (P = 0.000069). Our study showed a highly significant association between a COMT haplotype of two functional SNPs and aggressive behavior in schizophrenia. PMID- 18163387 TI - Association study of two polymorphisms of the serotonin-2A receptor gene and suicide attempts. AB - Serotonin (5-HT) receptors may have a role in suicidal behavior. Previous studies have shown an association between the T102C polymorphism of the 5-HT2a receptor gene and suicidal behavior. However, negative findings have also been reported. We examined the association between the T102C and C1354T (His452Tyr) polymorphisms of the 5-HT2a receptor gene and suicide attempts. Four hundred forty-one suicide attempters, 339 psychiatric patients, and 410 healthy controls were compared for genotypes of the T102C and C1354T (His452Tyr) polymorphisms. There were significant differences in the distribution of the three genotypes (TT, TC, and CC) of the T102C polymorphism in the three groups (controls, psychiatric patients, and suicide attempters). There was an excess of C/C genotypes in the suicide attempter group compared with the control group, but there were no significant differences between suicide attempters and psychiatric controls. We found no association between the C1354T polymorphism and suicide attempts. The C allele of the T102C polymorphism of the 5-HT2A receptor gene may be associated with biological susceptibility for suicidal behavior or psychiatric conditions. PMID- 18163388 TI - Parent of origin effects in attention/deficit hyperactivity disorder (ADHD): analysis of data from the international multicenter ADHD genetics (IMAGE) program. AB - There are conflicting reports suggesting that the parental origin of transmitted risk alleles may play a role in the etiology of attention deficit/hyperactivity disorder (ADHD). A recent report by Hawi and colleagues observed a generalized paternal over-transmission of alleles associated with ADHD. This was not replicated in more recent studies. Using data from a large multicenter study we examined the overall and gene-specific parent of origin effect in 554 independent SNPs across 47 genes. Transmission disequilibrium and explicit parent of origin test were performed using PLINK. Overall parent of origin effect was tested by Chi-square. There was no overall parent of origin effect in the IMAGE sample (chi(1)(2) = 1.82, P = 0.117). Five markers in three genes, DDC, TPH2, and SLC6A2 showed nominal association (P < 0.01) with ADHD combined subtype when restricted to maternal or paternal transmission only. Following the initial report by Hawi and co-workers three studies, including this one, found no evidence to support an overall parent of origin effect for markers associated with ADHD. We cannot however, exclude gene-specific parent of origin effect in the etiology ADHD. PMID- 18163390 TI - Parents' perspectives on having their children interviewed for research. AB - We describe parents' perspectives on research interviews with their children with single gene conditions. One hundred forty-two parents were interviewed between 2002 and 2003 in a larger study using a qualitative descriptive design. Two questions from the semi-structured interview guide were used to identify parents' perspectives about future interviews. Almost all of the parents said they would allow an interview with their children, but some parents specified stipulations. These stipulations included: focusing on age-appropriate information, limiting information with child, considering input from parents, and providing a child oriented environment. Knowing this information, researchers can prepare to work more collaboratively with parents and include them more fully in the research process. PMID- 18163389 TI - Family-based SNP association study on 8q24 in bipolar disorder. AB - Previous linkage studies have identified chromosome 8q24 as a promising positional candidate region to search for bipolar disorder (BP) susceptibility genes. We, therefore, sought to identify BP susceptibility genes on chromosome 8q24 using a family-based association study of a dense panel of SNPs selected to tag the known common variation across the region of interest. A total of 1,458 SNPs across 16 Mb of 8q24 were examined in 3,512 subjects, 1,954 of whom were affected with BP, from 737 multiplex families. Single-locus tests were carried out with FBAT and Geno-PDT, and multi-locus test were carried out with HBAT and multi-locus Geno-PDT. None of the SNPs were associated with BP in the single locus tests at a level that exceeded our threshold for study-wide significance (P < 3.00 x 10(-5)). However, there was consistent evidence at our threshold for the suggestive level (P < 7.00 x 10(-4)) from both the single locus and multi-locus tests of associations with SNPs in the genes ADCY8, ST3GAL1, and NSE2. Multi locus analyses suggested joint effects between ADCY8 and ST3GAL1 (P = 3.00 x 10( 4)), with at least one copy of the "high risk" allele required at both genes for association with BP, consistent with a jointly dominant-dominant model of action. These findings with ADCY8 and ST3GAL1 warrant further investigation in order to confirm the observed associations and their functional significance for BP susceptibility. PMID- 18163391 TI - PRODH gene is associated with executive function in schizophrenic families. AB - The aim of this study was to investigate the relationship between polymorphisms in the PRODH and COMT genes and selected neurocognitive functions. Six SNPs in PRODH and two SNPs in COMT were genotyped in 167 first-episode schizophrenic families who had been assessed by a set of 14 neuropsychological tests. Neuropsychological measures were selected as quantitative traits for association analysis. The haplotype of SNPs PRODH 1945T/C and PRODH 1852G/A was associated with impaired performance on the Tower of Hanoi, a problem-solving task mainly reflecting planning capacity. There was no significant evidence for association with any other neuropsychological traits for other SNPs or haplotypes of paired SNPs in the two genes. This study takes previous findings of association between PRODH and schizophrenia further by associating variation within the gene with performance on a neurocognitive trait characteristic of the illness. It fails to confirm previous reports of an association between COMT and cognitive function. PMID- 18163392 TI - Negotiating the challenges of multidisciplinary responses to sexual assault victims: sexual assault nurse examiner and victim advocacy programs. AB - The purpose of this study was to examine Sexual Assault Nurse Examiner (SANE) programs' relationships with victim advocacy organizations. A national telephone survey was conducted of randomly selected SANE programs, all of which served adults, and some of which also served adolescents and children. Nearly one-third of 231 participants reported that SANEs in their program had experienced conflicts related to roles and boundaries with victim advocacy organizations at some point during the program's history. The most frequently mentioned conflicts involved professional autonomy, control, or turf issues. Future researchers should address personnel, organizational, and structural factors that facilitate collaborative relationships between SANE and victim advocacy organizations. PMID- 18163393 TI - Gene copy number variation in schizophrenia. AB - Recent reports have highlighted the possibility that gene copy number variations play a role in the development of complex disorders and have suggested that some variations are very common in schizophrenic patients. We have carried out a comparative genomic hybridization screen using oligonucleotide probes of 891 candidate genes to look for very common copy number variance in schizophrenic patients. In addition we have developed a new approach for the detection and validation of putative copy number variation based upon established methods of allele quantification by DNA pooling and have used it to study 15 major candidates including dysbindin (DTNBP1), neuregulin (NRG1), RGS4 and DISC1. With the exception of positive control sequences, no copy number variations were found for any of the genes in any samples by the use of either technique. Our data for the genes studied are in line with the known existence and frequency of CNVs as reported by recent large scale studies and suggest that gene copy number variations are not more common in schizophrenics than controls, although large ethnic differences cannot be excluded. PMID- 18163394 TI - Bcl-xL prevents staurosporine-induced hepatocyte apoptosis by restoring protein kinase B/mitogen-activated protein kinase activity and mitochondria integrity. AB - Our study reports that staurosporine induces apoptosis in cultured rat hepatocytes in a dose- and time-dependent fashion. Staurosporine induced apparent cleavage of caspase-8, caspase-9, and caspase-3. The release of cytochrome c from mitochondria, and Bid activation were also detected in staurosporine-treated primary hepatocytes. These results suggest that mitochondria-mediated cell death signaling may be involved in staurosporine-induced hepatocyte apoptosis. Bcl-x(L) overexpression protected from "loss of" mitochondrial transmembrane potential and prevented staurosporine-induced caspase-3 and caspase-8 cleavage. Overexpression of constitutively active ERK and PKB inhibited staurosporine-induced caspase-3 activation and hepatocyte death. PI3K inhibitor (LY294002) and ERK inhibitor (PD98059) significantly reversed the protective effects of Bcl-x(L) on staurosporine-induced hepatocyte death. Our data suggest that Bcl-x(L) prevents staurosporine-induced hepatocyte apoptosis by modulating protein kinase B and p44/42 mitogen-activated protein kinase activity and disrupts mitochondria death signaling. PMID- 18163395 TI - Understanding ethnic disparities in the use of total joint arthroplasty: application of the health belief model. AB - OBJECTIVE: The Health Belief Model holds promise in understanding patient-related factors that may explain disparities in the use of total joint arthroplasty (TJA). We examined whether patients' health beliefs differ between African Americans and whites. METHODS: In a primary care clinic setting, 691 African Americans and whites with at least a moderately severe degree of osteoarthritis (OA) completed the Arthritis-related Health Belief Instrument. The instrument has 4 scales: perceived benefits of TJA, perceived barriers to obtaining TJA, perceived severity of arthritis, and perceived susceptibility of arthritis to worsen. RESULTS: The sample (40% women) consisted of 263 (38%) African Americans and 428 (62%) whites who were similar with respect to education, amount of insurance coverage, number of comorbidities, and self-report OA severity score. The African American group was younger, had less men, had more participants who reported an annual income<$15,000, and had a higher body mass index than whites. After controlling for confounders, African Americans were almost 50% (odds ratio [OR] 0.60, 95% confidence interval [95% CI] 0.42-0.86, P=0.005) as likely as whites to perceive that TJA is beneficial or helpful for their arthritis. Furthermore, African Americans were 70% (OR 1.7, 95% CI 1.18-2.44, P=0.004) more likely than whites to recognize barriers (e.g., risky, etc.) to TJA. Race was not associated with either the perceived severity or the perceived susceptibility of arthritis to worsen. CONCLUSION: Among patients with at least moderately severe OA, African Americans were significantly less likely than whites to perceive the benefits of TJA and more likely to recognize barriers to TJA. PMID- 18163396 TI - Sugar-sweetened soft drinks, diet soft drinks, and serum uric acid level: the Third National Health and Nutrition Examination Survey. AB - OBJECTIVE: Sugar-sweetened soft drinks contain large amounts of fructose, which may significantly increase serum uric acid levels and the risk of gout. Our objective was to evaluate the relationship between sugar-sweetened soft drink intake, diet soft drink intake, and serum uric acid levels in a nationally representative sample of men and women. METHODS: Using data from 14,761 participants age>or=20 years from the Third National Health and Nutrition Examination Survey (1988-1994), we examined the relationship between soft drink consumption and serum uric acid levels using linear regression. Additionally, we examined the relationship between soft drink consumption and hyperuricemia (serum uric acid level>7.0 mg/dl for men and >5.7 mg/dl for women) using logistic regression. Intake was assessed by a food-frequency questionnaire. RESULTS: Serum uric acid levels increased with increasing sugar-sweetened soft drink intake. After adjusting for covariates, serum uric acid levels associated with sugar sweetened soft drink consumption categories (<0.5, 0.5-0.9, 1-3.9, and >or=4 servings/day) were greater than those associated with no intake by 0.08, 0.15, 0.33, and 0.42 mg/dl, respectively (95% confidence interval 0.11, 0.73; P<0.001 for trend). The multivariate odds ratios for hyperuricemia according to the corresponding sweetened soft drink consumption levels were 1.01, 1.34, 1.51, and 1.82, respectively (P=0.003 for trend). Diet soft drink consumption was not associated with serum uric acid levels or hyperuricemia (multivariate P>0.13 for trend). CONCLUSION: These findings from a nationally representative sample of US adults suggest that sugar-sweetened soft drink consumption is associated with serum uric acid levels and frequency of hyperuricemia, but diet soft drink consumption is not. PMID- 18163397 TI - Reduced lower extremity vibratory perception in osteoarthritis of the knee. AB - OBJECTIVE: Vibratory perception threshold (VPT) assesses a distinct yet related sensory pathway that has been associated with neuropathic arthropathy but has not been assessed in knee osteoarthritis (OA). The purpose of this study was to evaluate VPT in subjects with knee OA to determine whether the lower extremity afferent deficits observed in knee OA involve more than just proprioception. METHODS: Twenty-seven individuals with symptomatic and radiographic knee OA were compared with 14 age-matched normal subjects. VPT was assessed using a biothesiometer. Five sites of the lower extremity were evaluated. VPT of OA subjects was compared with VPT of normal subjects. RESULTS: VPT of the OA subjects was significantly reduced at all 5 testing sites compared with normal subjects (P<0.05 at all sites). VPT scores (mean+/-SEM volts) for OA subjects and normal subjects were as follows: first metatarsophalangeal joint (15.0+/-1.9 versus 6.4+/-0.9), medial malleolus (22.0+/-2.2 versus 12.3+/-1.4), lateral malleolus (22.3+/-2.0 versus 10.4+/-0.8), medial femoral condyle (25.8+/-1.8 versus 15.9+/-1.9), and lateral femoral condyle (27+/-1.9 versus 18.9+/-2.4). CONCLUSION: This was the first study to evaluate VPT in OA and demonstrate that VPT is reduced at the lower extremity of subjects with knee OA. The noted deficits in VPT may have significant implications in the neuromechanical pathophysiology of OA. VPT is a simple and reliable technique to measure sensory deficits in subjects with OA of the knee. PMID- 18163398 TI - Validation of the systemic lupus erythematosus activity questionnaire in a large observational cohort. AB - OBJECTIVE: To examine the reliability, construct validity, and responsiveness of the Systemic Lupus Erythematosus Activity Questionnaire (SLAQ) in a large observational cohort of persons with systemic lupus erythematosus (SLE). METHODS: We evaluated the reliability of the SLAQ using Cronbach's alpha and principal factor analysis and ascertained construct validity by studying the association of the SLAQ with other clinically relevant, validated patient assessments of health. We estimated responsiveness by calculating standardized response means and analyzing the association of changes in SLAQ scores with changes in other patient assessments of health. RESULTS: The SLAQ had excellent reliability, as reflected by Cronbach's alpha (0.87) and principal factor analysis (one factor accounted for 92% of the variance). SLAQ scores were strongly correlated with other health indices, including the Short Form 12 Physical Component Summary and the Short Form 36 Physical Functioning subscale. Scores were significantly higher for respondents reporting a flare, more disease activity, hospitalization in the last year, concurrent use of immunosuppressive medication, and work disability. The SLAQ demonstrated a small to moderate degree of responsiveness; standardized response means were 0.66 and -0.37 for those reporting clinical worsening and improvement, respectively. Across a range of other patient assessments of disease status, the SLAQ had a response in the direction predicted by these other measures. CONCLUSION: The SLAQ demonstrates adequate reliability, construct validity, and responsiveness in our large, community-based cohort and appears to represent a promising tool for studies of SLE outside the clinical setting. PMID- 18163399 TI - Effect of length on laterally-wedged insoles in knee osteoarthritis. PMID- 18163400 TI - Remitting seronegative syndrome with pitting edema as the presenting symptom in a young, male patient with sarcoidosis: a case report. PMID- 18163404 TI - The provisional Paediatric Rheumatology International Trials Organisation/American College of Rheumatology/European League Against Rheumatism Disease activity core set for the evaluation of response to therapy in juvenile dermatomyositis: a prospective validation study. AB - OBJECTIVE: To validate a core set of outcome measures for the evaluation of response to treatment in patients with juvenile dermatomyositis (DM). METHODS: In 2001, a preliminary consensus-derived core set for evaluating response to therapy in juvenile DM was established. In the present study, the core set was validated through an evidence-based, large-scale data collection that led to the enrollment of 294 patients from 36 countries. Consecutive patients with active disease were assessed at baseline and after 6 months. The validation procedures included assessment of feasibility, responsiveness, discriminant and construct ability, concordance in the evaluation of response to therapy between physicians and parents, redundancy, internal consistency, and ability to predict a therapeutic response. RESULTS: The following clinical measures were found to be feasible, and to have good construct validity, discriminative ability, and internal consistency; furthermore, they were not redundant, proved responsive to clinically important changes in disease activity, and were associated strongly with treatment outcome and thus were included in the final core set: 1) physician's global assessment of disease activity, 2) muscle strength, 3) global disease activity measure, 4) parent's global assessment of patient's well-being, 5) functional ability, and 6) health-related quality of life. CONCLUSION: The members of the Paediatric Rheumatology International Trials Organisation, with the endorsement of the American College of Rheumatology and the European League Against Rheumatism, propose a core set of criteria for the evaluation of response to therapy that is scientifically and clinically relevant and statistically validated. The core set will help standardize the conduct and reporting of clinical trials and assist practitioners in deciding whether a child with juvenile DM has responded adequately to therapy. PMID- 18163405 TI - Do mood symptoms subdivide the schizophrenia phenotype? Association of the GMP6A gene with a depression subgroup. AB - Genetic studies of clinically defined subgroups of schizophrenia patients may reduce the phenotypic heterogeneity of schizophrenia and thus facilitate the identification of genes that confer risk to this disorder. Several latent class analyses have provided subgroups of psychotic disorders that show considerable consistency over these studies. The presence or absence of mood symptoms was found to contribute most to the delineations of these subgroups. In this study we used six previously published subtypes of psychosis derived from latent class analysis of a large sample of psychosis patients. In 280 schizophrenia patients and 525 healthy controls we investigated the associations of these subgroups with myelin related genes. After bonferroni correction we found an association of the glycoprotein M6A gene (GPM6A) with the subgroup of schizophrenia patients with high levels of depression (P-corrected = 0.006). Borderline association of the microtubulin associated protein tau (MAPT) with a primarily non-affective group of schizophrenia patients (P-corrected = 0.052) was also observed. GPM6A modulates the influence of stress on the hippocampus in animals. Thus our findings could suggest that GMP6A plays a role in the stress-induced hippocampal alterations that are found in psychiatric disorders in general and schizophrenia in particular. Overall, these finding suggests that investigating subgroups of schizophrenia based symptoms profile and particularly mood symptoms can facilitate genetic studies of schizophrenia. PMID- 18163406 TI - Missing data in randomized controlled trials of rheumatoid arthritis with radiographic outcomes: a simulation study. AB - OBJECTIVE: To assess the impact, in terms of statistical power and bias of treatment effect, of approaches to dealing with missing data in randomized controlled trials of rheumatoid arthritis with radiographic outcomes. METHODS: We performed a simulation study. The missingness mechanisms we investigated copied the process of withdrawal from trials due to lack of efficacy. We compared 3 methods of managing missing data: all available data (case-complete), last observation carried forward (LOCF), and multiple imputation. Data were then analyzed by classic t-test (comparing the mean absolute change between baseline and final visit) or F test (estimation of treatment effect with repeated measurements by a linear mixed-effects model). RESULTS: With a missing data rate close to 15%, the treatment effect was underestimated by 18% as estimated by a linear mixed-effects model with a multiple imputation approach to missing data. This bias was lower than that obtained with the case-complete approach (-25%) or LOCF approach (-35%). This statistical approach (combination of multiple imputation and mixed-effects analysis) was moreover associated with a power of 70% (for a 90% nominal level), whereas LOCF was associated with a power of 55% and a case-complete power of 58%. Analysis with the t-test gave qualitatively equivalent but poorer quality results, except when multiple imputation was applied. CONCLUSION: Our simulation study demonstrated multiple imputation, offering the smallest bias in treatment effect and the highest power. These results can help in planning trials, especially in choosing methods of imputation and data analysis. PMID- 18163407 TI - Comparison of Vancouver and International League of Associations for rheumatology classification criteria for juvenile psoriatic arthritis. AB - OBJECTIVE: The International League of Associations for Rheumatology (ILAR) criteria constitute the current international diagnostic standard for juvenile psoriatic arthritis (PsA), replacing the less-restrictive Vancouver criteria. The impact of this change on the population diagnosed with juvenile PsA is unknown. METHODS: We reviewed the records of patients seen in a pediatric rheumatology clinic with International Classification of Diseases, Ninth Revision diagnosis codes for psoriasis, PsA, or spondylarthritis. Characteristics of children who met the Vancouver and ILAR criteria were compared. RESULTS: Of 139 children meeting the Vancouver criteria for juvenile PsA, ILAR criteria excluded 80 (58%). Grounds for exclusion were insufficiently definitive rash (44%), a competing diagnosis of enthesitis-related arthritis (23%), family history of psoriasis limited to second-degree relatives (16%), fulfillment of criteria for >1 subtype of juvenile idiopathic arthritis (JIA) (5%), and HLA-B27 in a male with arthritis onset after age 6 (2%). Remaining patients were not homogeneous but could be divided into younger and older subpopulations differing in clinical features as described previously among patients identified under the Vancouver standard. Of excluded patients, 76% were reclassified as having other forms of JIA yet were phenotypically comparable with those retained. CONCLUSION: Despite apparently modest changes from previous criteria, ILAR definitions strikingly restrict the diagnosis of PsA in childhood. Similarity between excluded and included patients suggests that these restrictions may not reflect substantive clinical differences. To the extent that excluded patients become reclassified within JIA, current criteria risk compromising other ILAR categories while reducing the number of patients available for the study of juvenile PsA. PMID- 18163408 TI - Asking the experts: exploring the self-management needs of adolescents with arthritis. AB - OBJECTIVE: To explore the self-management needs of adolescents with juvenile idiopathic arthritis and the acceptability of a Web-based program of self management aimed at improving quality of life. METHODS: A descriptive qualitative design was used. A convenience sample of 36 adolescents (male and female) who varied in age, disease onset subtype, and disease severity were recruited from 4 Canadian tertiary care pediatric centers. Individual (n=25) and 3 focus-group (n=11) interviews were conducted with adolescents using semistructured interview guides. After each interview session, the audiotaped interview data were transcribed verbatim. NUD*IST 6.0 was used to assist with the sorting, organizing, and coding of the data. Data were organized into categories that reflected emerging themes. RESULTS: Adolescents articulated how they developed effective self-management strategies through the process of "letting go" from others who had managed their illness (health care professionals, parents) and "gaining control" over managing their illness on their own. The 2 strategies that assisted in this process were gaining knowledge and skills to manage the disease and experiencing understanding through social support. Five further subthemes emerged around skills to manage the disease, including knowledge and awareness about the disease, listening to and challenging care providers, communicating with the doctor, managing pain, and managing emotions. CONCLUSION: Adolescents were united in their call for more information, self-management strategies, and meaningful social support to better manage their arthritis. They believed that Web-based interventions were a promising avenue to improve accessibility and availability of these interventions. PMID- 18163409 TI - A randomized controlled trial of the people with arthritis can exercise program: symptoms, function, physical activity, and psychosocial outcomes. AB - OBJECTIVE: To evaluate the basic 8-week People with Arthritis Can Exercise (PACE) program for improvements in primary (symptoms, functioning, level of physical activity) and secondary (psychosocial) outcomes. METHODS: A total of 346 individuals with self-reported arthritis from 18 sites participated in a randomized controlled trial of PACE. Outcomes were measured at baseline and 8 weeks. The intervention group completed self-reported assessments at 3 and 6 months. Two-level multiple linear regression models were estimated to calculate adjusted outcome means in the intervention and control groups. A mixed-effects repeated-measures model was used to calculate adjusted means in the intervention group at 3 and 6 months. Both intent-to-treat (ITT) and as-treated (AT) analyses were conducted. RESULTS: At 8 weeks, the intervention group had improvements in the following outcomes: 2 symptom outcomes (pain, fatigue) and 1 psychosocial outcome (self-efficacy for managing arthritis) in the ITT analyses; 1 symptom outcome (pain), 1 function outcome (chair stands), and 1 psychosocial outcome (self-efficacy for arthritis management) in the AT analyses. In addition, completers who attended>or=9 classes had improvements in 3 symptom outcomes (pain, fatigue, stiffness), 2 function outcomes (10-pound lifts, chair stands), and 1 psychosocial outcome (self-efficacy for arthritis management) at 8 weeks. Relative to baseline, PACE participants maintained significant improvements in symptoms at 6 months, but declined in function and self-efficacy for exercise. CONCLUSION: If adults with arthritis attend a majority of PACE classes, they may expect improvements in symptoms, self-efficacy for arthritis management, and upper and lower extremity function. Achieving sustained improvement in outcomes may require continued participation in PACE. PMID- 18163410 TI - Hydroxychloroquine blood assay as a marker of nonadherence in patients with systemic lupus erythematosus: comment on the article by Koneru et al. PMID- 18163411 TI - A review and proposal for a core set of factors for prospective cohorts in low back pain: a consensus statement. PMID- 18163412 TI - Physical inactivity in patients with rheumatoid arthritis: data from twenty-one countries in a cross-sectional, international study. AB - OBJECTIVE: Regular physical activity is associated with decreased morbidity and mortality. Traditionally, patients with rheumatoid arthritis (RA) have been advised to limit physical exercise. We studied the prevalence of physical activity and associations with demographic and disease-related variables in patients with RA from 21 countries. METHODS: The Questionnaires in Standard Monitoring of Patients with Rheumatoid Arthritis (QUEST-RA) is a cross-sectional study that includes a self-report questionnaire and clinical assessment of nonselected consecutive outpatients with RA who are receiving usual clinical care. Frequency of physical exercise (>or=30 minutes with at least some shortness of breath, sweating) is queried with 4 response options: >or=3 times weekly, 1-2 times weekly, 1-2 times monthly, and no exercise. RESULTS: Between January 2005 and April 2007, a total of 5,235 patients from 58 sites in 21 countries were enrolled in QUEST-RA: 79% were women, >90% were white, mean age was 57 years, and mean disease duration was 11.6 years. Only 13.8% of all patients reported physical exercise>or=3 times weekly. The majority of the patients were physically inactive with no regular weekly exercise: >80% in 7 countries, 60-80% in 12 countries, and 45% and 29% in 2 countries, respectively. Physical inactivity was associated with female sex, older age, lower education, obesity, comorbidity, low functional capacity, and higher levels of disease activity, pain, and fatigue. CONCLUSION: In many countries, a low proportion of patients with RA exercise. These data may alert rheumatologists to motivate their patients to increase physical activity levels. PMID- 18163413 TI - Patellar taping and bracing for the treatment of chronic knee pain: a systematic review and meta-analysis. AB - OBJECTIVE: To evaluate the evidence for patellar taping and bracing in the management of chronic knee pain. METHODS: Randomized or quasi-randomized studies assessing patellar taping or bracing effects on chronic knee pain were sourced from 7 electronic databases (to November 2006), and assessed using the Physiotherapy Evidence Database scale. Weighted mean differences were determined, and pooled estimates of taping and bracing effects were obtained using random effects models. RESULTS: Of 16 eligible trials, 13 investigated patellar taping or bracing effects in individuals with anterior knee pain, and 3 investigated taping effects in individuals with knee osteoarthritis (OA). The methodologic quality of the taping studies was significantly higher than the bracing studies (mean+/-SD 4.8+/-2.1 versus 2.8+/-0.8; P<0.05). On a 100-mm scale, tape applied to exert a medially-directed force on the patella decreased chronic knee pain compared with no tape by 16.1 mm (95% confidence interval [95% CI] -22.2, -10.0; P<0.001) and sham tape by 10.9 mm (95% CI -18.4, -3.4; P<0.001). For anterior knee pain and OA, medially-directed tape decreased pain compared with no tape by 14.7 mm (95% CI -22.8, -6.9; P<0.001) and 20.1 mm (95% CI -26.0, -14.3; P<0.001), respectively. There was disputable evidence from low-quality studies for patellar bracing benefits. CONCLUSION: There was evidence that tape applied to exert a medially-directed force on the patella produces a clinically meaningful change in chronic knee pain. There was limited evidence to demonstrate the efficacy of patellar bracing. These outcomes were limited by the presence of high heterogeneity between study outcomes and significant publication bias. PMID- 18163414 TI - Daily spiritual experiences of older adults with and without arthritis and the relationship to health outcomes. AB - OBJECTIVE: Strategies to improve coping with chronic disease are increasingly important, especially with the aging US population. For many, spirituality serves as a source of strength and comfort. However, little is known about the prevalence of daily spiritual experiences (DSE) and how they may relate to physical and mental health. METHODS: We surveyed older adults age>50 years with chronic health conditions seen in a primary care setting about their DSE, health perceptions, pain, energy, and depression. RESULTS: Of 99 patients, 80% reported DSE most days and many times per day. Women had significantly lower DSE scores than men (reflecting more frequent DSE, mean+/-SD 37.3+/-15.0 versus 45.8+/-17.5; P=0.012). African American women reported the most frequent DSE and white men reported the least frequent DSE (mean+/-SD 35.9+/-13.6 versus 52.2+/-19.1). Frequent DSE were significantly associated with a higher number of comorbid conditions (P=0.003), although not with age, education, or employment status. Persons with arthritis reported significantly more DSE than those without arthritis (mean+/-SD 35.2+/-12.1 versus 47.1+/-18.6; P<0.001). After adjustment for age, race, sex, pain, and comorbid conditions, more frequent DSE were associated with increased energy (P<0.009) and less depression (P<0.007) in patients with arthritis. CONCLUSION: DSE are common among older adults, especially those with arthritis. Increased DSE may be associated with more energy and less depression. DSE may represent one pathway through which spirituality influences mental health in older adults. PMID- 18163415 TI - Participation restrictions among older adults with osteoarthritis: a mediated model of physical symptoms, activity limitations, and depression. AB - OBJECTIVE: Research emphasizes the negative impact of osteoarthritis (OA) on social participation, yet few studies have examined the roles of symptoms, activity limitations, and depression in this relationship. The present longitudinal study tested a model that hypothesizes that the relationship between physical symptoms and later participation restrictions among older adults with OA is mediated by activity limitations and depressive symptoms. METHODS: Participants were 184 community-dwelling senior adults (age>or=55 years) with a physician diagnosis of OA who were interviewed at 2 time points 18 months apart. Measures included demographic variables; a derived physical symptoms measure based on severity of pain, stiffness, and fatigue in the previous week; and depressive symptoms measured by the Center for Epidemiologic Studies Depression Scale. Measures of activity limitations and participation restrictions were derived by factor analysis of questions about difficulty in everyday life. Sequential multiple linear regression analyses controlling for demographic and illness-related variables were used to test for mediation. RESULTS: Severity of time 1 physical symptoms was associated with difficulties in participation 18 months later. Sequential introduction of variables showed that this relationship was partially mediated by time 1 activity limitations and time 1 depressive symptoms. When both of these variables were included in the model, the effect of symptoms severity was completely mediated. CONCLUSION: This study demonstrates the importance of taking into account both the physical (activity limitations) and psychological (depressive symptoms) consequences of OA symptoms and suggests that these factors act as a pathway to subsequent participation restrictions. PMID- 18163416 TI - Missing data in trials: do we have to keep carrying the last observation forward? PMID- 18163417 TI - Clinical remission and/or minimal disease activity in patients receiving adalimumab treatment in a multinational, open-label, twelve-week study. AB - OBJECTIVE: To evaluate the effect of adalimumab treatment on clinical remission and/or minimal disease activity (MDA) in 6,610 patients with active rheumatoid arthritis (RA) who were enrolled in the Research in Active RA trial, a multinational, open-label, 12-week study with an optional extension period. METHODS: Clinical remission was defined as a Disease Activity Score in 28 joints (DAS28)<2.6, Simplified Disease Activity Index (SDAI) score0.05). There was an increase in BNIP3 expression under conditions of hypoxia (0.1% 0(2)) but not with dihydrotestosterone stimulation or with Casodex treatment. CONCLUSIONS: These findings suggest that BNIP3 is directly regulated by hypoxia but that there may be a hormonal independent mechanism coordinating the expression of BNIP3 in prostate tumors. PMID- 18163428 TI - A molecular analysis of prokaryotic and viral DNA sequences in prostate tissue from patients with prostate cancer indicates the presence of multiple and diverse microorganisms. AB - BACKGROUND: Inflammation, both acute and chronic, is a common feature of prostate histology. While inflammation has been proposed to play an important role in both benign and malignant growth of the prostate, the stimuli for this inflammation remain poorly characterized. Infectious pathogens are potential stimuli for prostatic inflammation. METHODS: Universal eubacterial PCR was used to test 170 prostate tissue core samples from 30 cancer patients for 16S rDNA gene sequences. Positive PCR products (n=64, 37%) were cloned and sequenced. For comparison, tissue samples from 30 patients were cultured using standard clinical microbiological techniques. DNA samples from 200 additional patients were tested by organism-specific PCR for the presence of Chlamydia trachomatis, Propionibacterium acnes, Trichomonas vaginalis, BK virus, Epstein-Barr virus, human cytomegalovirus, human papillomavirus, and xenotropic murine leukemia related virus. RESULTS: 16S sequencing results indicated the presence of 83 distinct microorganisms. Microbiological culture isolated markedly fewer species. In general, organism-specific PCR failed to detect multiple organisms previously reported as common in the prostate. There was no significant association between the presence of particular species of bacteria and histologic evidence of acute or chronic inflammation. CONCLUSIONS: Most prostates from men undergoing prostatectomy (87%) contain bacterial DNA from one or more species. However, the majority of individual tissue core samples were negative, suggesting regional heterogeneity in the presence of bacteria and a lack of a generalized or ubiquitous prostatic flora. Culture results suggest either the "unculturable" nature of species present in the prostate or that 16S rDNA sequences were derived from non-viable bacteria. PMID- 18163429 TI - Genetic polymorphisms in CYP17, CYP3A4, CYP19A1, SRD5A2, IGF-1, and IGFBP-3 and prostate cancer risk in African-American men: the Flint Men's Health Study. AB - BACKGROUND: Association studies have examined the significance of several candidate genes based on biological pathways relevant to prostate carcinogenesis, including both the androgen and insulin-like growth factor pathways. Clinical and epidemiologic evidence suggest that androgens, specifically testosterone and dihydrotestosterone (DHT) are important not only in normal prostate growth but in the pathogenesis of prostate cancer. Similarly, the insulin-like growth factor-1 (IGF-1) signaling pathway regulates both cellular proliferation and apoptosis. Therefore, genes involved in the biosynthesis, activation, metabolism and degradation of androgens and the stimulation of mitogenic and antiapoptotic activities of prostate epithelial cells represent important candidates for affecting the development and progression of prostate cancer. METHODS: Using resources from the Flint Men's Health Study, a population-based case control study of African-American men aged 40-79, we evaluated the associations between selected single-nucleotide polymorphisms (SNPs) in the CYP17, CYP3A4, CYP19A1, SDR5A2, IGF1, and IGFBP3 genes and prostate cancer diagnosis in 473 men (131 prostate cancer cases and 342 disease-free controls). RESULTS: We found a significant association between prostate cancer and selected CYP17 SNP genotypes, with the heterozygous genotype conferring decreased risk. Suggestive evidence for association between IGF1 SNPs and prostate cancer were also found. No significant associations were observed between SNPs in the other genes and prostate cancer. CONCLUSIONS: These findings suggest that variation in or around CYP17 and/or IGF1 may be associated with prostate cancer development in the African-American population. Additional studies are needed to determine whether these polymorphisms are indeed associated with prostate cancer risk in African Americans. PMID- 18163430 TI - TGF-beta signaling and androgen receptor status determine apoptotic cross-talk in human prostate cancer cells. AB - BACKGROUND: A signaling interaction between transforming growth factor-beta (TGF beta) and androgens promotes apoptosis in human prostate cancer cells LNCaP TbetaRII (androgen-sensitive and TGF-beta responsive). This study investigated the contribution of androgen receptor (AR) in the combined effect of TGF-beta and dihydrotestosterone (DHT), on regulation of apoptosis and AR- and TGF-beta mediated transcriptional activity in human prostate cancer cells. METHODS: Transcriptional activation in response to TGF-beta (5 ng/ml) and DHT (1 nM) was evaluated using transient transfections and luciferase assays in human prostate cancer cells, LNCaP-TbetaRII and PC-3, overexpressing the wild type AR. The apoptotic response to DHT/TGFbeta treatment was correlated with AR cellular distribution and the AR interaction with TGF-beta intracellular effector Smad4. RESULTS: The results revealed that TGF-beta signaling induced AR-mediated transcriptional activation in two androgen-responsive promoters [probasin and prostate specific antigen (PSA)]. TGF-beta1 induced transcriptional activity enhanced by DHT in both cell lines (LNCaP-TbetaRII and PC-3-AR) via AR-Smad4 interaction. This interaction however does not exclusively drive TGF-beta mediated apoptosis as DHT failed to enhance such an effect in PC-3 AR (wt) cells. CONCLUSIONS: These results demonstrate that the AR status determines the sensitivity of prostate cancer cells to the apoptotic effects of TGF-beta1, thus providing a new insight into the mechanism via which TGF-beta cross-sections the AR axis toward the functional convergence of the two pathways in the development of androgen-independent prostate cancer. This study is potentially significant in defining the contribution of AR status to the emergence of androgen-independent prostate tumors. PMID- 18163431 TI - Effectiveness of teaching an early parenting approach within a community-based support service for adolescent mothers. AB - A single blind, pre-test, post-test design was used to test the effectiveness of the Keys to Caregiving Program in enhancing adolescent mother-infant interactions. Participants were sequentially allocated to groups in order of referral. The outcome was the enhancement of maternal and infant behaviors that exhibited mutual responsiveness as measured by the Nursing Child Assessment Teaching Scale. Issues with recruitment and collaboration with the community agencies made achieving a desirable sample size difficult. Pre-tests and post tests were completed for 13 participants. While the sample size was insufficient to confidently establish whether or not the Keys to Caregiving produced a between groups treatment effect, mothers within the treatment group evidenced significantly greater contingent responsiveness over time than those within the control group. PMID- 18163432 TI - Association of complement factor H Y402H gene polymorphism with Alzheimer's disease. AB - Alzheimer's disease (AD) and age-related macular degeneration (AMD) share several epidemiological and biochemical features. The present study aimed to assess the possible influence of the AMD-associated complement factor H (CFH) Y402H (1277T > C) polymorphism on the risk of AD. Caucasian subjects (n = 800) meeting the criteria for probable (n = 717) or definite (n = 83) AD and Caucasian non demented controls (n = 1265) were included in this multi-center case-control study, in which genotype and allele frequencies of the CFH 1277T > C polymorphism were determined and related to diagnosis, APOE genotype, Mini-Mental State Examination score (MMSE) and the cerebrospinal fluid (CSF) biomarkers total-tau (T-tau), phospho-tau(181) (P-tau(181)), and beta-amyloid(1-42) (Abeta(1-42)). The AMD-associated CFH genotypes (1277CC and 1277TC) were overrepresented in subjects with AD as compared to control individuals (P = 0.029). Positive C carrier status was associated with an odds ratio (OR) for AD of 1.24 (95% confidence interval [CI] 1.02-1.50). When APOE epsilon4 carrier status was included in the regression model, this association was even stronger (OR 1.34, 95% CI: 1.08-1.65, P = 0.007). Subgroup analysis showed that the association between CFH C allele positivity and AD was only evident for individuals carrying the APOE epsilon4 allele. Positive C carrier status was also associated with lower levels of CSF Abeta(1-42) selectively in the control group in an APOE epsilon4-independent manner (P = 0.003). In conclusion, the CFH 1277T > C polymorphism seems to influence the risk of AD and there appears to be an interaction between CFH 1277C and APOE epsilon4 alleles. The CFH 1277C allele may predispose patients for co morbidity in AD and AMD. PMID- 18163433 TI - Associations of ATF4 gene polymorphisms with schizophrenia in male patients. AB - Activating transcription factor 4 (ATF4) is considered as a positional candidate gene for schizophrenia due to its location at chromosome 22q13, a region linked to schizophrenia. Furthermore, as protein interaction partner of ATF4, disrupted in schizophrenia 1 (DISC1) and its signal pathway implicated in the pathophysiology of schizophrenia have been widely supported by a number of genetic and neurobiological studies. Our aim was to investigate whether ATF4 is associated with schizophrenia in case-control samples of Han Chinese subjects consisting of 352 schizophrenia patients and 357 healthy controls. We detected 18 single nucleotide polymorphisms (SNPs) in ATF4 locus, two of which were analyzed, including one insertion at the putative core promoter region (rs17001266, -/C) and one nonsynonymous variant in exon 1 (rs4894, C/A, Pro22Gln). Allele distributions of two SNPs showed significant associations with schizophrenia in male subjects (respectively, rs17001266: P = 0.021, OR = 1.58, 95% CI = 1.07 2.33; rs4894: P = 0.004, OR = 1.78, 95% CI = 1.19-2.67), but not in female subjects as well as the entire population. Two haplotypes CC and -A constructed of rs17001266-rs4894 also revealed significant associations with schizophrenia in male group (global P = 0.0097). These findings support that ATF4 gene may be involved in susceptibility to schizophrenia with sex-dependent effect in the Chinese Han population and suggest that further functional assays are needed to verify their relevance to the pathogenesis of schizophrenia. PMID- 18163434 TI - Regulation of NGFI-B/Nur77 gene expression in the rat ovary and in leydig tumor cells MA-10. AB - NR4A1, also called NGFI-B in the rat, Nur77 in the mouse and TR3 in humans, belongs to the orphan nuclear steroid hormone receptor superfamily and is one of the immediate-early genes. In the endocrine organs, including the gonads, NGFI B/Nur77 gene expression is rapidly induced by pituitary hormones. NGFI-B/Nur77 expression was found to be rapidly reduced by an estrogenic endocrine disrupter, diethylstilbestrol (DES) in theca interna cells of immature rat ovaries. DES treatment also triggered a rapid decrease of serum luteinizing hormone (LH) levels, suggesting that DES acts on the hypothalamo-pituitary axis to suppress LH secretion from the pituitary. The transcriptional regulation of NGFI-B/Nur77 by LH/human chorionic gonadotropin (hCG) or 8-bromoadenosine 3'-5'-cyclic monophosphate (8 Br-cAMP) was examined in mouse Leydig tumor cells MA-10. Luciferase assays using NGFI-B/Nur77 promoter constructs and electric mobility shift assays (EMSA) showed that NGFI-B/Nur77 gene expression was mediated through three of the four activator protein-1 (AP-1)-like sites, namely the -233 AP-1, 213 AP-1 and -69 AP-1 sites adjacent to the transcription start site of the NGFI B/Nur77 promoter. We also demonstrated here that both the Jun family and cAMP responsive element binding (CREB) proteins bind to the -233 AP-1 site, whereas the main binding protein to the -213 AP-1 site was CREB, and Jun family protein to the -69 AP-1 site, respectively. The rapid induction of NGFI-B/Nur77 gene expression by LH/hCG in MA-10 cells appears to be mediated by both CREB and Jun family proteins through the cAMP-protein kinase A (PKA) pathway. PMID- 18163435 TI - Prognostic significance of depth of invasion, vascular invasion and numbers of lymph node retrievals in combination for patients with stage II colorectal cancer undergoing radical resection. AB - BACKGROUND AND OBJECTIVES: To determine which aspects of tumor histology influenced the postoperative relapse and overall survival rates after radical resection of UICC stage II colorectal cancer (CRC). METHODS: Data were collected on 259 patients with stage II CRC who underwent radical resection in Kaohsiung Medical University Hospital between January 2002 and December 2005. RESULTS: A univariate analysis identified that the depth of invasion, the presence of vascular invasion, the presence of perineural invasion, and the number of examined lymph nodes were significant prognostic factors for postoperative relapse. A combination of depth, vascular invasion, and numbers of lymph node retrieval as predictors of postoperative relapse showed that the more predictors that are involved, the higher chance that postoperative relapse would occur. Furthermore, T4 depth of tumor invasion, the presence of vascular invasion, and the number of examined lymph nodes <12 were considerably correlated to the poorer overall survival rates by survival analyses. CONCLUSIONS: This study has revealed that the depth of invasion, the presence of vascular invasion, and number of examined lymph nodes, may prominently affect the prognosis of stage II CRC patients after radical resection. The increasing risk of postoperative relapse is proportionate to numbers of these three parameters. PMID- 18163436 TI - Comments on Neurophysiological effects of mobile phone electromagnetic fields on humans: a comprehensive review. PMID- 18163437 TI - Psychomotor performance is not influenced by brief repeated exposures to mobile phones. AB - The present study investigated the presence of a cumulative effect of brief and repeated exposures to a GSM mobile phone (902.40 MHz, 217 Hz modulated; peak power of 2 W; average power of 0.25 W; SAR = 0.5 W/kg) on psychomotor functions. To this end, after each of 3 15-min exposures, both an acoustic simple reaction time task (SRTT) and a sequential finger tapping task (SFTT) were administered to 24 subjects. The present study was unable to detect the cumulative effects of brief and repeated EMF exposure on human psychomotor performance, although there was a non-statistical trend to shorter reaction times. In summary, these data show an absence of effects with these particular exposure conditions; however, possible cognitive effects induced by different signal characteristics cannot be excluded. PMID- 18163438 TI - Characterization of secretory leukocyte protease inhibitor as an inhibitor of implantation serine proteinases. AB - We have recently identified and characterized two implantation serine proteinase genes, ISP1 and ISP2, which give rise to a dimeric proteinase, ISP that facilitates embryo invasion during peri-implantation period. As many proteinases have cognate serpins that regulate their proteolytic activity, we have been investigating anti-tryptases, expressed during this window of implantation. Here, we report the differential expression of secretory leukocyte protease inhibitor (SLPI) in uterine endometrium around the implantation period. The co-localization of SLPI and ISP suggests the possibility that SLPI is an ISP serpin and that expression of SLPI may lead to a reduction in ISP activity. The expression of SLPI is down regulated during the window of embryo-uterine receptivity. Our results are consistent with a model suggesting that the drop in SLPI expression may help to refine the opening of the window of implantation, by allowing the proteolytic activity of embryo invasive serine proteinases such as the ISPs. PMID- 18163439 TI - Pulsed electric field reduces the permeability of potato cell wall. AB - The effect of the application of pulsed electric fields to potato tissue on the diffusion of the fluorescent dye FM1-43 through the cell wall was studied. Potato tissue was subjected to field strengths ranging from 30 to 500 V/cm, with one 1 ms rectangular pulse, before application of FM1-43 and microscopic examination. Our results show a slower diffusion of FM1-43 in the electropulsed tissue when compared with that in the non-pulsed tissue, suggesting that the electric field decreased the cell wall permeability. This is a fast response that is already detected within 30 s after the delivery of the electric field. This response was mimicked by exogenous H2O2 and blocked by sodium azide, an inhibitor of the production of H2O2 by peroxidases. PMID- 18163440 TI - In vitro effect of pulsed 900 MHz GSM radiation on mitochondrial membrane potential and motility of human spermatozoa. AB - Ejaculated, density purified, human spermatozoa were exposed to pulsed 900 MHz GSM mobile phone radiation at two specific absorption rate levels (SAR 2.0 and 5.7 W/kg) and compared with controls over time. Change in sperm mitochondrial membrane potential was analysed using flow cytometry. Sperm motility was determined by computer assisted sperm analysis (CASA). There was no effect of pulsed 900 MHz GSM radiation on mitochondrial membrane potential. This was also the case for all kinematic parameters assessed at a SAR of 2.0 W/kg. However, over time, the two kinematic parameters straight line velocity (VSL) and beat cross frequency (BCF) were significantly impaired (P < 0.05) after the exposure at SAR 5.7 W/kg and no exposure by time interaction was present. This result should not be ascribed to thermal effects, due to the cooling methods employed in the RF chamber and temperature control within the incubator. PMID- 18163441 TI - Enhanced secretion of prostaglandin E2 from osteoblasts by exposure to a strong static magnetic field. AB - Exposure to static magnetic fields (SMFs) has been reported to promote osteoblast differentiation in vitro, and increase bone formation in vivo and in clinical studies. Prostaglandins respond early to exogenous mechanical loading, and play an important role in bone formation. In this study, we investigated whether exposure to a strong SMF affects prostaglandin E(2) (PGE(2)) secretion from a mouse osteoblastic cell line, MC3T3-E1. We also investigated the PGE(2) synthesizing enzyme, cyclooxygenase 2 (Cox-2), and translocation of the transcription factor nuclear factor kappa B (NF-kappaB), which is involved in the induction of Cox-2 expression. In the SMF exposures, experiments were performed at the 10 T-exposure position, at which the magnetic flux density was highest, and at the 6 T-exposure position, at which the magnetic field gradient was highest (41.7 T/m). PGE(2) secretion was not affected by exposure at the 10 T exposure position compared to sham-exposure, but was enhanced at the 6 T-exposure position (about 1.5-fold). Similarly, Cox-2 expression and NF-kappaB translocation were not enhanced at the 10 T-exposure position, but increased at the 6 T-exposure position (about twofold, two- to threefold, respectively). These findings suggested that exposure to a high magnetic field gradient induced secretion of PGE(2) and expression of the Cox-2 protein, which was mediated through increased translocation of NF-kappaB. PMID- 18163442 TI - Isolation and characterization of the spermatid-specific Smrp1 gene encoding a novel manchette protein. AB - The manchette, which is the structure that appears around the nuclei of elongated spermatids, is assumed to be involved in nuclear shaping during spermiogenesis and the transport of various proteins between the nucleus and sperm tail. In this report, we describe the molecular cloning and characterization of a mouse spermatid-specific manchette-related protein 1 (Smrp1) from a spermatid-specific subtracted mouse testis cDNA library. The isolated Smrp1 cDNA clones could be divided into three variants based on sequence analysis. Computer-assisted analysis showed that these variants were splice variants from a single locus of the mouse genome. The three putative proteins consisted of 296, 260, and 175 amino acids, respectively. Although 155 amino acids of the N terminus were common to the three proteins, they were distinguished by their C-terminal regions. Western blot analyses using specific antisera showed that SMRP1 expression was specific to the testes and that only the 261-amino-acid form was translated into protein. Immunohistochemistry revealed that SMRP1 was localized to the cytoplasm of step 9-12 elongated spermatids. The protein appeared in a cap formation that covered the caudal sides of the elongated nuclei. This localization pattern coincided with that of the manchette. SMRP1 may play an important role as a functional protein that co-operates with manchette proteins. PMID- 18163443 TI - Goldsinny wrasse (Ctenolabrus rupestris) is an extreme vtgAa-type pelagophil teleost. AB - During oocyte maturation in the goldsinny wrasse (Ctenolabrus rupestris) extensive proteolysis of yolk proteins generates a large pool of free amino acids that drive hydration of the pelagic egg. By cloning hepatic vitellogenins (vtg) and using mass spectrometry, N-terminal microsequencing, and Western immunoblotting to identify the yolk proteins (Yp), we show that multiple forms of vitellogenin mRNAs (vtgAa, vtgAb, and vtgC) are expressed in the liver, but only a single major class of the Yps derived from vtgAa predominates in the oocytes. Some Yps derived from vtgAb and vtgC appear also to be incorporated in the oocytes and eggs, but only at background levels. During oocyte hydration the vtgAa-derived lipovitellin heavy chain (LvH-Aa) and its cleavage variants are completely degraded leaving only a processed lipovitellin light chain (LvL-Aa) fragment as the major yolk protein for embryonic development. The maturational cleavage site of the LvL-Aa is identified as two amino acids downstream from the conserved Tyr(1168) of VtgAa in Atlantic halibut. In addition, although a beta' component (approximately 18 kDa) is present in the oocytes, it is not fully degraded during the hydration process. PMID- 18163444 TI - Cre-loxP system as a versatile tool for conferring increased levels of tissue specific gene expression from a weak promoter. AB - Attempts to image reporter gene expression driven by weak promoters are often hampered by the poor transcriptional activity of such promoters. Most tissue specific promoters are weak compared with stronger but constitutively expressing viral promoters. In this study, we validated methods of enhancing the transcriptional activity of weak promoters using a Cre-loxP system in vitro and in vivo. We constructed a tester vector, pCTL, which carries a strong systemic cytomegalovirus enhancer/chicken beta-actin promoter (CAG), loxP-flanked CAT, and firefly luciferase (luc) cDNAs. Herpes simplex virus-thymidine kinase (HSV-tk) promoter was used as a weak and systemic promoter and ligated to Cre for construction of pTC. Luc activity was higher (about 10-fold enhancement) in co transfected (with pCTL and pTC) than in singly (with HSV-tk promoter-driven luc expression vector pTL) transfected NIH3T3 cells. In vivo electroporation-mediated gene delivery of both pCTL and pTC into murine oviductal epithelium yielded results (about 16-fold enhancement) similar to those obtained with in vitro transfected NIH3T3 cells. To evaluate tissue-specific enhancement of gene expression, podocyte (glomerular visceral epithelial cell)-specific nephrin promoter was ligated to the Cre gene or luc cDNA to create pNC and pNL, respectively. We achieved 2.4-fold improvement of luc gene expression in the mouse kidney in vivo when pCTL and pNC were co-transfected via the tail vein via the lipoplex method. The combination of a weak tissue-specific promoter with the Cre-loxP system could thus be used to enhance the strength of tissue-specific promoters in vitro and in vivo. PMID- 18163445 TI - Effect of temporary nuclear arrest by phosphodiesterase 3-inhibitor on morphological and functional aspects of in vitro matured mouse oocytes. AB - The present study aimed to analyze detailed morphological and functional characteristics of mouse in vitro matured oocytes after a pre-maturation culture (PMC) by temporary nuclear arrest with the specific phosphodiesterase 3-inhibitor (PDE3-I) Cilostamide. In a first experiment the lowest effective dose of Cilostamide was determined. Cumulus-oocyte complexes (COCs), isolated from small antral follicles, were exposed to different concentrations of Cilostamide (ranging from 0 (control) to 10 microM) for 24 hr. Afterwards, oocytes were removed from PDE3-I-containing medium and underwent in vitro maturation (IVM) for 16-18 hr. A concentration of 1 microM Cilostamide was the lowest effective dose for maximum level of inhibition and reversibility of meiosis inhibition. This concentration was used in further experiments to evaluate oocyte quality following IVM in relation to different parameters: kinetics of meiotic progression, metaphase II (MII) spindle morphology, aneuploidy rate, fertilization, and embryonic developmental rates. The results were compared to nonarrested (in vitro control) and in vivo matured oocytes (in vivo control). Following withdrawal of the inhibitor, the progression of meiosis was more synchronous and accelerated in arrested when compared to nonarrested oocytes. A PMC resulted in a significant increase in the number of oocytes constituting a MII spindle of normal morphology. None of the oocytes exposed to PDE3-I showed numerical chromosome alterations. In addition, fertilization and embryonic developmental rates were higher in the PMC group compared to in vitro controls, but lower than in vivo controls. These results provide evidence that induced nuclear arrest by PDE3-I is a safe and reliable method to improve oocyte quality after IVM. PMID- 18163446 TI - The utility of SELENBP1 gene expression as a biomarker for major psychotic disorders: replication in schizophrenia and extension to bipolar disorder with psychosis. AB - While microarray studies are generating novel insights into the etiology of major psychiatric disorders, the validation of microarray-identified candidate genes and their role in the causality of these disorders has been less often studied. We have previously demonstrated, by microarray, up-regulation of SELENBP1 in the brain and blood of patients with schizophrenia. The main aim of the current study was to validate this finding using quantitative real-time PCR (QPCR) in an independent brain cohort that included patients with bipolar disorder. Our sample consisted of mRNAs from the dorsolateral prefrontal cortex (dlPFC) of 34 schizophrenic patients, 33 bipolar disorder patients (including 20 with psychotic history), and 34 normal control subjects. QPCR was employed to assess gene expression changes, with C(T) values analyzed using an ANCOVA approach. The results demonstrated that SELENBP1 mRNA was upregulated in schizophrenic brains versus controls (P = 0.046) and, in addition, that SELENBP1 gene expression was strongly positively correlated with presence of psychosis across diagnoses (P < 0.001, increased by 12%). Based on these findings, we conclude that elevated SELENBP1 is a possibly consistent feature in the schizophrenic brain and that this finding could underlie some commonalities of psychosis across the boundaries of diagnoses. Future studies should exploit DNA-based methods and molecular investigations on the role of SELENBP1 in order to gain insights into the nature of its influence on schizophrenia and psychotic symptoms. PMID- 18163447 TI - Health outcomes associated with potentially inappropriate medication use in older adults. AB - The purpose of this study was to examine the prevalence of potentially inappropriate medication use (PIMs) among community-dwelling older adults and the association between PIMs and health care outcomes. Participants were 17,971 individuals age 65 years and older. PIM use was defined by the Beers criteria. Drug-related problems (DRPs) were defined using ICD-9 codes. Forty percent of the 17,971 individuals filled at least 1 PIM prescription, and 13% filled 2 or more PIM prescriptions. Overall DRP prevalence among those with at least 1 PIM prescription was 14.3% compared to 4.7% in the non-PIM group (p < .001). In conclusion, preventing PIM use may be important for decreasing medication-related problems, which are increasingly being recognized as requiring an integrated interdisciplinary approach. PMID- 18163448 TI - The ubiquitin-proteasome pathway and enhanced activity of NF-kappaB in gastric carcinoma. AB - BACKGROUND AND OBJECTIVES: NF-kappa B, ubiquitin and proteasome have been shown be important factors in oncogenesis. The aim of this study was to determine whether NF-kappa B could be a sensitive biomarker for gastric carcinoma. METHODS: Tumor and adjacent mucosal tissue specimens in 92 patients with gastric carcinoma were studied. The expression of NF-kappa B was evaluated by immunohistochemistry. The expression of I kappa B alpha, ubiquitin in cytoplasm and NF-kappa B in nucleoplasm was assayed by Western blot. DNA binding-activity of NF-kappa B was confirmed by electrophoretic mobility shift assay (EMSA). Fluorogenic technique was performed to measure the 26S proteasome activity. RESULTS: NF-kappa B positive expression in tumor tissues (82.4%) was significant higher than that in adjacent mucosal tissues (32.7%, P < 0.05). The increase of NF-kappa B activation was accompanied by the increases of ubiqutin, 26S proteasome activation and a degradation of I kappa B alpha but not the ubiquitin-conjugated I kappa B alpha/NF-kappa B complex in gastric carcinoma. NF-kappa B expression was significantly increased in patients with lymph node metastasis, TNM stage III/IV or with the habit of high intake of pickled vegetables. CONCLUSIONS: This study demonstrates that the constitutive activation of NF-kappa B is likely due to the activation of ubiquitin-proteasome pathway and NF-kappa B can be used as a prognostic biomarker. PMID- 18163449 TI - Variation in GABA-A subunit gene copy number in an autistic patient with mosaic 4 p duplication (p12p16). AB - Autism has been associated with chromosomal aberrations, including duplications at chromosome 4, and the identification of genetic factors contributing to the etiology of this disease is the focus of much research. Here we report a Japanese girl with mosaic of chromosome 4p duplication, mos 46,XX,dup(4)(p12p16)[54]/46,XX[6], who was diagnosed with autism at 3 years of age. Fluorescence in situ hybridization (FISH) with probes covering the region spanning a cluster of the gamma aminobutyric acid A (GABA-A) receptor subunit genes in the proximal short arm of chromosome 4 demonstrated total three signals for the GABRG1, GABRA4, and GABRA2 genes, but only two signals for GABRB1. This suggests that aberrant copy number of the GABA-A receptor subunit genes may contribute to the etiology of autism in this patient. PMID- 18163450 TI - A linkage analysis of cigarette and alcohol consumption in an unselected Mexican American population. AB - The use of alcohol and tobacco is highly prevalent. Studying the rate of consumption in a non-selected population could contribute to the elucidation of pathways involved in addiction or to the development of prevention programs. The San Antonio Family Heart Study has approximately 1,400 members with longitudinal data and did not select the proband with regard to exposure status. The goal of this study was to perform genome-wide linkage analysis of the rate of alcohol and cigarette consumption in a "normal" population. We used SOLAR to perform variance components based analysis of the transformed maximal rate of consumption. Despite estimated heritabilities of 0.52 (P < 0.001) for cigarette and 0.39 (P < 0.001) for alcohol consumption, univariate linkage analyses produced only suggestive LOD scores, however the second suggestive linkage peak for the alcohol phenotype was present at 148 cM on chromosome 10, in the exact vicinity of the peak for the cigarette phenotype. In a bivariate analyses, the environmental correlation between alcohol and cigarette consumption was not significantly different from zero (rho(e) = -0.15, P = 0.18) and the overall genetic correlation was not different from zero (rho(g) = 0.16, P = 0.34). The results from the bivariate linkage analysis found a maximum LOD score of 3.82 (genome-wide P = 0.0054) at 151 cM on chromosome 10, at the location of the overlapping peaks from the univariate analyses. PMID- 18163451 TI - Subthalamic nucleus deep brain stimulation improves deglutition in Parkinson's disease. AB - Relatively little is known about the role of the basal ganglia in human deglutition. Deep brain stimulation (DBS) affords us a model for examining deglutition in humans with known impairment of the basal ganglia. The purpose of this study was to examine the effects of subthalamic nuclei (STN) DBS on the oral and pharyngeal stages of deglutition in individuals with Parkinson's Disease (PD). It was hypothesized that DBS would be associated with improved deglutition. Within participant, comparisons were made between DBS in the ON and OFF conditions using the dependent variables: pharyngeal transit time, maximal hyoid bone excursion, oral total composite score, and pharyngeal total composite score. Significant improvement occurred for the pharyngeal composite score and pharyngeal transit time in the DBS ON condition compared with DBS OFF. Stimulation of the STN may excite thalamocortical or brainstem targets to sufficiently overcome the bradykinesia/hypokinesia associated with PD and return some pharyngeal stage motor patterns to performance levels approximating those of "normal" deglutition. However, the degree of hyoid bone excursion and oral stage measures did not improve, suggesting that these motor acts may be under the control of different sensorimotor pathways within the basal ganglia. PMID- 18163452 TI - The variability of levodopa response in Parkinson's disease: is sensitization reversible? PMID- 18163453 TI - For better or worse: The effect of levodopa on speech in Parkinson's disease. AB - While the beneficial effect of levodopa on traditional motor control tasks have been well documented over the decades, its effect on speech motor control has rarely been objectively examined and the existing literature remains inconclusive. This paper aims to examine the effect of levodopa on speech in patients with Parkinson's disease. It was hypothesized that levodopa would improve preparatory motor set related activity and alleviate hypophonia. Patients fasted and abstained from levodopa overnight. Motor examination and speech testing was performed the following day, pre-levodopa during their "off" state, then at hourly intervals post-medication to obtain the best "on" state. All speech stimuli showed a consistent tendency for increased loudness and faster rate during the "on" state, but this was accompanied by a greater extent of intensity decay. Pitch and articulation remained unchanged. Levodopa effectively upscaled the overall gain setting of vocal amplitude and tempo, similar to its well-known effect on limb movement. However, unlike limb movement, this effect on the final acoustic product of speech may or may not be advantageous, depending on the existing speech profile of individual patients. PMID- 18163454 TI - Excessive dopamine neuron loss in progressive supranuclear palsy. AB - Progressive supranuclear palsy (PSP) and Parkinson's disease (PD) differ in their response to dopaminergic replacement therapies, despite having a similar degree of neuronal degeneration in the dopaminergic substantia nigra. We observed more widespread dopamine neuron loss in the extranigral A10 midbrain cell groups in PSP compared with PD. These cell groups innervate subcortical and cortical regions and may be required for adequate response to levodopa therapy. PMID- 18163455 TI - Clinical and neurophysiological characterization of myoclonus in complex regional pain syndrome. AB - The origin of myoclonus in patients with complex regional pain syndrome (CRPS) is unknown. Eight patients with CRPS related myoclonus were clinically evaluated and studied with intermuscular and corticomuscular coherence analysis. Jerks were present at rest, aggravated during action and were frequently associated with tremulousness or dystonia. Electromyography demonstrated a burst duration ranging from 25 to 240 ms with burst frequencies varying from <1 jerk/s during rest to 20 Hz during action. Coherence studies showed increased intermuscular coherence in 4 patients in the 6 to 12 Hz band, as reported in patients with enhanced physiological tremor. In 2 patients side-to-side coherence was observed, pointing to a central oscillatory drive. Significant coherence entrainment was detected in 5 patients. We conclude that the characteristics of myoclonus in CRPS are different from other forms of myoclonus. PMID- 18163456 TI - Staged unilateral or bilateral STN-DBS? PMID- 18163457 TI - Increased temporal blood flow associated with visual hallucinations in Parkinson's disease with dementia. PMID- 18163458 TI - Sucrose octasulfate regulates fibroblast growth factor-2 binding, transport, and activity: potential for regulation of tumor growth. AB - The antithrombotic activity of heparin has largely been credited with the success found in some cancer treatment by heparin. There are, however, many potent growth factors involved in tumor and blood vessel growth that bind to heparin with high affinity and their regulation by heparin may play a role in heparin's efficacy. We therefore chose to study the activity of a heparin analog, sucrose octasulfate (SOS), which has been similarly shown to interact with heparin-binding growth factors. Using mouse melanoma and lung carcinoma models, we demonstrate in vivo inhibition of tumor growth by SOS. SOS, however, showed little effect in coagulation assays indicating that this activity was not a primary mechanism of action for this molecule. Studies were then performed to assess the effect of SOS on basic fibroblast growth factor (FGF-2) activity, a growth factor which promotes tumor and blood vessel growth and is produced by B16 melanoma cells. SOS potently inhibited FGF-2 binding to endothelial cells and stripped pre-bound FGF 2 from cells. SOS also regulated FGF-2 stimulated proliferation. Further, SOS facilitated FGF-2 diffusion through Descemet's membrane, a heparan sulfate-rich basement membrane from the cornea, suggesting a possible role in FGF-2 clearance. Our results suggest that molecules such as SOS have the potential to remove growth factors from tumor microenvironments and the approach offers an attractive area for further study. PMID- 18163459 TI - Celecoxib-induced growth inhibition in SW480 colon cancer cells is associated with activation of protein kinase G. AB - Although it is often assumed that the antitumor effects of nonsteroidal anti inflammatory drugs (NSAIDs) are due to inhibition of cyclooxgenase (COX) activity, specifically COX-2, there is accumulating evidence that COX-2 independent mechanisms can also play an important role. Studies with sulindac sulfone (Aptosyn) and related derivatives have revealed a novel pathway of tumor growth inhibition and apoptosis mediated by activation of the guanosine 3',5' monophosphate (cGMP)-dependent enzyme protein kinase G (PKG). The present study indicates that concentrations of the NSAIDs celecoxib, indomethacin, and meclofenamic acid that inhibit growth of SW480 human colon cancer cells inhibit subcellular cGMP-phosphodiesterase (PDE) enzymatic activity and in intact cells induce a two- to threefold increase in intracellular levels of cGMP. This is associated with phosphorylation of the protein VASP, a marker of PKG activation, activation of JNK1 and a decrease in cellular levels of cyclin D1; effects seen with other agents that cause activation of PKG in these cells. On the other hand even a high concentration of the COX-2 specific inhibitor rofecoxib (500 microM) did not inhibit growth of SW480 cells. Nor did rofecoxib inhibit cGMP-PDE activity or cause other changes related to PKG activation in these cells. Since activation of the PKG pathways by celecoxib, indomethacin, and meclofenamic acid in this cell culture system required high concentrations of these compounds, it remains to be determined whether activation of this pathway contributes to the in vivo antitumor effects of specific NSAIDs. PMID- 18163460 TI - Core biopsy for diagnosis and tissue procurement. PMID- 18163461 TI - Outcome after surgical treatment of suspected gastrointestinal stromal tumors involving the duodenum: is limited resection appropriate? AB - INTRODUCTION: Present surgical opinion is divided regarding the optimal method for the treatment of duodenal gastrointestinal stromal tumor (GIST) with some supporting the selective use of limited resection (LR) versus others who prefer pancreaticoduodenectomy (PD). METHODS: A retrospective review of 22 patients who underwent surgery for suspected GIST involving the duodenum. RESULTS: There were 15 GISTs, 1 leiomyosarcoma and 6 other non-GIST benign submucosal tumors. Seven patients underwent LR and seven underwent PD for GIST. The median follow-up was 42 (range, 2-174) months. Patients who underwent LR versus PD had similar mean disease-specific survival [144 (95% CI, 92-196) vs. 130 (95% CI, 82-127) months, P = 0.808] and recurrence rates (14% vs. 29%, P = 0.515). All recurrences occurred at distant sites. Comparison between LR versus PD demonstrated that LR was associated with a significantly shorter operation time [125 (range, 50-305) vs. 350 (range, 210-465) min., P = 0.001] but similar morbidity rate (23% vs. 43%, P = 0.357). Comparison between GIST and other benign tumors demonstrated that size was the only statistically significant distinguishing factor [8.5 (range, 2.5-18.0) vs. 2.5 (range, 1.5-8.0) cm, P = 0.014]. CONCLUSION: Benign non GIST tumors may be distinguished from duodenal GIST as they are smaller in size. LR is a viable treatment option for suspected GIST involving the duodenum. PMID- 18163463 TI - Modification to the Lampariello approach to evaluate reactive oxygen species production by flow cytometry. AB - The aim of this article is to perform a statistical analysis of reactive oxygen species (ROS) cytometric data. It is demonstrated that the classical parametric and nonparametric statistical tests are not suitable to examine these data; the Kolmogorov-Smirnov test and the modification proposed by Lampariello are shown to be too sensitive with respect to the experimental bias (due to procedure or to the instrument) and variability in the ROS production within the repeated samples. Several approaches are examined and discussed. Modifications of the Lampariello's procedure are proposed to include the variability within samples. The validity of the proposed approach is verified by analyzing repeated measurements of ROS formation in cultured human lymphocytes untreated or treated with ferrous sulfate. The proposed approach is successful in considering the "intersample" variability in the ROS data analysis and keeps a good level of validity. Nevertheless, this procedure is not user-friendly and needs to be handled by an expert operator. PMID- 18163464 TI - Cell cycle studies based upon quantitative image analysis. AB - When cell cycle studies are performed following cell cycle synchronization, it is possible that critical properties of an actively cycling cell will be overlooked. For this reason past studies have not revealed critical aspects of cell cycle control; such as how a cell determines when to exit the cell cycle, or how rapidly it should cycle. To address these challenging questions we have developed a procedure to quantitate fluorescent stains in a monolayer culture, where nuclear fluorescence and cell cycle history can be assessed with accuracy on a cell by cell basis. The cell cycle position of each cell can be determined by analyzing DNA and BrdU levels. The behavior of cells in a given cell cycle position can then be studied by quantitating up to two other stained markers. When the microinjection of siRNA, neutralizing antibodies, and expression plasmids are coupled with quantitative image analysis, these cell cycle studies can be conducted following alterations in the expression levels of selected cellular targets. With these techniques we have discovered critical aspects of cell cycle control; including how cyclin D1 levels vary through the cell cycle, the molecular mechanisms governing these changes, and the biological implications of changes in cyclin D1 concentration in various cell cycle stages. Our studies with cyclin D1, coupled with similar studies of p27Kip1, form the basis of an entirely new model of cell cycle control proposed here. This model explains how cell cycle progression is terminated, and how the length of the cell cycle is regulated. PMID- 18163465 TI - Histamine-induced actin polymerization in human eosinophils: an imaging approach for histamine H4 receptor. AB - Image-based screening, a new and flexible tool in the drug discovery cascade, is amenable to many different targets. This article describes a particular use of the Cellomics ArrayScan in developing a functional screen for histamine H(4) receptor (H(4)R) antagonists that have potential utility in inflammatory diseases of the airways such as asthma, with H(4)R being expressed on a wide variety of immune cells including eosinophils. Exposure to histamine causes eosinophils to migrate from the bloodstream into the tissue where they contribute to inflammation. Migration is manifested through rearrangements of the actin cytoskeleton and phalloidin, a biological peptide, selectively binds F-actin over G-actin and can be used to detect these cytoskeletal changes mediating inflammatory function. A fluorescent conjugate of phalloidin was used to visualize histamine-induced actin polymerization in human eosinophils on the Cellomics ArrayScan. Inhibition of this phenomenon by commercially available histamine receptor antagonists was measured. The selective H(4)R antagonist JNJ7777120 inhibited histamine-induced actin polymerization in eosinophils most potently. This assay illustrates that this phenomenon is mediated through the H(4)R and that the image-based format has enhanced screening utility for identifying selective H(4)R antagonists over traditional flow cytometry methods. PMID- 18163466 TI - Automated classification of bacterial particles in flow by multiangle scatter measurement and support vector machine classifier. AB - Biological microparticles, including bacteria, scatter light in all directions when illuminated. The complex scatter pattern is dependent on particle size, shape, refraction index, density, and morphology. Commercial flow cytometers allow measurement of scattered light intensity at forward and perpendicular (side) angles (2 degrees or= 1 units at the last available followup, using a modified Whole-Organ MRI Score. We used logistic regression adjusted for alignment, body mass index, Kellgren/Lawrence score, sex, and age. RESULTS: One hundred thirty-nine knees (53.8%) had central BMLs, of which 129 had type 1 BMLs (96 abutted the ACL and had no coexistent type 2 features) and 25 had type 2 BMLs (often overlapped with type 1). Type 1 lesions were associated with ACL tears (odds ratio [OR] 5.9, 95% confidence interval [95% CI] 2.2-16.2) but not with cartilage loss (OR 1.6, 95% CI 0.8-3.1), while medial type 2 BMLs were related to medial cartilage loss (OR 6.1, 95% CI 1.0-35.2). CONCLUSION: Central BMLs that abutted the ACL were highly prevalent and strongly related to ACL pathology, suggesting a role of enthesopathy in OA. Only BMLs with medial extension were related to ipsilateral cartilage loss. PMID- 18163485 TI - Atacicept in patients with rheumatoid arthritis: results of a multicenter, phase Ib, double-blind, placebo-controlled, dose-escalating, single- and repeated-dose study. AB - OBJECTIVE: Atacicept is a recombinant fusion protein that binds and neutralizes B lymphocyte stimulator and a proliferation-inducing ligand. The purpose of this study was to investigate the tolerability, pharmacokinetics, and pharmacodynamics of atacicept treatment in patients with rheumatoid arthritis (RA) and to collect exploratory data on clinical outcomes. METHODS: In this multicenter, phase Ib, randomized, placebo-controlled, dose-escalating trial, 73 patients were enrolled into 6 escalating-dose cohorts. Patients received atacicept or placebo as single doses (70, 210, or 630 mg) or as repeated doses given at 2-week intervals (3 doses of 70 mg, 3 doses of 210 mg, or 7 doses of 420 mg), followed by 10 weeks of trial assessments, with a followup assessment at 3 months after the final dose. RESULTS: Atacicept was well tolerated, with few differences between treatment groups and no obvious safety concerns. The pharmacokinetics profile was nonlinear, but was consistent and predictable across all doses and regimens. Treatment-related decreases in immunoglobulin (particularly IgM) and rheumatoid factor levels were evident, and a clear decrease in anti-citrullinated protein antibodies was observed in the cohort that received 7 doses of 420 mg. The B cell response was biphasic, with an initial transient increase (dominated by memory B cells) followed by a dose-related decrease (dominated by mature B cells). Clinical assessments showed trends toward improvement with the 3-month treatment. Little effect on the erythrocyte sedimentation rate or C-reactive protein levels was seen. CONCLUSION: Atacicept was well tolerated both systemically and locally. The results demonstrated that the biologic activity of atacicept was consistent with its mechanism of action. PMID- 18163486 TI - Life cell quantification of mitochondrial membrane potential at the single organelle level. AB - Mitochondrial membrane potential (Deltapsi) is key to mitochondrial function and cellular survival. Here, we aimed to develop an automated protocol allowing sensitive quantification of Deltapsi in living cells at the level of individual mitochondria. Human skin fibroblasts were stained with the fluorescent cation tetramethyl rhodamine methyl ester (TMRM), which is sequestered by mitochondria according to their Deltapsi. Cells were visualized by videomicroscopy and the acquired images were processed to generate a mitochondria-specific mask. The latter was superimposed on the original image to allow quantification of TMRM fluorescence. Following validation, our approach revealed that mitochondria with different Deltapsi coexisted within the same cell. Furthermore, our method allowed reproducible detection of small (<10%) reductions in TMRM intensity induced by the complex III inhibitor antimycin A. Mitochondrial uncoupling by p trifluoromethoxy carbonyl cyanide phenyl hydrazone (FCCP) greatly reduced mitochondrial TMRM fluorescence. Under these conditions faithful mask calculation and TMRM intensity analysis were still possible using a mitochondria-targeted green fluorescence protein (mitoAcGFP1), expressed in the cells using baculoviral transfection. PMID- 18163487 TI - Deformation-based nuclear morphometry: capturing nuclear shape variation in HeLa cells. AB - The empirical characterization of nuclear shape distributions is an important unsolved problem with many applications in biology and medicine. Numerous genetic diseases and cancers have alterations in nuclear morphology, and methods for characterization of morphology could aid in both diagnoses and fundamental understanding of these disorders. Automated approaches have been used to measure features related to the size and shape of the cell nucleus, and statistical analysis of these features has often been performed assuming an underlying Euclidean (linear) vector space. We discuss the difficulties associated with the analysis of nuclear shape in light of the fact that shape spaces are nonlinear, and demonstrate methods for characterizing nuclear shapes and shape distributions based on spatial transformations that map one nucleus to another. By combining large deformation metric mapping with multidimensional scaling we offer a flexible approach for elucidating the intrinsic nonlinear degrees of freedom of a distribution of nuclear shapes. More specifically, we demonstrate approaches for nuclear shape interpolation and computation of mean nuclear shape. We also provide a method for estimating the number of free parameters that contribute to shape as well as an approach for visualizing most representative shape variations within a distribution of nuclei. The proposed methodology can be completely automated, is independent of the dimensionality of the images, and can handle complex shapes. Results obtained by analyzing two sets of images of HeLa cells are shown. In addition to identifying the modes of variation in normal HeLa nuclei, the effects of lamin A/C on nuclear morphology are quantitatively described. PMID- 18163488 TI - Abnormal tumor necrosis factor receptor I cell surface expression and NF-kappaB activation in tumor necrosis factor receptor-associated periodic syndrome. AB - OBJECTIVE: Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is an autosomal-dominant autoinflammatory condition caused by mutations in the TNFRSF1A gene. The cellular mechanisms by which mutations in this gene trigger inflammation are currently unclear. Because NF-kappaB is the major intracellular signaling component inducing secretion of proinflammatory cytokines, we sought to determine whether differences in the clinical phenotype of patients with TRAPS may be attributable to variable effects of TNFRSF1A mutations on TNFRI expression, localization, or NF-kappaB activity. METHODS: Peripheral blood mononuclear cells were obtained from patients (following informed consent), and cellular nuclear and cytosolic fractions were generated by subcellular fractionation. Localization of IkappaBalpha and NF-kappaB was determined by Western blotting of the resultant fractions. NF-kappaB subunit activity was determined by enzyme-linked immunosorbent assay analysis and confirmed by electrophoretic mobility shift assay. Subcellular localization of TNFRI was determined by immunofluorescence confocal microscopy or by immunoblotting following affinity isolation of plasma membrane by subcellular fractionation. RESULTS: Cells from patients with the fully penetrant C73R mutation had marked activation of the proinflammatory p65 subunit of NF-kappaB. In contrast, cells from patients with the low-penetrant R92Q mutation displayed high levels of DNA binding by the p50 subunit, an interaction previously linked to repression of inflammation. Interestingly, although cells from patients with the C73R mutation have no TNFRI shedding defect, there was nonetheless an unusually high concentration of functional TNFRI at the plasma membrane. CONCLUSION: High levels of TNFRI at the cell surface in patients with the C73R mutation hypersensitizes cells to stimulation by TNF, leading to increased NF-kappaB p65 subunit activation and an exaggerated proinflammatory response. PMID- 18163490 TI - Antiinflammatory therapy for rheumatoid arthritis? PMID- 18163491 TI - Differences in synovial tissue infiltrates between anti-cyclic citrullinated peptide-positive rheumatoid arthritis and anti-cyclic citrullinated peptide negative rheumatoid arthritis. AB - OBJECTIVE: To compare synovial tissue infiltrates from patients with anti-cyclic citrullinated peptide (anti-CCP)-positive rheumatoid arthritis (RA) with those from patients with anti-CCP-negative RA. METHODS: Synovial tissue samples were obtained arthroscopically from the inflamed knee joints of 57 patients with RA (34 of whom were anti-CCP positive) and examined for several histologic features along with immunohistologic expression of cell markers. Joint damage was assessed using the Kellgren/Lawrence (K/L) scale (range 0-4) on standard anteroposterior radiographs. In 31 patients (18 of whom were anti-CCP positive), synovial tissue was available from an earlier time point, allowing analysis of temporal changes. RESULTS: Synovial tissue from anti-CCP-positive patients was characterized by a higher mean number of infiltrating lymphocytes (61.6 versus 31.4/high-power field [hpf] [400x]; P=0.01), less extensive fibrosis (mean score of 1.2 versus 2.0; P=0.04), and a thinner synovial lining layer (mean score of 2.1 versus 3.3; P=0.002) compared with synovial tissue from anti-CCP-negative patients. Anti-CCP positive patients expressed more CD3, CD8, CD45RO, and CXCL12. More anti-CCP positive patients had a K/L score >1 compared with anti-CCP-negative patients. The difference in the mean lymphocyte counts was already present a mean of 3.8 years before the index biopsy (76.7 lymphocytes/hpf and 26.7 lymphocytes/hpf in anti-CCP-positive patients and anti-CCP-negative patients, respectively; P=0.008) and was independent of disease duration and K/L score. CONCLUSION: Synovitis in patients with anti-CCP-positive RA differs from that in patients with anti-CCP- negative RA, notably with respect to infiltrating lymphocytes, and is associated with a higher rate of local joint destruction. PMID- 18163492 TI - Regional differences in chondrocyte metabolism in osteoarthritis: a detailed analysis by laser capture microdissection. AB - OBJECTIVE: To determine the change in metabolic activity of chondrocytes in osteoarthritic (OA) cartilage, considering regional difference and degree of cartilage degeneration. METHODS: OA cartilage was obtained from knee joints with end-stage OA, at both macroscopically intact areas and areas with various degrees of cartilage degeneration. Control cartilage was obtained from age-matched donors. Using laser capture microdissection, cartilage samples were separated into superficial, middle, and deep zones, and gene expression was compared quantitatively in the respective zones between OA and control cartilage. RESULTS: In OA cartilage, gene expression changed markedly with the site. The expression of cartilage matrix genes was highly enhanced in macroscopically intact areas, but the enhancement was less obvious in the degenerated areas, especially in the upper regions. In contrast, in those regions, the expression of type III collagen and fibronectin was most enhanced, suggesting that chondrocytes underwent a phenotypic change there. Within OA cartilage, the expression of cartilage matrix genes was significantly correlated with SOX9 expression, but not with SOX5 or SOX6 expression. In OA cartilage, the strongest correlation was observed between the expression of type III collagen and fibronectin, suggesting the presence of a certain link(s) between their expression. CONCLUSION: The results of this study revealed a comprehensive view of the metabolic change of the chondrocytes in OA cartilage. The change of gene expression profile was most obvious in the upper region of the degenerated cartilage. The altered gene expression at that region may be responsible for the loss of cartilage matrix associated with OA. PMID- 18163493 TI - Arthritogenic antibodies specific for a major type II collagen triple-helical epitope bind and destabilize cartilage independent of inflammation. AB - OBJECTIVE: To investigate the significance and pathogenic potential of a highly conserved major type II collagen triple-helical epitope-specific antibody (U1; amino acids 494-504) in vivo and in vitro in patients with early rheumatoid arthritis (RA) and in experimental animal models of collagen-induced arthritis (CIA). METHODS: U1-specific antibodies in sera from patients with early RA (with or without joint erosions) were analyzed. Disease progression in the CIA models in mice and rats with anti-U1 antibodies was compared. The pathogenicity of binding of monoclonal antibodies (mAb) UL1 and CIIF4 to the U1 epitope and the F4 epitope (aa 926-936), respectively, was compared in vivo and on chondrocyte cultures and preformed cartilage in vitro, using Fourier transform infrared microspectroscopy analysis. In addition, UL1-induced proteoglycan depletion in vivo in the presence and absence of the complement factor C5 was analyzed. RESULTS: Increased levels of U1 antibodies were observed in patients with early RA, especially in association with joint erosions. A significant correlation of U1-specific antibodies with disease progression was found in rats and mice with CIA. UL1 mAb induced, whereas CIIF4 mAb inhibited, the progression of arthritis. Similarly, UL1, but not CIIF4, impaired matrix synthesis on chondrocyte cultures and adversely affected preformed cartilage. Furthermore, UL1 induced significant proteoglycan depletion in vivo 3 days after injection, even in the absence of C5. CONCLUSION: Antibody epitope specificity contributes significantly to the development of arthritis, and the early pathogenic events operate independent of inflammation both in vitro and in vivo. PMID- 18163494 TI - Fibrodysplasia ossificans progressiva seen on three-dimensional computed tomography. PMID- 18163495 TI - Association of serum nitrate and nitrite levels with longitudinal assessments of disease activity and damage in systemic lupus erythematosus and lupus nephritis. AB - OBJECTIVE: Reactive intermediate production is an essential component of the innate immune response that is induced during disease activity in murine lupus. This study was undertaken to determine whether a marker of systemic nitric oxide (NO) production correlates with prospectively studied disease activity in human systemic lupus erythematosus (SLE) and lupus nephritis patients. METHODS: Eighty three SLE patients and 40 control subjects were studied longitudinally. The SLE group included 23 patients with lupus nephritis documented by renal biopsy and 26 with a history of lupus nephritis. During each visit, following a 24-hour low nitrate diet, traditional markers of disease activity and damage were determined. Serum nitrate plus nitrite (NOx) levels were determined by chemiluminescence detection. RESULTS: NOx levels were higher in SLE patients than in controls during the first visit. In univariate longitudinal analyses, NOx levels were associated with SLE Disease Activity Index scores. In multivariate analyses, NOx levels were associated with serum levels of C3 and creatinine and the urinary protein:creatinine ratio. Among patients with lupus nephritis, those with proliferative lesions had higher NOx levels, and higher NOx levels were associated with accumulation of renal damage and lack of response to therapy. CONCLUSION: This is the first study to prospectively demonstrate longitudinal associations between serum NOx levels and markers of SLE and lupus nephritis disease activity. The more pronounced association with proliferative lupus nephritis and with longitudinal response to lupus nephritis therapy provides a rationale for the study of reactive intermediates as biomarkers of disease activity and therapeutic targets in proliferative lupus nephritis. PMID- 18163497 TI - Estimates of the prevalence of arthritis and other rheumatic conditions in the United States. Part II. AB - OBJECTIVE: To provide a single source for the best available estimates of the US prevalence of and number of individuals affected by osteoarthritis, polymyalgia rheumatica and giant cell arteritis, gout, fibromyalgia, and carpal tunnel syndrome, as well as the symptoms of neck and back pain. A companion article (part I) addresses additional conditions. METHODS: The National Arthritis Data Workgroup reviewed published analyses from available national surveys, such as the National Health and Nutrition Examination Survey and the National Health Interview Survey. Because data based on national population samples are unavailable for most specific rheumatic conditions, we derived estimates from published studies of smaller, defined populations. For specific conditions, the best available prevalence estimates were applied to the corresponding 2005 US population estimates from the Census Bureau, to estimate the number affected with each condition. RESULTS: We estimated that among US adults, nearly 27 million have clinical osteoarthritis (up from the estimate of 21 million for 1995), 711,000 have polymyalgia rheumatica, 228,000 have giant cell arteritis, up to 3.0 million have had self-reported gout in the past year (up from the estimate of 2.1 million for 1995), 5.0 million have fibromyalgia, 4-10 million have carpal tunnel syndrome, 59 million have had low back pain in the past 3 months, and 30.1 million have had neck pain in the past 3 months. CONCLUSION: Estimates for many specific rheumatic conditions rely on a few, small studies of uncertain generalizability to the US population. This report provides the best available prevalence estimates for the US, but for most specific conditions more studies generalizable to the US or addressing understudied populations are needed. PMID- 18163498 TI - Early-onset degeneration of the intervertebral disc and vertebral end plate in mice deficient in type IX collagen. AB - OBJECTIVE: Type IX collagen is an important component of the intervertebral disc extracellular matrix. Mutations in type IX collagen are associated with premature disc degeneration in mice and a predisposition to disc disorders in humans. The aim of this study was to assess the prevalence and timeline of intervertebral disc degeneration in mice homozygous for an inactivated Col9a1 gene. METHODS: Intact spine segments were harvested from wild-type (WT) and type IX collagen knockout (Col9a1(-/-)) mice at 3, 6, and 12 months of age. Sagittal spine sections were evaluated for evidence of histologic changes, by 2 blinded graders, using a semiquantitative grading method. RESULTS: There was evidence of more degeneration of the disc and end plate in the spines of Col9a1(-/-) mice compared with those of WT controls, at most time points. These findings were significant for the disc region at 3 and 6 months (P<0.01) and at 12 months (P<0.10) and for the end plate region only at 6 months (P<0.10). Degenerative changes in the disc consisted of cellular changes and mucous degeneration. Degeneration in the end plates was associated with more cell proliferation, cartilage disorganization, and new bone formation. CONCLUSION: A deletion mutation for type IX collagen is associated with connective tissue changes characteristic of musculoskeletal degeneration in bony and cartilaginous tissue regions. Some of the observed changes were similar to cartilage changes in osteoarthritis, while others were more similar to disc degenerative changes in humans. The finding of premature onset of intervertebral disc degeneration in this mouse model may be useful in studies of the pathology and treatment of human disc degeneration. PMID- 18163499 TI - Association of microsomal prostaglandin E synthase 1 deficiency with impaired fracture healing, but not with bone loss or osteoarthritis, in mouse models of skeletal disorders. AB - OBJECTIVE: Prostaglandin E synthase (PGES) functions as the terminal enzyme in the biosynthesis of prostaglandin E(2) (PGE(2)) and is a potent regulator of bone and cartilage metabolism. Among the 3 isozymes of PGES, microsomal PGES-1 (mPGES 1) is known to play the most critical role in the production of PGE(2) in pathophysiologic events. This study investigated the roles of mPGES-1 under normal physiologic and pathophysiologic conditions in the skeletons of mPGES-1 deficient (mPGES-1(-/-)) mice. METHODS: Skeletons of mPGES-1(-/-) mice and their wild-type littermates were compared by radiologic and histologic analyses. Four models of skeletal disorders were created: bone loss induced by ovariectomy, bone loss induced by hind limb unloading, osteoarthritis (OA) induced by instability in the knee joint, and bone fracture by osteotomy at the tibial midshaft. Expression of the PGES enzymes was examined by immunohistochemistry and real-time reverse transcription-polymerase chain reaction. The cellular mechanism of fracture healing was examined in ex vivo cultures of costal cartilage chondrocytes. RESULTS: Microsomal PGES-1(-/-) mice had unaffected skeletal phenotypes under normal physiologic conditions. In the bone fracture model, fracture healing was impaired by the mPGES-1 deficiency, with half of the mice remaining in a non-bone union state even after 21 days; normal fracture healing was restored by adenoviral reintroduction of mPGES-1. The other skeletal disorders were not affected by the mPGES-1 deficiency. In vivo and ex vivo analyses revealed an impaired proliferation of chondrocytes in cartilage with the mPGES-1 deficiency, at an early stage of fracture healing. CONCLUSION: In these mouse models of skeletal disorders, mPGES-1 was indispensable for bone repair through chondrocyte proliferation, but was not essential for the skeleton under normal physiologic conditions, nor did it play a role in the pathophysiologic conditions of bone loss due to ovariectomy, bone loss due to unloading, or stress induced OA. PMID- 18163500 TI - Is it safe to readminister tumor necrosis factor alpha antagonists following tuberculosis flare? PMID- 18163502 TI - Sphingomyelinase decreases type II collagen expression in bovine articular cartilage chondrocytes via the ERK signaling pathway. AB - OBJECTIVE: Ceramide, a mediator of proinflammatory cytokine signaling, induces cartilage degradation and reduces type II collagen synthesis in articular cartilage. The accumulation of ceramide is associated with arthritis in Farber's disease. The aim of this study was to investigate the mechanism of ceramide induced down-regulation of type II collagen. METHODS: Bovine articular chondrocytes were stimulated with sphingomyelinase (SMase) to increase levels of endogenous ceramide. Components of the ERK pathway were inhibited by Raf-1 kinase inhibitor and the MEK inhibitor, PD98059. Cell extracts were analyzed by Western blotting for ERK-1/2, SOX9, c-Fos, and type II collagen, and the level of c-fos messenger RNA (mRNA) was analyzed by quantitative polymerase chain reaction. Localization of ERK-1/2, SOX9, and c-Fos was assessed by immunocytochemistry and confocal microscopy. RESULTS: SMase treatment of chondrocytes caused sustained phosphorylation of ERK-1/2 throughout the cytoplasm and nucleus that was reduced by inhibitors of Raf-1 kinase and MEK-1/2. SMase treatment of chondrocytes also induced translocation of c-Fos to the nucleus and phospho-SOX9 to the cytoplasm and increased expression of c-fos mRNA. Type II collagen expression, which was down-regulated by SMase treatment, was restored by the MEK-1/2 inhibitor, PD98059. CONCLUSION: SMase down-regulates type II collagen in articular chondrocytes via activation of the ERK signaling cascade, redistribution of SOX9, and recruitment of c-Fos. This new mechanism for cartilage degradation provides potential targets for future treatment of arthritic disease. PMID- 18163503 TI - Interleukin-1beta up-regulation of Smad7 via NF-kappaB activation in human chondrocytes. AB - OBJECTIVE: We have previously shown that interleukin-1beta (IL-1beta) impairs transforming growth factor beta (TGFbeta) signaling through TGFbeta receptor type II (TGFbetaRII) down-regulation and Smad7 up-regulation. This mechanism could account for the reduced responsiveness of osteoarthritic chondrocytes to TGFbeta and the cartilage breakdown linked to this disease. The aim of this study was to investigate the molecular mechanism underlying the IL-1beta-induced stimulation of Smad7 in human articular chondrocytes. METHODS: Human articular chondrocytes were treated with IL-1beta in the presence of TGFbeta1, pyrrolidine dithiocarbamate (a repressor of the NF-kappaB pathway), or cycloheximide. Then, steady-state messenger RNA and protein levels were estimated by real-time reverse transcription-polymerase chain reaction and immunocytology. In addition, transient transfections of p65 expression vector or p65-targeted short hairpin RNA were performed to define the effect of NF-kappaB on Smad7 expression. RESULTS: TGFbetaRII overexpression restored the TGFbeta response of human articular chondrocytes. However, this effect was transient, implying that a secondary mechanism was responsible for the alteration of the TGFbeta response with long-term exposure to IL-1beta. Moreover, IL-1beta caused a late induction of the inhibitory Smad7. This effect was direct, since it did not require de novo synthesis. In addition, we established, by experiments with gain/loss of function, that the up-regulation of Smad7 by IL-1beta is mediated through the NF kappaB pathway, especially the p65 subunit. CONCLUSION: These findings clarify the regulatory process of IL-1beta on Smad7 expression. Understanding the molecular basis of IL-1beta induction of Smad7 and the reduction of chondrocyte responsiveness to TGFbeta provides new insights into the molecular mechanisms of osteoarthritis and may facilitate the identification of novel approaches for its treatment. PMID- 18163504 TI - A biomarker-based mathematical model to predict bone-forming potency of human synovial and periosteal mesenchymal stem cells. AB - OBJECTIVE: To develop a biomarker-based model to predict osteogenic potency of human mesenchymal stem cells (MSCs) from synovial membrane and periosteum. METHODS: MSC populations were derived from adult synovium and periosteum. Phenotype analysis was performed by fluorescence-activated cell sorting and real time reverse transcriptase-polymerase chain reaction (RT-PCR). Telomere lengths were determined by Southern blot analysis. In vitro osteogenesis was assessed quantitatively by measurements of alkaline phosphatase activity and calcium deposits. To investigate bone formation in vivo, MSCs were seeded onto osteoinductive scaffolds and implanted subcutaneously in nude mice. Bone was assessed by histology, and the human origin investigated by in situ hybridization for human Alu genomic repeats. Quantitation was achieved by histomorphometry and real-time RT-PCR for human osteocalcin. Analysis at the single-cell level was performed with clonal populations obtained by limiting dilution. Multiple regressions were used to explore the incremental predictive value of the markers. RESULTS: Periosteal MSCs had significantly greater osteogenic potency than did synovial MSCs inherent to the single cell. Bone was largely of human origin in vivo. Within the same tissue type, there was variability between different donors. To identify predictors of osteogenic potency, we measured the expression levels of osteoblast lineage genes in synovial and periosteal clonal MSCs prior to osteogenic treatment. We identified biomarkers that correlated with osteogenic outcome and developed a mathematical model based on type I collagen and osteoprotegerin expression that predicts the bone-forming potency of MSC preparations, independent of donor-related variables and tissue source. CONCLUSION: Our findings indicate that our quality-control mathematical model estimates the bone-forming potency of MSC preparations for bone repair. PMID- 18163505 TI - High N-terminal pro-brain natriuretic peptide levels and low diffusing capacity for carbon monoxide as independent predictors of the occurrence of precapillary pulmonary arterial hypertension in patients with systemic sclerosis. AB - OBJECTIVE: To evaluate predictors of pulmonary arterial hypertension (PAH) in a prospective cohort of patients with systemic sclerosis (SSc). METHODS: Routine clinical assessments as well as measurements of the diffusing capacity for carbon monoxide/alveolar volume (DLCO/VA) ratio and N-terminal pro-brain natriuretic peptide (NT-proBNP) level were performed in a prospective cohort of 101 SSc patients who did not have PAH or severe comorbidities. After a planned 36-month followup, we evaluated the predictive value of these parameters for the development of precapillary PAH, as demonstrated by cardiac catheterization, disease progression, and death. Criteria for cardiac catheterization were a systolic pulmonary artery pressure (PAP) of >40 mm Hg on echocardiography, a DLCO value of <50% without pulmonary fibrosis, and unexplained dyspnea. RESULTS: Eight patients developed PAH, 29 had disease progression, and 10 died during a median followup of 29 months. Kaplan-Meier analysis identified the following baseline parameters as being predictors of PAH: DLCO/VA ratio <70% or <60% (P<0.01 for each comparison), elevated plasma NT-proBNP level (>97th percentile of normal; P = 0.005), echocardiographically estimated systolic PAP >40 mm Hg (P=0.08), and erythrocyte sedimentation rate >28 mm/hour (P=0.015). In multivariate analyses, an elevated baseline NT-proBNP level (hazard ratio [HR] 9.97 [95% confidence interval (95% CI) 1.69-62.42]) and a DLCO/VA ratio <60% (HR 36.66 [95% CI 3.45 387.6]) were predictors of the occurrence of PAH during followup. An increased NT proBNP level together with a decreased DLCO/VA ratio of <70% was highly predictive of the occurrence of PAH during followup (HR 47.20 [95% CI 4.90 450.33]). CONCLUSION: This prospective study identified a decreased DLCO/VA ratio and an increased NT-proBNP as predictors of PAH in SSc. Use of these markers should result in improved PAH risk stratification and allow earlier initiation of therapy. PMID- 18163506 TI - Intravenous immunoglobulins for relapses of systemic vasculitides associated with antineutrophil cytoplasmic autoantibodies: results of a multicenter, prospective, open-label study of twenty-two patients. AB - OBJECTIVE: To evaluate at 9 months and 24 months the safety and efficacy of intravenous immunoglobulins (IVIGs) administered for 6 months to treat relapses of Wegener's granulomatosis (WG) or microscopic polyangiitis (MPA) occurring either under treatment or during the year following discontinuation of corticosteroids and/or immunosuppressants. METHODS: Patients received IVIGs (0.5 gm/kg/day for 4 days) as additional therapy administered monthly for 6 months and were assessed every 3-6 months. Corticosteroids could be maintained or reintroduced at the time of relapse; immunosuppressants could be continued but could not be reintroduced. At months 9 (end point) and 24 (followup), the following information was collected: complete or partial remission, relapse as assessed with the Birmingham Vasculitis Activity Score (BVAS) 2005, and tolerance and safety of IVIG therapy. RESULTS: Twenty-two Caucasian patients (7 men and 15 women) were studied: 19 had WG, and 3 had MPA. Their median age was 53 years (range 19-75 years), and their median duration of systemic vasculitis was 27 months (range 7-109 months). Their median BVAS 2005 score was 11 (range 3-25). At study entry, 21 patients were ANCA positive, and 21 patients were taking steroids and/or immunosuppressants. All patients experiencing relapse were treated with the same drug(s) plus IVIGs. All patients initially responded to IVIG therapy. By month 9, 13 patients had complete remission, 1 had partial remission, 7 had relapse, and 1 had treatment failure. In 8 of the 14 patients who had remission, the response persisted at month 24. Seven patients experienced minor side effects. CONCLUSION: IVIGs induced complete remissions of relapsed ANCA associated vasculitides in 13 of 22 patients at month 9. Because of the good safety and tolerance profiles of IVIGs, these agents can be included in a therapeutic strategy with other drugs used to treat relapses of WG or MPA. PMID- 18163507 TI - Uridine supplementation antagonizes zidovudine-induced mitochondrial myopathy and hyperlactatemia in mice. AB - OBJECTIVE: Zidovudine is an antiretroviral nucleoside analog reverse transcriptase inhibitor that induces mitochondrial myopathy by interfering with the replication of mitochondrial DNA (mtDNA). Because zidovudine inhibits thymidine kinases, the mechanism of mtDNA depletion may be related to an impairment of the de novo synthesis of pyrimidine nucleotides, which are required building blocks of mtDNA. This study was undertaken to determine whether mitochondrial myopathy is a class effect of antiretroviral nucleoside analogs, and whether the muscle disease can be prevented by treatment with uridine as a pyrimidine nucleotide precursor. METHODS: BALB/c mice were treated with zidovudine or zalcitabine. Some of the mice were cotreated with mitocnol, a dietary supplement with high uridine bioavailability. Mice hind limb muscles were examined after 10 weeks. RESULTS: Zidovudine induced muscle fiber thinning, myocellular fat deposition, and abnormalities of mitochondrial ultrastructure. In mice treated with zidovudine, organelles contained low mtDNA copy numbers and reduced cytochrome c oxidase activity. The expression of the mtDNA-encoded cytochrome c oxidase I subunit, but not of nucleus-encoded mitochondrial proteins, was impaired. Zidovudine also increased the levels of myocellular reactive oxygen species and blood lactate. Uridine supplementation attenuated or normalized all pathologic abnormalities and had no intrinsic effects. Zalcitabine did not elicit muscle toxicity. CONCLUSION: Our findings indicate that zidovudine, but not zalcitabine, induces mitochondrial myopathy, which is substantially antagonized by uridine supplementation. These results provide proof of the importance of pyrimidine pools in the pathogenesis of zidovudine myopathy. Since uridine supplementation is tolerated well by humans, this treatment strategy should be investigated in clinical trials. PMID- 18163508 TI - Differing results of trials of glucosamine for pain in arthritis: comment on the article by VLAD et al. PMID- 18163509 TI - Maintenance of infliximab treatment in ankylosing spondylitis: results of a one year randomized controlled trial comparing systematic versus on-demand treatment. AB - OBJECTIVE: Continuous treatment with the anti-tumor necrosis factor alpha (anti TNFalpha) antibody infliximab is efficacious in ankylosing spondylitis (AS), whereas treatment discontinuation results in disease relapse, with variable delay. This study was undertaken to compare the efficacy of continuous treatment with infliximab with that of a treatment regimen adapted to symptom recurrence. Methotrexate (MTX) in combination with infliximab was also tested. METHODS: Patients with active AS were randomly assigned at week 0 to receive infliximab every 6 weeks (continuous treatment) or upon symptom recurrence (on-demand treatment), following infusions at weeks 4, 6, and 10. Patients in the on-demand group were randomly assigned to receive either MTX in combination with infliximab or infliximab alone. Patients were monitored for 1 year. The primary end point was the proportion of patients who met the ASsessment in AS International Working Group criteria for 20% improvement (ASAS20) at week 58. RESULTS: Of 247 patients, 124 were assigned to receive infliximab every 6 weeks and 123 to receive on demand treatment. Among the latter, 62 received MTX, and 61 received infliximab alone. A greater proportion of patients receiving infliximab every 6 weeks fulfilled ASAS20 response criteria at week 58 than did patients receiving on demand treatment (75% versus 46%; P<0.0001). Patients in the continuous treatment group received more infliximab infusions after week 10 than did those in the on demand group (mean+/-SD 5.8+/-2.2 versus 3.5+/-2; P<0.0001). Addition of MTX did not significantly affect the proportion of patients with an ASAS20 response at week 58, nor the number of infliximab infusions administered. CONCLUSION: These findings indicate that continuous treatment of AS with infliximab is more efficacious than on-demand treatment, and that the addition of MTX to infliximab provides no significant benefit. PMID- 18163510 TI - Differentially regulated expression of growth differentiation factor 5 and bone morphogenetic protein 7 in articular cartilage and synovium in murine chronic arthritis: potential importance for cartilage breakdown and synovial hypertrophy. AB - OBJECTIVE: To examine whether the endogenous expression of growth differentiation factor 5 (GDF-5) and bone morphogenetic protein 7 (BMP-7) is altered in the cartilage and synovium of human tumor necrosis factor alpha (TNFalpha)-transgenic (hTNFtg) mice with chronic arthritis, and to investigate the response of hTNFtg chondrocytes as well as fibroblast-like synoviocytes (FLS) to these morphogens in vitro. METHODS: Analyses were performed in hTNFtg mice with chronic destructive arthritis and in wild-type (WT) mice as controls. Expression of GDF-5 and BMP-7 in the articular cartilage and synovium was examined by real-time polymerase chain reaction and immunohistochemistry. Human TNFtg cartilage explants, chondrocytes, and FLS monolayer cultures were assessed for basal matrix biosynthesis as well as growth factor responsiveness, using (35)S-sulfate incorporation assays. In addition, the DNA content/cell proliferation rate was measured. RESULTS: The expression of GDF-5 and BMP-7 was decreased in articular cartilage from hTNFtg mice, whereas expression of both morphogens was increased in arthritic synovium from hTNFtg mice, as compared with the levels in WT controls. Isotope incorporation revealed a marked reduction of matrix synthesis in hTNFtg cartilage as well as a decrease in responsiveness to GDF-5 and BMP-7. The DNA content did not change in arthritic cartilage as compared with WT cartilage. In hTNFtg FLS, growth factor stimulation increased the rate of cell proliferation and the production of extracellular matrix. CONCLUSION: In this murine model of TNFalpha-mediated arthritis, the expression of GDF-5 and BMP-7 is regulated differentially in articular cartilage and synovium. In articular cartilage, the down-regulation of GDF-5 and BMP-7, which function to maintain matrix integrity, could potentially compromise tissue repair, whereas in synovium, the increased expression of GDF-5 and BMP-7 might contribute to synovial hypertrophy. PMID- 18163511 TI - Cannabinoid-mediated antinociception is enhanced in rat osteoarthritic knees. AB - OBJECTIVE: To determine whether local administration of the cannabinoid 1 (CB(1)) receptor agonist arachidonyl-2-chloroethylamide (ACEA) can modulate joint nociception in control rat knee joints and in experimental osteoarthritis (OA). METHODS: OA was induced in male Wistar rats by intraarticular injection of 3 mg of sodium mono-iodoacetate, with a recovery period of 14 days. Electrophysiologic recordings were made of knee joint primary afferent nerve fibers in response to normal rotation and noxious hyperrotation of the joint both before and after close intraarterial injection of different doses of ACEA. RESULTS: Local application of the CB(1) agonist significantly reduced the firing rate of afferent nerve fibers by up to 50% in control knee joints (n=19) and up to 62% in OA knee joints (n=29; P<0.01). Coadministration of the CB(1) receptor antagonist AM251 or the transient receptor potential vanilloid 1 (TRPV-1) ion channel antagonist SB366791 significantly reduced the desensitizing effect of ACEA. The CB(1) receptor antagonist AM251 by itself had no effect in the control joint but significantly increased the firing rate of afferent nerve fibers in the OA joint. CONCLUSION: These findings indicate that activation of peripheral CB(1) receptors reduces the mechanosensitivity of afferent nerve fibers in control and OA knee joints. Blockade of either the CB(1) receptor or the TRPV-1 channel significantly reduced the efficacy of ACEA, which suggests that both receptors are involved in cannabinoid-mediated antinociception. The increased nerve activity observed following CB(1) receptor antagonism suggests a tonic release of endocannabinoids during OA. As such, peripheral CB(1) receptors may be important targets in controlling OA pain. PMID- 18163512 TI - Differential requirements for IKKalpha and IKKbeta in the differentiation of primary human osteoarthritic chondrocytes. AB - OBJECTIVE: Osteoarthritic (OA) chondrocytes behave in an intrinsically deregulated manner, characterized by chronic loss of healthy cartilage and inappropriate differentiation to a hypertrophic-like state. IKKalpha and IKKbeta are essential kinases that activate NF-kappaB transcription factors, which in turn regulate cell differentiation and inflammation. This study was undertaken to investigate the differential roles of each IKK in chondrocyte differentiation and hypertrophy. METHODS: Expression of IKKalpha or IKKbeta was ablated in primary human chondrocytes by retro-transduction of specific short-hairpin RNAs. Micromass cultures designed to reproduce chondrogenesis with progression to the terminal hypertrophic stage were established, and anabolism and remodeling of the extracellular matrix (ECM) were investigated in the micromasses using biochemical, immunohistochemical, and ultrastructural techniques. Cellular parameters of hypertrophy (i.e., proliferation, viability, and size) were also analyzed. RESULTS: The processes of ECM remodeling and mineralization, both characteristic of terminally differentiated hypertrophic cells, were defective following the loss of IKKalpha or IKKbeta. Silencing of IKKbeta markedly enhanced accumulation of glycosaminoglycan in conjunction with increased SOX9 expression. Ablation of IKKalpha dramatically enhanced type II collagen deposition independent of SOX9 protein levels but in association with suppressed levels of runt-related transcription factor 2. Moreover, IKKalpha-deficient cells retained the phenotype of cells in a pre-hypertrophic-like state, as evidenced by the smaller size and faster proliferation of these cells prior to micromass seeding, along with the enhanced viability of their differentiated micromasses. CONCLUSION: IKKalpha and IKKbeta exert differential roles in ECM remodeling and endochondral ossification, which are events characteristic of hypertrophic chondrocytes and also complicating factors often found in OA. Because the effects of IKKalpha were more profound and pleotrophic in nature, our observations suggest that exacerbated IKKalpha activity may be responsible, at least in part, for the characteristic abnormal phenotypes of OA chondrocytes. PMID- 18163513 TI - Prevalence of malignancy in psoriatic arthritis. AB - OBJECTIVE: To determine the prevalence and types of malignancy in a large cohort of patients with psoriatic arthritis (PsA), and to compare this rate with that in the general population. METHODS: A cohort analysis of patients who were followed up prospectively from 1978 to 2004 at the University of Toronto Psoriatic Arthritis Clinic was performed. Patients were followed up at 6-12-month intervals according to a standard protocol, which included recording of malignancy, and tracked on a computer database. The cohort was linked with a provincial database to find malignancies that may have been missed by the protocol or developed after patients were lost to followup. Data were presented and analyzed using descriptive statistics and the Cox regression model with robust estimate of variance. Rates of first malignancy in the cohort were compared with rates in the population to derive standardized incidence ratios (SIRs). RESULTS: Of the 665 patients included, 68 (10.2%) developed a malignancy at an average age of 62.4 years. The most frequently seen malignancies were breast (20.6%), lung (13.2%), and prostate (8.8%) cancer. The SIR for all cancers was 0.98 (95% confidence interval 0.77-1.24). Overall cancer type-specific SIRs were 0.69 (95% CI 0.26 1.83) for hematologic and 0.88 (95% CI 0.46-1.69) for lung cancer. In females, the SIR for breast cancer was 1.55 (95% CI 0.92-2.62), and in males, the SIR for prostate cancer was 0.65 (95% CI 0.29-1.44). CONCLUSION: Overall, 10.2% of patients in the Toronto PsA cohort developed cancer. The most frequent cancers were breast, lung, and prostate cancer. The incidence of malignancy in the large PsA cohort did not differ from that in the general population. PMID- 18163514 TI - T cell dependence of chronic destructive murine arthritis induced by repeated local activation of Toll-like receptor-driven pathways: crucial role of both interleukin-1beta and interleukin-17. AB - OBJECTIVE: The pathogenesis of rheumatoid arthritis is often linked to bacterial infections. The present study was undertaken to develop a mouse model of chronic destructive arthritis induced by repeated intraarticular (IA) exposure to bacterial cell wall fragments and to investigate the cytokine dependence of this model. METHODS: Mice that were deficient in various cytokines were injected IA with cell wall fragments of Streptococcus pyogenes on days 0, 7, 14, and 21. The development of chronic destructive arthritis was compared between groups of mice lacking different cytokines, to assess which cytokines were crucial for development of chronic destructive arthritis. RESULTS: Repeated exposure of a joint to S pyogenes cell wall fragments resulted in the development of chronic destructive arthritis. In mice deficient in recombination-activating gene 2, streptococcal cell wall (SCW)-directed T cell reactivity was found and chronic arthritis did not develop, implicating T cells in the generation of chronic SCW induced arthritis. Interleukin-17 (IL-17) receptor-deficient mice showed a reduction of joint destruction in the chronic stage, implicating a detrimental role of the recently discovered IL-17-producing T helper cells (Th17 cells). IL 23 expression was apparent during the late stages of arthritis. Joint swelling was no longer dependent on tumor necrosis factor alpha (TNFalpha) after the last flare, and pronounced cartilage damage was found after 28 days in TNFalpha deficient mice. In contrast, IL-1beta-deficient mice were fully protected against joint swelling and cartilage and bone destruction during the late stages of disease. CONCLUSION: These findings indicate that the TNFalpha dependence of arthritis is lost during the erosive stage, when Th17 cells become crucial. IL 1beta dependence remains strong, consistent with its pivotal role in the generation of Th17 cells. PMID- 18163515 TI - Index to ring finger length ratio and the risk of osteoarthritis. AB - OBJECTIVE: To determine the relationship between the index to ring finger (2D:4D) length ratio and the risk of knee and hip osteoarthritis (OA). METHODS: We conducted a case-control study, in which cases with persistent symptoms and radiographic evidence of knee or hip OA were compared with controls with no symptoms and no radiographic evidence of knee or hip OA. Hand radiographs were visually classified as type 1 (index finger longer than the ring finger), type 2 (index finger equal to the ring finger), or type 3 (index finger shorter than the ring finger). The 2D:4D phalangeal and metacarpal length ratios were measured separately. The odds ratio (OR) and 95% confidence interval (95% CI) were calculated and adjusted for possible confounding factors using a logistic regression model. RESULTS: Of 2,049 cases, 1,013 had radiographic evidence of knee OA and 995 had hip OA. Of 1,123 controls, 836 had no knee OA and 1,050 had no hip OA. The type 3 finger pattern was associated with knee OA (OR 1.94, 95% CI 1.54-2.44), and the risk was greater in women (OR 3.05, 95% CI 2.08-4.47) than in men (OR 1.45, 95% CI 1.08-1.95). There was a dose-response relationship between both 2D:4D phalangeal and metacarpal length ratios and the risk of knee OA. The risk of hip OA was inconsistent. CONCLUSION: Compared with types 1 and 2, the type 3 "male" pattern 2D:4D length ratio is associated with OA, especially knee OA. The risk is independent of other major OA risk factors. PMID- 18163516 TI - Rupture of Brodie's abscess. PMID- 18163518 TI - The effects of rituximab on immunocompetency in patients with autoimmune disease. PMID- 18163519 TI - Antibodies against citrullinated vimentin in rheumatoid arthritis: higher sensitivity and extended prognostic value concerning future radiographic progression as compared with antibodies against cyclic citrullinated peptides. AB - OBJECTIVE: The Sa autoantigen can be found in inflamed synovium of patients with rheumatoid arthritis (RA), and at least part of the humoral RA-specific anti-Sa response is directed against citrullinated vimentin. This study was undertaken to evaluate the sensitivity, specificity, and prognostic value of determination of levels of antibodies against modified citrullinated vimentin (anti-MCV) as compared with antibodies against cyclic citrullinated peptides (anti-CCP) in an inception cohort of patients with early RA. METHODS: Clinical data, radiographs, and measurements of levels of anti-MCV and anti-CCP antibodies were obtained in 273 patients with early RA at baseline, after 3 months, and after 1, 2, 3, and 5 years. Autoantibodies were also analyzed in 100 healthy controls. RESULTS: Of the 273 patients, 193 (70.7%) were anti-MCV positive and 158 (57.9%) were anti-CCP positive at the time of diagnosis, with nearly equal specificities (95% and 96%, respectively). Forty (14.7%) were anti-MCV positive only, and 5 (1.8%) were anti CCP positive only. Anti-MCV-positive and anti-MCV-negative patients had similar disease activity at baseline, but presence of anti-MCV was predictive of subsequent high disease activity and continued radiographic progression. Changes in anti-MCV level showed stronger correlation with changes in clinical parameters than did changes in anti-CCP level. The subgroup of patients who were anti-MCV positive and anti-CCP negative showed a higher rate of radiographic destruction than did patients who were negative for both anti-MCV and anti-CCP. CONCLUSION: These findings show that when patients with early RA are compared with healthy controls, analysis of anti-MCV yields greater sensitivity and unchanged specificity as compared with analysis of anti-CCP. Anti-MCV also appears to perform better than anti-CCP in identifying poor radiographic prognosis in patients with early RA. PMID- 18163520 TI - Expression of mucin 3 and mucin 5AC in arthritic synovial tissue. AB - OBJECTIVE: Rheumatoid arthritis (RA) is a chronic inflammatory disease that is characterized by hypertrophy of the synovial tissue, leukocyte infiltration, angiogenesis, and ultimately joint destruction. Mucins (MUCs) are a family of heavily glycosylated proteins that protect epithelial membranes and are used as ligands for cell adhesion. MUC gene expression has been found to be altered in many cancers and inflammatory states. This study was undertaken to examine its expression in synovial tissue (ST) and role in arthritis. METHODS: We performed immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction to determine expression patterns of MUC1, MUC2, MUC3, and MUC5AC in RA, osteoarthritic (OA), and normal human ST. RESULTS: MUC3 was expressed in synovial lining cells, macrophages, and fibroblasts. Significantly more RA (n=12) and OA (n=13) synovial lining cells expressed MUC3 than did normal synovial lining cells (n=7) (22% and 24% versus 0.4%, respectively; P<0.05). Additionally, macrophages in RA and OA ST expressed significantly more MUC3 than did macrophages in normal ST (50% and 51% versus 10%, respectively; P<0.05). MUC5AC was expressed at low levels in synovial lining cells, macrophages, and endothelial cells in RA and OA ST, and was barely expressed in normal ST. MUC1 and MUC2 proteins were not detected in ST. Messenger RNA (mRNA) for MUC3 and MUC5AC was detected in ST, and mRNA for MUC3 was detected in cultured ST fibroblasts. CONCLUSION: These data demonstrate up-regulated MUC expression by ST cells and suggest a novel role of MUC3 and MUC5AC in the pathogenesis of arthritis. PMID- 18163521 TI - Comparison of the clinical efficacy and safety of subcutaneous versus oral administration of methotrexate in patients with active rheumatoid arthritis: results of a six-month, multicenter, randomized, double-blind, controlled, phase IV trial. AB - OBJECTIVE: To compare the efficacy and safety of subcutaneous (SC) versus oral administration of methotrexate (MTX) in patients with active rheumatoid arthritis (RA). METHODS: MTX-naive patients with active RA (Disease Activity Score in 28 joints >or= 4) were eligible for the study if they had not previously taken biologic agents and had not taken disease-modifying antirheumatic drugs for 2 weeks prior to randomization. Patients were randomly assigned to receive 15 mg/week of MTX either orally (2 7.5-mg tablets plus a dummy prefilled syringe; n=187 patients) or SC (prefilled syringe containing 10 mg/ml plus 2 dummy tablets; n=188 patients) for 24 weeks. At week 16, patients who did not meet the American College of Rheumatology criteria for 20% improvement (ACR20) were switched from 15 mg of oral MTX to 15 mg of SC MTX and from 15 mg of SC MTX to 20 mg of SC MTX for the remaining 8 weeks, still in a blinded manner. The primary outcome was an ACR20 response at 24 weeks. RESULTS: At week 24, significantly more patients treated with SC MTX than with oral MTX showed ACR20 (78% versus 70%) and ACR70 (41% versus 33%) responses. Patients with a disease duration >or= 12 months had even higher ACR20 response rates (89% for SC administration and 63% for oral). In 52 of the ACR20 nonresponders (14%), treatment was switched at week 16. Changing from oral to SC MTX and from 15 mg to 20 mg of SC MTX resulted in 30% and 23% ACR20 response rates, respectively, in these patients. MTX was well tolerated. The rate of adverse events was similar in all groups. CONCLUSION: This 6-month prospective, randomized, controlled trial is the first to examine oral versus SC administration of MTX. We found that SC administration was significantly more effective than oral administration of the same MTX dosage. There was no difference in tolerability. PMID- 18163522 TI - Risks and relative risks of Wegener's granulomatosis among close relatives of patients with the disease. AB - OBJECTIVE: The etiology of Wegener's granulomatosis (WG) supposedly involves interplay between genetic susceptibility and environmental triggers. However, little is known about whether WG actually clusters in families. Information on the degree of familial aggregation in WG is of clinical relevance, because patients with WG often want to know whether their diagnosis puts their closest relatives at increased risk of the disease. The aim of this study was to investigate the risk of WG in relatives of patients with WG. METHODS: Using Swedish nationwide registers on morbidity, family structure, and vital status, we compared the occurrence of WG (register-based plus chart review) among 6,670 first-degree relatives and 428 spouses of 1,944 Swedish patients with WG with the occurrence among 68,994 first-degree relatives and 4,812 spouses of 19,655 control subjects from the general population. Relative risks were estimated using the Cox proportional hazards regression model. RESULTS: Two of the 6,670 first degree relatives of patients with WG and 13 of the 68,994 first-degree relatives of their population controls had WG, resulting in a relative risk of 1.56 (95% confidence interval 0.35-6.90). None of the 428 spouses of patients had WG. CONCLUSION: In absolute terms, the occurrence of WG among close biologic and nonbiologic relatives of patients with WG is low. In terms of relative risk, our results provide strong evidence against a pronounced increase in familial risk such as that noted for systemic lupus erythematosus, irritable bowel disease, and multiple sclerosis but are compatible with familial aggregation of a magnitude similar to that for rheumatoid arthritis. PMID- 18163523 TI - Regulation of a novel alphaN-catenin splice variant in schizophrenic smokers. AB - The alphaN-catenin (CTNNA2) gene represents a promising candidate gene for schizophrenia based upon previous genetic linkage, expression, and mouse knockout studies. CTNNA2 is differentially regulated by smoking in schizophrenic patients. In this report, the genomic structure of a primate-specific alphaN-catenin splice variant (alphaN-catenin III) is described. A comparison of alphaN-catenin III mRNA expression across postmortem hippocampi from schizophrenic and non-mentally ill smokers and non-smokers revealed a significant decrease in expression among patient non-smokers compared to all other groups. The recent evolutionary divergence of this gene, as well as the differences in gene expression in postmortem brain of schizophrenic non-smokers, supports the role of alphaN catenin III as a novel disease susceptibility gene. PMID- 18163524 TI - Differential expression of CYP6A5 and CYP6A5v2 in pyrethroid-resistant house flies, Musca domestica. AB - Two cytochrome P450 alleles, CYP6A5 and CYP6A5v2, were isolated from a pyrethroid resistant house fly stain, ALHF. The two alleles shared 98% similarity in amino acid sequence. To understand the importance of these two alleles in resistance and examine the expression profile of the two alleles between resistant and susceptible strains, quantitative real-time PCR (qRT-PCR) was performed and compared with the Northern blot analysis. We found that qRT-PCR was an efficient method to characterize the expression profiles between these two sequence-closely related P450 genes between resistant and susceptible houses flies. One of them, CYP6A5v2, was constitutively overexpressed in ALHF house flies compared with susceptible house fly strains. Moreover, this gene was predominantly expressed in the abdominal tissues of ALHF, in which the primary detoxification organs of insects are located. However, there was no significant difference in the expression of CYP6A5 between ALHF and susceptible house flies. The genetic linkage analysis was conducted to determine the possible link between the constitutively overexpressed CYP6A5v2 and insecticide resistance. CYP6A5v2 was mapped on autosome 5, which is correlated with the linkage of resistance in ALHF. Taken together, the study suggests the importance of CYP6A5v2 in increasing metabolic detoxification of insecticides in ALHF. The distinct expression of CYP6A5 and CYP6A5v2 in resistant and susceptible house flies implies the functional difference of theses two genes in house flies and suggests that they are two recently diverged P450 genes presented in a single organism. PMID- 18163525 TI - cDNA cloning, characterization, and developmental expression of the 20S proteasome alpha5 subunit in the Mediterranean fruit fly Ceratitis capitata. AB - In the present study, we report the cDNA cloning, characterization, and developmental expression of the 20S proteasome alpha5 subunit from the Mediterranean fruit fly Ceratitis capitata (medfly). Using an RT-PCR fragment that corresponds to the amino-terminal region of the Drosophila melanogaster 20S proteasome alpha5 subunit, we isolated a 987-bp cDNA that encodes the complete coding region of the medfly ortholog, which was named CcPSMA5. CcPSMA5 consists of 241 amino acids and has a predicted molecular weight of 26.4 kDa and pI 4.75. Comparison of the CcPSMA5 amino acid sequence with the sequences of all known 20S proteasome alpha5 subunits from different organisms indicated that the medfly 20S proteasome alpha5 subunit has the strongest homology to that of Drosophila. In situ hybridization showed that the CcPSMA5 gene is mapped in the region 44B of chromosome 4. Northern blot hybridization analysis showed that the CcPSMA5 mRNA has a size of approximately 1.2 kb. High levels of the CcPSMA5 mRNA were detected in freshly laid eggs, indicating that they were maternally deposited. The mRNA expression pattern during medfly development suggests that the CcPSMA5 gene is upregulated before mid-embryogenesis and at the onset of metamorphosis. PMID- 18163526 TI - Novel cytochrome P450s, CYP6BB1 and CYP6P10, from the salt marsh mosquito Aedes sollicitans (Walker) (Diptera: Culicidae). AB - Based on the conserved heme-binding region and the charge pair consensus of insect cytochrome P450s, two novel full-length P450 cDNAs, CYP6BB1 and CYP6P10, were cloned from the salt marsh mosquito Aedes sollicitans (Walker). CYP6BB1 and CYP6P10 had open reading frames of 1,518 and 1,521 nucleotides encoding 506 and 507 amino acid residue proteins, respectively. Several alleles with amino acid substitutions were found both in CYP6BB1 and CYP6P10. The deduced proteins are typical microsomal P450s sharing signature sequences with other insect CYP6 P450s. Sequence analysis showed that both CYP6BB1 and CYP6P10 shared highest sequence identities with P450 CYP6P4, 56% and 65%, respectively. Phylogenetic analysis showed both CYP6BB1 and CYP6P10 were grouped into the clade containing several P450s from subfamily CYP6P. Real-time RT-PCR analysis showed CYP6BB1 but not CYP6P10 transcription in females was significantly increased 24 h after a blood meal. Neither CYP6BB1 nor CYP6P10 were life stage or gender specific. Protein expression experiments are needed to determine the functions of these proteins. PMID- 18163527 TI - Comparison of two acetylcholinesterase gene cDNAs of the lesser mealworm, Alphitobius diaperinus, in insecticide susceptible and resistant strains. AB - Two cDNAs encoding different acetylcholinesterase (AChE) genes (AdAce1 and AdAce2) were sequenced and analyzed from the lesser mealworm, Alphitobius diaperinus. Both AdAce1 and AdAce2 were highly similar (95 and 93% amino acid identity, respectively) with the Ace genes of Tribolium castaneum. Both AdAce1 and AdAce2 have the conserved residues characteristic of AChE (catalytic triad, intra-disulfide bonds, and so on). Partial cDNA sequences of the Alphitobius Ace genes were compared between two tetrachlorvinphos resistant (Kennebec and Waycross) and one susceptible strain of beetles. Several single nucleotide polymorphisms (SNPs) were detected, but only one non-synonymous mutation was found (A271S in AdAce2). No SNPs were exclusively found in the resistant strains, the A271S mutation does not correspond to any mutations previously reported to alter sensitivity of AChE to organophosphates or carbamates, and the A271S was found only as a heterozygote in one individual from one of the resistant A. diaperinus strains. This suggests that tetrachlorvinphos resistance in the Kennebec and Waycross strains of A. diaperinus is not due to mutations in either AChE gene. The sequences of AdAce1 and AdAce2 provide new information about the evolution of these important genes in insects. PMID- 18163528 TI - Purification and characterization of two cysteine peptidases of the Mediterranean fruit fly Ceratitis capitata during metamorphosis. AB - In holometabolous insects, there is a complete body remodeling from larva to adult. We determined in Ceratitis capitata that the transition from pre-pupa to pupa, 40 to 48 h after puparium formation (h APF), is a key moment of metamorphosis; when salivary glands, intestine, fat body, and muscles are in different stages of cell death. At 44-46 h APF, muscles from segments 1-3 (thoracic region) appeared fully disintegrated, whereas posterior muscles just started death processes. To understand some of the biochemical events eventually involved in histolytic processes during early metamorphosis, two cysteine peptidases coined "Metamorphosis Associated Cysteine Peptidase" (MACP-I and MACP II) were purified to homogeneity from 40-46-h APF insects. Both enzymes were inhibited by Ep-475, a specific inhibitor of papain-like cysteine-peptidases. MACP-I is a single chain protein with an apparent molecular mass of 80 kDa and includes several isoforms with pI values of pH 6.25-6.35, 6.7, and 7.2. The enzyme has an optimum pH of 5.0 and its pH stability ranges from pH 4.0 to 6.0. The molecular weight and N-terminal sequence suggest that MACP-I might be a novel enzyme. MACP-II is an acidic single chain protein with a pI of pH 5.85 and an apparent molecular mass of 30 kDa. The enzyme is labile with a maximum stability in the pH range of 4.0 to 6.0 and an optimum pH among 5.0 to 6.0. MAPCP-II characteristics suggest it is a cathepsin B-like enzyme. PMID- 18163529 TI - Sample size requirements for indirect association studies of gene-environment interactions (G x E). AB - Association studies accounting for gene-environment interactions (G x E) may be useful for detecting genetic effects. Although current technology enables very dense marker spacing in genetic association studies, the true disease variants may not be genotyped. Thus, causal genes are searched for by indirect association using genetic markers in linkage disequilibrium (LD) with the true disease variants. Sample sizes needed to detect G x E effects in indirect case-control association studies depend on the true genetic main effects, disease allele frequencies, whether marker and disease allele frequencies match, LD between loci, main effects and prevalence of environmental exposures, and the magnitude of interactions. We explored variables influencing sample sizes needed to detect G x E, compared these sample sizes with those required to detect genetic marginal effects, and provide an algorithm for power and sample size estimations. Required sample sizes may be heavily inflated if LD between marker and disease loci decreases. More than 10,000 case-control pairs may be required to detect G x E. However, given weak true genetic main effects, moderate prevalence of environmental exposures, as well as strong interactions, G x E effects may be detected with smaller sample sizes than those needed for the detection of genetic marginal effects. Moreover, in this scenario, rare disease variants may only be detectable when G x E is included in the analyses. Thus, the analysis of G x E appears to be an attractive option for the detection of weak genetic main effects of rare variants that may not be detectable in the analysis of genetic marginal effects only. PMID- 18163530 TI - More respect for anatomy in hernia repair, please! PMID- 18163531 TI - Gene expression analysis on sections of zebrafish regenerating fins reveals limitations in the whole-mount in situ hybridization method. AB - The caudal fin of adult zebrafish is used to study the molecular mechanisms that govern regeneration processes. Most reports of gene expression in regenerating caudal fins rely on in situ hybridization (ISH) on whole-mount samples followed by sectioning of the samples. In such reports, expression is mostly confined to cells other than those located between the dense collagenous structures that are the actinotrichia and lepidotrichia. Here, we re-examined the expression of genes by performing ISH directly on cryo-sections of regenerates. We detected expression of some of these genes in cell types that appeared to be non expressing when ISH was performed on whole-mount samples. These results demonstrate that ISH reagents have a limited capacity to penetrate between the regenerating skeletal matrices and suggest that ISH performed directly on fin sections is a preferable method to study gene expression in fin regenerates. PMID- 18163532 TI - Impaired placental trophoblast lineage differentiation in Alkbh1(-/-) mice. AB - E. coli AlkB has been intensively studied since 1983, but the in vivo roles of its mammalian homologue Alkbh1 are unknown. We, therefore, created null mice for Alkbh1. Alkbh1 mRNA is expressed at highest levels in the trophoblast lineages of the developing placenta. Alkbh1(-/-) placentas have decreased expression of differentiated trophoblast markers including Tpbp, Gcm1, and Pl-1, and increased expression of the trophoblast stem cell marker Eomes. Alkbh1 localizes to nuclear euchromatin, and interacts strongly with Mrj, an essential placental gene that mediates gene repression by recruitment of class II histone deacetylases (HDACs). Competition experiments show Alkbh1 and HDAC4 binding to Mrj are mutually exclusive, which causes decreased HDAC activity and increased target gene expression. Our study demonstrates Alkbh1 performs important functions in placental trophoblast lineage differentiation and participates in mechanisms of transcriptional regulation. PMID- 18163533 TI - In vivo imaging and evaluation of different biomatrices for improvement of stem cell survival. AB - Therapeutic effects from injection of stem cells are often hampered by acute donor cell death as well as migration away from damaged areas. This is likely due to the fact that injected cells do not have the physical and biochemical cues for ordered engrafment. Here we evaluate 3 common biomatrices (Matrigel, Collagen I, Purmatrix) that has the potential of providing suitable scaffolds needed to enhance stem cell survival. The longitudinal fate of transplanted stem cells was monitored by reporter imaging techniques. PMID- 18163534 TI - Novel FRET-based assay to detect reverse transcriptase activity using modified dUTP analogues. AB - We have developed a novel continuous assay to measure reverse transcriptase (RT) polymerase activity. The assay uses fluorescence energy transfer measurements to detect the incorporation of complementary pairs of fluorescently labeled deoxyuridine into cDNA product. The fluorescently labeled dUTP substrates were prepared using commercially available reagents with a simple coupling reaction. The fluorescent dye pairs have significant spectral overlap which allows FRET interaction between dyes incorporated into the cDNA. Using a polyA/oligo dT primer/template, the assay can readily detect DNA polymerase activity from any viral reverse transcriptase enzyme. The reaction proceeds linearly over time, and the rate is proportional to the enzyme concentration. We used the assay to compare the thermostability of a number of wild-type and mutant viral RT enzymes. Our results indicate that the wild-type AMV (avian myeloblastosis virus) enzyme is slightly more stable at 43 degrees C than the HIV-1 (human immunodeficiency virus) or MMLV (Moloney murine leukemia virus) enzymes. The thermostability of the RT enzyme was dramatically increased by the presence of primer/template with the enzyme. We also used the assay to study the effects of inhibitors on HIV-1 RT polymerase activity. This assay may be highly useful for the identification and characterization of potent RT inhibitors which could be candidates for development as therapeutic antiviral agents. PMID- 18163535 TI - Near-infrared fluorescent labeled peptosome for application to cancer imaging. AB - Nonionic amphiphilic copolypeptides, which were composed of hydrophilic poly(sarcosine) and hydrophobic poly(gamma-methyl L-glutamate) blocks, were synthesized with varying chain lengths of the blocks. The polypeptides having a suitable hydrophilic and hydrophobic balance were found to form vesicular assemblies of 100 nm size in buffer, which was evidenced by the TEM observation, the DLS analysis, and the encapsulation experiment. The genuine peptide vesicles, peptosomes, were labeled with a near-infrared fluorescence (NIRF) probe. In vivo retention in blood experiment showed long circulation of the peptosome in rat blood as stable as the PEGylated liposome. NIRF imaging of a small cancer on mouse by using the peptosome as a nanocarrier was successful due to the EPR effect of the peptosome. Peptosome is shown here as a novel excellent nanocarrier for molecular imaging. PMID- 18163536 TI - Evaluation of maleimide derivative of DOTA for site-specific labeling of recombinant affibody molecules. AB - Affibody molecules are a new class of small (7 kDa) scaffold affinity proteins, which demonstrate promising properties as agents for in vivo radionuclide targeting. The Affibody scaffold is cysteine-free and therefore independent of disulfide bonds. Thus, a single thiol group can be engineered into the protein by introduction of one cysteine. Coupling of thiol-reactive bifunctional chelators can enable site-specific labeling of recombinantly produced Affibody molecules. In this study, the use of 1,4,7,10-tetraazacyclododecane-1,4,7-tris-acetic acid 10-maleimidoethylacetamide (MMA-DOTA) for 111 In-labeling of anti-HER2 Affibody molecules His 6-Z HER2:342-Cys and Z HER2:2395-Cys has been evaluated. The introduction of a cysteine residue did not affect the affinity of the proteins, which was 29 pM for His 6-Z HER2:342-Cys and 27 pM for Z HER2:2395-Cys, comparable with 22 pM for the parental Z HER2:342. MMA-DOTA was conjugated to DTT reduced Affibody molecules with a coupling efficiency of 93% using a 1:1 molar ratio of chelator to protein. The conjugates were labeled with 111 In to a specific radioactivity of up to 7 GBq/mmol, with preserved binding for the target HER2. In vivo, the non-His-tagged variant 111 In-[MMA-DOTA-Cys61]-Z HER2:2395-Cys demonstrated appreciably lower liver uptake than its His-tag-containing counterpart. In mice bearing HER2-expressing LS174T xenografts, 111 In-[MMA-DOTA Cys61]-Z HER2:2395-Cys showed specific and rapid tumor localization, and rapid clearance from blood and nonspecific compartments, leading to a tumor-to-blood ratio of 18 +/- 8 already 1 h p.i. Four hours p.i., the tumor-to-blood ratio was 138 +/- 8. Xenografts were clearly visualized already 1 h p.i. PMID- 18163537 TI - Synthesis, characterization, antitumor activity of pluronic mimicking copolymer micelles conjugated with doxorubicin via acid-cleavable linkage. AB - Pluronic mimicking poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymer having multiple hydroxyl groups in the PPO middle segment (core-functionalized Pluronic: CF-PLU) was synthesized for conjugation of doxorubicin (DOX). DOX was conjugated on the multiple hydroxyl groups of CF-PLU via an acid-labile hydrazone linkage (CF-PLU-DOX). In aqueous solution, CF-PLU-DOX copolymers self-assembled to form a core/shell-type micelle structure consisting of a hydrophobic DOX-conjugated PPO core and a hydrophilic PEO shell layer. The conjugated DOX from CF-PLU-DOX micelles was released out more rapidly at pH 5 than pH 7.4, indicating that the hydrazone linkage was cleaved under acidic condition. CF-PLU-DOX micelles exhibited greatly enhanced cytotoxicity for MCF-7 human breast cancer cells compared to naked DOX, while CF PLU copolymer itself showed extremely low cytotoxicity. Flow cytometry analysis revealed that the extent of cellular uptake for CF-PLU-DOX micelles was greater than free DOX. Confocal image analysis also showed that CF-PLU-DOX micelles had a quite different intracellular distribution profile from free DOX. CF-PLU-DOX micelles were mainly distributed in the cytoplasm, endosomal/lysosomal vesicles, and nucleus, while free DOX was localized mainly within the nucleus, suggesting that CF-PLU-DOX micellar formulation might be advantageously used for overcoming the multidrug resistance (MDR) effect, which gradually develops in many tumor cells during repeated drug administration. PMID- 18163538 TI - Acridine-based agents with topoisomerase II activity inhibit pancreatic cancer cell proliferation and induce apoptosis. AB - A series of substituted 9-aminoacridines is evaluated for antiproliferative activity toward pancreatic cancer cells. The results indicate that the compounds inhibit cell proliferation by inducing a G1-S phase arrest. A model is also developed that explains the molecular basis to inhibition through a DNA "threading" mechanism. We conclude that the drug-DNA complex formed blocks topoisomerase II binding and activity leading to catalytic inhibition of the enzyme and the induction of apoptosis and programmed cell death. PMID- 18163539 TI - Influence of postharvest hot water treatment on nutritional and functional properties of kumquat (Fortunella japonica Lour. Swingle Cv. Ovale) fruit. AB - The present study investigated the influence of a hot water dip (HWD) for 2 min at 50 degrees C, a standard and effective treatment for postharvest decay control of citrus fruit, on the nutritional and health-related properties of kumquats. The results show that most of the parameters examined, including titratable acidity, soluble solids content, maturity index, glucose, fructose, sucrose, ascorbic acid, dehydroascorbic acid, alpha- and gamma-tocopherols, beta-carotene, zeaxantin, rhoifolin, and antioxidant activity, were not significantly affected by treatment. The levels of beta-cryptoxanthin, narirutin, and total flavonoids increased after HWD, whereas lutein and total phenols decreased. The concentration of the essential oil and the relative percentage of the individual components of the essential oil were not affected by HWD except for the minor compound p-menta-1,5-dien-1-ol, which increased after HWD. After storage, lower levels of glucose, total sugars, beta-carotene, beta-cryptoxanthin and lutein were recorded in HWD fruit. A decrease in antioxidant activity and increases in alpha-tocopherol and total vitamin E were found both in control and HWD fruit. The influence of HWD at 50 degrees C for 2 min on individual nutraceuticals and health-related properties was thus generally low and may depend on storage conditions. PMID- 18163540 TI - Chitosan film as rhBMP2 carrier: delivery properties for bone tissue application. AB - Tissue engineering approaches need biomaterials with suitable properties to provide an appropriate environment for cell attachment and growth. The performance of these biomaterials can be greatly enhanced through the incorporation of bioactive agents. For this reason, we developed chitosan films with cell-attachment ability, rhBMP-2 carrier capacity, and good in vivo performance, and we employ them as covering for implantable materials. In this work, we have tried to explain how the rh-BMP2 is delivered to the surroundings from the development chitosan films. Protein diffusion from film, film stability versus in vitro dissolution, and biodegradation were evaluated to study rhBMP-2 delivery. Our results show that chitosan film has sufficiently good features to be used as an rhBMP-2 carrier. A low diffusion rate was observed, which was sufficient to quickly induce an in vitro differentiation stimulus, although heavily activated films retain more than 80-85% of the protein on the film. On the other hand, we estimated that chitosan film dissolution due to initial acidification in the wound environment is no more than 15-20%. We also estimated chitosan film response to lysozyme and concluded that degradation via this process proceeded at a slow kinetic rate. In addition, rhBMP-2 in vitro activity after film processing, as well as in vivo film behavior, were studied. We confirm that rhBMP-2 remains active on the film and after release, both in vitro and in vivo. These results support the conclusion that the developed chitosan film allows sustained release of the rhBMP-2 osteoinductive protein and could be used as an activated coat for implant and surgical prosthesis. PMID- 18163541 TI - Ionic liquid-based preparation of cellulose-dendrimer films as solid supports for enzyme immobilization. AB - Surface-active cellulose films for covalent attachment of bioactive moieties were achieved by codissolution of cellulose with polyamidoamine (PAMAM) dendrimers in an ionic liquid followed by regeneration of the composite as a film. Different generations of PAMAM were used for the formation of cellulose-dendrimer composites, as well as films with the dendrimer covalently bonded to the cellulose by means of the linker 1,3-phenylene diisocyanate. Surface characterization, thermal stability, and utility for immobilization of laccase were determined. The presence of the dendrimer amino groups was confirmed by detailed characterization of the films' surfaces. These modified films exhibit acceptable thermal stability, comparable to that of other regenerated cellulose films, but the number of active functional groups on the surface is much smaller than the theoretical amount expected. Films made with 1,3-phenylene diisocyanate as linker for covalently bound cellulose and dendrimers exhibit a better performance for immobilization of laccase than those prepared by simple mixing of the cellulose and dendrimer. In general, a linear correspondence between the dendrimer generation within the films and the specific activity of immobilized laccase in such films was not observed. PMID- 18163542 TI - Reactions of glyceraldehyde 3-phosphate dehydrogenase sulfhydryl groups with bis electrophiles produce DNA-protein cross-links but not mutations. AB - The environmental contaminant 1,2-dibromoethane and diepoxybutane, an oxidation product of the important industrial chemical butadiene, are bis-functional electrophiles and are known to be mutagenic and carcinogenic. One mechanism by which bis-electrophiles can exert their toxic effects is through the induction of genotoxic and mutagenic DNA-peptide cross-links. This mechanism has been shown in systems overexpressing the DNA repair protein O6 -alkylguanine DNA alkyltransferase (AGT) or glutathione S-transferase and involves reactions with nucleophilic cysteine residues. The hypothesis that DNA-protein cross-link formation is a more general mechanism for genotoxicity by bis-electrophiles was investigated by screening nuclear proteins for reactivity with model monofunctional electrophiles. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was identified as a candidate because of the nucleophilicity of two cysteine residues (Cys152 and Cys246) in reaction screens with model electrophiles (Dennehy, M. K. et al. (2006) Chem. Res. Toxicol. 19, 20-29). Incubation of GAPDH with bis-electrophiles resulted in inhibition of its catalytic activity, but only at high concentrations of diepoxybutane. In vitro assays indicated DNA-GAPDH cross-link formation in the presence of diepoxybutane, and bis-electrophile reactivity at Cys246 was confirmed using mass spectral analysis. In contrast to AGT, overexpression of human GAPDH in Escherichia coli did not enhance mutagenesis by diepoxybutane. We propose that the lack of mutational enhancement is in part due to the inherently lower reactivity of GAPDH toward bis electrophiles as well as the reduced DNA binding ability relative to AGT, preventing the in vivo formation of DNA-protein cross-links and enhanced mutagenesis. PMID- 18163543 TI - Essential role of the AH receptor in the dysfunction of heme metabolism induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - The dysfunction of hepatic heme synthesis by 2,3,7,8-tetrachlordibenzo- p-dioxin (TCDD) in mice, enhanced by iron, leads to accumulation of uroporphyrins I and III (uroporphyria) and resembles the human disorder porphyria cutanea tarda (PCT) precipitated by alcohol and estrogenic drugs. Although consequences of TCDD are considered entirely dependent on the aryl hydrocarbon receptor (AHR), this is not proven for uroporphyria. Administration of TCDD (75 microg/kg) caused uroporphyria in susceptible C57BL/6J mice with high-affinity AHR after 5 weeks (>600-fold increase in hepatic uroporphyrins). Transcriptomics showed significant modified gene expressions for intermediary, heme, and iron metabolism as well as for oxidative stress and cell injury. Resistant low-affinity AHR DBA/2 mice (no increase in porphyrins) showed far fewer changes. At this dose of TCDD, persistent up-regulation of some traditional AH battery genes occurred in both strains. Essentiality of AHR was demonstrated with C57BL/6 Ahr knockout mice. Elevation of hepatic uroporphyrins was 964-fold in Ahr (+/+) mice, lower in Ahr (+/-) (60-fold), but undetectable with Ahr (-/-) . Consistent with an oxidative mechanism, iron overload enhanced porphyria as well as general liver injury in Ahr (+/+) and Ahr (+/-) mice but had no interactive effect in Ahr (-/-) . In contrast, when iron-treated mice received, instead of TCDD, the heme precursor 5 aminolevulinic acid (ALA), causing uroporphyia in Ahr (+/+) mice (242-fold rise in uroporphyrins), elevation of uroporphyrins I and III (42-fold) also occurred in Ahr (-/-) mice and was seemingly associated with AHR-independent expression of Cyp1a2. The findings prove that AHR is a key factor in porphyria induced in mice by TCDD. However, in other models of human PCT, participation of AHR may not be an essential requirement. PMID- 18163544 TI - Binding and hydrolysis of soman by human serum albumin. AB - Human plasma and fatty acid free human albumin were incubated with soman at pH 8.0 and 25 degrees C. Four methods were used to monitor the reaction of albumin with soman: progressive inhibition of the aryl acylamidase activity of albumin, the release of fluoride ion from soman, 31P NMR, and mass spectrometry. Inhibition (phosphonylation) was slow with a bimolecular rate constant of 15 +/- 3 M(-1) min (-1). MALDI-TOF and tandem mass spectrometry of the soman-albumin adduct showed that albumin was phosphonylated on tyrosine 411. No secondary dealkylation of the adduct (aging) occurred. Covalent docking simulations and 31P NMR experiments showed that albumin has no enantiomeric preference for the four stereoisomers of soman. Spontaneous reactivation at pH 8.0 and 25 degrees C, measured as regaining of aryl acylamidase activity and decrease of covalent adduct (pinacolyl methylphosphonylated albumin) by NMR, occurred at a rate of 0.0044 h (-1), indicating that the adduct is quite stable ( t1/2 = 6.5 days). At pH 7.4 and 22 degrees C, the covalent soman-albumin adduct, measured by MALDI-TOF mass spectrometry, was more stable ( t1/2 = 20 days). Though the concentration of albumin in plasma is very high (about 0.6 mM), its reactivity with soman (phosphonylation and phosphotriesterase activity) is too slow to play a major role in detoxification of the highly toxic organophosphorus compound soman. Increasing the bimolecular rate constant of albumin for organophosphates is a protein engineering challenge that could lead to a new class of bioscavengers to be used against poisoning by nerve agents. Soman-albumin adducts detected by mass spectrometry could be useful for the diagnosis of soman exposure. PMID- 18163545 TI - Methylation of dimethyltin in mice and rats. AB - Organotins are widely used as stabilizers of polyvinyl chloride and as catalysts or biocides. It is well known that dimethyltin (DMT) is less neurotoxic than trimethyltin (TMT). A Korean worker who was exposed to DMT compounds showed neurological symptoms similar to those of TMT encephalopathy, in association with high levels of both DMT and TMT in the urine and blood. The case suggested the possibility of the methylation of DMT in humans. Here, we investigated whether TMT is detected in the urine of mice and rats exposed only to DMT dichloride (DMTC). Three Slc:ICR mice and three Slc:Wistar rats were placed in individual metabolic cages, and one day later, they were injected intraperitoneally with DMTC (10 mg/kg body weight (wt); 5.4 mgSn/kg body wt; 45.5 micromol/kg body wt) over 4 consecutive days. Twenty-four hour urine samples were collected every evening for 11 consecutive days starting at baseline (before treatment). Speciation analyses of methyltin compounds in urine were performed using a combination of high performance liquid chromatograph-inductively coupled plasma mass spectrometry. High concentrations of DMT and time-dependent increase in TMT concentrations were found in both mice and rats during the 4-day treatment, and their concentrations decreased gradually after the cessation of treatment. The chemical compound of the detected peak was confirmed to be TMT by liquid chromatography-tandem mass spectrometry. Neither DMT nor TMT was detected in the samples collected at baseline. Our results indicate urinary excretion of TMT in mice and rats injected with DMTC, confirming the production of TMT in vivo, probably through methylation of DMT. PMID- 18163546 TI - Reversal of arsenic-induced hepatic apoptosis with combined administration of DMSA and its analogues in guinea pigs: role of glutathione and linked enzymes. AB - Arsenicosis, due to contaminated drinking water in the Indo-Bangladesh region, is a serious health hazard in terms of morbidity and mortality. Reactive oxygen species (ROS) generated due to arsenic toxicity have been attributed as one of the initial signals that impart cellular toxicity, which is controlled by the internal antioxidant glutathione (GSH). In the present study, we investigated (i) the role of GSH and its linked enzymes, glutathione peroxidase and glutathione reductase, in reversing chronic arsenic toxicity using a thiol chelating agent, meso-2,3-dimercaptosuccinic acid (DMSA), or one of its analogues individually or in combination; (ii) if alterations in the carbon side chain of DMSA increased efficacy; and (iii) whether the combination therapy enhance arsenic removal from hepatic tissue and prevent hepatic apoptosis. Results indicated that chronic arsenic exposure led to a ROS-mediated, mitochondrial-driven, caspase-dependent apoptosis in hepatic cells with a significant increase in glutathione disulfide (GSSG) levels and decreased glutathione reductase levels. Monotherapy with DMSA and its analogues did show minimal recovery postchelation. However, the combination of DMSA with long carbon chain analogues like monoisoamyl DMSA (MiADMSA) or monocyclohexyl DMSA (MchDMSA) showed a better efficacy in terms of reducing the arsenic burden as well as reversing altered biochemical variables indicative of oxidative stress and apoptosis. We also observed that GSH and its linked enzymes, especially glutathione reductase, play a vital role in scavenging ROS, maintaining GSH pools, and providing clinical recoveries. On the basis of the above observations, we recommend that combinational therapy of DMSA and its long carbon chain analogues MiADMSA or MchDMSA would be more effective in arsenic toxicity. PMID- 18163547 TI - Preparation of ribonuclease S domain-swapped dimers conjugated with DNA and PNA: modulating the activity of ribonucleases. AB - Obtaining highly specific and active ribonuclease activities is an important goal with numerous medical and biochemical applications. As a step toward more active and specific ribonucleases, we describe the preparation and the enzymatic and structural properties of RNase S monomers and dimers conjugated to DNA and PNA molecules. Poly(dT)n (2'-oligodeoxyribonucleotides, n = 8, 15) and t8 peptide nucleic acid (PNA) chains have been conjugated to the S-peptide of ribonuclease S. Monomers and dimers of the conjugated enzyme have been obtained and characterized by 1H NMR spectroscopy, showing that DNA or PNA conjugation does not alter the native structure of ribonuclease S. The oligonucleotide-conjugated RNase S monomer and dimer show significant activity against single-stranded RNA and very low/negligible hydrolysis of double-stranded poly(A).poly(U). In contrast, the t8-conjugated RNase S monomer and dimer show substantial activity against both ssRNA and dsRNA. These results highlight the importance of positive charges near but not in the active site in enhancing activity against dsRNA and reveal the promise of PNA-RNase conjugates for modulating RNase activity. PMID- 18163548 TI - Fragment-based discovery of mexiletine derivatives as orally bioavailable inhibitors of urokinase-type plasminogen activator. AB - Fragment-based lead discovery has been applied to urokinase-type plasminogen activator (uPA). The (R)-enantiomer of the orally active drug mexiletine 5 (a fragment hit from X-ray crystallographic screening) was the chemical starting point. Structure-aided design led to elaborated inhibitors that retained the key interactions of (R)-5 while gaining extra potency by simultaneously occupying neighboring regions of the active site. Subsequent optimization led to 15, a potent, selective, and orally bioavailable inhibitor of uPA. PMID- 18163550 TI - A Drosophila model to identify polyamine-drug conjugates that target the polyamine transporter in an intact epithelium. AB - Polyamine transport is elevated in many tumor types, suggesting that toxic polyamine-drug conjugates could be targeted to cancer cells via the polyamine transporter (PAT). We have previously reported the use of Chinese hamster ovary (CHO) cells and its PAT-deficient mutant cell line, CHO-MG, to screen anthracene polyamine conjugates for their PAT-selective targeting ability. We report here a novel Drosophila-based model for screening anthracene-polyamine conjugates in a developing and intact epithelium ( Drosophila imaginal discs), wherein cell-cell adhesion properties are maintained. Data from the Drosophila assay are consistent with previous results in CHO cells, indicating that the Drosophila epithelium has a PAT with vertebrate-like characteristics. This assay will be of use to medicinal chemists interested in screening drugs that use PAT for cellular entry, and it offers the possibility of genetic dissection of the polyamine transport process, including identification of a Drosophila PAT. PMID- 18163551 TI - Novel prodrugs of tegafur that display improved anticancer activity and antiangiogenic properties. AB - New and more potent prodrugs of the 5-fluorouracyl family derived by hydroxymethylation or acyloxymethylation of 5-fluoro-1-(tetrahydro-2-furanyl) 2,4(1H,3H)-pyrimidinedione (tegafur, 1) are described. The anticancer activity of the butyroyloxymethyl-tegafur derivative 3 and not that of tegafur was attenuated by the antioxidant N-acetylcysteine, suggesting that the increased activity of the prodrug is in part mediated by an increase of reactive oxygen species. Compound 3 in an in vitro matrigel assay was found to be a more potent antiangiogenic agent than tegafur. In vivo 3 was significantly more potent than tegafur in inhibiting 4T1 breast carcinoma lung metastases and growth of HT-29 human colon carcinoma tumors in a mouse xenograft. In summary, the multifunctional prodrugs of tegafur display selectivity toward cancer cells, antiangiogenic activity, and anticancer activities in vitro and in vivo, superior to those of tegafur. 5-fluoro-1-(tetrahydro-2-furanyl)-2,4(1 H,3 H) pyrimidinedione (tegafur, 1), the oral prodrug of 5-FU, has been widely used for treatment of gastrointestinal malignancies with modest efficacy. The aim of this study was to develop and characterize new and more potent prodrugs of the 5-FU family derived by hydroxymethylation or acyloxymethylation of tegafur. Comparison between the effect of tegafur and the new prodrugs on the viability of a variety of cancer cell lines showed that the IC50 and IC90 values of the novel prodrugs were 5-10-fold lower than those of tegafur. While significant differences between the IC50 values of tegafur were observed between the sensitive HT-29 and the resistant LS-1034 colon cancer cell lines, the prodrugs affected them to a similar degree, suggesting that they overcame drug resistance. The increased potency of the prodrugs could be attributed to the antiproliferative contribution imparted by formaldehyde and butyric acid, released upon metabolic degradation. The anticancer activity of the butyroyloxymethyl-tegafur derivative 3 and not that of tegafur was attenuated by the antioxidant N-acetylcysteine, suggesting that the increased activity of the prodrug is in part mediated by an increase of reactive oxygen species. Compound 3 in an in vitro matrigel assay was found to be a more potent antiangiogenic agent than tegafur. In vivo 3 was significantly more potent than tegafur in inhibiting 4T1 breast carcinoma lung metastases and growth of HT-29 human colon carcinoma tumors in a mouse xenograft. In summary, the multifunctional prodrugs of tegafur display selectivity toward cancer cells, antiangiogenic activity and anticancer activities in vitro and in vivo, superior to those of tegafur. PMID- 18163552 TI - Interspecific rice hybrid of Oryza sativa x Oryza nivara reveals a significant increase in seed protein content. AB - Wild species offer a potential reservoir of genetic variation for crop improvement. Besides the valuable genes for disease resistance that the wild species have provided for rice improvement, recent studies have shown that these wild species could also provide favorable alleles for the improvement of yield and yield-related traits. The present study reports yet another potential of wild relatives of rice, which involves the improvement of seed protein content. A significant increase in seed protein content was observed in an interspecific hybrid between Oryza sativa ssp. indica and the wild species Oryza nivara. The hybrid showed a protein content of 12.4%, which was 28 and 18.2% higher than those of the parents O. nivara and IR 64, respectively. The increase in protein content was dependent on the genetic background of the rice variety used in the hybridization. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of seed storage proteins demonstrated that a significant increase in prolamins and glutelins was mainly responsible for the elevated protein content of the hybrid. Amino acid analysis of seed proteins revealed that the hybrid had net gains of 19.5% in lysine and 19.4% in threonine over the O. nivara parent on a seed dry weight basis. Molecular analysis indicated that the increase in protein content of the hybrid was not a result of chromosomal rearrangements or transposable element activation, at least in the chromosomal regions containing seed storage protein genes. A preliminary genetic analysis of the F 2 segregating population showed that the inheritance of the increased protein content was polygenic in nature. The development of this interspecific hybrid offers a great potential for selecting new rice cultivars that combine the high yield and superior cooking quality of IR 64 with improved seed protein content. PMID- 18163553 TI - Organically fertilized onions (Allium cepa L.): effects of the fertilizer placement method on quercetin content and soil nitrogen dynamics. AB - Field-cured onions cv. Hyskin ( Allium cepa L.) supplied with organic nitrogen fertilizer were studied. The fertilizer was applied by broadcasting and harrowing, broadcasting and rotary cultivation, or placement between rows. Nitrogen dynamics were monitored throughout the growing season by soil sampling. Variation in quercetin content in the onion scales was analyzed by HPLC. The organically fertilized onions were compared with inorganically fertilized onions grown in the same field. Inoculation with arbuscular mycorrhizal fungi (AMF) in the row at sowing or during commercial transplant production was tested but did not significantly affect mycorrhizal root colonization levels in the field. Onions that received no fertilizer at all or that had fertilizer placed between rows had better establishment, probably due to more favorable soil nitrogen concentrations for seedling emergence. Broadcast application led to higher nitrogen concentration in the root zone, resulting in fewer but larger individual onions. Quercetin levels were not significantly altered as a result of nitrogen fertilizer source (inorganic or organic), application method, or mycorrhizal inoculation. However, variation between years was significant, with quercetin levels in 2004 almost twice as high as those in 2005. PMID- 18163554 TI - Impact of surface-active compounds on physicochemical and oxidative properties of edible oil. AB - The physical properties of lipids can have a major influence on lipid oxidation reactions. Edible oils contain surface-active compounds and water that can form physical structures such as reverse micelles. This study used the fluorescence probe, 5-dodecanoylaminofluorescein (DAF), to study both the physical and the chemical properties of stripped corn oil containing oleic acid and phosphatidylcholine. The fluorescence intensity of DAF increased with increasing water concentration in the edible oil. The addition of oleic acid decreased DAF fluorescence due to the ability of the free fatty acid to decrease the pH of the aqueous phase of the bulk oil. Phosphatidylcholine increased DAF fluorescence due to its ability to increase DAF exposure to the aqueous phase. Oleic acid had no impact on interactions between DAF and water-soluble peroxyl radicals, while phosphatidylcholine decreased peroxyl radical degradation of DAF. These results suggest that DAF could be a useful analytical tool to study the impact of the aqueous environment of bulk oil on lipid oxidation. PMID- 18163555 TI - Flavor enhancement of chicken broth from boiled celery constituents. AB - The flavor-enhancing effects of the volatile constituents in celery were investigated. The test samples were prepared by adding celery fractions to chicken broth at a concentration that distinct odors of them were not detected, and the samples were sensorially evaluated for the perceived intensities of 8 terms such as "thick," "impactful," "mild," "lasting," "satisfied," "complex," "refined," and "clarified," which are considered to be the elements of the complex flavor and for 3 terms such as "sweet," "salty," and "umami" taste. A comparison of effects between the volatile and nonvolatile fractions of celery revealed that the volatile compounds in celery enhanced the complex flavor of chicken broth more than the nonvolatile compounds. Among the characteristic odorants of celery, three phthalides, namely, sedanenolide, 3- n-butylphthalide, and sedanolide, were shown to contribute to the complex flavor of chicken broth, and sedanenolide was most effective. The three phthalides enhanced perceived intensities of "umami" and "sweet" despite their no taste properties in addition to the complex flavor. PMID- 18163556 TI - Online RP-HPLC-DPPH screening method for detection of radical-scavenging phytochemicals from flowers of Acacia confusa. AB - Acacia confusa is traditionally used as a medicinal plant in Taiwan. In this study, phytochemicals and antioxidant activities of extracts from flowers of A. confusa were investigated for the first time. In addition, a rapid screening method, online RP-HPLC-DPPH system, for individual antioxidants in complex matrices was developed. Accordingly, six antioxidants including gallic acid ( 1), myricetin 3-rhamnoside ( 2), quercetin 3-rhamnoside ( 3), kaempferol 3-rhamnoside ( 4), europetin 3-rhamnoside ( 5), and rhamnetin 3-rhamnoside ( 6) were detected using the developed screening method. Of these, compounds 2, 3, and 5 were found to be major bioactive phytochemicals, and their contents were determined as 11.3, 6.7, and 8.7 mg/g of crude extract, respectively. By comparison with quercetin, a well-known antioxidant, these compounds had the order of compound 2 > compound 5 > quercetin > compound 3 for DPPH radical-scavenging activity. Their IC 50 values were 3.0, 3.2, 4.5, and 7.4 microM, respectively. Moreover, the same order was observed for superoxide radical-scavenging activity, and their IC50 values were 2.6, 2.7, 4.3, and 5.3 microM, respectively. However, for lipid peroxidation, quercetin, an aglycon, showed the best inhibitory activity. The IC50 values of quercetin, compound 2, compound 5, and compound 3 were 46.7, 88.5, 90.7, and 124.6 microM, respectively. These results indicated that a rhamnoside at the C3 position of flavonoids had a negative effect on radical-scavenging activity and antilipid peroxidation. In contrast, the number of hydroxyl groups on the B-ring exhibited a positive relationship with their inhibitory activities. PMID- 18163557 TI - Fate of resveratrol and piceid through different hop processings and storage times. AB - Trans-Piceid and trans-resveratrol contents of hop cones, hop pellets, CO2 extracts, and spent hop from American varieties (harvest 2004) were determined by reverse-phase high-performance liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry [RP-HPLC-APCI(+)-MS/MS]. Pelletization induced strong stilbene degradation in some cultivars. Similarly, 1 year of storage at 4 degrees C led to a huge loss of trans-piceid, especially in the case of hop cones (much faster than in model media, although well protected from light and oxygen). Therefore, after 8 months of storage, the overall stilbene content was in the same range whatever the conditioned form. Absent in fresh hop cones or pellets, cis-resveratrol was released from cis-piceid in all stored samples. On the other hand, no delta-viniferin was detected despite it is present in light-protected model media spiked with trans-piceid. Because supercritical carbon dioxide proved inefficient for recovering resveratrol and piceid from pellets, spent hop emerged as the most interesting material for subsequent specific stilbene extraction. PMID- 18163558 TI - Identification of characteristic aroma components of Thai fried chili paste. AB - Three forms of Thai fried chili pastes (CP) were prepared, consisting of an unheated CP (UH-CP), a CP heated at 100 degrees C for 25 min (H25-CP, typical product), and a CP excessively heated for 50 min (H50-CP). The potent odorants in the CPs were investigated by two gas chromatography-olfactometry methods: dynamic headspace dilution analysis (DHDA) and aroma extract dilution analysis (AEDA). DHDA revealed that the predominant odorants in heated CPs were mainly sulfur containing compounds, followed by lipid-derived compounds, Strecker aldehydes, and Maillard reaction products. Dimethyl sulfide, allyl mercaptan, 2- (or 3-) methylbutanal, ally methyl sulfide, 2,3-butanedione, 3,3'-thiobis(1-propene), and methyl propyl disulfide were among the most potent headspace odorants detected by DHDA. By AEDA, 2-vinyl-4 H-1,3-dithiin and diallyl trisulfide had the highest FD factors in H25-CP. On the basis of their high FD factors by both GCO methods, the predominant odorants in H25-CP were 3-vinyl-4 H-1,2-dithiin, allyl methyl disulfide, and allyl methyl trisulfide. Furthermore, dimethyl trisulfide and diallyl disulfide had the highest odor activity values in H25-CP, suggesting that these were also potent odorants in CP. In addition, methional, 3-methylbutanoic acid, 4-hydroxy-2,5-dimethyl-3-(2 H)-furanone, and 3-hydroxy-4,5-dimethyl-2( 5H) furanone (sotolon) were indicated as potent thermally derived odorants of H25-CP. PMID- 18163559 TI - Microwave-assisted efficient extraction of different parts of Hippophae rhamnoides for the comparative evaluation of antioxidant activity and quantification of its phenolic constituents by reverse-phase high-performance liquid chromatography (RP-HPLC). AB - The outcome of different extraction procedures (microwave, ultrasound, Soxhlet, and maceration) on the antioxidant activity of seeds, leaves, pulp, and fruits of Hippophae rhamnoides (sea buckthorn or SBT) was investigated by two different bioassays: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. The SBT extracts were found to possess strong antioxidant activity measured in terms of TEAC (2.03-182.13 and 6.97-282.75 mg/g) with ABTS and DPPH assays, respectively. In general, the antioxidant capacity of microwave-assisted extracts was found to be significantly higher than those obtained by ultrasound-assisted extraction (UAE) and maceration while being slightly higher than Soxhlet extracts. Further, microwave extracts of seeds were found to possess maximum antioxidant capacity followed by leaves, fruits, and pulp. Also, the chemical composition of extracts, studied in terms of the total phenolic content, was found to be in the range of 1.9-23.5 mg/g Gallic acid equivalent (GAE), which indicates a strong correlation between antioxidant activity and phenolic content present in the SBT. In addition, some of its bioactive phenolic constituents, such as rutin ( 1), quercetin-3- O-galactoside ( 2), quercetin ( 3), myricetin ( 4), kaempferol ( 5), and isorhamnetin ( 6), were also quantified in different extracts by reverse-phase high-performance liquid chromatography (RP-HPLC). PMID- 18163560 TI - New volatile sulfur-containing constituents in a simultaneous distillation extraction extract of red bell peppers (Capsicum annuum). AB - An extract of red bell peppers ( Capsicum annuum) was prepared by simultaneous distillation-extraction (SDE, Likens-Nickerson). In addition to the already known (3 E)-3-hepten-2-one ( 1), the unsaturated C9-ketones 1-nonen-4-one ( 2), (2 E)-2 nonen-4-one ( 3), and (2 E,5 E)-2,5-nonadien-4-one ( 4), 2-methoxy-3 isobutylpyrazine ( 5), and heptane-2-thiol ( 6), we identified 19 new thiols (the aliphatic saturated and unsaturated thiols 14- 16, and 22- 27, the mercapto ketones 12 and 13, the mercapto-alcohols 17, 18, and 30, the dithiols 19 and 28, the methylthio-thiols 20 and 21, and the thiophene-thiol 31) and the two new dithiolanes 10 and 29. All of them are structurally related to the unsaturated C7 and C9-ketones 1- 4. The free thiols were enriched using Affi-Gel 501 ( p aminophenyl-mercuric acetate grafted on an agarose gel). The new compounds were confirmed by syntheses and were organoleptically evaluated. PMID- 18163561 TI - Specific and sensitive enzyme-linked immunosorbent assays for analysis of residual allergenic food proteins in commercial bottled wine fined with egg white, milk, and nongrape-derived tannins. AB - Regulations introduced by the Food Standards Australia New Zealand in December 2002 require all wine and wine product labels in Australia to identify the presence of a processing aid, additive or other ingredient, which is known to be a potential allergen. The objective of this study was to establish sensitive assays to detect and measure allergenic proteins from commonly used processing aids in final bottled wine. Sensitive and specific enzyme-linked immunosorbent assays (ELISA) were developed and established for the proteins casein, ovalbumin, and peanut. Lower limits of detection of these proteins were 8, 1, and 8 ng/mL, respectively. A panel of 153 commercially available bottled Australian wines were tested by these ELISA, and except for two red wines known to contain added whole eggs, residuals of these food allergens were not detected in any wine. These findings are consistent with a lack of residual potentially allergenic egg-, milk , or nut-derived processing aids in final bottled wine produced in Australia according to good manufacturing practice at a concentration that could cause an adverse reaction in egg, milk, or peanut/tree-nut allergic adult consumers. PMID- 18163562 TI - Identification of urinary and intestinal bacterial metabolites of ellagitannin geraniin in rats. AB - Hydrolyzable tannins, including ellagitannins, occur in foods such as berries and nuts. Various biological activities, including antioxidant, antiviral, and antitumor activities, have been noted and reported for ellagitannins, but the absorption and metabolism of purified ellagitannins are poorly understood. We describe herein the characterization of urinary and intestinal microbial metabolites in rats after the ingestion of ellagitannins. Urine samples were collected after oral administration of ellagitannins such as geraniin ( 1), corilagin ( 2), and their related polyphenols. The suspension of rat intestinal microflora was anaerobically incubated with ellagitannins. Each sample was separated by column chromatography and/or preparative HPLC to give seven metabolites, M1- M7. The structures of these metabolites were determined on the basis of spectroscopic data and chemical evidence. These compounds, except for M1, were characterized as ellagitannin metabolites for the first time. Furthermore, among four major metabolites ( M1- M4) in urine, M2 showed an antioxidant activity comparable to intact geraniin and related polyphenols. PMID- 18163563 TI - Lignan content of selected foods from Japan. AB - Lignans constitute a group of phytochemicals widely distributed in the human diet. Once ingested, most dietary lignans are metabolized by the gut microflora to enterolactone and enterodiol, also known as enterolignans. Together with isoflavones and coumestrol, enterolignans have been traditionally classified as phytoestrogens, plant-derived compounds that exhibit estrogen-like activity. In addition to a higher intake of vegetables, many studies have suggested that the presence of phytoestrogens in the Japanese diet is one of the factors that might explain the reduced incidence of certain chronic diseases in Japanese populations compared with Western countries. Being a vegetable-based diet, exposure to dietary lignans should be high, but to assess this exposure, a lignan food database is required. Stable isotope-dilution gas chromatography coupled with mass spectrometry was used to quantify six plant lignans in 86 food items commonly consumed in Japan. These data will complement the previous databases and most importantly expand the knowledge of occurrence of lignans in food to Eastern diets. PMID- 18163564 TI - Surface hydrophobicity of physicochemically and enzymatically treated milk proteins in relation to techno-functional properties. AB - Surface hydrophobicity (SH) of milk proteins treated physicochemically (by heating and Maillard reaction) or modified enzymatically (by transglutaminase, lactoperoxidase, laccase, and glucose oxidase) was assessed in relation to their techno-functional properties. Heat-treatment increased SH of whey protein isolate and decreased SH of sodium caseinate and bovine serum albumin. Maillard reaction of milk proteins caused time-depended decreases of SH. Only for total milk protein reacting with glucose and lactose elevated SH-values were detected. Protein modification with transglutaminase, laccase, and lactoperoxidase strongly increased the SH of whey protein isolate and total milk protein. Incubation with glucose oxidase elevated SH values of sodium caseinate, whey protein isolate, and total milk protein. When correlating SH with techno-functional properties, a positive correlation was observed between SH and foam formation, and a negative correlation was observed between SH and foam stability as well as emulsion stability. No clear correlation was detected between SH and emulsifying activity, surface tension, viscosity, and heat stability of enzymatically and physicochemically treated milk proteins. PMID- 18163565 TI - Broccoli: a unique vegetable that protects mammalian hearts through the redox cycling of the thioredoxin superfamily. AB - Epidemiological evidence indicates several health benefits of the consumption of broccoli, especially related to chemoprevention. Because broccoli contains high amounts of selenium and glucosinolates (particularly glucoraphanin and isothiocyanate sulforaphane), which can produce redox-regulated cardioprotective protein thioredoxin (Trx), it was reasoned that consumption of broccoli could be beneficial to the heart. To test this hypothesis, a group of rats were fed broccoli (slurry made with water) through gavaging; control animals were gavaged water only. After 30 days, the rats were sacrificed; isolated hearts perfused via working mode were made ischemic for 30 min followed by 2 h of reperfusion. The results demonstrated significant cardioprotection with broccoli as evidenced by improved postischemic ventricular function, reduced myocardial infarct size, and decreased cardiomyocyte apoptosis accompanied by reduced cytochrome c release and increased pro-caspase 3 activities. Ischemia/reperfusion reduced both RNA transcripts and protein levels of the thioredoxin superfamily including Trx1, Trx2, glutaredoxin Grx1, Grx2, and peroxiredoxin (Prdx), which were either restored or enhanced with broccoli. Broccoli enhanced the expression of Nrf2, a cytosolic suppressor of Keap1, suggesting a role of antioxidant response element (ARE) in the induction of Trx. Additionally, broccoli induced the expression of another cardioprotective protein, heme oxygenase (HO)-1, which could be transactivated during the activation of Trx. Examination of the survival signal revealed that broccoli caused the phosphorylation of Akt and the induction of Bcl2 in concert with the activation of redox-sensitive transcription factor NF kappa B and Src kinase, indicating a role of Akt, Bcl2, and cSrc in the generation of survival signal. Taken together, the results of the present study indicate that the consumption of broccoli triggers cardioprotection by generating a survival signal through the activation of several survival proteins and by redox cycling of thioredoxins. PMID- 18163566 TI - Tissue depletion of amoxicillin and its major metabolites in pigs: influence of the administration route and the simultaneous dosage of clavulanic acid. AB - A residue depletion study of amoxicillin (AMO) and its major metabolites, amoxicilloic acid (AMA) and amoxicillin diketopiperazine-2',5'-dione, was performed after a single oral (p.o.) and intravenous (i.v.) administration of amoxicillin (20 mg kg (-1)) and amoxicillin/clavulanic acid (20 and 5 mg kg (-1)) to pigs. Animals were slaughtered 12, 36, 48, 60, 72, and 84 h after dosing. Tissue samples were analyzed using liquid chromatography-tandem mass spectrometry. Kidney samples contained high concentrations of amoxicilloic acid metabolite, which depleted much slower from tissues than amoxicillin, both after p.o. (t1/2AMO = 4.5 h vs t1/2AMA = 8 h) and i.v. (t1/2AMO = 4 h vs t1/2AMA = 8 h) administration. Moreover, after oral administration, significantly higher amoxicilloic acid concentrations were measured in liver and kidney than after i.v. administration. The coadministration of amoxicillin with clavulanic acid provoked no significant differences in amoxicilloic acid tissue concentrations as compared to an amoxicillin dosing. The prolonged presence of residues of amoxicilloic acid in edible tissues can play an important role in food safety, because the compound could give rise to a possible health risk, although it is not included in the maximum residue limit legislation. PMID- 18163567 TI - Porcine skeletal muscle troponin is a good source of peptides with Angiotensin-I converting enzyme inhibitory activity and antihypertensive effects in spontaneously hypertensive rats. AB - In the search for novel peptides that inhibit the angiotensin I-converting enzyme (ACE), porcine skeletal troponin was hydrolyzed with pepsin, and the products were subjected to various types of chromatography to isolate active peptides. Glu Lys-Glu-Arg-Glu-Arg-Gln (EKERERQ) and Lys-Arg-Gln-Lys-Tyr-Asp-Ile (KRQKYDI) were identified as active peptides, and their 50% inhibitory concentrations were found to be 552.5 and 26.2 microM, respectively. These are novel ACE inhibitory peptides, and the activity of KRQKYDI was the strongest among previously reported troponin-originated peptides. KRQKYDI was slowly hydrolyzed by treatment with ACE, and kinetic studies indicated that this peptide was a competitive inhibitor of the enzyme. When KRQKYDI was administered orally to spontaneously hypertensive rats (SHR) at a dose of 10 mg/kg, a temporary antihypertensive activity was observed at 3 and 6 h after administration. PMID- 18163568 TI - Investigations into inhibitor type and mode, simulated gastrointestinal digestion, and cell transport of the angiotensin I-converting enzyme-inhibitory peptides in Pacific hake (Merluccius productus) fillet hydrolysate. AB - Fish protein hydrolysate (FPH) produced by incubation of Pacific hake fillet with 3.00% Protamex at pH 6.5 and 40 degrees C for 125 min demonstrated in vitro ACE inhibitory activity (IC50 = 165 microg/mL), which was enhanced by ultrafiltration through a 10 kDa molecular weight cutoff membrane (IC50 = 44 microg/mL). However, after simulated gastrointestinal digestion, FPH and ultrafiltrate had similar ACE inhibitory activity (IC 50 = 90 microg/mL), indicating that FPH peptides act as "pro-drug type" inhibitors and that enrichment by ultrafiltration may be unnecessary. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry confirmed that the molecular weights of major peaks were <1 kDa regardless of ultrafiltration. ACE-inhibitory activities of digested hydrolysates were not significantly affected by preincubation with ACE ( P > 0.05) and exhibited a competitive inhibitory mode. A permeability assay using fully differentiated colorectal adenocarcinoma (Caco-2) cells showed an apical to basolateral transport of peptides that ranged from approximately 2 to 20% after 2 h at 37 degrees C. Pacific hake fillet hydrolysates are a potentially bioavailable source of ACE-inhibitory peptides awaiting further in vivo study. PMID- 18163569 TI - Establishment of a highly sensitive sandwich enzyme-linked immunosorbent assay specific for ovomucoid from hen's egg white. AB - A highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) kit was established for quantifying ovomucoid from hen's egg white, which has been considered as one of the major allergen in egg white. The detection limit reached 0.041 ng/mL, and linearity ranged from 0.1 to 6.25 ng/mL. Intra- and interassay coefficient variations were all lower than 5% at three concentrations (0.5, 2.5, and 5 ng/mL). No cross-reactivity was observed with bovine serum, horse serum, goat serum, human serum, duck egg white, goose egg white, quail egg white, and pigeon egg white, but a low level of cross-reactivity was found with chicken serum. The ELISA kit was established on the basis of two monoclonal antibodies (mAbs) recognizing different epitopes of ovomucoid. However, these mAbs were generated using commercially purified ovalbumin as immunogen. Studies on the relative allergenicity and antigenicity of egg white protein have been performed by many researchers, but there were controversial opinions reported previously because of the impurity of each egg white protein used in various studies. In the present work we measured the degree of ovomucoid contamination in commercially purified ovalbumin sample, and the value was about 11%. We also determined the ovomucoid residue in influenza vaccine samples for the first time. These data showed that the ELISA kit we established could serve as an effective method for precisely quantifying concentrations of ovomucoid in the egg industry and as a useful tool for the research of allergenicity and antigenicity of hen's egg proteins. PMID- 18163570 TI - Characterization and purification of glycosaminoglycans from crude biological samples. AB - Chondroitin sulfate (CS) is a glycosaminoglycan derived from cartilage and commonly used to treat osteoarthritis, psoriasis, and other conditions. The dimethylmethylene blue (DMMB) assay has been used often to measure glycosaminoglycan levels in relatively pure samples. In this study, we verified the accuracy of the DMMB assay in measuring CS levels in unpurified extract from bovine trachea and shark cartilage, despite potential interference from salts, proteins, and DNA. We found that the glycosaminoglycan signal obtained was due to CS and not to other glycosaminoglycan species. This was confirmed using fluorophore-assisted carbohydrate electrophoresis, which also revealed that the majority of the CS was monosulfated at the C4 or C6 position. Finally, we used anion-exchange chromatography to purify the bovine extract and obtained complete recovery of the glycosaminoglycans, with no contaminating protein. The results of this study should be very useful for future purification and analysis of this common supplement. PMID- 18163571 TI - Functional mimicry of the active site of glutathione peroxidase by glutathione imprinted selenium-containing protein. AB - For imitating the active site of antioxidant selenoenzyme glutathione peroxidase (GPx), an artificial enzyme selenosubtilisin was employed as a scaffold for reconstructing substrate glutathione (GSH) specific binding sites by a bioimprinting strategy. GSH was first covalently linked to selenosubtilisin to form a covalent complex GSH-selenosubtilisin through a Se-S bond, then the GSH molecule was used as a template to cast a complementary binding site for substrate GSH recognition. The bioimprinting procedure consists of unfolding the conformation of selenosubtilisin and fixing the new conformation of the complex GSH-selenosubtilisin. Thus a new specificity for naturally occurring GPx substrate GSH was obtained. This bioimprinting procedure facilitates the catalytic selenium moiety of the imprinted selenosubtilisin to match the reactive thiol group of GSH in the GSH binding site, which contributes to acceleration of the intramolecular catalysis. These imprinted selenium-containing proteins exhibited remarkable rate enhancement for the reduction of H2O2 by GSH. The average GPx activity was found to be 462 U/micromol, and it was approximately 100 times that for unimprinted selenosubtilisin. Compared with ebselen, a well-known GPx mimic, an activity enhancement of 500-fold was observed. Detailed steady state kinetic studies demonstrated that the novel selenoenzyme followed a ping pong mechanism similar to the naturally occurring GPx. PMID- 18163572 TI - Structural and mechanical properties of UV-photo-cross-linked poly(N-vinyl-2 pyrrolidone) hydrogels. AB - Biocompatible poly( N-vinyl-2-pyrrolidone) (PVP) hydrogels have been produced by UV irradiation of aqueous polymer mixtures, using a high-pressure mercury lamp. The resulting materials have been characterized by a combination of experimental techniques, including rheology, small-angle neutron scattering (SANS), electron paramagnetic resonance (EPR), and pulsed gradient spin-echo nuclear magnetic resonance (PGSE-NMR), to put in evidence the relationship between the microstructural properties and the macrofunctional behavior of the gels. Viscoelastic measurements showed that UV photo-cross-linked PVP hydrogels present a strong gel mechanical behavior and viscoelastic moduli values similar to those of biological gels. The average distance between the cross-linking points of the polymer network was estimated from the hydrogels elastic modulus. However, SANS measurements showed that the network microstructure is highly inhomogeneous, presenting polymer-rich regions more densely cross-linked, surrounded by a water rich environment. EPR and PGSE-NMR data further support the existence of these water-rich domains. Inclusion of a third component, such as glycerol, in the PVP aqueous mixture to be irradiated has been also investigated. A small amount of glycerol (<3% w/w) can be added keeping satisfactory properties of the hydrogel, while higher amounts significantly affect the cross-linking process. PMID- 18163573 TI - In situ cross-linking of elastin-like polypeptide block copolymers for tissue repair. AB - Rapid cross-linking of elastin-like polypeptides (ELPs) with hydroxymethylphosphines (HMPs) in aqueous solution is attractive for minimally invasive in vivo implantation of biomaterials and tissue engineering scaffolds. In order to examine the independent effect of the location and number of reactive sites on the chemical cross-linking kinetics of ELPs and the mechanical properties of the resulting hydrogels, we have designed ELP block copolymers comprised of cross-linkable, hydrophobic ELP blocks with periodic Lys residues (A block) and aliphatic, hydrophilic ELP blocks with no cross-linking sites (B block); three different block architectures, A, ABA, and BABA were synthesized in this study. All ELP block copolymers were rapidly cross-linked with HMPs within several minutes under physiological conditions. The inclusion of the un-cross linked hydrophilic block, its length relative to the cross-linkable hydrophobic block, and the block copolymer architecture all had a significant effect on swelling ratios of the cross-linked hydrogels, their microstructure, and mechanical properties. Fibroblasts embedded in the ELP hydrogels survived the cross-linking process and remained viable for at least 3 days in vitro when the gels were formed from an equimolar ratio of HMPs and Lys residues of ELPs. DNA quantification of the embedded cells indicated that the cell viability within triblock ELP hydrogels was statistically greater than that in the monoblock gels at day 3. These results suggest that the mechanical properties of ELP hydrogels and the microenvironment that they present to cells can be tuned by the design of the block copolymer architecture. PMID- 18163574 TI - Gold nanoparticles induce surface morphological transformation in polyurethane and affect the cellular response. AB - Nanocomposites from a hexamethylene diisocyanate (HDI)-based polyester-type waterborne polyurethane (PU) containing different amounts (17.4-174 ppm) of gold (Au) nanoparticles (approximately 5 nm) were prepared. The microstructure and physiochemical properties of the nanocomposites were characterized. The cell attachment and proliferation, platelet activation, and bacterial adhesion on the nanocomposites were evaluated. Gold nanoparticles in small amounts induced significant changes in surface morphology and domain structures, from hard segment lamellae to soft segment micelles. These changes resembled the morphological transformation among different mesophases occurred in diblock copolymers. Better cellular proliferation, lower platelet activation, and reduced bacterial adhesion were demonstrated for the PU nanocomposite with 43.5 or 65 ppm of Au than the pure PU or the nanocomposite containing a different amount of Au. The different cellular response on PU-Au nanocomposites was attributed to the extensively modified surface morphology and phase separation in the presence of a small amount of Au nanoparticles. PMID- 18163575 TI - The physical action of cellulases revealed by a quartz crystal microbalance study using ultrathin cellulose films and pure cellulases. AB - The effects of fungal cellulases on model cellulose films were studied using a high-resolution quartz crystal microbalance (QCM) sensitive to minute changes of the nanometer thick model cellulose films. It was found that endoglucanases not only produce new end groups but also cause a swelling of the cellulose film. The cellobiohydrolases degraded the films quickly, which was detected as a rapid decrease in the remaining amount of cellulose on the QCM crystal. However, changing viscoelastic properties of the films also indicated a softening of the film during the degradation. A defined mixture of selected cellulases caused a significantly higher rate of degradation than only cellobiohydrolases. Cellulase synergism is discussed with the endoglucanase swelling effects and film softening added. PMID- 18163576 TI - Synthesis, characterization, and DNA adsorption studies of ampholytic model conetworks based on cross-linked star copolymers. AB - Five model conetworks based on cross-linked star ampholytic copolymers were synthesized by group transfer polymerization. The ampholytic copolymers were based on two hydrophilic monomers: the positively ionizable 2 (dimethylamino)ethyl methacrylate (DMAEMA) and the negatively ionizable methacrylic acid (MAA). Ethylene glycol dimethacrylate was used as the cross linker. These five ampholytic model conetworks were isomers based on equimolar DMAEMA-MAA copolymer stars of different architectures: heteroarm (two), star block (two), and statistical. The two networks based on the homopolymer stars were also synthesized. The MAA units were introduced via the polymerization of tetrahydropyranyl methacrylate and the acid hydrolysis of the latter after network formation. All the precursors to the (co)networks were characterized in terms of their molecular weights using gel permeation chromatography (GPC). The mass of the extractables from the (co)networks was measured and characterized in terms of molecular weight and composition using GPC and proton nuclear magnetic resonance (1H NMR) spectroscopy, respectively. The degrees of swelling (DS) of all the ampholytic conetworks were measured as a function of pH and were found to present a minimum at a pH value which was taken as the isoelectric point, pI. The DS and the pI values did not present a dependence on conetwork architecture. Finally, DNA adsorption studies onto the ampholyte conetworks indicated that DNA binding was governed by electrostatics. PMID- 18163577 TI - Reactivity of wheat gluten protein during mechanical mixing: radical and nucleophilic reactions for the addition of molecules on sulfur. AB - Mechanical properties of gluten-based biomaterials, such as break stress, were known to be influenced by temperature and shear stresses applied during processing. It is well documented in literature that these processing parameters promoted wheat gluten protein aggregation. Exchange between disulfide bonds and thiol groups oxidation are the postulated mechanisms that lead to gluten protein solubility loss in sodium dodecyl sulfate buffers. Both nucleophilic and radical reactions were postulated to act during gluten aggregation. To graft molecules on gluten, a study was carried out to explore the reactivity of its thiol and disulfide groups during thermomechanical mixing. A range of reactants able to react via radical or nucleophilic pathways with thiol groups were synthesized. Reactivity between gluten and functions was quantified by gluten solubility measurements. This investigation and literature observations allowed proposal of a general gluten aggregation mechanism during mixing. PMID- 18163578 TI - Biodegradable foam plastics based on castor oil. AB - In this work, a simple but effective approach was proposed for preparing biodegradable plastic foams with a high content of castor oil. First of all, castor oil reacted with maleic anhydride to produce maleated castor oil (MACO) without the aid of any catalyst. Then plastic foams were synthesized through free radical initiated copolymerization between MACO and diluent monomer styrene. With changes in MACO/St ratio and species of curing initiator, mechanical properties of MACO foams can be easily adjusted. In this way, biofoams with comparable compressive stress at 25% strain as commercial polyurethane (PU) foams were prepared, while the content of castor oil can be as high as 61 wt %. The soil burial tests further proved that the castor oil based foams kept the biodegradability of renewable resources despite the fact that some petrol-based components were introduced. PMID- 18163579 TI - Characterization of a gel in the cell wall to elucidate the paradoxical shrinkage of tension wood. AB - Wood behavior is characterized by high sensibility to humidity and strongly anisotropic properties. The drying shrinkage along the fibers, usually small due to the reinforcing action of cellulosic microfibrils, is surprisingly high in the so-called tension wood, produced by trees to respond to strong reorientation requirements. In this study, nitrogen adsorption-desorption isotherms of supercritically dried tension wood and normal wood show that the tension wood cell wall has a gel-like structure characterized by a pore surface more than 30 times higher than that in normal wood. Syneresis of the tension wood gel explains its paradoxical drying shrinkage. This result could help to reduce technological problems during drying. Potential applications in biomechanics and biomimetics are worth investigating, considering that, in living trees, tension wood produces tensile growth stresses 10 times higher than that of normal wood. PMID- 18163580 TI - FT-IR imaging spectroscopy of phase separation in blends of poly(3 hydroxybutyrate) with poly(L-lactic acid) and poly(epsilon-caprolactone). AB - The detection of phase separation and identification of miscibility in biopolymer blends is an important aspect for the improvement of their physical properties. In this article, the phase separation in blends of poly(3-hydroxybutyrate) (PHB) with poly(L-lactic acid) (PLA) and poly(epsilon-caprolactone) (PCL), respectively, has been studied as a function of the blend composition by FT-IR imaging spectroscopy. For both polymer blend systems, a miscibility gap has been found around the 50:50% (w/w) composition of the two components. Furthermore, the separating phases have been identified as blends of the two polymer components and their compositions could be determined from calibrations based on the spectra of the blends in the compositional range of miscibility. The data derived from FT IR spectroscopic imaging were corroborated by additional DSC analyses and mechanical stress-strain measurements of polymer blend films, which exhibited a characteristic fracture behavior as a function of PHB composition. PMID- 18163581 TI - Observation of the three-dimensional structure of actin bundles formed with polycations. AB - Three-dimensional structures of actin bundles formed with polycations were observed by using transmission electron microtomography and atomic force microscopy. We found, for the first time, that the cross-sectional morphology of actin bundles depends on the polycation species and ionic strength, while it is insensitive to the degree of polymerization and concentration of polycation. Actin bundles formed with poly-N-[3-(dimethylamino)propyl] acrylamide methyl chloride quaternary show a ribbon-like cross-sectional morphology in low salt concentrations that changes to cylindrical cross-sectional morphology with hexagonal packing of the actin filaments in high salt concentrations. Contrastingly, actin bundles formed with poly-L-lysine show triangular cross sectional morphology with hexagonal packing of the actin filaments. These variations in cross-sectional morphology are discussed in terms of anisotropy in the electrostatic energy barrier. PMID- 18163582 TI - Benzoic acid derivatives, acetophenones, and anti-inflammatory constituents from Melicope semecarpifolia. AB - Two new benzoic acid derivatives, (E)-3-acetyl-6-(3,7-dimethylocta-2,6-dienyloxy) 2,4-dihydroxybenzoic acid (1) and (E)-3-acetyl-4-(3,7-dimethylocta-2,6-dienyloxy) 2,6-dihydroxybenzoic acid (2), and three new acetophenones, (E)-1-(5-(3,7 dimethylocta-2,6-dienyloxy)-7-hydroxy-2,2-dimethyl-2 H-chromen-8-yl)ethanone (3), (E)-1-(5-(3,7-dimethylocta-2,6-dienyloxy)-7-hydroxy-2-methyl-2-(4-methylpent-3 enyl)-2 H-chromen-8-yl)ethanone (4), and (R,E)-1-(5-(3,7-dimethylocta-2,6 dienyloxy)-3,7-dihydroxy-2,2-dimethylchroman-8-yl)ethanone (5), have been isolated from the fruits of Melicope semecarpifolia, together with eight known compounds. The structures were determined through in-depth NMR and mass spectrometric analyses. Among the isolated compounds, 2-(1'-geranyloxy)-4,6,beta trihydroxyacetophenone (8), 4-(1'-geranyloxy)-2,6,beta-trihydroxyacetophenone (9), 5-hydroxy-3,7,3',4'-tetramethoxyflavone (10), 5,4'-dihydroxy-3,7,3' trimethoxyflavone (11), and 5,4'-dihydroxy-3,7-dimethoxyflavone (12) exhibited potent inhibition (IC50<4 microg/mL) on superoxide anion generation and elastase release by human neutrophils in response to fMet-Leu-Phe/cytochalasin B. PMID- 18163583 TI - Fe2+-catalyzed formation of nitriles and thionamides from intact glucosinolates. AB - The ratio of isothiocyanates to nitriles formed upon the hydrolysis of glucosinolates is a key factor determining the physiological effect of glucosinolate-containing plants and materials. A micellar electrokinetic capillary chromatography (MECC) method was used to study the nonenzymatic Fe2+ catalyzed transformation of glucosinolates. At room temperature, pH 5, and in the presence of only 2 molar excess of Fe2+ all glucosinolate was degraded in 24 h. At all molar excess Fe2+ tested, nitriles were the major compounds formed. Thionamides were also formed from glucosinolates that contained a side chain hydroxylated at C-2; in this case, trace amounts of oxazolidine-2-thione were also detected. The presence of Fe3+ had no effect. The nonenzymatic Fe2+ catalyzed transformation of glucosinolates involves the binding of Fe2+ to the glucosinolate to form a complex. PMID- 18163584 TI - Symplocamide A, a potent cytotoxin and chymotrypsin inhibitor from the marine Cyanobacterium Symploca sp. AB - Investigation of a Symploca sp. from Papua New Guinea has led to the isolation of symplocamide A (1), a potent cancer cell cytotoxin, which also inhibits serine proteases with a 200-fold greater inhibition of chymotrypsin over trypsin. The complete stereostructure of symplocamide A was determined by detailed NMR and MS analysis as well as chiral HPLC analysis of the component amino acid residues. The presence of several unusual structural features in symplocamide A provides new insights into the pharmacophore model for protease selectivity in this drug class and may underlie the potent cytotoxicity of this compound to H-460 lung cancer cells (IC50=40 nM) as well as neuro-2a neuroblastoma cells (IC50=29 nM). PMID- 18163585 TI - Taxanes from Shells and Leaves of Corylus avellana. AB - Paclitaxel is an effective antineoplastic agent originally extracted in low yield from the bark of Taxus brevifolia. Although it was generally considered a particular metabolite of Taxus sp., paclitaxel was recently found in hazel cell cultures. The aim of the present work was to verify whether hazel differentiated tissues could be used as a commercial source of paclitaxel and other taxanes. Thus, shells and leaves of hazel plants were analyzed by ELISA and HPLC-MS. Both shell and leaf extracts contained taxanes. Among these, paclitaxel, 10 deacetylbaccatin III, baccatin III, paclitaxel C, and 7-epipaclitaxel were identified and quantified. Hazel extracts also showed biological activity, inhibiting metaphase to anaphase transition in a human tumor cell line. The level of total taxanes in leaves was higher than in shells collected in the same period from the same plants. However, the finding of these compounds in shells, which are considered discarded material and are mass produced by many food industries, is of interest for the future availability of paclitaxel and other antineoplastic compounds. PMID- 18163586 TI - Polydiscamides B-D from a marine sponge Ircinia sp. as potent human sensory neuron-specific G protein coupled receptor agonists. AB - Polydiscamides B, C, and D (1-3) were isolated from a sponge Ircinia sp. The structures of 1 to 3 were elucidated by the comparison of their NMR and HRESIMS spectroscopic data with that of a structurally related compound, polydiscamide A. All compounds showed potent agonist activity against human sensory neuron specific G protein couple receptor (SNSR), a receptor involved in the modulation of pain, and they are the first examples of nonendogenous human SNSR agonists. PMID- 18163587 TI - Pim2 inhibitors from the Papua New Guinean plant Cupaniopsis macropetala. AB - Bioassay-guided fractionation of an organic extract from the leaves of Cupaniopsis macropetala resulted in the isolation of a new alkaloid, galloyl tyramine ( 1), together with the known flavonoid glycoside quercitrin ( 2). The structure of 1 was determined following 1D and 2D NMR, IR, UV, and MS data analysis. Compounds 1 and 2 displayed IC 50 values of 161 and 25 microM, respectively, in a Pim2 enzyme assay. PMID- 18163588 TI - Gentisyl alcohol derivatives from the marine-derived fungus Penicillium terrestre. AB - Nine new gentisyl alcohol derivatives, namely, the trimeric terrestrol A (8), dimeric terrestrols B-H (1-7), and a monomeric derivative (12), together with four known analogues (9-11, 13) were isolated from the marine-derived fungus Penicillium terrestre. The structures of the new compounds were elucidated by spectroscopic methods including one- and two-dimensional NMR as well as low- and high-resolution mass spectrometric analysis. These new compounds (1-8, 12) showed cytotoxic effects on HL-60, MOLT-4, BEL-7402, and A-549 cell lines with IC50 values in the range 5-65 microM. Compound 6 also showed moderate inhibitory activity against protein tyrosine kinases (Src and KDR). Furthermore, all new compounds exhibited moderate radical scavenging activity against DPPH with IC50 values in the range 2.6-8.5 microM. PMID- 18163589 TI - Antileukemic activity of genistein, a major isoflavone present in soy products. AB - Soy has been used in traditional medicine for the treatment of various diseases, including cancer. The isoflavones present in soy have been shown in animal models to have cancer-preventing activity. However, the therapeutic effects of isoflavones against cancer are still unclear. We have evaluated the in vitro and in vivo antileukemic activity of genistein (1), a major isoflavone present in soy. We observed that it produced a dose- and time-dependent antineoplastic activity against myeloid and lymphoid leukemic cell lines. In addition, genistein treatment of the leukemic cells reactivated tumor suppressor genes that were silenced by aberrant DNA methylation. A genistein-enriched diet produced a moderate, but significant, antileukemic effect in mice. The limited extent of this in vivo response may have been due to the rapid metabolic inactivation of genistein in mice. Due to the longer half-life of genistein in humans, a soy enriched diet has the potential to produce plasma levels of this isoflavone in the range of the concentrations used in vitro that produced an antileukemic activity. PMID- 18163590 TI - Isolation of cytotoxic metabolites from targeted peruvian amazonian medicinal plants. AB - The antiproliferative bioassay-guided fractionation of five Peruvian plants, Doliocarpus dentatus, Picramnia sellowii, Strychnos mitscherlichii, Iryanthera juruensis, and Croton alnifolius, led to the isolation and identification of their different major cytotoxic constituents, betulinic acid (1), nataloe-emodin (2), bisnordihydrotoxyferine (4), 2',4'-dihydroxy-6'-methoxy-3,4 methylenedioxydihydrochalcone (5), and 2',4'-dihydroxy-4,6' dimethoxydihydrochalcone (6) and 12-O-tetradecanoylphorbol-13-acetate (7), respectively. Eight human tumor cell lines and two nontumorigenic cell lines were used in this investigation. Their in vitro activity against Mycobacterium tuberculosis is also reported. PMID- 18163592 TI - Alkaloids from stems and leaves of Stemona japonica and their insecticidal activities. AB - Five new alkaloids, 6beta-hydroxystemofoline (1), 16-hydroxystemofoline (2), neostemofoline (3), protostemodiol (4), and 13-demethoxy-11(S*),12(R*) dihydroprotostemonine (5), along with 10 known alkaloids, were isolated from stems and leaves of Stemona japonica. Their structures were elucidated by 1D and 2D NMR and other spectroscopic studies. The insecticidal activity of the agonist 16-hydroxystemofoline (2) and antagonist 13-demethoxy-11(S*),12(R*) dihydroprotostemonine (5) was demonstrated by electrophysiological in vitro tests on the insect nicotinic acetylcholine receptor and by in vivo screenings against relevant agricultural insect pests. PMID- 18163591 TI - Antiproliferative cassane diterpenoids of Cordyla madagascariensis ssp. madagascariensis from the Madagascar rainforest. AB - Bioassay-guided fractionation of an ethanol extract of a Madagascar collection of the fruits of Cordyla madagascariensis ssp. madagascariensis led to the isolation of the four new cassane diterpenoids 1-4. The 1H and 13C NMR spectra of all compounds were fully assigned using a combination of 2D NMR experiments, including COSY, HSQC, HMBC, and ROESY sequences. All of the isolates were tested against the A2780 human ovarian cancer cell line, and compounds 1 and 2 showed mild antiproliferative activity with IC50 values of 10 and 36 microM, respectively. PMID- 18163593 TI - Microwave-assisted fluorous synthesis of 2-aryl-substituted 4-thiazolidinone and 4-thiazinanone libraries. AB - A straightforward two-step protocol for the synthesis of 2-aryl-substituted 4 thiazolidinone and 4-thiazinanone libraries has been developed. The one-pot, three-component reactions of fluorous benzaldehydes with amines and mercaptoacetic acid or mecaptopropanoic acid produce the heterocyclic systems. Intermediates purified by fluorous solid-phase extraction are subject to microwave-assisted palladium-catalyzed coupling reactions to simultaneously cleave the fluorous tag and introduce the biaryl and thioaryl functional groups to the 2-position of 4-thiazolidinones and 4-thiazinanones. PMID- 18163594 TI - Studies towards the synthesis of methionine aminopeptidase inhibitors: diversification utilizing a ROMP-derived coupling reagent. AB - Efforts to synthesize potential methionine aminopeptidase inhibitors is described. Preliminary SAR and docking studies served as a guide to design the compound libraries. "Chromatography-free" synthesis of various heterocyclic amides was realized by using a high-load, soluble coupling reagent derived via ring-opening metathesis polymerization (ROMP). Subsequent microwave-assisted Suzuki reactions with ortho-substituted arylboronic acids, followed by chromatographic purification afforded a 55-member library in high yields and purities. While the biological testing was not satisfactory, concurrent X-ray crystallography studies revealed key structural features essential for inhibition of methionine aminopeptidase, which directed fruitful results reported in the accompanying manuscript. In addition, in silico Lipinksi profiles and ADME properties of the library are also reported. PMID- 18163595 TI - Ionic immobilization, diversification, and release: application to the generation of a library of methionine aminopeptidase inhibitors. AB - Development of an ionic immobilization, diversification, and release method for the generation of methionine aminopeptidase inhibitors is reported. This method involves the immobilization of 5-bromofuran-2-carboxylic acid and 5 bromothiophene-2-carboxylic acid onto PS-BEMP, followed by Suzuki reaction on a resin-bound intermediate and subsequent release to provide products in moderate yields and excellent purities. Compound potencies were evaluated on the Co(II), Mn(II), Ni(II), and Fe(II) forms of Escherichia coli MetAP1. The furoic-acid analogs were found to be Mn(II) selective with IC 50 values in the low micromolar range. Qualitative SAR analysis, supplemented by molecular modeling studies, provides valuable information on structural elements responsible for potency and selectivity. PMID- 18163596 TI - Frequency-coded chemical sensors. AB - We have developed a new method to intelligently sample analytes and introduce the analytes to sensors. The method automatically adjusts sampling duration according to the sensors' response to the analytes and converts the amplitude of the sensor output to a frequency output, giving us another opportunity to reduce noise in the signal. It also addresses some of the common sensor issues such as response time, saturation, chemical dynamic range, and sensor protection, saving precious detection time, protecting sensors, and enabling sensitive sensors built for low concentration detection to be used for high-concentration detection as well. We have put together a system using a tuning fork chemical sensor as a sample sensor to demonstrate the feasibility and benefits of the new sensing technique. PMID- 18163597 TI - Elimination of systematic mass measurement errors in liquid chromatography-mass spectrometry based proteomics using regression models and a priori partial knowledge of the sample content. AB - The high mass measurement accuracy and precision available with recently developed mass spectrometers is increasingly used in proteomics analyses to confidently identify tryptic peptides from complex mixtures of proteins, as well as post-translational modifications and peptides from nonannotated proteins. To take full advantage of high mass measurement accuracy instruments, it is necessary to limit systematic mass measurement errors. It is well known that errors in m/z measurements can be affected by experimental parameters that include, for example, outdated calibration coefficients, ion intensity, and temperature changes during the measurement. Traditionally, these variations have been corrected through the use of internal calibrants (well-characterized standards introduced with the sample being analyzed). In this paper, we describe an alternative approach where the calibration is provided through the use of a priori knowledge of the sample being analyzed. Such an approach has previously been demonstrated based on the dependence of systematic error on m/z alone. To incorporate additional explanatory variables, we employed multidimensional, nonparametric regression models, which were evaluated using several commercially available instruments. The applied approach is shown to remove any noticeable biases from the overall mass measurement errors and decreases the overall standard deviation of the mass measurement error distribution by 1.2-2-fold, depending on instrument type. Subsequent reduction of the random errors based on multiple measurements over consecutive spectra further improves accuracy and results in an overall decrease of the standard deviation by 1.8-3.7-fold. This new procedure will decrease the false discovery rates for peptide identifications using high-accuracy mass measurements. PMID- 18163598 TI - Study of hydrogen and deuterium emission characteristics in laser-induced low pressure helium plasma for the suppression of surface water contamination. AB - An experimental study was conducted in search of the experimental condition required for the much needed suppression of spectral interference caused by surface water in hydrogen analysis using laser-induced low-pressure helium plasma spectroscopy. The problem arising from the difficulty in distinguishing hydrogen emission from hydrogen impurity inside the sample and that coming from the water molecules was overcome by taking advantage of similar emission characteristics shared by hydrogen and deuterium demonstrated in this experiment by the distinct time-dependent and pressure-dependent variations of the D and H emission intensities from the D-doped zircaloy-4 samples. This similarity allows the study of H impurity emission in terms of D emission from the D-doped samples and thereby separating it from the H emission originating from the water molecules. Employing this strategy has allowed us to achieve the large suppression of water induced spectral interference from the previous minimum of 400 microg/g to the current value of 30 microg/g when a laser beam of 34 mJ under tight focusing condition was employed. Along with this favorable result, this experimental condition has also provided a much better (about 6-fold higher) spatial resolution, although these results were achieved at the expense of reducing the linear calibration range from the previous 4 300 microg/g to the present 200 microg/g. PMID- 18163599 TI - Theoretical study and rate constant computation on the reaction HFCO + OH --> CFO + H2O. AB - The potential energy surface, including the geometries and frequencies of the stationary points, of the reaction HFCO + OH is calculated using the MP2 method with 6-31+G(d,p) basis set, which shows that the direct hydrogen abstraction route is the most dominating channel with respect to addition and substitution channels. For the hydrogen abstraction reaction, the single-point energies are refined at the QCISD(T) method with 6-311++G(2df,2pd) basis set. The calculated standard reaction enthalpy and barrier height are -17.1 and 4.9 kcal mol(-1), respectively, at the QCISD(T)/6-311++G(2df,2pd)//MP2/6-31+G(d,p) level of theory. The reaction rate constants within 250-2500 K are calculated by the improved canonical variational transition state theory (ICVT) with small-curvature tunneling (SCT) correction at the QCISD(T)/6-311++G(2df,2pd)//MP2/6-31+G(d,p) level of theory. The fitted three-parameter formula is k = 2.875 x 10(-13) (T/1000)1.85 exp(-325.0/T) cm(3) molecule(-1) s(-1). The results indicate that the calculated ICVT/SCT rate constant is in agreement with the experimental data, and the tunneling effect in the lower temperature range plays an important role in computing the reaction rate constants. PMID- 18163600 TI - Mass analyzed threshold ionization spectroscopy of o-, m-, and p dichlorobenzenes. Influence of the chlorine position on vibrational spectra and ionization energy. AB - For the first time, vibrational spectra of the 35Cl2 and 35Cl37Cl isotopomers of o-, m-, and p-dichlorobenzene cations in the electronic ground state have been measured via S1 intermediate states by mass analyzed threshold ionization (MATI) spectroscopy. Additionally, ab initio calculations at DFT (density functional theory), CIS (configuration interaction singles), and CASSCF (complete active space self-consistent field) levels of theory have been conducted to compare experimental findings with theory. From the MATI spectra, adiabatic ionization energies of the ortho, meta, and para isomers have been determined to be the same for each pair of investigated isotopomers to 73,237 +/- 6, 72,191 +/- 6, and 73,776 +/- 6 cm(-1), respectively. Several vibrational modes, including fundamentals, combinations, and progressions have been assigned by comparing the experimental and theoretical results. The appearance of overtone progressions involving the 7a mode could be explained by a geometry change of all three isomers during ionization in the direction of this mode by retraining the symmetry of the molecules. Although the general spectral features of the investigated isotopomers are similar, frequencies of some vibrations are slightly different up to a few wavenumbers depending on the involvement of the chlorine atoms in the molecular motion. PMID- 18163601 TI - Photophysics and nonlinear optical properties of tetra- and octabrominated silicon naphthalocyanines. AB - The effect of the number of bromide substituents on the photophysical and nonlinear optical properties of the tetrabrominated naphthalocyanine Br4(tBu2PhO)4NcSi[OSi(Hex)3]2 (1) and the octabrominated naphthalocyanine Br8NcSi[OSi(Hex)3]2 (2) has been investigated through various spectroscopic techniques. Absorption and emission of 1 and 2 have been studied at room temperature and 77 K to determine the spectral properties of the ground and the excited states and the lifetimes and quantum yields of formation of the excited states. There is a moderate increase of the quantum yield of the triplet excited state formation (PhiT = 0.10 vs 0.13) and a decrease of the triplet excited-state lifetime (tauT approximately 70 vs 50 mus) from 1 to 2. These can be attributed to the stronger heavy atom effect produced by the larger number of peripheral bromide substituents in 2 considering that an excited state with a triplet manifold is involved in the excitation dynamics of both complexes 1 and 2. The quantum yields of the singlet oxygen formation (PhiDelta) upon irradiation of 1 and 2 at 355 nm were also evaluated, and a value of PhiDelta(1) = PhiDelta(2) = 0.16 was obtained. In addition to that, octabrominated complex 2 displays a larger decrease of nonlinear optical transmission for nanosecond pulses at 532 nm with respect to the tetrabrominated complex 1. The nanosecond Z-scan experiments reveal that 1 and 2 exhibit both a reverse saturable absorption and a nonlinear refraction at 532 nm. However, both the sign and the magnitude of the nonlinear refraction change from 1 to 2. For picosecond Z-scan in the visible spectral region, these two complexes exhibit only reverse saturable absorption, and the excited-state absorption cross-section increases at longer wavelengths. PMID- 18163602 TI - Infrared spectroscopy of discrete uranyl anion complexes. AB - The Free-Electron Laser for Infrared Experiments (FELIX) was used to study the wavelength-resolved multiple photon photodissociation of discrete, gas-phase uranyl (UO22+) complexes containing a single anionic ligand (A), with or without ligated solvent molecules (S). The uranyl antisymmetric and symmetric stretching frequencies were measured for complexes with general formula [UO2A(S)n]+, where A was hydroxide, methoxide, or acetate; S was water, ammonia, acetone, or acetonitrile; and n = 0-3. The values for the antisymmetric stretching frequency for uranyl ligated with only an anion ([UO2A]+) were as low or lower than measurements for [UO2]2+ ligated with as many as five strong neutral donor ligands and are comparable to solution-phase values. This result was surprising because initial DFT calculations predicted values that were 30-40 cm(-1) higher, consistent with intuition but not with the data. Modification of the basis sets and use of alternative functionals improved computational accuracy for the methoxide and acetate complexes, but calculated values for the hydroxide were greater than the measurement regardless of the computational method used. Attachment of a neutral donor ligand S to [UO2A]+ produced [UO2AS]+, which produced only very modest changes to the uranyl antisymmetric stretch frequency, and did not universally shift the frequency to lower values. DFT calculations for [UO2AS]+ were in accord with trends in the data and showed that attachment of the solvent was accommodated by weakening of the U-anion bond as well as the uranyl. When uranyl frequencies were compared for [UO2AS]+ species having different solvent neutrals, values decreased with increasing neutral nucleophilicity. PMID- 18163603 TI - Modeling anti-Trypanosoma cruzi activity of N-oxide containing heterocycles. AB - In the present study a systematic approach was used to model the anti-T. cruzi activity of a series of N-oxide containing heterocycles belonging to four chemical families with a wide structural diversity. The proposed mode of action implies the reduction of the N-oxide moiety; however, the biochemical mechanism underlying the anti-T. cruzi activity is still unkown. For structural representation two types of descriptors were analyzed: quantum chemical (AM1) global descriptors and properties coded by radial distribution function (RDF). Both types of descriptors point to the relevance of electronic properties. The local-RDF (LRDF) identified an electrophilic center at 4.1-4.9 A from the oxygen atom of the N-oxide moiety, although other properties are required to explain the biological activity. While the mode of action of N-oxide containing heterocycles is still unknown, the results obtained here strengthen the importance of the electrophilic character of the molecule and the possible participation of the heterocycle in a reduction process. The ability of these descriptors to distinguish among activity classes was assessed using Kohonen neural networks, and the best clustering descriptors were later used for model building. Different learning algorithms were used for model development, and stratified 10-fold cross validation was used to evaluate the performance of each classifier. The best results were obtained using k-nearest neighbors (k-NN) and decision tree (J48) methods combined with global descriptors. Since tree-based methods are easily translated into classification rules, the J48 model is a useful tool in the de novo construction of new N-oxide containing heterocycle lead structures. PMID- 18163604 TI - Surface structure relaxation of poly(methyl methacrylate). AB - Surface structure relaxations caused by temperature changes at the free surface of poly(methyl methacrylate) were studied using IR-visible sum-frequency generation (SFG). A polarization-rotating technique was introduced to enhance the sensitivity of SFG for monitoring the surface structure relaxations during a cooling process. A new surface structure relaxation was observed at 67 degrees C. This temperature does not match any known structure relaxation temperatures for the bulk and is 40 degrees C below the bulk glass transition temperature. As expected for a free-surface phenomenon, the surface relaxation temperature was found to be independent of film thickness in the range of 0.1-0.5 microm. PMID- 18163605 TI - Fluorescence emission of disperse Red 1 in solution at room temperature. AB - In this article, we report the fluorescence emission of Disperse Red 1 in solution at room temperature and pumping at 532 nm with a 25 mW diode laser. We have measured its fluorescence quantum yield in methanol, ethylene glycol, glycerol, and phenol obtaining values as high as 10(-3) in the aliphatic alcohols. The excitation spectra of Disperse Red 1 in all four solvents as well as its excitation anisotropy in glycerol are presented. Applying a Gaussian decomposition method to the absorption spectra along with the support from the excitation spectra, the positions of the different transitions in this pseudo stilbene azobenzene dye were determined. Solvatochromic and isomerization constraint effects are discussed. Calculations using density functional theory at TD-B3LYP/6-31G*//HF/6-31G* level were performed to interpret the experimental observations. PMID- 18163606 TI - Prediction of partition coefficients by multiscale hybrid atomic-level/coarse grain simulations. AB - Coarse-grain models are becoming an increasingly important tool in computer simulations of a wide variety of molecular processes. In many instances it is, however, desirable to describe key portions of a molecular system at the atomic level. There is therefore a strong interest in the development of simulation methodologies that allow representations of matter with mixed granularities in a multiscale fashion. We report here a strategy to conduct mixed atomic-level and coarse-grain simulations of molecular systems with a recently developed coarse grain model. The methodology is validated by computing partition coefficients of small molecules described in atomic detail and solvated by water or octane, both of which are represented by coarse-grain models. Because the present coarse-grain force field retains electrostatic interactions, the simplified solvent particles can interact realistically with the all-atom solutes. The partition coefficients computed by this approach rival the accuracy of fully atomistic simulations and are obtained at a fraction of their computational cost. The present methodology is simple, robust and applicable to a wide variety of molecular systems. PMID- 18163607 TI - Time-resolving analysis of cryotropic gelation of water/poly(vinyl alcohol) solutions via small-angle neutron scattering. AB - The structural transformations occurring in initially homogeneous aqueous solutions of poly(vinyl alcohol) (PVA) through application of freezing (-13 degrees C) and thawing (20 degrees C) cycles is investigated by time resolving small-angle neutron scattering (SANS). These measurements indicate that formation of gels of complex hierarchical structure arises from occurrence of different elementary processes, involving different length and time scales. The fastest process that could be detected by our measurements during the first cryotropic treatment consists of the crystallization of the solvent. However, solvent crystallization is incomplete, and an unfrozen liquid microphase more concentrated in PVA than the initial solution is also formed. Crystallization of PVA takes place inside the unfrozen liquid microphase and is slowed down because of formation of a microgel fraction. Water crystallization takes place in the early 10 min of the treatment of the solution at subzero temperatures, and although below 0 degrees C the PVA solutions used for preparation of cryogels should be below the spinodal curve, occurrence of liquid-liquid phase separation could not be detected in our experiments. Upon thawing, ice crystals melt, and transparent gels are obtained that become opaque in approximately 200 min, due to a slow and progressive increase of the size of microheterogeneities (dilute and dense regions) imprinted during the fast freezing by the crystallization of water. During the permanence of these gels at room temperature (for hours), the presence of a high content of water (higher than 85% by mass) prevents further crystallization of PVA. Crystallization of PVA, in turn, is resumed by freezing the gels at subzero temperatures, after water crystallization and consequent formation of an unfrozen microphase. The kinetic parameters of PVA crystallization during the permanence of these gels at subzero temperatures are the same shown by PVA during the first freezing step of the solutions. PMID- 18163608 TI - Chromophore/DNA interactions: femto- to nanosecond spectroscopy, NMR structure, and electron transfer theory. AB - The mechanism of photoinduced hole injection into DNA has been studied using an integrated approach that combines NMR structural analysis, time-resolved spectroscopy, and quantum-chemical calculations. A covalently linked acridinium derivative, the protonated 9-amino-6-chloro-2-methoxyacridine (X+), is replacing a thymine and separated from either guanine (G) or the easier to oxidize 7 deazaguanine (Z) by one adenine.thymine (A.T) base pair. The key features of this donor/acceptor system are the following: (i) In more than 95% of the duplexes, X+ is located in a central, coplanar position between the neighboring A.T base pairs with its long axis in parallel showing minimal twist and tilt angles (<15 degrees). The complementary adenine base is turned out into the extrahelical space. In a minority of less than 5%, X+ is found to be still attached to the duplex. X+ is most probably associated with one of the phosphates, since it is neither intercalated between more remote base pairs nor bound to sugars or grooves. This minority characterized by an excited state lifetime >10 ns gives rise to a small background signal in time-resolved measurements and contributes predominantly to steady-state fluorescence spectra. (ii) Although the intercalation mode of X+ is well defined, the NMR structure reveals that there are two conformations of X+ with respect to the arrangement of its methoxy substituent. In one conformation, the methoxy group is in the plane of the chromophore, while, in the other extraplanar conformation, the methoxy group forms an angle of 70 degrees with the acridinium ring. The fluorescence decay of 5'-ZAX and 5'-GAX tracts can be fitted to a biexponential function with similar amplitudes, reflecting the oxidation dynamics of G and Z, with the slower rate being determined by larger thermal activation energy. The attribution of biexponential electron transfer (ET) dynamics to the bimodal orientation of the methoxy group at the acridinium is supported by quantum-chemical calculations. These predict a larger free energy change for hole transfer in the nonplanar conformation as compared to the planar one, whereas the difference in the electronic couplings is negligible. (iii) Kinetic studies of the directionality of the 1(X+)* induced hole injection reveal similarly fast decay components in both directions of the duplex, that is, in 5'-ZAX and 5'-XAZ, with the amplitude of the fast component being significantly reduced in 5'-XAZ. The NMR structure shows that local structural deviations from B-DNA are much more pronounced in the 3'-5' direction than in the 5'-3' direction. According to quantum-chemical calculations, the directionality of charge injection is not a universal feature of the DNA duplex but depends critically on the rotation angle of the aromatic plane of the acridinium within the pi stack. The arrangement of X+ in 5'-ZAX and 5'-XAZ corresponds to a conformation with weak directionality of the electronic couplings. The increased disorder in the 3'-5'direction favors slow hole transfer components at the expense of the fast ones. (iv) A comparison of the hole transfer in 5'-GAX and 5'-ZAG shows that classical Marcus theory can explain the ratio of the charge shift rates of more than 2 orders of magnitude on the basis of a free energy difference between G and Z of 0.3 eV. Both NMR structures and quantum-chemical calculations justify the appreciable neglect of differences of electronic couplings as well as in the reorganization energy in 5'-GAX and 5' ZAG. Despite the attractive concept for the behavior of floppy DNA oligonucleotides, in this acridinium/DNA system, there is no evidence for conformational gating, that is, for fluctuations in the electronic couplings that permit the ET to occur. PMID- 18163609 TI - Effect of nonpolar solvents on the solute rotation and solvation dynamics in an imidazolium ionic liquid. AB - Recognizing the potential of the mixed solvent systems comprising ionic liquid as one of the constituents in real applications, the steady-state and time-resolved fluorescence behavior of C153 has been studied in neat 1-butyl-3 methylimidazolium hexafluorophosphate and its mixtures with nonpolar solvents, namely, toluene and 1,4-dioxane. No significant effect of the cosolvent on the steady-state absorption or fluorescence spectra of C153 in ionic liquid has been observed. Time-resolved fluorescence anisotropy measurements show a decrease of the rotational correlation time of C153 with gradual addition of the cosolvent. Solvation dynamics in ionic liquid-cosolvent mixtures is found to be biphasic, and a decrease of the average solvation time is observed with increasing amount of the cosolvent in solution. The time-zero spectrum of C153 is found to shift toward higher energy with gradual addition of the nonpolar solvent, suggesting that the probe molecule experiences a more nonpolar environment at the early stage of the dynamics in mixed solvents. The blue shift of the time-zero spectrum caused by the addition of the nonpolar solvent results in a larger Stokes shift of the time-dependent spectra due to solvent relaxation in mixed solvents. A comparison of the time-dependent spectral data of the ionic liquid-toluene and ionic liquid-dioxane systems shows that, while a small amount of toluene can significantly affect the dynamics, comparatively, a larger amount of dioxane is required to bring about the same effect. This is explained in terms of favorable interactions between toluene and the imidazolium ring system leading to a more effective solubilization of toluene in the cybotactic region of the probe. PMID- 18163610 TI - Designing carbon nanotube membranes for efficient water desalination. AB - The transport of water and ions through membranes formed from carbon nanotubes ranging in diameter from 6 to 11 A is studied using molecular dynamics simulations under hydrostatic pressure and equilibrium conditions. Membranes incorporating carbon nanotubes are found to be promising candidates for water desalination using reverse osmosis, and the size and uniformity of tubes that is required to achieve a desired salt rejection is determined. By calculating the potential of mean force for ion and water translocation, we show that ions face a large energy barrier and will not pass through the narrower tubes studied ((5,5) and (6,6) "armchair" type tubes) but can pass through the wider (7,7) and (8,8) nanotubes. Water, however, faces no such impediment due to the formation of stable hydrogen bonds and crosses all of the tubes studied at very large rates. By measuring this conduction rate under a hydrostatic pressure difference, we show that membranes incorporating carbon nanotubes can, in principle, achieve a high degree of desalination at flow rates far in excess of existing membranes. PMID- 18163611 TI - Salts of methylated 5-aminotetrazoles with energetic anions. AB - 1-methyl-5-aminotetrazole (4, MAT) can easily be protonated by strong acids, yielding known but largely uninvestigated 1-methyl-5-aminotetrazolium nitrate (4a) and perchlorate (4b). Methylation, rather than protonation, of 4 with iodomethane followed by the exchange of the iodide (5a) for nitrate (5b), perchlorate (5c), azide (5d), and dinitramide (5e) yields a new family of energetic methylated aminotetrazole salts. In all cases, stable salts were obtained and fully characterized by vibrational (IR, Raman) spectroscopy, multinuclear NMR spectroscopy, mass spectrometry, elemental analysis, and X-ray structure determination. Compounds 4a, 4b, and 5c crystallize in the monoclinic space group P2(1)/n, whereas compounds 5b and 5e crystallize in the orthorhombic space group P2(1)2(1)2(1) and 5d in the orthorhombic Fddd. Initial safety testing (impact, friction, and electrostatic sensitivity) and thermal stability measurements (DSC) were also carried out. The MAT salts all exhibit good thermal stabilities (decomposition above 150 degrees C). The constant volume energies of combustion (DeltacU) of 4a, 5b, 5d, and 5e were determined to be -2510(10) cal/g, -3190(30) cal/g, -4500(100) cal/g, and -2570(70) cal/g, respectively, experimentally using oxygen bomb calorimetry. From the experimentally determined density, chemical composition and energies of formation (back calculated from the heats of combustion), the detonation pressures and velocities of 4a (8100 m/s, 25.6 GPa), 5b (7500 m/s, 20.2 GPa), 5d (8200 m/s, 21.7 GPa), and 5e (7500 m/s, 21.2 GPa) were predicted using the EXPLO5 code. PMID- 18163612 TI - Synthesis, structure, and bonding of Sc4MgxCu15-xGa approximately 7.5 (x=0, 0.5). Two incommensurately modulated scandium substitution derivatives of cubic Mg2Cu6Ga5. AB - The substitution of scandium for magnesium in Mg2Cu6Ga5 (Mg2Zn11-type) yields an irrational superstructure phase that includes the refined compositions, Sc4Mg0.50(2)Cu14.50(2)Ga7.61(2) and Sc4Cu14.76(2)Ga7.51(2). These crystallize in Cmmm, a=approximately 8.31 A, b=approximately 21.72 A, c=approximately 8.30 A. The structures feature Sc2 dimers, Cu6 octahedra, a 3D CuGa (Cu12Ga2) framework, and arachno gallium-centered Cu4Ga6 icosahedra that are condensed into zigzag chains. The arrangement of these building blocks exhibits a topologic relationship to Mg2Cu6Ga5. Further studies reveal that the quaternary compound exhibits incommensurate modulations along a, with q=(0.694, 0, 0). Structure refinements with superspace group Xmmm(a00)000 led to saw-tooth modulations for two fractional or mixed sites that avoid short Cu-Ga distances. Band structure analyses reveal that the Fermi surface and bonding are sensitive to the incommensurately modulated atoms. PMID- 18163613 TI - Counterions influence reactivity of metal ions with cysteinyldopa model compounds. AB - Cysteinyldopas are naturally occurring conjugates of cysteine and dopa (3,4 dihydroxy-l-phenylalanine) that are precursors to red pheomelanin pigments. Metal ions are known to influence pheomelanogenesis in vitro and may be regulatory factors in vivo. Cydo (3-[(2-amino-ethyl)sulfanyl]-4,6-di-tert-butylbenzene-1,2 diol) and CarboxyCydo (2-amino-3-(4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) propionic acid) are model compounds of cysteinyldopa that retain its metal binding functionalities but cannot polymerize due to the presence of blocking tert-butyl groups. Cydo reacts readily with zinc(II) acetate or nickel(II) acetate to form a cyclized 1,4-benzothiazine (zine) intermediate that undergoes ring contraction to form benzothiazole (zole) unless it is stabilized by coordination to a metal ion. The crystal structure of [Ni(zine)2] is reported. The acetate counteranion is required for the zinc-promoted reactivity, as neither zinc(II) sulfate nor zinc(II) chloride alone promotes the transformation. The counterion is less important for redox-active copper and iron, which both readily promote the oxidation of Cydo to zine and zole species; Cu(II) complexes of both zine and zole have been characterized by X-ray crystallography. In the case of CarboxyCydo, a 3-carboxy-1,4-benzothiazine intermediate decarboxylates to form [Cu(zine)2] under basic conditions, but in the absence of base forms a mixture of products that includes the carboxylated dimer 2,2'-bibenzothiazine (bi-zine). These products are consistent with species implicated in the pheomelanogenesis biosynthetic pathway and emphasize how metal ions, their counteranions, and reaction conditions can alter pheomelanin product distribution. PMID- 18163614 TI - Oxygen-centered hexatantalum tetradecaimido cluster complexes. AB - The syntheses and characterization of several octahedral hexatantalum cluster compounds of formula (ArN)14Ta6O are described (Ar=Ph, p-MeC6H4, p-MeOC6H4, p-t BuC6H4, p-BrC6H4, m-ClC6H4). Treatment of Bn3Ta=N-t-Bu (Bn=CH2C6H5) or pentakis(dimethylamido)tantalum with an excess of the appropriate aniline and stoichiometric water or tantalum oxide afforded varying yields of arylimido clusters. The structures of two species were confirmed by X-ray diffraction (XRD), while the identity of the central oxygen atom was elucidated by electrospray mass spectrometry (MS) using 17O/18O-enriched material. The title species are very air- and moisture-sensitive but quite thermally stable in solution. Experimentally determined optical properties and oxidation/reduction potentials, as well as some computational results, indicate that they possess an electronic structure wherein the highest occupied molecular orbitals are ligand centered, while the lowest unoccupied orbitals are metal-centered and delocalized throughout the tantalum cage. Whereas chemical oxidation resulted in cluster decomposition, reduction with decamethylcobaltocene yielded stable salts of formula [Cp*2Co][(ArN)14Ta6O] (Ar=Ph, Ar=p-MeC6H4). Small-molecule reactivity studies on one of these clusters showed that its imido functionalities are moderately reactive toward oxide donors but inert with respect to metallaheterocycle-forming processes. Clean imido/oxo exchange was observed with aldehydes and ketones, leading cleanly to organic imines with no soluble byproducts being observed. This exchange was also observed with a rhenium oxo compound (generating an imidorhenium complex as the only soluble species). All 14 imido groups were transferred in these reactions, and no mixed-ligand cluster intermediates were ever observed. PMID- 18163615 TI - Structure of the molybdenum site of Escherichia coli trimethylamine N-oxide reductase. AB - We report a structural characterization of the molybdenum site of recombinant Escherichia coli trimethylamine N-oxide (TMAO) reductase using X-ray absorption spectroscopy. The enzyme active site shows considerable similarity to that of dimethyl sulfoxide (DMSO) reductase, in that, like DMSO reductase, the TMAO reductase active site can exist in multiple forms. Examination of the published crystal structure of TMAO oxidase from Shewanella massilia indicates that the postulated Mo coordination structure is chemically impossible. The presence of multiple active site structures provides a potential explanation for the anomalous features reported from the crystal structure. PMID- 18163616 TI - Six-coordinate nitrato complexes of iron(III) porphyrins. AB - The interaction of tetrahydrofuran (THF) with thin films of the nitrato complexes Fe(III)(Por)(eta(2)-O(2)NO) [Por = meso-tetraphenylporphyrinato (TPP) and meso tetratolylporphyrinato (TTP) dianion] at low temperature leads to the formation of the six-coordinate nitrato complex Fe(Por)(THF)(NO(3)), which was characterized by IR and UV-visible spectroscopies. Formation of the THF adduct was accompanied by nitrate linkage isomerization from bidentate to monodentate coordination. The iron(III) center remains in a high spin state in contrast with the previously observed low-spin nitratonitrosyl complex Fe(TPP)(NO)(eta(10 ONO(2)). Upon warming, THF dissociates to restore the initial five-coordinate bidentate nitrato complex. PMID- 18163617 TI - Local electronic structure of dicarba-closo-dodecarboranes C2B10H12. AB - We report nonresonant inelastic X-ray scattering (NRIXS) measurement of core shell excitations from both B 1s and C 1s initial states in all three isomers of the dicarba-closo-dodecarboranes C2B10H12. First, these data yield an experimental determination of the angular-momentum-projected final local density of states (l-DOS). We find low-energy resonances with distinctive local s- or p type character, providing a more complete experimental characterization of bond hybridization than is available from dipole-transition limited techniques, such as X-ray absorption spectroscopies. This analysis is supported by independent density functional theory and real-space full multiple scattering calculation of the l-DOS which yield a clear distinction between tangential and radial contributions. Second, we investigate the isomer sensitivity of the NRIXS signal and compare and contrast these results with prior electron energy loss spectroscopy measurements. This work establishes NRIXS as a valuable tool for borane chemistry, not only for the unique spectroscopic capabilities of the technique but also through its compatibility with future studies in solution or in high-pressure environments. In addition, this work also establishes the real space full multiple scattering approach as a useful alternative to traditional approaches for excited states calculations of aromatic polyhedral boranes and related systems. PMID- 18163618 TI - Interplay among tetrahedrane, butterfly diradical, and planar rhombus structures in the chemistry of the binuclear iron carbonyl phosphinidene complexes Fe2(CO)6(PX)2. AB - Density functional theory studies on a series of Fe2(CO)6(PX)2 derivatives show the tetrahedrane to be the most stable for the alkyl (X = Me, tBu), P-H (X = H), and chloro (X = Cl) derivatives. However, butterfly diradical and planar rhombus structures are found to be more stable than tetrahedranes for the amino (X = NH2, NMe2, and NiPr2) and aryloxy (R = 2,6-tBu2-4-Me-C6H2O) derivatives. For the chloro (X = Cl) and methoxy (X = OMe) derivatives energetically accessible bishomotetrahedrane Fe2(CO)6P2(mu-X)2 isomers are observed in which the X substituents on the phosphorus atoms interact with the iron atom to form two direct Fe-X bonds at the expense of two of the four Fe-P bonds. In addition, the global minimum for the hydroxy (X = OH) derivative is an unusual FeP-butterfly structure with a central Fe-P bond as well as two external Fe-P bonds, one external P-P bond, and one external Fe=Fe double bond. Comparison of calculated with experimental nu(CO) frequencies shows that low-temperature Nujol matrix photolysis of (iPr2NP)2COFe2(CO)6 leads to a planar rhombus rather than a tetrahedrane isomer of Fe2(CO)6(PNiPr2)2. PMID- 18163619 TI - Catalysis of 3-pyrrolidinecarboxylic acid and related pyrrolidine derivatives in enantioselective anti-Mannich-type reactions: importance of the 3-acid group on pyrrolidine for stereocontrol. AB - The development of enantioselective anti-selective Mannich-type reactions of aldehydes and ketones with imines catalyzed by 3-pyrrolidinecarboxylic acid and related pyrrolidine derivatives is reported in detail. Both (3R,5R)-5-methyl-3 pyrrolidinecarboxylic acid and (R)-3-pyrrolidinecarboxylic acid efficiently catalyzed the reactions of aldehydes with alpha-imino esters under mild conditions and afforded anti-Mannich products with high diastereo- and enantioselectivities (anti/syn up to 99:1, up to >99% ee). For the reactions of ketones with alpha-imino esters, (R)-3-pyrrolidinecarboxylic acid was an efficient catalyst (anti/syn up to >99:1, up to 99% ee). Evaluation of a series of pyrrolidine-based catalysts indicated that the acid group at the beta-position of the pyrrolidine ring of the catalyst played an important role in forwarding the carbon-carbon bond formation and in directing anti-selectivity and enantioselectivity. PMID- 18163620 TI - Metal-coupled folding of Cys2His2 zinc-finger. AB - Zinc-fingers, which widely exist in eukaryotic cell and play crucial roles in life processes, depend on the binding of zinc ion for their proper folding. To computationally study the zinc-coupled folding of the zinc-fingers, charge transfer and metal induced protonation/deprotonation effects have to be considered. Here, by attempting to implicitly account for such effects in classical molecular dynamics and performing intensive simulations with explicit solvent for the peptides with and without zinc binding, we investigate the folding of the Cys2His2-type zinc-finger motif and the coupling between the peptide folding and zinc binding. We find that zinc ion not only stabilizes the native structure but also participates in the whole folding process. It binds to the peptide at an early stage of folding and directs or modulates the folding and stabilizations of the component beta-hairpin and alpha-helix. Such a crucial role of zinc binding is mediated by the packing of the conserved hydrophobic residues. We also find that the packing of the hydrophobic residues and the coordination of the native ligands are coupled. Meanwhile, the processes of zinc binding, mis ligation, ligand exchange, and zinc induced secondary structure conversion as well as the water behavior due to the involvement of zinc ion are characterized. Our results are in good agreement with related experimental observations and provide significant insight into the general mechanisms of the metal cofactor dependent protein folding and other metal-induced conformational changes of biological importance. PMID- 18163621 TI - Solid-state 19F NMR spectroscopy reveals that Trp41 participates in the gating mechanism of the M2 proton channel of influenza A virus. AB - The integral membrane protein M2 of influenza A virus assembles as a tetrameric bundle to form a proton-conducting channel that is activated by low pH. The side chain of His37 in the transmembrane alpha-helix is known to play an important role in the pH activation of the proton channel. It has also been suggested that Trp41, which is located in an adjacent turn of the helix, forms part of the gating mechanism. Here, a synthetic 25-residue peptide containing the M2 transmembrane domain was labeled with 6F-Trp41 and studied in lipid membranes by solid-state 19F NMR. We monitored the pH-dependent differences in the 19F dipolar couplings and motionally narrowed chemical shift anisotropies of this 6F-Trp41 residue, and we discuss the pH activation mechanism of the H+ channel. At pH 8.0, the structural parameters implicate an inactivated state, while at pH 5.3 the tryptophan conformation represents the activated state. With the aid of COSMOS force field simulations, we have obtained new side-chain torsion angles for Trp41 in the inactivated state (chi1 = -100 degrees +/- 10 degrees , chi2 = +110 degrees +/- 10 degrees ), and we predict a most probable activated state with chi1 = -50 degrees +/- 10 degrees and chi2 = +115 degrees +/- 10 degrees . We have also validated the torsion angles of His37 in the inactivated state as chi1 = -175 degrees +/- 10 degrees and chi2 = -170 degrees +/- 10 degrees . PMID- 18163622 TI - Oxidations of N-(3-indoleethyl) cyclic aliphatic amines by horseradish peroxidase: the indole ring binds to the enzyme and mediates electron-transfer amine oxidation. AB - Although oxidations of aromatic amines by horseradish peroxidase (HRP) are well known, typical aliphatic amines are not substrates of HRP. In this study, the reactions of N-benzyl and N-methyl cyclic amines with HRP were found to be slow, but reactions of N-(3-indoleethyl) cyclic amines were 2-3 orders of magnitude faster. Analyses of pH-rate profiles revealed a dominant contribution to reaction by the amine-free base forms, the only species found to bind to the enzyme. A metabolic study on a family of congeneric N-(3-indoleethyl) cyclic amines indicated competition between amine and indole oxidation pathways. Amine oxidation dominated for the seven- and eight-membered azacycles, where ring size supports the change in hybridization from sp3 to sp2 that occurs upon one electron amine nitrogen oxidation, whereas only indole oxidation was observed for the six-membered ring congener. Optical difference spectroscopic binding data and computational docking simulations suggest that all the arylalkylamine substrates bind to the enzyme through their aromatic termini with similar binding modes and binding affinities. Kinetic saturation was observed for a particularly soluble substrate, consistent with an obligatory role of an enzyme-substrate complexation preceding electron transfer. The significant rate enhancements seen for the indoleethylamine substrates suggest the ability of the bound indole ring to mediate what amounts to medium long-range electron-transfer oxidation of the tertiary amine center by the HRP oxidants. This is the first systematic investigation to document aliphatic amine oxidation by HRP at rates consistent with normal metabolic turnover, and the demonstration that this is facilitated by an auxiliary electron-rich aromatic ring. PMID- 18163623 TI - Formation and characterization of self-organized TiO2 nanotube arrays by pulse anodization. AB - This paper describes TiO2 nanotube arrays prepared by anodic oxidation of Ti substrates using pulse voltage waveforms. Voltages were pulsed between 20 and -4 V or between 20 and 0 V with varying durations from 2 to 16 s at the lower limit of the pulse waveform. Ammonium fluoride or sodium fluoride (and mixtures of both) was used as the electrolyte with or without added medium modifier (glycerol, ethylene glycol, or poly (ethylene glycol) (PEG 400)) in these experiments. The pulse waveform was optimized to electrochemically grow TiO2 nanotubes and chemically etch their walls during its cathodic current flow regime. The resultant TiO2 nanotube arrays showed a higher quality of nanotube array morphology and photoresponse than samples grown via the conventional continuous anodization method. Films grown with a 20 V/-4 V pulse sequence and pulse duration of 2 s at its negative voltage limit afforded a superior photoresponse compared to other pulse durations. Specifically, the negative voltage limit of the pulse (-4 V) and its duration promote the adsorption of NH4+ species that in turn inhibits chemical attack of the growing oxide nanoarchitecture by the electrolyte F- species. The longer the period of the pulse at the negative voltage limit, the thicker the nanotube walls and the shorter the nanotube length. At variance, with 0 V as the low voltage limit, the longer the pulse duration, the thinner the oxide nanotube wall, suggesting that chemical attack by fluoride ions is not counterbalanced by NH3/NH4+ species adsorption, unlike the interfacial situation prevailing at -4 V. Finally, the results from this study provide useful evidence in support of existing mechanistic models for anodic growth and self-assembly of oxide nanotube arrays on the parent metal surface. PMID- 18163624 TI - Stacking interactions and the twist of DNA. AB - The importance of stacking interactions for the Twist and stability of DNA is investigated using the fully ab initio van der Waals density functional (vdW-DF). Our results highlight the role that binary interactions between adjacent sets of base pairs play in defining the sequence-dependent Twists observed in high resolution experiments. Furthermore, they demonstrate that additional stability gained by the presence of thymine is due to methyl interactions with neighboring bases, thus adding to our understanding of the mechanisms that contribute to the relative stability of DNA and RNA. Our mapping of the energy required to twist each of the 10 unique base pair steps should provide valuable information for future studies of nucleic acid stability and dynamics. The method introduced will enable the nonempirical theoretical study of significantly larger pieces of DNA or DNA/amino acid complexes than previously possible. PMID- 18163625 TI - Addition of adamantylidene to La2@C78: isolation and single-crystal X-ray structural determination of the monoadducts. AB - Thermal and photochemical reactions of La2@C78 with 2-admantane-2,3-[3H] diazirine are investigated. Four isomers of the monoadduct (La2@C78Ad) synthesized by the photoreaction are isolated by HPLC and characterized by mass, UV-vis-NIR absorption, cyclic voltammogram and differential pulse voltammogram spectroscopy, proton and 13C NMR spectroscopic analysis, single-crystal X-ray diffraction analysis, and theoretical approaches. The addition reactions occur at both the [5,6] and [6,6] positions. X-ray and theoretical studies indicate that one of the monoadduct isomers has an open structure with two La atoms on the C3 axis of the D3h cage of La2@C78. PMID- 18163626 TI - Electrical conductance of conjugated oligomers at the single molecule level. AB - We determine and compare, at the single molecule level and under identical environmental conditions, the electrical conductance of four conjugated phenylene oligomers comprising terminal sulfur anchor groups with simple structural and conjugation variations. The comparison shows that the conductance of oligo(phenylene vinylene) (OPV) is slightly higher than that of oligo(phenylene ethynylene) (OPE). We find that solubilizing side groups do neither prevent the molecules from being anchored within a break junction nor noticeably influence the conductance value. PMID- 18163627 TI - Redox-switched oxidation of dihydrogen using a non-innocent ligand. AB - Organometallic complexes containing non-innocent ligands of the type Cp*Ir(tBAFPh)(1), where H2tBAFPh is 2-(2-trifluoromethyl)anilino-4,6-di-tert butylphenol, were found to activate H2 in a redox-switchable manner. The 16e- complex 1 was inert with respect to H2, CO, as well as conventional basic substrates until oxidation. Oxidation of 16-electron 1 with 1 equiv of Ag+ resulted in ligand-centered oxidation affording salts of [1]+, which were characterized by crystallographically, EPR, and elemental analyses. [1]+ was reduced to 1 in the presence of H2 and the sterically hindered base, 2,6 (tBu)2C5H3N, via a pathway that is first-order in both metal and dihydrogen. Compound [1]+ forms adducts with MeCN, which inhibits catalysis. The catalytic oxidation of H2 was established by electrochemical methods to be associated with the monocation. PMID- 18163628 TI - Metal-organic framework from an anthracene derivative containing nanoscopic cages exhibiting high methane uptake. AB - A microporous metal-organic framework, PCN-14, based on an anthracene derivative, 5,5'-(9,10-anthracenediyl)di-isophthalate (H4adip), was synthesized under solvothermal reaction conditions. X-ray single crystal analysis revealed that PCN 14 consists of nanoscopic cages suitable for gas storage. N2-adsorption studies of PCN-14 at 77 K reveal a Langmuir surface area of 2176 m2/g and a pore volume of 0.87 cm3/g. Methane adsorption studies at 290 K and 35 bar show that PCN-14 exhibits an absolute methane-adsorption capacity of 230 v/v, 28% higher than the DOE target (180 v/v) for methane storage. PMID- 18163629 TI - Length dependence of the coil <--> beta-sheet transition in a membrane environment. AB - The most abundant structural element in protein aggregates is the beta-sheet. Designed peptides that fold into a beta-sheet structure upon binding to lipid membranes are useful models to elucidate the thermodynamic characteristics of the random coil <-->beta-structure transition. Here, we examine the effect of strand length on the random coil <--> beta-sheet transition of the (KIGAKI)n peptide with the total chain length varying between 7 and 30 amino acids. The beta-sheet content of the peptides in the presence and absence of membranes was measured with circular dichroism spectroscopy. The peptides were titrated with small unilamellar lipid vesicles, and the thermodynamic binding parameters were determined with isothermal titration calorimetry (ITC). Membrane binding includes at least two processes, namely (i) the transfer of the peptide from the aqueous phase to the lipid surface and (ii) the conformational change from a random coil conformation to a beta-sheet structure. CD spectroscopy and ITC analysis demonstrate that beta-sheet formation depends cooperatively on the peptide chain length with a distinct increase in beta-structure for n > 10-12. Binding to the lipid membrane is an entropy-driven process as the binding enthalpy is always endothermic. The contribution of the beta-sheet folding reaction to the overall process was determined with analogues of the KIGAKI repeat where two adjacent amino acids were replaced by their D-enantiomers. The folding reaction for peptides with n >or= 12 is characterized by a negative free folding energy of DeltaG(degree)beta approximately equal -0.15 kcal/mol per amino acid residue. The folding step proper is exothermic with DeltaH(degree)(beta) approximately equal 0.2 to -0.6 kcal/mol per residue and counteracted by a negative entropy term TDeltaS(degree)(beta) = -0.1 to -0.5 kcal/mol per residue, depending on the chain length (18 100 times) than a model compound containing only one pi-prismand-like cavity, and the single silver cation hops intramolecularly between the two adjoined cavities in duplexiphane. PMID- 18163638 TI - Enantiopure anthrylene-ethynylene cyclic tetramer and racemization via rotation of anthracene unit about acetylenic axes. AB - Four anthracene and four acetylene units are used to construct a chiral pi conjugate macrocycle, the chirality of which is due to the restricted rotation about acetylenic axes. Enantiomers were readily resolved by chiral HPLC and racemized slowly even at 70 degrees C. PMID- 18163639 TI - Photorelease of carboxylic acids mediated by visible-light-absorbing gold nanoparticles. AB - Visible-light-absorbing citrate-stabilized gold nanoparticles and tryptophan dithiane-conjugate-stabilized gold nanoparticles have been used to mediate electron transfer between dithiothreitol (DTT), a good electron donor, and an N methylpicolinium ester in aqueous solution. Quantitative yield of the free carboxylate has been obtained with quantum yields of release, Phirel, ranging from 0.5 to 4.5. PMID- 18163640 TI - Walsucochins A and B with an unprecedented skeleton isolated from Walsura cochinchinensis. AB - Walsucochins A (1) and B (2) with an unprecedented skeleton were isolated from Walsura cochinchinensis. Their structures including absolute configuration were elucidated by spectral methods. A biosynthetic pathway of 1 and 2 was postulated. Both 1 and 2 exhibited significant cell protecting activities against H2O2 induced PC12 cell damage. PMID- 18163641 TI - A methodical approach to ultra-scale-down of process sequences: application to casein removal from the milk of transgenic animals. AB - Scale-down models of individual operations are widely used in biopharmaceutical process development to obtain information about the performance of production scale equipment on the basis of inexpensive and efficient laboratory-scale tests, for the purposes of validation or optimization or characterization studies. We have investigated the ability of scale-down models of whole process sequences to provide reliable information for process scale-up from laboratory- to pilot scales of operation. Using the example of the recovery of a protein from transgenic milk, we have conducted an a priori scale-down analysis of a projected pilot-scale process sequence. A systematic approach was developed to ensure that all critical aspects of process behavior were included in the scale-down model, resulting in the creation of an accurate and reliable scale-down representation of the pilot-scale process. The data from scale-down process trials conducted at 70 and 200 mL scales of operation served to highlight crucial factors determining process performance, and proved reliable in predicting the performance of the pilot-scale process over a scaling factor of 1000. PMID- 18163642 TI - Proteomic profiling of secreted proteins from CHO cells using Surface-Enhanced Laser desorption ionization time-of-flight mass spectrometry. AB - Chinese hamster ovary (CHO) cells are the most commonly used host cell line for the production of recombinant biopharmaceuticals. These biopharmaceuticals are typically secreted from CHO cells and purified from harvested cell culture media. The purpose of this study was to investigate changes in the secreted proteome of CHO cells over the various stages of the growth cycle using Surface Enhanced Laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). Conditioned media samples were collected each day over a 6 day growth period from CHO-K1 cells grown in low serum (0.5% FBS) conditions in monolayer culture. Samples were profiled on a number of ProteinChip arrays with different chromatographic surfaces. From this study, 24 proteins were found to be differentially regulated at different phases of the growth cycle in CHO-K1 cells, when profiled on two chromatographic surfaces, Q10 (anionic) and IMAC30 (metal affinity) ProteinChip arrays. PMID- 18163643 TI - Palladium-catalyzed carbonylative cross-coupling with imines: a multicomponent synthesis of imidazolones. AB - The palladium-catalyzed coupling of imines, chloroformates, organotin reagents, and carbon monoxide leads to the one-pot formation of ketocarbamates in good yields. These products can further be converted to highly substituted imidazolones via a cyclocondensation reaction. Overall, this methodology provides an alternative approach to imidazolones from five simple and readily available building blocks via a one-pot, multicomponent process. PMID- 18163644 TI - The in, out asymmetric pseudo-triple helical form of a D3h diaza-macropentacycle. AB - A sterically encumbered [N(2)S(6)] macropentacycle (5) related to diazamacrobicycles and cryptands has been synthesized in 53% yield by the [1+1] condensation reaction between functionalized macrocyclic and macrotricyclic precursors. A macrononacycle (18) resulting from the corresponding [2+2] condensation was isolated in 7% yield from the reaction mixture. Both compounds showed broad features in their room-temperature (1)H NMR spectra, but their maximal average symmetry (D(3h) and D(2h), respectively) was achieved at high temperature (380 K). At low temperature (200 K, CD(2)Cl(2) solution), the macropentacycle is "frozen" to a single asymmetric (C1) conformation on the (1)H NMR time scale, which has also the molecular structure observed in the solid state by X-ray crystallography: pseudo-triple helical ( not equalC(3)) shape, io (in, out) form resulting from the endo/exo configuration at the nitrogen bridgehead atoms, and similar orientations of the tosyl substituents. The solution dynamics of the molecule can be described by coupled bridgehead nitrogen inversion, triple helix symmetrization, and reversal of triple helix handedness, with DeltaGc = 54.2 kJ mol(-1) in CD(2)Cl(2) at 300 K. Adoption of the io form by macropentacycle 5 in the crystal and in solution at low-temperature most probably results from the steric crowding and strain introduced by the [15]ane-N(2)S(2) macrocyclic bridging subunits. PMID- 18163645 TI - Studies on the synthesis of apoptolidin A. 1. Synthesis of the C(1)-C(11) fragment. AB - A synthesis of the C(1)-C(11) fragment of apoptolidin A has been accomplished by a convergent route involving the stereoselective glycosidation of 9 and the Suzuki cross-coupling reaction of bromodienoate 7 and the vinylborane generated via chemoselective hydroboration of diyne 6 with diisopinocampheylborane. PMID- 18163646 TI - Studies on the synthesis of apoptolidin A. 2. Synthesis of the disaccharide unit. AB - Disaccharide 3 correspoinding to the disaccharide unit of apoptolidin A has been synthesized via the regio- and stereoselective TBS-OTf-promoted beta glycosidation reaction of 2,6-dideoxy-2-iodo-beta-glucopyranosyl acetate (5) and p-methoxybenzyl 2,6-dideoxy-2-iodo-3-C-methyl-alpha-mannopyranoside (11). PMID- 18163647 TI - Direct entry to peptidyl ketones via SmI2-mediated C-C bond formation with readily accessible N-peptidyl oxazolidinones. AB - In this work, a new method for the preparation of peptidyl ketones is presented employing a SmI(2)/H(2)O-mediated coupling of N-peptidyl oxazolidinones with electron-deficient alkenes. The requisite peptide imides were easily prepared by solution-phase peptide synthesis starting from an N-acyl oxazolidinone derivative of an amino acid. Importantly, they could be used directly in the C-C bond forming step without the need for further functionalization. Coupling of these peptide derivatives with a second peptide possessing an N-terminal acryloyl group leads to ketomethylene isosteres of glycine-containing peptides. This method represents an alternative means for ligating two small peptides through a C-C bond-forming step. PMID- 18163648 TI - Dynamic structure of disulfide-removed linear analogs of tachyplesin-I in the lipid bilayer from solid-state NMR. AB - Tachyplesin-I (TP-I) is a 17-residue beta-hairpin antimicrobial peptide containing two disulfide bonds. Linear analogs of TP-I where the four Cys residues were replaced by aromatic and aliphatic residues, TPX4, were found to have varying degrees of activities, with the aromatic analogs similarly potent as TP-I. Understanding the different activities of the linear analogs should give insight into the mechanism of action of TP-I. To this end, we have investigated the dynamic structures of the active TPF4 and the inactive TPA4 in bacteria mimetic anionic POPE/POPG bilayers and compared them with the wild-type TP-I using solid-state NMR spectroscopy. 13C isotropic chemical shifts and backbone (phi, psi) torsion angles indicate that both TPF4 and TPA4 adopt beta-strand conformations without a beta-turn at key residues. 1H spin diffusion from lipid chains to the peptide indicates that the inactive TPA4 binds to the membrane water interface, similar to the active TP-I. Thus, neither the conformation nor the depth of insertion of the three peptides correlates with their antimicrobial activities. In contrast, the mobility of the three peptides correlates well with their activities: the active TP-I and TPF4 are both highly mobile in the liquid crystalline phase of the membrane while the inactive TPA4 is completely immobilized. The different mobilities are manifested in the temperature-dependent 13C and 15N spectra, 13C-1H and 15N-1H dipolar couplings and 1H rotating-frame spin-lattice relaxation times. The dynamics of TP-I and TPF4 are both segmental and global. Combined, these data suggest that TP-I and TPF4 disrupt the membrane by large-amplitude motion in the plane of the membrane. The loss of this motion in TPA4 due to aggregation significantly weakens its activity because a higher peptide concentration is required to disturb lipid packing. Thus molecular motion, rather than structure, appears to be the key determinant for the membrane disruptive activities of tachyplesins. PMID- 18163649 TI - The mechanism for isopenicillin N synthase from density-functional modeling highlights the similarities with other enzymes in the 2-His-1-carboxylate family. AB - Isopenicillin N synthase (IPNS) catalyzes a key step in the biosynthesis of the important beta-lactam antibiotics penicillins and cephalosporins. Density functional calculations with the B3LYP functional are used to propose a detailed mechanism for this reaction. The results support the general scheme outlined from experimental observations, with formation of a four-membered beta-lactam ring followed by formation of a five-membered thiazolidine ring. However, an alternative mechanism for the heterolytic O-O bond cleavage and beta-lactam ring formation steps is proposed. The former part involves protonation of the distal oxygen by an iron-bound water ligand. This mechanism highlights the strong similarities that exist between IPNS and other enzymes of the 2-histidine-1 carboxylate family, especially pterin-dependent amino acid hydroxylases and alpha keto acid-dependent dioxygenases. Both activation of the cysteine beta-C-H bond by an iron-bound superoxo radical and activation of the valine beta-C-H bond by a ferryl-oxo species show reaction barriers close to the experimentally measured one. These results are in agreement with kinetic isotope experiments that suggest both C-H bond activation steps to be partially rate limiting. The ring formation sequence is determined by the relative strengths of the two C-H bonds. Only the ferryl-oxo intermediate is capable of activating the stronger valine beta-C-H bond. PMID- 18163650 TI - Role of Phe1010 in light-induced structural changes of the neo1-LOV2 domain of Adiantum. AB - Phototropin (phot) is a blue-light sensor protein that elicits several photo responses in plants. Phototropin has two flavin mononucleotide (FMN)-binding domains, LOV1 and LOV2, in its N-terminal half. The C-terminal half is a blue light-regulated Ser/Thr kinase. Various functional studies have reported that only LOV2 is responsible for the kinase activity, whereas the X-ray crystallographic structures of the LOV1 and LOV2 domains are almost identical. How does such a functional difference emerge? Our previous FTIR study of the LOV domains of Adiantum neochrome1 (neo1) showed that light-induced protein structural changes are small and temperature independent for neo1-LOV1, whereas the structural changes are large and highly temperature dependent for neo1-LOV2, which involve loops, alpha-helices, and beta-sheets. These observations successfully explained the different functions in terms of protein structural changes. They also suggested the presence of some crucial amino acids responsible for greater protein structural changes in the LOV2 domain. Here, we focused on phenylalanine-1010 (Phe1010) in neo1-LOV2, where FMN is sandwiched between Phe1010 and the reactive cysteine. Phenylalanine at this position is conserved for LOV2 domains, while the corresponding amino acid is leucine for LOV1 domains in almost all plant phototropins. We observed that unlike wild-type LOV2, the FTIR spectra of F1010L LOV2 exhibited no temperature dependence in the alpha helical and beta-sheet regions and that spectral changes in amide-I of these regions were significantly reduced, which was similar to LOV1. Thus, the replacement of phenylalanine with leucine converts neo1-LOV2 into neo1-LOV1 in terms of protein structural changes that must be related to the different functions. We will discuss the roles of phenylalanine and leucine in the LOV2 and LOV1 domains, respectively. PMID- 18163651 TI - Crystal structure of a trapped phosphate intermediate in vanadium apochloroperoxidase catalyzing a dephosphorylation reaction. AB - The crystal structure of the apo form of vanadium chloroperoxidase from Curvularia inaequalis reacted with para-nitrophenylphosphate was determined at a resolution of 1.5 A. The aim of this study was to solve structural details of the dephosphorylation reaction catalyzed by this enzyme. Since the chloroperoxidase is functionally and evolutionary related to several acid phosphatases including human glucose-6-phosphatase and a group of membrane-bound lipid phosphatases, the structure sheds light on the details of the dephosphorylation catalyzed by these enzymes as well. The trapped intermediate found is bound to the active site as a metaphosphate anion PO3-, with its phosphorus atom covalently attached to the Nepsilon2 atom of His496. An apical water molecule is within hydrogen-bonding distance to the phosphorus atom of the metaphosphate, and it is in a perfect position for a nucleophilic attack on the metaphosphate-histidine intermediate to form the inorganic phosphate. This is, to our knowledge, the first structural characterization of a real reaction intermediate of the inorganic phosphate group release in a dephosphorylation reaction. PMID- 18163652 TI - Investigation of mechanical properties of insulin crystals by atomic force microscopy. AB - Mechanical properties of protein crystals and aggregates depend on the conformational and structural properties of individual protein molecules as well as on the packing density and structure within solid materials. An atomic force microscopy (AFM)-based approach is developed to measure the elastic modulus of small protein crystals by nanoindentation and is applied to measure the elasticity of insulin crystals. The top face of the crystals deposited on mica substrates is identified as the (001) face. Insulin crystals exhibit a nearly elastic response during the compression cycle. The elastic modulus measured on the top face has asymmetric distribution with a significant width. This width is related to the uncertainty in the deflection sensitivity. A model that takes into account the distribution of the sensitivity values is used to correct the elastic modulus. Measurements performed in aqueous buffer on several crystals at different locations with three different AFM probes give a mean elastic modulus of 164 +/- 10 MPa. This value is close to the static elastic moduli of other protein crystals measured by different techniques that are usually measured in the range from 100 MPa to 1 GPa. The measured modulus of insulin crystals falls between the elastic modulus values of insulin amyloid fibrils measured previously at two orthogonal directions (a modulus of 14 MPa was measured by compressing the fibril in the direction perpendicular to the fibril axis, and a modulus of 3.3 GPa was measured in the direction along the fibril axis). This comparison indicates the heterogeneous structure of fibrils in the direction perpendicular to the fibril axis, with a packing density of the amyloid fibril core that is higher than the average packing density in insulin crystals. The mechanical wear of insulin crystals is detected during AFM measurements. In nanoindentation experiments on insulin crystal, the compressive load by the AFM tip ( approximately 1 nN, corresponding to a pressure of around 5 MPa) occasionally removes protein molecules from the top or the second top layer of insulin crystal in a sequential manner. The molecular model of this surface damage is proposed. In addition, the removal of the multiple layers of molecules is observed during the AC-mode imaging in aqueous buffer. The number of removed layers depends on the scan size. PMID- 18163653 TI - Surface modification of poly(dimethylsiloxane) with a perfluorinated alkoxysilane for selectivity toward fluorous tagged peptides. AB - Poly(dimethylsiloxane) (PDMS) and similar polymers have proved to be of widespread interest for use in microfluidic and similar microanalytical devices. Surface modification of PDMS is required to extend the range of applications for devices made of this polymer, however. Here we report on the grafting of perfluorooctyltriethoxysilane via hydrolysis onto an oxidized PDMS substrate in order to form a fluorinated microchannel. Such a fluorinated device could be used for separating fluorous tagged proteins or peptides, similar to that which has been recently demonstrated in a capillary electrophoresis system or in an open tubular capillary column. The modified polymer is characterized using chemical force titrations, contact angle measurements, and X-ray photoelectron spectroscopy (XPS). We also report on a novel means of performing electroosmotic measurements on this material to determine the surface zeta potential. As might be expected, contact angle and chemical force titration measurements indicate the fluorinated surface to be highly hydrophobic. XPS indicates that fluorocarbon groups segregate to the surface of the polymer over a period of days following the initial surface modification, presumably driven by a lower surface free energy. One of the most interesting results is the zeta potential measurements, which show that significant surface charge can be maintained across a wide range of pH on this modified polymer, sufficient to promote electroosmotic flow in a microfluidic chip. Matrix-assisted time-of-flight mass spectrometry (MALDI-TOF MS) measurements show that a fluorous-tagged peptide will selectively adsorb on the fluorinated PDMS in aqueous solution, demonstrating that the fluorinated polymer could be used in devices designed for the enrichment or enhanced detection of fluorous-labeled proteins and peptides. PMID- 18163654 TI - Deprotecting thioacetyl-terminated terphenyldithiol for assembly on gallium arsenide. AB - We characterize the assembly of terphenyldithiol (TPDT) on gallium arsenide (GaAs) from ethanol (EtOH) and tetrahydrofuran (THF) as a function of ammonium hydroxide (NH4OH) concentration. NH4OH facilitates the conversion of thioacetyl end groups of the TPDT precursor to thiolates in the assembly solution. The final structure of TPDT assembled on GaAs is sensitive not only to the assembly solvent but also to NH4OH concentration. In the presence of low concentrations of NH4OH (1 mM), TPDT assemblies from EtOH are oriented upright. The same assemblies are less upright when adsorption is carried out at higher NH4OH concentrations. In THF, TPDT does not adsorb significantly on GaAs at low NH4OH concentrations. The surface coverage and structural organization of these assemblies improve with increasing NH4OH concentrations, although these assemblies are never as organized as those from EtOH. The difference in the final structure of TPDT assemblies is attributed to differences in the thiolate fraction in the assembly solution at the point of substrate immersion. PMID- 18163655 TI - Effect of contact geometry on the pull-off force evaluated under high-vacuum and humid atmospheric conditions. AB - The effect of condensed water on pull-off forces under high vacuum (HV) and 0 to 83% relative humidity (RH) N2 atmospheric conditions was evaluated for different contact geometries using atomic force microscopy (AFM). The pull-off force was measured using two types of contact geometry: contact between hemispherical asperities and a flat silicon probe on an AFM cantilever (called a spherical-flat contact) and between a flat silicon substrate and a flat nickel probe on an AFM cantilever (called a flat-flat contact). The hemispherical asperities were fabricated using a focused ion beam (FIB) system, and each peak had a radius of curvature of between 70 and 610 nm. The flat nickel probe was fabricated by friction-induced wear. Measurement results showed that for the spherical-flat contact the pull-off force was proportional to the radius of curvature of the asperity peak and was slightly lower in HV than in humid 14% RH N2. For the flat flat contact in HV, with increasing contact time, the pull-off force increased in HV but decreased in humid 62 and 83% RH N2. The pull-off force in HV was lower than that in humid N2 when the contact time was less than 10 s but was higher when the contact time was longer than 30 s. The estimated adhesion force based on the Laplace pressure from the contact geometry agreed reasonably well with the measured pull-off force. PMID- 18163656 TI - Nanostructure of gel-derived aluminosilicate materials. AB - In the present work, aluminosilicate aerogels prepared under various conditions were compared with respect to their nanostructures and porosity. The purpose of this investigation was to find a suitable way to predict the final product structure and to tailor a required texture. Several Al and Si precursors (Al nitrate, Al isopropoxide, Al acetate, tetraethoxysilane (TEOS), and sodium silicate) were used in our examinations; the solvent content (water and alcohols), surfactants, as well as the catalysts were varied. In addition, the aerogels were subjected to various heat treatments. Hybrid aerogels were synthesized by the addition of different polymers (poly(acrylic acid), polyvinyl acetate, and polydimethylsiloxane). Aluminosilicate and hybrid aerogel structures were investigated by 27Al MAS NMR, SAXS, SEM, and porosity measurements. Loose fractal structures with a good porosity and high Al incorporation can be achieved from TEOS and Al nitrate or isopropoxide via a sol-gel preparation route. The use of Al acetate led to compact aerogel structures independently of the Si precursor, the pH, and the catalyst. PMID- 18163657 TI - Crystallization of carbamazepine pseudopolymorphs from nonionic microemulsions. AB - Crystallization of carbamazepine (CBZ), an antiepileptic drug, precipitated from confined spaces of nonionic microemulsions was investigated. The study was aimed to correlate the structure of the microemulsion [water-in-oil (W/O), bicontinuous, and oil-in-water (O/W)] with the crystalline structure and morphology of solid CBZ. The precipitated CBZ was studied by DSC, TGA, powder XRD, single-crystal XRD, SEM, and optical microscopy. The results suggest that the microstructure of the microemulsions influences the crystallization process and allows crystallizing polymorphs that exhibit different crystal structure and habits. W/O nanodroplets orient the crystallizing CBZ molecules to form a prismlike anhydrous polymorphic form with monoclinic unit cell and P21/n space group. Bicontinuous structures lead to platelike dihydrate crystals with orthorhombic unit cell and Cmca space group. The O/W nanodroplets cause the formation of needlelike dihydrate crystals with monoclinic unit cell and P21/c space group. The morphological features of solid CBZ remain predetermined by the basic symmetry and parameters of its unit cell. Precipitation of CBZ pseudopolymorphs from supersaturated microemulsion is discussed in terms of oriented attachment that provides perfect packing of numerous separately nucleated ordered nuclei of CBZ into microscale platelets and then into macroscopic crystals. Crystallization from microemulsion media enabling one to obtain the drug (CBZ) with predicted structure and morphology should be of great significance for pharmaceutical applications. PMID- 18163658 TI - New method for fabrication of loaded micro- and nanocontainers: emulsion encapsulation by polyelectrolyte layer-by-layer deposition on the liquid core. AB - A novel approach to the emulsion encapsulation was developed by combining the advantages of direct encapsulation of a liquid colloidal core with the accuracy and multifunctionality of layer-by-layer polyelectrolyte deposition. Experimental data obtained for the model oil-in-water emulsion confirm unambiguously the alternating PE assembly in the capsule shell as well as the maintenance of the liquid colloidal core. Two different mechanisms of capsule destruction upon interaction with the solid substrate were observed and qualitatively explained. The proposed method can be easily generalized to the preparation of oil-filled capsules in various oil/water/polyelectrolyte systems important in the field of pharmacy, medicine, and food industry. PMID- 18163659 TI - Using breath figure patterns on structured substrates for the preparation of hierarchically structured microsieves. AB - Microsieves are advanced filtration membranes characterized by a uniform pore size, a high pore density, and a thickness smaller than the pore diameter. The uniform pore size provides a high selectivity; the small thickness gives rise to a high flux and allows efficient removal of any filter cake by backflushing. However, microsieves are sensitive to mechanical stress. Thus, they need either an external macroporous support or a hierarchical structure that provides an integrated supportive structure. We prepare microsieves with a hierarchical pore structure by creating breath figure patterns within layers of solutions of polymers in a volatile solvent that are spread out on top of structured supports. For the formation of breath figure patterns, the volatile solvent is evaporated in a moist atmosphere. This cools the surface to such an extent that dew droplets form on the thin film, partially penetrate into the layer, and create a concave imprint in the final solid polymer layer. This procedure is usually done on flat surfaces; in our case the spreading of the polymer solution is done on a support decorated with protrusions. In this procedure, the dew droplets touch the protrusions of the structured support before the polymer solution vitrifies. At the same time, the trenches of the structured substrate are filled with polymer much deeper than the penetration depth of the dew droplets. After the separation of the vitrified layer from the substrate, we obtain thin polymer membranes with a hierarchical structure consisting of an ultrathin active separation layer with submicrometer pores and a supporting layer with larger pores. PMID- 18163660 TI - Bubble colloidal AFM probes formed from ultrasonically generated bubbles. AB - Here we introduce a simple and effective experimental approach to measuring the interaction forces between two small bubbles (approximately 80-140 microm) in aqueous solution during controlled collisions on the scale of micrometers to nanometers. The colloidal probe technique using atomic force microscopy (AFM) was extended to measure interaction forces between a cantilever-attached bubble and surface-attached bubbles of various sizes. By using an ultrasonic source, we generated numerous small bubbles on a mildly hydrophobic surface of a glass slide. A single bubble picked up with a strongly hydrophobized V-shaped cantilever was used as the colloidal probe. Sample force measurements were used to evaluate the pure water bubble cleanliness and the general consistency of the measurements. PMID- 18163661 TI - On total internal reflection investigation of nanoparticles by integrated micro fluidic system. AB - We report on a novel sensor for characterization of nanoparticles colloidal suspensions. We employ a diffraction grating under total internal reflection for investigation of nanodisperse fluids passing through an integrated microfluidic channel. Dispersions containing polymeric, metallic, and ferromagnetic nanoparticles are studied. Using this device, we can accurately determine in real time the specific refractive index for the nanoparticle suspension and the nanoparticle concentration. The nanoparticle concentrations can be calculated with a resolution of 0.3-0.5 wt% for polymeric nanoparticles, 0.03-0.05 wt% for metallic nanoparticles, and 0.05-0.1 wt% for ferromagnetic nanoparticles. This translates to an effective refractive index that can be determined with an accuracy of 7 x 10(-4) for the polymeric and 2 x 10(-4) for the metallic and ferromagnetic dispersions. PMID- 18163662 TI - Botryolides A-E, decarestrictine analogues from a fungicolous Botryotrichum sp. (NRRL 38180). AB - Four new decarestrictine analogues (botryolides A-D; 1- 4), a biosynthetically related gamma-lactone (botryolide E; 5), and the known compounds decarestrictine D ( 6) and sterigmatocystin have been isolated from cultures of a fungicolous isolate of Botryotrichum sp. (NRRL 38180). The structures of these compounds were determined by analysis of 2D NMR and ESIMS data. The relative configurations of 1 5 were established on the basis of NMR data and/or X-ray diffraction analysis, while the absolute configuration of 1 was assigned using the modified Mosher method. PMID- 18163663 TI - Evidence for an energy level substructure of molecular states in helium droplets. AB - The pure tunneling inversion transition of ammonia embedded in (4)He droplets was investigated in the microwave frequency range. We observed a spectrum that consists of a sharp peak, only 15 MHz wide, on top of a broad feature. The peculiar line shape could be simulated with an empirical model and is a clear experimental evidence for an energy level substructure of molecular states in doped helium droplets. PMID- 18163664 TI - The electronic nonadiabatic coupling term: can it be ignored in dynamic calculations? AB - Whereas the search for the degeneracy points which are better known as conical intersections (or ci-points) is usually carried out with a lot of devotion, the nonadiabatic coupling terms (NACTs) which together with the adiabatic potential energy surfaces appear in the nuclear Born-Oppenheimer-Schrodinger equation are ignored in most dynamical calculations. In the present article we consider two well known frameworks, namely, the semiclassical surface hopping method and the vibrational coupling model Hamiltonian that avoid the NACTs and examine to what extent, this procedure is justified. PMID- 18163665 TI - Calculation of the magnetic circular dichroism B term from the imaginary part of the Verdet constant using damped time-dependent density functional theory. AB - A time-dependent density functional theory (TDDFT) formalism with damping for the calculation of the magnetic optical rotatory dispersion and magnetic circular dichroism (MCD) from the complex Verdet constant is presented. For a justification of such an approach, we have derived the TDDFT analog of the sum over-states formula for the Verdet constant. The results of the MCD calculations by this method for ethylene, furan, thiophene, selenophene, tellurophene, and pyrrole are in good agreement with our previous theoretical sum-over-states MCD spectra. For the pi-->pi(*) transition of propene, we have obtained a positive Faraday B term. It is located between the two negative B terms. This finding is in agreement with experiment in the range of 6-8 eV. PMID- 18163666 TI - Two-photon absorption of [2.2]paracyclophane derivatives in solution: a theoretical investigation. AB - The two-photon absorption of a class of [2.2]paracyclophane derivatives has been studied using quadratic response and density functional theories. For the molecules investigated, several effects influencing the two-photon absorption spectra have been investigated, such as side-chain elongation, hydrogen bonding, the use of ionic species, and solvent effects, the latter described by the polarizable continuum model. The calculations have been carried out using a recent parallel implementation of the polarizable continuum model in the DALTON code. Special attention is given to those aspects that could explain the large solvent effect on the two-photon absorption cross sections observed experimentally for this class of compounds. PMID- 18163667 TI - Uniqueness of the iterative solution of the optimized effective potential equation. AB - The optimized effective potential (OEP) equation can be used in a numerically efficient self-consistent form to solve for the density functional exchange and correlation potentials, as shown in a recent paper of Kummel and Perdew [Phys. Rev. Lett. 90, 43004 (2003)]. The uniqueness of an iterative solution of the OEP equation has not yet been adequately addressed. In this paper, it is shown that no nonconstant multiplicative potentials that can contaminate an iterative solution of the OEP equation exist and, hence, that formally the exact exchange correlation potential determined form of the OEP equation is unique to within a constant. PMID- 18163668 TI - Multireference self-consistent-field energies without the many-electron wave function through a variational low-rank two-electron reduced-density-matrix method. AB - The variational two-electron reduced-density-matrix (2-RDM) method allows for the computation of accurate ground-state energies and 2-RDMs of atoms and molecules without the explicit construction of an N-electron wave function. While previous work on variational 2-RDM theory has focused on calculating full configuration interaction energies, this work presents the first application toward approximating multiconfiguration self-consistent-field (MCSCF) energies via low rank restrictions on the 1- and 2-RDMs. The 2-RDM method with two- or three particle N-representability conditions reduces the exponential active-space scaling of MCSCF methods to a polynomial scaling. Because the first-order algorithm [Mazziotti, Phys. Rev. Lett. 93, 213001 (2004)] represents each form of the 1- and 2-RDMs by a matrix factorization, the RDMs are readily defined to have a low rank rather than a full rank by setting the matrix factors to be rectangular rather than square. Results for the potential energy surfaces of hydrogen fluoride, water, and the nitrogen molecule show that the low-rank 2-RDM method yields accurate approximations to the MCSCF energies. We also compute the energies along the symmetric stretch of a 20-atom hydrogen chain where traditional MCSCF calculations, requiring more than 17x10(9) determinants in the active space, could not be performed. PMID- 18163669 TI - Cavity ring-down spectroscopy of jet-cooled silane isotopologues in the Si-H stretch overtone region. AB - Absorption spectra of silane in the region of the first overtone of the Si-H stretch vibration have been recorded in a seeded supersonic jet expansion by cavity ring-down spectroscopy as well as in a static gas cell at room temperature by photoacoustic spectroscopy. Spectral simplification due to strong rotational cooling in the jet expansion enables us to clearly resolve and assign the rovibrational transitions of the (2000) and (1100) bands of the three isotopologues, (28)SiH(4), (29)SiH(4), and (30)SiH(4), in their natural isotopic abundance. Interconversion between different nuclear spin species of SiH(4) is found to be absent during the jet expansion. Isotope shifts for (29)SiH(4) and (30)SiH(4) relative to (28)SiH(4) are measured and found to be suitable for selective vibrational excitation of any of three silane isotopologues by pulsed laser excitation in a jet expansion. PMID- 18163670 TI - Interactions of transition metal atoms in high-spin states: Cr2, Sc-Cr, and Sc Kr. AB - The high-spin van der Waals states are examined for the following dimers: Cr(2) ((13)Sigma(g)(+)), Sc-Cr ((8)Sigma(+), (8)Pi, (8)Delta), and Sc-Kr ((2)Sigma(+), (2)Pi, (2)Delta). These three systems offer a wide range of van der Waals interactions: anomalously strong, intermediate, and typically weak. The single reference [coupled cluster with single, double, and noniterative triple excitations, RCCSD(T)] method is used in the calculations for all three systems. In addition, a range of configuration-interaction based methods is applied in Cr(2) and Sc-Cr. The three dimers are shown to be bound by the dispersion interaction of varying strength. In a related effort, the dispersion energy and its exchange counterpart are calculated using the newly developed open-shell variant of the symmetry-adapted perturbation theory (SAPT). The restricted open shell time-dependent Hartree-Fock linear response function is used in the calculations of the dispersion energy in Sc-Cr and Sc-Kr calculations, while the restricted open-shell time-dependent density functional linear response function is used for Cr(2). A hybrid method combining the repulsive restricted open-shell Hartree-Fock (or complete active space self-consistent field) interaction energy with the dispersion and exchange-dispersion terms is tested against the RCCSD(T) results for the three complexes. The Cr(2) ((13)Sigma(g)(+)) complex has the well depth of 807.8 cm(-1) at the equilibrium distance of 6.18a(0) and the dissociation energy of 776.8 cm(-1). The octet-state Sc-Cr is about four times more strongly bound with the order of well depths of (8)Delta>(8)Pi>(8)Sigma(+) and a considerable anisotropy. The enhanced bonding is attributed to the unusually strong dispersion interaction. Sc-Kr ((2)Sigma(+), (2)Pi, (2)Delta) is a typical van der Waals dimer with well depths in the range of 81 cm(-1) ((2)Delta), 84 cm(-1) ((2)Sigma(+)), and 86 cm(-1) ((2)Pi). The hybrid model based on SAPT leads to results which are in excellent qualitative agreement with RCCSD(T) for all three interactions. PMID- 18163671 TI - Low-energy electron attachment to SF6. I. Kinetic modeling of nondissociative attachment. AB - Low-energy electron-molecule collisions are analyzed by kinetic modeling within the framework of statistical unimolecular rate theory. Nondissociative electron attachment to SF(6) is used to illustrate the approach. An internally consistent representation is provided for attachment cross sections and rate coefficients in relation to detachment lifetimes, and both thermal and specific rate coefficients for detachment. By inspecting experimental data, the contributions of intramolecular vibrational redistribution and vibrationally inelastic collisions can be characterized quantitatively. This allows for a prediction of attachment rate coefficients as a function of electron and gas temperature as well as gas pressure over wide ranges of conditions. The importance of carefully controlling all experimental parameters, including the carrier gas pressure, is illustrated. The kinetic modeling in Part II of this series is extended to dissociative electron attachment to SF(6). PMID- 18163672 TI - Low-energy electron attachment to SF6. II. Temperature and pressure dependences of dissociative attachment. AB - Low-energy electron-molecule collisions, leading to dissociative attachment through metastable anionic states, are kinetically modeled within the framework of statistical unimolecular rate theory. The reaction e(-)+SF(6)-->SF(5)(-)+F is used as an illustrative example. The modeling is applied to new measurements of branching fractions for SF(5)(-) formation in the bath gas He between 360 and 670 K at 1 and 2 Torr, and between 490 and 620 K over the range of 0.3-9 Torr. The analysis of the data follows the previous kinetic modeling of the nondissociative electron attachment, e(-)+SF(6)-->SF(6)(-), from Part I of this series. Experimental results from the present work and the literature on branching fractions and total cross sections for anion formation as functions of electron energies, bath gas temperatures and pressures, as well as observation times are analyzed. The assumption of a participation of the electronic ground state of SF(6)(-) alone suffices to model the available experimental data. A value of the dissociation energy of SF(6)(-) into SF(5)(-)+F of E(0,dis)=1.61(+/-0.05) eV is determined, which may be compared to the electron affinity of SF(6), EA=1.20(+/ 0.05) eV, such as derived in Part III of this series. PMID- 18163673 TI - Low-energy electron attachment to SF6. III. From thermal detachment to the electron affinity of SF6. AB - The thermal attachment of electrons to SF(6) is measured in a flowing-afterglow Langmuir-probe apparatus monitoring electron concentrations versus axial position in the flow tube. Temperatures between 300 and 670 K and pressures of the bath gas He in the range of 0.3-9 Torr are employed. Monitoring the concentrations of SF(6)(-) and SF(5)(-), the latter of which does not detach electrons under the applied conditions, an onset of thermal detachment and dissociation of SF(6) at temperatures above about 530 K is observed. Analysis of the mechanism allows one to deduce thermal detachment rate coefficients. Thermal dissociation rate coefficients for the reaction SF(6)(-)-->SF(5)(-)+F can only be estimated by unimolecular rate theory based on the results from Part I and II of this series. Under the applied conditions they are found to be smaller than detachment rate coefficients. Combining thermal attachment and detachment rates in a third-law analysis, employing calculated vibrational frequencies of SF(6) and SF(6)(-), leads to the electron affinity (EA) of SF(6)(-). The new value of EA=1.20(+/ 0.05) eV is significantly higher than previous recommendations which were based on less direct methods. PMID- 18163674 TI - Stability of small Pdn (n=1-7) clusters on the basis of structural and electronic properties: a density functional approach. AB - Density functional calculations within the generalized gradient approximation have been used to investigate the lowest energy electronic and geometric structures of neutral, cationic, and anionic Pd(n) (n=1-7) clusters in the gas phase. In this study, we have examined three different spin multiplicities (M=1, 3, and 5) for different possible structural isomers of each neutral cluster. The calculated lowest energy structures of the neutral clusters are found to have multiplicities, M=1 for Pd(1), Pd(3), Pd(5), Pd(6), and Pd(7), while M=3 for Pd(2) and Pd(4). We have also determined the lowest energy states of cationic and anionic Pd(n) (n=1-7) clusters, formed from the most stable neutral clusters, in three spin multiplicities (M=2, 4, and 6). Bond length, coordination number, binding energy, fragmentation energy, bond dissociation energy, ionization potential, electron affinity, chemical hardness, and electric dipole moment of the optimized clusters are compared with experimental and other theoretical results available in the literature. Based on these criteria, we predict the four atom palladium cluster to be a magic-number cluster. PMID- 18163675 TI - Quantum wave packet dynamics of N(2D)+H2 reaction. AB - The quantum wave packet dynamics of the title reaction within the coupled state approximation is examined here and initial state-selected reaction probabilities, integral reaction cross sections, and thermal rate constants are reported. The ab initio potential energy surface of the electronic ground state (1(2)A(")) of the system recently reported by Ho et al. [J. Chem. Phys., 119, 3063 (2003)] is employed in this investigation. All partial wave contributions up to the total angular momentum J=55 were necessary to obtain converged integral reaction cross sections up to a collision energy of 1.0 eV. Thermal rate constants are calculated from the reaction cross sections and compared with the available theoretical and experimental results. Typical resonances formed during the course of the reaction and elucidating the insertion type mechanism for the product formation are calculated. Vibrational energy levels supported by the deep well (approximately 5.5 eV) of the 1(2)A(") potential energy surface of NH(2) are also calculated for the total angular momentum J=0. A statistical analysis of the spacing between the adjacent levels of this energy spectrum is performed and the extent of irregularity in the spectral sequence is assessed. PMID- 18163676 TI - Analogy of silicon clusters with deltahedral boranes: how far can it go? Reexamining the structure of Sin and Sin 2-, n=5-13 clusters. AB - Silicon clusters of 5 up to 13 atoms, Si(n), n=5-13, and their dianions are studied in the light of an anticipated analogy with the corresponding isoelectronic boranes suggested recently by Zdetsis [J. Chem. Phys. 127, 014314 (2007)]. It is demonstrated that this analogy is a fruitful and powerful concept which allows the straightforward determination of the structures of silicon clusters, based on the structure of corresponding closo-boranes, meeting the requirements of well known structural rules. All lowest-lying structures of Si(n), n=5-13 clusters, have been obtained through a systematic way on the basis of this analogy. For magic clusters, such as Si(6) and Si(10), characterized by special stability, the analogy to boranes seems to be much stronger. PMID- 18163677 TI - Boron 1s photoelectron spectrum of 11BF3: vibrational structure and linewidth. AB - The boron 1s photoelectron spectrum of (11)BF(3) has been measured at a photon energy of 400 eV and a resolution of about 55 meV. The pronounced vibrational structure seen in the spectrum has been analyzed to give the harmonic and anharmonic vibrational frequencies of the symmetric stretching mode, 128.1 and 0.15 meV, as well as the change in equilibrium BF bond length upon ionization, 5.83 pm. A similar change in bond length has been observed for PF(3) and SiF(4), but a much smaller change for CF(4). Theoretical calculations for BF(3) that include the effects of electron correlation give results that are in reasonable accord with the experimental values. The Lorentzian (lifetime) width of the boron 1s core hole in BF(3) is found to be 72 meV, comparable to the value of 77 meV that has been reported for CF(4). PMID- 18163678 TI - Infrared spectra of SF6-.(H2O)n (n=1-3): incipient reaction and delayed onset of water network formation. AB - We present data on the microsolvation of an extended charge distribution with SF(6)(-) as a model system. Infrared spectroscopy, aided by ab initio calculations, shows that the first two water molecules attach to the ion by a combination of single ionic H bonds, sharing one of the F atoms, and weak electrostatic interactions with other F atoms in the ion. No water-water bonds are formed at the dihydrate level, which is an unusual observation, given the strong propensity of water to form H-bonded networks. The onset of water networks occurs with the addition of the third water molecule. Moreover, the attachment of the first two water molecules considerably weakens the SF bond of the F atom involved in bonding to both ligands, indicating a possible mechanism for water induced reactions. PMID- 18163679 TI - Nonadiabatic dynamics of charge transfer in diatomic anion clusters. AB - We have studied the photodissociation and recombination dynamics of the diatomic anions X(2)(-) and XY(-) designed to mimic I(2)(-) and ICl(-), respectively, by using a one-electron model in size-selected N(2)O clusters. The one-electron model is composed of two nuclei and an extra electron moving in a two-dimensional plane including the two nuclei. The main purpose of this study is to explain the salient features of various dynamical processes of molecular ions in clusters using a simple theoretical model. For heteronuclear diatomic anions, a mass disparity and asymmetric electron affinity between the X and Y atoms lead to different phenomena from the homonuclear case. The XY(-) anion shows efficient recombination for a smaller cluster size due to the effect of collision-mediated energy transfer and an inherent potential wall on excited state at asymptotic region, while the recombination for the X(2)(-) anion is due to rearrangement of solvent configuration and faster nonadiabatic transitions. The results of the present study illustrate the microscopic details of the electronically nonadiabatic processes which control the photodissociation dynamics of molecular ions in clusters. PMID- 18163680 TI - Absolute asymmetric synthesis from an isotropic racemic mixture of chiral molecules with the help of their laser orientation-dependent selection. AB - We analyzed the absolute asymmetric synthesis (AAS) of enantiomers from an isotropic racemic mixture of chiral molecules, which employs the laser electrodipole interaction, and revealed a set of basic symmetry-based conditions on the parameters of field-molecule interaction. Using these conditions, we developed a novel scenario of the AAS (through selective photodestruction of the enantiomers of a given type) based on the joint action of the strong multicomponent femtosecond and picosecond laser pulses. Key mechanism of this scenario is the partly modified scheme of laser orientation-dependent selection of molecules proposed by us earlier [D. V. Zhdanov et al., JETP 130, 387 (2006)]. Calculations made on example of chiral molecule SiHNaClF show rather high efficiency and stability of the proposed AAS scenario with respect to the parameters of the incident laser pulses and even feasibility of its realization at room temperature. PMID- 18163681 TI - Multireference theoretical investigation on selectivity of the bond fissions in photodissociation of acetyl cyanide. AB - The selectivity of the C-CH(3) and C-CN bond fissions upon excitation of acetyl cyanide at 193 nm has been investigated at the theoretical level of multistate complete active space self-consistent field second order perturbation. The calculated results indicated that the initially excited S(3) state relaxes to S(2) via ultrafast internal conversion. The S(2) state could dissociate via two pathways. One, adiabatically dissociates to CH(3)CO(X)+CN(A). The other one internally converts to S(1) before S(1) intersystem crossing to T(1). The T(1) state subsequently dissociates to two groups of products: CH(3)(X)+OCCN(X) and CH(3)CO(X)+CN(X). The experimentally observed preference branching of CN elimination over CH(3) one and bond selectivity are the results of the competition between the adiabatic and nonadiabatic dynamics of the S(2) state. PMID- 18163682 TI - Microwave spectrum of the H2DO+ ion: inversion-rotation transitions and inversion splitting. AB - Inversion-rotation spectral lines of the monodeuterated hydronium ion, H(2)DO(+), have been observed by a source-modulation spectrometer in the millimeter- to submillimeter-wave region. The ion was generated by a hollow-cathode discharge in a gas mixture of H(2)O and D(2)O. Nine inversion-rotation lines were measured precisely for the lowest pair of inversion doublets in the frequency region from 210 to 720 GHz. The measured lines were analyzed to derive rotational constants in the inversion-doublet states and inversion splitting. The inversion splitting in the ground state was determined to be 1,215,866(410) MHz, that is, 40.5569(137) cm(-1), where the numbers in parentheses give probable uncertainties estimated from the Jacobian matrix of the assumed centrifugal distortion constants of the inversion-doublet states. The determined inversion splitting is off by -0.58 cm(-1) from the predicted value of 41.14 cm(-1) by Rayamaki et al. using high-order coupled cluster ab initio calculations [J. Chem. Phys. 118, 10929 (2003)], and by 0.039 cm(-1) from the observed value of 40.518(10) cm(-1) by Dong and Nesbitt using high-resolution jet-cooled infrared spectroscopy [J. Chem. Phys. 125, 144311 (2006)] beyond the quoted uncertainty. The most astronomically important transition 0(00)(-)-1(0)(+) for the ortho species was measured at 673,257.024(31) MHz, which could be used as a radioastronomical probe investigating interstellar chemistry of deuterium fractionation in space. PMID- 18163683 TI - Collisional quenching of rotations in lithium dimers by ultracold helium: the Li2(a3Sigma u+) and Li2+(X2Sigma g+) targets. AB - Quantum coupled-channel scattering calculations have been carried out at ultralow energies (down to 10(-5) K) for rotational quenching of ionic and spin-stretched states of the lithium dimer in collision with He atoms. Marked cross section differences, to be related to changes in their interactions with He, have been observed with respect to the singlet case while little changes in the collisional behavior are seen to occur upon ionization when the spin-stretched target is considered. Both effects originate from an interplay of structural and dynamical features of the colliding partners which are analyzed in detail. PMID- 18163684 TI - An approximate theory of the ozone isotopic effects: rate constant ratios and pressure dependence. AB - The isotopic effects in ozone recombination reactions at low pressures are studied using an approximate theory which yields simple analytic expressions for the individual rate constant ratios, observed under "unscrambled" conditions. It is shown that the rate constant ratio between the two competing channels XYZ- >X+YZ and XYZ-->XY+Z is mainly determined by the difference of the zero-point energies of diatomic molecules YZ and XY and by the efficiency of the deactivation of the newly formed excited ozone molecules, whereas the mass independent fractionation depends on a "nonstatistical" symmetry factor eta and the collisional deactivation efficiency. Formulas for the pressure effects on the enrichment and on the rate constant ratios are obtained, and the calculated results are compared with experiments and more exact calculations. In all cases, ratios of isotope rates and the pressure dependence of enrichments, the agreement is good. While the initial focus was on isotope effects in the formation of O(3), predictions are made for isotope effects on ratios of rate constants in other reactions such as O+CO-->CO(2), O+NO-->NO(2), and O+SO-->SO(2). PMID- 18163685 TI - Nature of the metal-nonmetal transition in metal-ammonia solutions. I. Solvated electrons at low metal concentrations. AB - Using a theory of polarizable fluids, we extend a variational treatment of an excess electron to the many-electron case corresponding to finite metal concentrations in metal-ammonia solutions (MAS). We evaluate dielectric, optical, and thermodynamical properties of MAS at low metal concentrations. Our semianalytical calculations based on a mean-spherical approximation correlate well with the experimental data on the concentration and temperature dependencies of the dielectric constant and the optical absorption spectrum. The properties are found to be mainly determined by the induced dipolar interactions between localized solvated electrons, which result in the two main effects: the dispersion attractions between the electrons and a sharp increase in the static dielectric constant of the solution. The first effect creates a classical phase separation for the light alkali metal solutes (Li, Na, K) below a critical temperature. The second effect leads to a dielectric instability, i.e., polarization catastrophe, which is the onset of metallization. The locus of the calculated critical concentrations is in a good agreement with the experimental phase diagram of Na-NH(3) solutions. The proposed mechanism of the metal-nonmetal transition is quite general and may occur in systems involving self-trapped quantum quasiparticles. PMID- 18163686 TI - Electronic polarization effect on low-frequency infrared and Raman spectra of aprotic solvent: molecular dynamics simulation study with charge response kernel by second order Moller-Plesset perturbation method. AB - Low-frequency infrared (IR) and depolarized Raman scattering (DRS) spectra of acetonitrile, methylene chloride, and acetone liquids are simulated via molecular dynamics calculations with the charge response kernel (CRK) model obtained at the second order Moller-Plesset perturbation (MP2) level. For this purpose, the analytical second derivative technique for the MP2 energy is employed to evaluate the CRK matrices. The calculated IR spectra reasonably agree with the experiments. In particular, the agreement is excellent for acetone because the present CRK model well reproduces the experimental polarizability in the gas phase. The importance of interaction induced dipole moments in characterizing the spectral shapes is stressed. The DRS spectrum of acetone is mainly discussed because the experimental spectrum is available only for this molecule. The calculated spectrum is close to the experiment. The comparison of the present results with those by the multiple random telegraph model is also made. By decomposing the polarizability anisotropy time correlation function to the contributions from the permanent, induced polarizability and their cross term, a discrepancy from the previous calculations is observed in the sign of permanent induce cross term contribution. The origin of this discrepancy is discussed by analyzing the correlation functions for acetonitrile. PMID- 18163687 TI - When is H2O not water? AB - We have combined a computational search strategy with first-principles density functional-theory calculations to identify metastable phases of H(2)O under pressure. The most stable structures consist of water molecules, while the most energetic metastable phases consist of oxygen and hydrogen molecules. In between lie many other metastable phases, consisting of various combinations of a few small molecules. It may be possible to synthesize some of these metastable phases, and we use our results to understand the nature of the crystalline metastable phase of H(2)O recently synthesized by Mao et al. [Science 314, 636 (2006)]. PMID- 18163688 TI - Quantum dynamics of an excited alkali atom in a noble gas cluster: lithium attached to a helium cluster. AB - An alkali atom-noble gas cluster system is considered as a model for solvation effects in optical spectra, within a quantum-classical description based on the density operator of a many-atom system and its partial Wigner transform. This leads to an eikonal-time-dependent molecular orbital treatment suitable for a time-dependent description of the coupling of light emission and atom dynamics in terms of the time-dependent electric dipole of the whole system. As an application, we consider an optically excited lithium atom as the dopant in a helium cluster at 0.5 K. We describe the motions of the excited Li atom interacting with a cluster of He atoms and calculate the time-dependent electric dipole of the Li-He(99) system during the dynamics. The electronic Hamiltonian is taken as a sum of three-body Li-He diatomic potentials including electronic polarization and repulsion, with l-dependent atomic pseudopotentials for Li and He, while we use a modified pair potential for He-He. The calculations involve the coupling of 12 quantum states with 300 classical degrees of freedom. We present results for the dynamics and spectra of a Li atom interacting with a model cluster surface of He atoms and also interacting with a droplet of He. We have found that the Li atom is attracted or repulsed from the He surface, depending on the orientation of its 2p orbitals. The spectra and dynamics of Li inside and at the surface of a cluster are found to be strongly dependent on its electronic states, its velocity direction, and whether light is present during emission or not. PMID- 18163689 TI - Supersonic molecular beam studies of dissociative adsorption of H2 on Ru(0001). AB - We examined reactivity of H(2) on Ru(0001) using molecular beam techniques and we compared our results to experimental results for similar systems. The dissociative adsorption of H(2) on Ru(0001) is similar to that on Pt(111) and Ni(111), although on ruthenium nonactivated adsorption is strongly suggested. However, we find no clear signature of a steering- or precursor-based mechanism that favors nonactivated reaction paths at low kinetic energy. In comparison to Pd(111) and Rh(111) our results indicate that a universal mechanism enhancing reactivity at low energy does not have a mass dependence. In addition, we have compared our results to predictions of reactivity for H(2) on Ru(0001) from six dimensional dynamical calculations using two different generalized gradient approximation functionals. It leads us to conclude that the PW91 functional yields a more accurate value for the minimum energy path but does not impose enough corrugation in the potential. The revised-Perdew-Burke-Ernzerhof (RPBE) functional appears to behave slightly better at higher energies, but we find significant quantitative disagreement. We show that the difference is not due to different energy resolutions between experiment and theory. However, it may be due to a dependence of the reactivity on rotational state or on omission of relevant dimensions in the theoretical description. PMID- 18163690 TI - Self-catalytic solution for single-crystal nanowire and nanotube growth. AB - Vast majority of nanowires is grown by the chemical vapor deposition (CVD), molecular beam epitaxy (MBE), metal-organic CVD (MOCVD), or the laser ablation method via the vapor-liquid-solid (VLS) route. Others are grown via the oxide assisted route. In this investigation a self-catalytic synthesis route based on VLS formalism and suitable for the CVD, MBE, MOCVD, or the laser ablation method has been described. Various issues pertaining to growth kinetics, nanowire alignment, diameter distribution, and nanotube formation have been addressed. The strength of the self-catalytic route has been highlighted. As this route does not make use of foreign element catalytic agents to mediate the synthesis, it suffers from difficulties. Attempts have been made to elucidate means to overcome these difficulties. Attempts have also been made to explain the means to separate the nanowires thus produced from the substrate/scaffold, and to control their physicochemical characteristics. PMID- 18163691 TI - Growth-related properties and postgrowth phenomena in organic molecular thin films. AB - The problem of monitoring the structural and morphological evolutions of thin films of organic molecular materials during their growth by organic molecular beam epitaxy and in the postgrowth stage is addressed here by a combination of in situ optical reflectance anisotropy measurements, ex situ optical and morphological investigations, and theoretical simulation of the material optical response. For alpha-quaterthiophene, a representative material in the class of organic molecular semiconductors, the results show that molecules crystallize in the first stage of growth in metastable structures, even when deposition is carried out at room temperature. In the postdeposition stage, the film structure evolves within a few days to the known equilibrium structure of the low temperature polymorph. When deposition is carried out at low substrate temperatures, an evolution of the film morphology is also demonstrated. PMID- 18163692 TI - Tuning LDA+U for electron localization and structure at oxygen vacancies in ceria. AB - We examine the real space structure and the electronic structure (particularly Ce4f electron localization) of oxygen vacancies in CeO(2) (ceria) as a function of U in density functional theory studies with the rotationally invariant forms of the LDA+U and GGA+U functionals. The four nearest neighbor Ce ions always relax outwards, with those not carrying localized Ce4f charge moving furthest. Several quantification schemes show that the charge starts to become localized at U approximately 3 eV and that the degree of localization reaches a maximum at approximately 6 eV for LDA+U or at approximately 5.5 eV for GGA+U. For higher U it decreases rapidly as charge is transferred onto second neighbor O ions and beyond. The localization is never into atomic corelike states; at maximum localization about 80-90% of the Ce4f charge is located on the two nearest neighboring Ce ions. However, if we look at the total atomic charge we find that the two ions only make a net gain of (0.2-0.4)e each, so localization is actually very incomplete, with localization of Ce4f electrons coming at the expense of moving other electrons off the Ce ions. We have also revisited some properties of defect-free ceria and find that with LDA+U the crystal structure is actually best described with U=3-4 eV, while the experimental band structure is obtained with U=7-8 eV. (For GGA+U the lattice parameters worsen for U>0 eV, but the band structure is similar to LDA+U.) The best overall choice is U approximately 6 eV with LDA+U and approximately 5.5 eV for GGA+U, since the localization is most important, but a consistent choice for both CeO(2) and Ce(2)O(3), with and without vacancies, is hard to find. PMID- 18163693 TI - Formation of supercooled liquid solutions from nanoscale amorphous solid films of methanol and ethanol. AB - Molecular beam techniques are used to create layered nanoscale composite films of amorphous methanol and ethanol at 20 K. The films are then heated, and temperature programed desorption and infrared spectroscopy are used to observe the mixing, desorption, and crystallization behavior from the initially unmixed amorphous layers. We find that the initially unmixed amorphous layers completely intermix to form a deeply supercooled liquid solution after heating above T(g). Modeling of the desorption kinetics shows that the supercooled liquid films behave as ideal solutions. The desorption rates from the supercooled and crystalline phases are then used to derive the binary solid-liquid phase diagram. Deviations from ideal solution desorption behavior are observed when the metastable supercooled solution remains for longer times in regions of the phase diagram when thermodynamically favored crystallization occurs. In those cases, the finite lifetime of the metastable solutions results in the precipitation of crystalline solids. Finally, in very thick films at temperatures and compositions where a stable liquid should exist, we unexpectedly observe deviations from ideal solution behavior. Visual inspection of the sample indicates that these apparent departures from ideality arise from dewetting of the liquid film from the substrate. We conclude that compositionally tailored nanoscale amorphous films provide a useful means for preparing and examining deeply supercooled solutions in metastable regions of the phase diagram. PMID- 18163694 TI - Biatomic substrates for bulk-molecule interfaces: the PtCo-oxygen interface. AB - We develop an ab initio procedure for materials participating in electron transfer reactions, to consider biatomic backgrounds (e.g., substrates, catalysts, electrodes), thus the precise effects of the continuum and long-range interactions as well as the effects of the discrete and local nature of the chemistry can be combined to study molecules under such biatomic backgrounds. We test this new procedure by studying the reactivity of molecular oxygen on bimetallic clusters of platinum and cobalt. The reaction of molecular oxygen on the surface of the L1(2) phase of Co(3)Pt yields three distinct chemisorbed precursors, two of which are energetically almost degenerate. One precursor is formed across a bridge site with the molecule parallel to the surface, the second one is formed at the bridge with the molecule again parallel to the surface, and the third is formed in the threefold fcc hollow site, with the molecule slightly tilted in a top-hollow-bridge geometry. PMID- 18163695 TI - White light emission from single layer poly (n-vinylcarbazole) polymeric light emitting devices by mixing singlet and triplet excimer emissions. AB - White light electroluminescence (EL) was obtained by mixing emission from singlet and triplet excimers from a single poly (n-vinylcarbazole) (PVK) spin cast layer after irradiation of the solution with UV light. With increased UV light irradiation, the intensity from the triplet excimer (red-630 nm) of PVK increased compared with that of the singlet excimer (blue-460 nm) due to an increased population of both adjacent benzene rings being aligned with one another (fully overlapping) versus only one of the adjacent benzene rings being aligned (partially overlapping). The emission color changed from blue to white with increased UV irradiation time while the EL brightness and current density decreased and the turn-on voltage increased. PMID- 18163696 TI - Density functional study of the interaction between small Au clusters, Au(n) (n=1 7) and the rutile TiO2 surface. II. Adsorption on a partially reduced surface. AB - We use density functional theory to examine the electronic structure of small Au(n) (n=1-7) clusters, supported on a rutile TiO(2)(110) surface having oxygen vacancies on the surface (a partially reduced surface). Except for the monomer, the binding energy of all Au clusters to the partially reduced surface is larger by approximately 0.25 eV than the binding energy to a stoichiometric surface. The bonding site and the orientation of the cluster are controlled by the shape of the highest occupied molecular orbitals (HOMOs) of the free cluster (free cluster means a gas-phase cluster with the same geometry as the supported one). The bond is strong when the lobes of the HOMOs overlap with those of the high-energy states of the clean oxide surface (i.e., with no gold) that have lobes on the bridging and the in-plane oxygen atoms. In other words, the cluster takes a shape and a location that optimizes the contact of its HOMOs with the oxygen atoms. Fivefold coordinated Ti atoms located at a defect site (5c-Ti(*)) participate in the binding only when a protruding lobe of the singly occupied molecular orbital (for odd n) or the lowest unoccupied molecular orbital (for even n) of the free Au(n) cluster points toward a 5c-Ti(*) atom. The oxygen vacancy influences the binding energy of the clusters (except for Au(1)) only when they are in direct contact with the defect. The desorption energy and the total charge on clusters that are close to, but do not overlap with, the vacancy differ little from the values they have when the cluster is adsorbed on a stoichiometric surface. The behavior of Au(1) is rather remarkable. The atom prefers to bind directly to the vacancy site with a binding energy of 1.81 eV. However, it also makes a strong bond (1.21 eV) with any 5c-Ti atom even if that atom is far from the vacancy site. In contrast, the binding of a Au monomer to the 5c-Ti atom of a surface without vacancies is weak (0.45 eV). The presence of the vacancy activates the 5c Ti atoms by populating states at the bottom of the conduction band. These states are delocalized and have lobes protruding out of the surface at the location of the 5c-Ti atoms. It is the overlap of these lobes with the highest orbital of the Au atom that is the major reason for the bonding to the 5c-Ti atom, no matter how far the latter is from the vacancy. The energy for breaking an adsorbed cluster into two adsorbed fragments is smaller than the kinetic energy of the mass selected clusters deposited on the surface in experiments. However, this is not sufficient for breaking the cluster upon impact with the surface, since only a fraction of the available energy will go into the reaction coordinate for breakup. PMID- 18163697 TI - Nucleation in electrochemical growth of the Ag(100) crystal face: determining the nucleus size via the nucleation theorem. AB - We employ the nucleation theorem for a model-independent determination of the size of the two-dimensional (2D) Ag nucleus with the aid of experimental data for the nucleation-mediated electrochemical growth of the Ag(100) crystal face in aqueous solution of AgNO(3) at 318 K. These data are for the stationary rate of 2D nucleation, for the initial portion of the potentiostatic current transient pertaining to atomically smooth face, and for the galvanostatic current corresponding to stationary growth of the face. It turns out that the 2D nucleus is constituted of 17-64 Ag atoms when the overpotential is in the range of 12 22.4 mV. Upon expressing the overpotential in terms of supersaturation, it is found that the experimental data for the size of the 2D Ag nucleus are in conformity with existing simulation data for the size of the 2D nucleus on the (100) face of Kossel crystal (the simulation nucleus contains 1-30 atoms). It is found as well that the Gibbs-Thomson equation of the classical theory of 2D nucleation describes very well the supersaturation dependence of the size of both the Ag and the simulation nucleus. PMID- 18163698 TI - Water wettability of close-packed metal surfaces. AB - We propose a new microscopic criterion to determine surface wetting: water wets the surface whenever its overlayer has a larger adsorption energy than three dimensional clusters on the bare or water-covered substrate. This conceptually intuitive criterion is validated by detailed first-principles calculations of the energetics of layers and clusters of water on different metal surfaces. This criterion resolves naturally the current discrepancy between theory and experiment on the wetting behavior of undissociated water on Ru(0001), as well as the hydrophobic nature of the Au(111) surface. It also explains the Stranski Krastanov ice growth on Pt(111) observed experimentally. PMID- 18163699 TI - Chemical imaging of single 4,7,12,15-tetrakis[2.2]paracyclophane by spatially resolved vibrational spectroscopy. AB - Single 4,7,12,15-tetrakis[2.2]paracyclophane were deposited on NiAl(110) surface at 11 K. Two adsorbed species with large and small conductivities were detected by the scanning tunneling microscope (STM). Their vibrational properties were investigated by inelastic electron tunneling spectroscopy (IETS) with the STM. Five vibrational modes were observed for the species with the larger conductivity. The spatially resolved vibrational images for the modes show striking differences, depending on the coupling of the vibrations localized on different functional groups within the molecule to the electronic states of the molecule. The vibrational modes are assigned on the basis of ab initio calculations. No IETS signal is resolved from the species with the small conductivity. PMID- 18163700 TI - Efficient knot group identification as a tool for studying entanglements of polymers. AB - A technique is presented for the identification of the knot group of knots, links, and other embedded graphs as a tool in numerical studies of entanglements of polymers. With this technique, the knot group is simultaneously more discriminating and easier to calculate than the knot invariants that have been used in such studies in the past. It can be applied even in cases of very complex knot projections with hundreds of crossings. Starting from an arbitrary projection of an embedded graph, we generate a sequence of representations, any one of which is a full and complete representation of the knot group. Any two knot groups are isomorphic if they have identical representations. Therefore, we compare the sequence of representations of any given knot or link against a previously determined lookup table, and if the group of the knot or link is represented in this table we eventually find a match and identify the knot group. PMID- 18163701 TI - Development of knotting during the collapse transition of polymers. AB - A dynamic Monte Carlo simulation of the collapse transition of polymer chains is presented. The chains are represented as self-avoiding walks on the simple cubic lattice with a nearest-neighbor contact potential to model the effect of solvent quality. The knot state of the chains is determined using the knot group procedure presented in the accompanying paper. The equilibrium knot spectrum and the equilibrium rms radius of gyration as functions of the chain length and the contact potential are reported. The collapse transition was studied following quenches from good-to poor-solvent conditions. Our results confirm the prediction that the newly formed globule is not yet at equilibrium, since it has not yet achieved its equilibrium knot spectrum. For our model system, the relaxation of the knot spectrum is about an order of magnitude slower than that of the radius of gyration. The collapse transition is also studied for a model in which both ends of the chain remain in good-solvent conditions. Over the time scale of these simulations, knot formation is frustrated in this inhomogeneous model, verifying that the mechanism of knotting is the tunneling of chain ends in and out of the globule. PMID- 18163702 TI - Formation of nonextinct ring-banded textures and multistacked lamella of tetra aniline-block-poly(L-lactide) rod-coil diblock oligomer films induced by solvent vapor treatment. AB - The self-assembly processes of the rod-coil diblock oligomer thin film of tetra aniline (TANI)-block-poly(L-lactide) (PLLA) with different film thicknesses induced in the coil-selective solvent of acetone vapor at room temperature were studied. The morphologies of the oligomer films were determined by the film thickness. For the thicker film (232 nm), the nonextinct concentric ring-banded textures could form. While for the thinner and appropriate film (about 6 nm), multistacked diamond-shaped appearances with the periodic thickness being about 8.5 nm(6-nm-thick extended PLLA chain and 2.5-nm-thick p-pi conjugating TANI bimolecular layer) formed. The possible formation models of those two regular morphologies were presented in detail. PMID- 18163703 TI - Modified interfacial statistical associating fluid theory: a perturbation density functional theory for inhomogeneous complex fluids. AB - A density functional theory based on Wertheim's first order perturbation theory is developed for inhomogeneous complex fluids. The theory is derived along similar lines as interfacial statistical associating fluid theory [S. Tripathi and W. G. Chapman, J. Chem. Phys. 122, 094506 (2005)]. However, the derivation is more general and applies broadly to a range of systems, retaining the simplicity of a segment density based theory. Furthermore, the theory gives the exact density profile for ideal chains in an external field. The general avail of the theory has been demonstrated by applying the theory to lipids near surfaces, lipid bilayers, and copolymer thin films. The theoretical results show excellent agreement with the results from molecular simulations. PMID- 18163704 TI - Effect of packing parameter on phase diagram of amphiphiles: an off-lattice Gibbs ensemble approach. AB - We determine the phase diagram of several amphiphilic molecules as a function of the amphiphilic parameter alpha defined as the ratio of the volume of hydrophilic to hydrophobic segments using the Gibbs ensemble Monte Carlo method supplemented by configurational bias scheme. Specifically, we study amphiphilic molecules h(1)t(7), h(2)t(6), and h(3)t(5), for which alpha=0.14, 0.33, and 0.60 respectively, and demonstrate that the former two exhibit phase separation while h(3)t(5) forms micelles, supporting the contention that alpha=0.5 is the border line for phase separation and micellization, as observed in previous lattice Monte Carlo studies [Panagiotopoulos et al., Langmuir 18, 2940 (2002)]. Further, we study the phase separation in amphiphilic molecules as a function of the packing parameter by varying the size of the hydrophilic head for each molecule. We find that a larger hydrophilic head lowers the critical temperature T(c), and raises the critical density rho(c). PMID- 18163705 TI - A dynamic density functional theory for particles in a flowing solvent. AB - We present a dynamic density functional theory (dDFT) which takes into account the advection of the particles by a flowing solvent. For potential flows, we can use the same closure as in the absence of solvent flow. The structure of the resulting advected dDFT suggests that it could be used for nonpotential flows as well. We apply this dDFT to Brownian particles (e.g., polymer coils) in a solvent flowing around a spherical obstacle (e.g., a colloid) and compare the results with direct simulations of the underlying Brownian dynamics. Although numerical limitations do not allow for an accurate quantitative check of the advected dDFT both show the same qualitative features. In contrast to previous works which neglected the deformation of the flow by the obstacle, we find that the bow wave in the density distribution of particles in front of the obstacle as well as the wake behind it are reduced dramatically. As a consequence, the friction force exerted by the (polymer) particles on the colloid can be reduced drastically. PMID- 18163706 TI - Mediation of long-range attraction selectively between negatively charged colloids on surfaces by solvation. AB - We propose a mean-field analytical model to account for the observed asymmetry in the ability to form long-range attraction by the negatively charged colloidal particles and not their equivalently charged positive counterpart. We conjecture that this asymmetry is due to solvation effects, and we phenomenologically capture its physics by considering the relative strength of this water-induced short-range repulsion between the different charge species. We then apply our model to the colloidal system of negatively charged disks that are neutralized by a sea of counterions and strongly absorbed to an interface in a compressible binary system. We demonstrate the resulting coexistence between a dilute isotropic ionic phase and a condensed hexagonal lattice phase as a function of density and interaction strength. PMID- 18163707 TI - Phase separation kinetics of polyelectrolyte solutions. AB - The kinetics of phase separation of aqueous solutions of sodium-poly(styrene sulfonate) (NaPSS) containing barium chloride (BaCl(2)) is studied by static and dynamic light scattering. We report a novel mechanism of phase separation, where an enrichment of polymer aggregates of well-defined size occurs in the very early stage of nucleation, which is then followed by a growth process in the formation of the new phase. In the latter stage, the polymer aggregates formed in the early stage act as the templating nuclei. Even in the homogeneous phase at higher temperatures above the upper critical phase boundary, polymer aggregates are present in agreement with previously reported results. Upon rapidly cooling the system below the phase boundary, the number concentration of the aggregates increases first by maintaining their size to be relatively monodisperse, before the growth process takes over at later times. The size and fractal dimension of aggregates in the homogeneous phase and the early nucleation stage of phase separation and the dependence of nucleation time and growth rate on quench depth and salt concentration are determined. The hydrodynamic radius (R(H)) of the unaggregated chains is of the order of 1-10 nm depending on the molecular weight of NaPSS, while R(H) of aggregates is of the order of 100 nm independent of the molecular weight of NaPSS. Unaggregated chains follow good solution behavior with a fractal dimension of 5/3 while the fractal dimension of aggregates is larger than 3.5 suggesting the branched nature of aggregates. Nucleation time is sensitive to quench depth and salt concentration. Increasing a quench depth or increasing BaCl(2) concentration shortens the nucleation time. After the nucleation time, during the growth period, the size of aggregates grows linearly with time, with growth rate being higher for deeper quench depths and higher BaCl(2) concentrations. The mechanism of phase separation of aqueous solutions of NaPSS and BaCl(2) is seen to proceed by utilizing the already-existing aggregates to nucleate the new phase, in marked contrast to hitherto known results on phase separation in uncharged polymer systems. PMID- 18163708 TI - Depletion induced isotropic-isotropic phase separation in suspensions of rod-like colloids. AB - When non-adsorbing polymers are added to an isotropic suspension of rod-like colloids, the colloids effectively attract each other via depletion forces. We performed Monte Carlo simulations to study the phase diagram of such rod-polymer mixture. The colloidal rods were modeled as hard spherocylinders; the polymers were described as spheres of the same diameter as the rods. The polymers may overlap with no energy cost, while the overlap of polymers and rods is forbidden. Large amounts of depletant cause phase separation of the mixture. We estimated the phase boundaries of isotropic-isotropic coexistence both in the bulk and in confinement. To determine the phase boundaries we applied the grand canonical ensemble using successive umbrella sampling [J. Chem. Phys. 120, 10925 (2004)], and we performed a finite size scaling analysis to estimate the location of the critical point. The results are compared with predictions of the free volume theory developed by Lekkerkerker and Stroobants [Nuovo Cimento D 16, 949 (1994)]. We also give estimates for the interfacial tension between the coexisting isotropic phases and analyze its power-law behavior on the approach of the critical point. PMID- 18163709 TI - Complete phase behavior of the symmetrical colloidal electrolyte. AB - We computed the complete phase diagram of the symmetrical colloidal electrolyte by means of Monte Carlo simulations. Thermodynamic integration, together with the Einstein-crystal method, and Gibbs-Duhem integration were used to calculate the equilibrium phase behavior. The system was modeled via the linear screening theory, where the electrostatic interactions are screened by the presence of salt in the medium, characterized by the inverse Debye length, kappa (in this work kappasigma=6). Our results show that at high temperature, the hard-sphere picture is recovered, i.e., the liquid crystallizes into a fcc crystal that does not exhibit charge ordering. In the low temperature region, the liquid freezes into a CsCl structure because charge correlations enhance the pairing between oppositely charged colloids, making the liquid-gas transition metastable with respect to crystallization. Upon increasing density, the CsCl solid transforms into a CuAu like crystal and this one, in turn, transforms into a tetragonal ordered crystal near close packing. Finally, we have studied the ordered-disordered transitions finding three triple points where the phases in coexistence are liquid-CsCl disordered fcc, CsCl-CuAu-disordered fcc, and CuAu-tetragonal-disordered fcc. PMID- 18163710 TI - Helix formation inside a nanotube: possible influence of backbone-water hydrogen bonding by the confining surface through modulation of water activity. AB - Recent molecular dynamics simulations of Sorin and Pande [J. Am. Chem. Soc. 128, 6316 (2006)] in explicit solvent found that helix formation of an alanine peptide is disfavored inside a nanotube relative to that in bulk solution. Here, we present a theory to quantitatively rationalize their simulation results. The basic idea is that the nonpolar inner surface of the nanotube creates a depletion layer and raises the activity of the confined water. The raised water activity, in turn, stabilizes the coil state through hydrogen bonding with the backbone amides and carbonyls. We account for the influence of water activity on helix formation within the Lifson-Roig theory. With physically reasonable parameters, the dependence of the helical content on the diameter of the nanotube obtained in the simulations is well reproduced. PMID- 18163711 TI - Base-pair interactions in the gas-phase proton-bonded complexes of C+G and C+GC. AB - Interactions involved in the formation of gas-phase proton-bonded molecular complexes of cytosine (C) and guanine (G) were theoretically investigated for the case of C(+)G and C(+)GC using B3LYP density functional theory. In this study, particular focus was on the dimeric interaction of proton-bonded C(+)G, where a proton bond and a hydrogen bond are cooperatively involved. The dimer interaction energy in terms of dissociation energy (D(e)) was predicted to be 41.8 kcal/mol. The lowest (frozen) energy structure for the C(+)G dimeric complex was found to be CH(+)...G rather than C...H(+)G in spite of the lower proton affinity of the cytosine moiety, which was more stable by 3.3 kcal/mol. The predicted harmonic vibrational frequencies and bond lengths suggest that the combined contributions of proton and hydrogen bonding may determine the resultant stability of each complex structure. In contrast to the dimer case, in the case of the isolated C(+)GC triplet, the two minimum energy structures of CH(+)...GC and C...H(+)GC were predicted to be almost equivalent in total energy. The dissociation energy (D(e)) for the C(+)G pairing in the C(+)GC triplet was 43.7 kcal/mol. Other energetics are also reported. As for the proton-transfer reaction in the proton bond axis, the forward proton-transfer barriers for the dimer and trimer complexes were also predicted to be very low, 3.6 and 1.5 kcal/mol (DeltaE(e)(PT)), respectively. PMID- 18163712 TI - Perspective: tools of modern magnetic materials research: Vector and Bragg magneto-optical Kerr effect for the analysis of nanostructured magnetic arrays [Rev. Sci. Instrum. 78, 121301 (2007)]. PMID- 18163713 TI - Invited article: Vector and Bragg Magneto-optical Kerr effect for the analysis of nanostructured magnetic arrays. AB - Experimental and theoretical aspects of obtaining the magnetic information carried by laser beams diffracted from an array of micro- or nanosized magnetic objects are reviewed. We report on the fundamentals of vector magneto-optic Kerr effect (MOKE), Bragg-MOKE, and second-order effects in the Kerr signal in longitudinal Kerr geometry as well as on an experimental setup used for vector and Bragg-MOKE experiments. The vector and Bragg-MOKE technique in combination with micromagnetic simulation is a reliable tool for measuring the complete magnetization vector and for characterizing the reversal mechanism of lateral magnetic nanostructures. We discuss the Bragg-MOKE effect for three standard domain configurations during the magnetization reversal process and present the expected behavior of the magnetic hysteresis loops. PMID- 18163714 TI - A hyperspectral fluorescence lifetime probe for skin cancer diagnosis. AB - The autofluorescence of biological tissue can be exploited for the detection and diagnosis of disease but, to date, its complex nature and relatively weak signal levels have impeded its widespread application in biology and medicine. We present here a portable instrument designed for the in situ simultaneous measurement of autofluorescence emission spectra and temporal decay profiles, permitting the analysis of complex fluorescence signals. This hyperspectral fluorescence lifetime probe utilizes two ultrafast lasers operating at 355 and 440 nm that can excite autofluorescence from many different biomolecules present in skin tissue including keratin, collagen, nicotinamide adenine dinucleotide (phosphate), and flavins. The instrument incorporates an optical fiber probe to provide sample illumination and fluorescence collection over a millimeter-sized area. We present a description of the system, including spectral and temporal characterizations, and report the preliminary application of this instrument to a study of recently resected (<2 h) ex vivo skin lesions, illustrating its potential for skin cancer detection and diagnosis. PMID- 18163715 TI - KMC-1: a high resolution and high flux soft x-ray beamline at BESSY. AB - The crystal monochromator beamline KMC-1 at a BESSY II bending magnet covers the energy range from soft (1.7 keV) to hard x-rays (12 keV) employing the (n,-n) double crystal arrangement with constant beam offset. The monochromator is equipped with three sets of crystals, InSb, Si (111), and Si (422) which are exchangeable in situ within a few minutes. Beamline and monochromator have been optimized for high flux and high resolution. This could be achieved by (1) a windowless setup under ultrahigh-vacuum conditions up to the experiment, (2) by the use of only three optical elements to minimize reflection losses, (3) by collecting an unusually large horizontal radiation fan (6 mrad) with the toroidal premirror, and (4) the optimization of the crystal optics to the soft x-ray range necessitating quasibackscattering crystal geometry (theta(Bragg,max)=82 degrees) delivering crystal limited resolution. The multipurpose beamline is in use for a variety of user facilities such as extended x-ray absorption fine structure, ((Bio-)EXAFS) near-edge x-ray absorption fine structure (NEXAFS), absorption and fluorescence spectroscopy. Due to the windowless UHV setup the k edges of the technologically and biologically important elements such as Si, P, and S are accessible. In addition to these experiments this beamline is now extensively used for photoelectron spectroscopy at high kinetic energies. Photon flux in the 10(11)-10(12) photons/s range and beamline resolving powers of more than E/DeltaE approximately 100.000 have been measured at selected energies employing Si (nnn) high order radiation in quasibackscattering geometry, thus photoelectron spectroscopy with a total instrumental resolution of about 150 meV is possible. This article describes the design features of the beamline and reports some experimental results in the above mentioned fields. PMID- 18163716 TI - Phase-locked scanning interferometer for frequency stabilization of multiple lasers. AB - We report a simple scheme for stabilizing and tuning the length of a conventional piezo-driven optical cavity against the resonant transmission of a master laser. In contrast with other schemes, we drive the piezo at its mechanical resonance of 5 kHz over an amplitude equivalent to one free spectral range and use a feedback circuit that incorporates phase-sensitive detection of the master-laser transmission. The bandwidth of our cavity-lock circuit is limited only by the resonance frequency of the cavity piezo and is 1.4 kHz. The stabilized mean cavity length reaches in 30 s a minimum Allan deviation of approximately 10 kHz (a length stability of 20 parts per trillion) equaling that of the polarization stabilized He-Ne we use as our master laser. Here, we investigate the mechanical characteristics of the cavity, describe the lock circuit and its measured performance, and provide simple analytical relations between the phase-sensitive signal and cavity displacement. Our setup economizes the cost and amount of equipment necessary for stabilizing multiple continuous-wave lasers operating at different wavelengths. PMID- 18163717 TI - High-intensity xenon plasma discharge lamp for bulk-sensitive high-resolution photoemission spectroscopy. AB - We have developed a highly brilliant xenon (Xe) discharge lamp operated by microwave-induced electron cyclotron resonance (ECR) for ultrahigh-resolution bulk-sensitive photoemission spectroscopy (PES). We observed at least eight strong radiation lines from neutral or singly ionized Xe atoms in the energy region of 8.4-10.7 eV. The photon flux of the strongest Xe I resonance line at 8.437 eV is comparable to that of the He Ialpha line (21.218 eV) from the He-ECR discharge lamp. Stable operation for more than 300 h is achieved by efficient air cooling of a ceramic tube in the resonance cavity. The high bulk sensitivity and high-energy resolution of PES using the Xe lines are demonstrated for some typical materials. PMID- 18163718 TI - Compact soft x-ray spectrometer for plasma diagnostics at the Heidelberg Electron Beam Ion Trap. AB - A compact flat-field soft x-ray grazing-incidence grating spectrometer equipped with a cryogenically cooled back-illuminated charge-coupled device camera was built and implemented at the Heidelberg Electron Beam Ion Trap. The instrument spans the spectral region from 1 to 37 nm using two different gratings. In slitless operation mode, it directly images a radiation source, in this case ions confined in an electron beam ion trap, with high efficiency and reaching hereby a resolving power of lambda/Deltalambda approximately =130 at 2 nm and of lambda/Deltalambda approximately =600 at 28 nm. Capable of automatized operation, its low noise and excellent stability make it an ideal instrument not only for spectroscopic diagnostics requiring wide spectral coverage but also for precision wavelength measurements. PMID- 18163719 TI - Recent progress in vacuum-ultraviolet polarization modulation spectroscopy using polarizing undulator at the TERAS BL5 beamline. AB - Polarization modulation spectroscopy using an Onuki-type undulator is a useful technique for circular dichroism study in the vacuum-ultraviolet region. We have been developing the vacuum-ultraviolet circular dichroism (vuv-CD) spectroscopy in TERAS BL5 beamline at AIST. This paper describes recent improvements in our instrumentation and methods of analysis to achieve precise and absolute measurements. The CD signal is usually accompanied by experimental artifacts, and elimination of all possible artifacts is the key issue for making reliable measurements. After improving beamline optical system, light flux monitor, and undulator operation method, the base line shift of the CD spectrum is suppressed less than 3x10(-4). Sample manipulation and data processing procedures are also described and absolute CD spectrum can be obtained even for linear anisotropic sample. These progresses lead to more quantitative comparison of experimental with calculation on vuv-CD spectrum. PMID- 18163720 TI - Direct measurement of the matched spot size in a slow capillary discharge optical waveguide. AB - This communication presents direct method for experimental determining the matched spot size in a plasma optical waveguide, created in a slow capillary discharge. It can be used for Laser Wakefield Acceleration experiments in addition to interferometry for fast control of optical properties of discharge plasma. The measurements are done by means of the comparison of the laser beam size at the entrance and at the exit of the plasma channel. They are direct in the sense that the interpretation is made in terms of the refractive index without usage of the information about electron density distribution. The method can be used for matched spot size measurement in conditions of the nonlinear effects (transmission of high power laser pulses). PMID- 18163721 TI - Mid-infrared optical coherence tomography. AB - A time domain optical coherence tomography (OCT) system is described that uses mid-infrared light (6-8 microm). To the best of our knowledge, this is the first OCT system that operates in the mid-infrared spectral region. It has been designed to characterize bioengineered tissues in terms of their structure and biochemical composition. The system is based upon a free-space Michelson interferometer with a germanium beam splitter and a liquid nitrogen cooled HgCdTe detector. A key component of this work has been the development of a broadband quantum cascade laser source (InGaAs/AlInAs containing 11 different active regions of the three well vertical transition type) that emits continuously over the 6-8 microm wavelength range. This wavelength range corresponds to the so called "mid-infrared fingerprint region" which exhibits well-defined absorption bands that are specifically attributable to the absorbing molecules. Therefore, this technology provides an opportunity for optical coherence molecular imaging without the need for molecular contrast agents. Preliminary measurements are presented. PMID- 18163722 TI - Portable x-ray fluorescence spectrometer for coating thickness measurement. AB - A handheld x-ray spectrometer has been realized and tested. The purpose of the device is to measure the thickness of coated samples in the range of 1-1500 nm in an industrial environment. Accuracy of approximately 3% has been achieved in this range with a measurement time of 1 min. Automated software has been implemented to allow utilization by a nonspecialist operator. An automated calibration procedure, based on measurements of reference samples, is used. PMID- 18163723 TI - Determination of neutral carbon concentration in electron cyclotron resonance generated plasma discharges. AB - Carbon containing plasmas play an important role not only in plasma technology but also in thermonuclear fusion research. In order to understand and control the processes taking place in the plasma, the knowledge of the carbon ground state density is of major importance. It can be determined by absorption and emission spectroscopy. Detailed measurements were performed in the past to determine the silicon ground state density by means of spectroscopy of the self-absorbed spectral lines of the silicon ground state multiplet at 251 nm. The same procedure was applied for the determination of the carbon concentration, for which the carbon multiplet at 165 nm was analyzed and compared to a simulated spectrum. The ground state density was determined by two independent methods. PMID- 18163724 TI - Design and operating characteristics of new type cold cathode ion source. AB - In this work, the design and performance of new type ion source are described. The discharge mechanism of the source is based on creating an arc discharge through a saddle electric field inside the discharge tube. The saddle electric field is created by immersing an annular anode inside the discharge tube covered from the upper and lower ends with two flanges. These two flanges act as cathodes. The discharge tube is surrounded by a solenoid coil which produces an axial magnetic field (up to 400 G) measured at the center of the source. Measurements have been performed to find out the influence of arc power, pressure, discharge voltage, magnetic field, and extracting voltage on the ion source properties. The source yields an argon ion current of approximately 0.6 mA and electron current of approximately 4 mA at normal operating conditions (extraction voltage V(ex)=7 kV, pressure of 5.5x10(-4) Torr, V(arc)=400 V, I(arc)=1 A, B=200 G). It showed an energy spread of 20 eV at a discharge voltage of 400 V and an extraction voltage of 3 kV. PMID- 18163725 TI - Femtosecond synchronism of x-rays and visible/infrared light in an x-ray free electron laser. AB - A way is proposed to obtain ultrashort pulses of intense infrared/visible light in few-femtosecond synchronism with x-rays from an x-ray free-electron laser (XFEL). It makes use of the recently proposed emittance-slicing technique [Emma et al., Phys. Rev. Lett. 92, 074801 (2004)] to both restrict the duration of self amplified spontaneous emission (SASE) to a few femtoseconds and to lead to a coherence enhancement of near-infrared transition undulator radiation (CTUR). The x-rays and the near-infrared light originate within the XFEL undulator from the same slice of electrons within a bunch and are therefore perfectly synchronized with each other. An example of realizing the scheme at the Linac Coherent Light Source is presented. A few side issues are explored briefly, such as the magnitude of the velocity term versus the acceleration term in the Lienard Wiechert fields and the possible use of the CTUR as a diagnostic tool for the SASE process itself. PMID- 18163726 TI - Fine positioning of a poloidal probe array. AB - Multipoint detection is an essential requirement for investigating plasma turbulence which is a highly nonlinear phenomenon in space and time. We have fabricated an array of 64-channel poloidal probes surrounding the linear cylindrical plasma named LMD-U in order to study turbulence properties, particularly the nonlinear mode couplings, in the domain of poloidal wave number and frequency. However, misalignments of probe tips produce spurious modes, which do not exist in the real plasma, to distort the precise wave number measurements. The paper presents the description of the 64-channel poloidal probe array with means to adjust the probe positions, with discussion on the effects of the misalignments on the wave number measurements. PMID- 18163727 TI - Two-dimensional signal reconstruction: the correlation sampling method. AB - An accurate approach for reconstructing a time-dependent two-dimensional signal from non-synchronized time series recorded at points located on a grid is discussed. The method, denoted as correlation sampling, improves the standard conditional sampling approach commonly employed in the study of turbulence in magnetoplasma devices. Its implementation is illustrated in the case of an artificial time-dependent signal constructed using a fractal algorithm that simulates a fluctuating surface. A statistical method is also discussed for distinguishing coherent (i.e., collective) from purely random (noisy) behavior for such two-dimensional fluctuating phenomena. PMID- 18163728 TI - A new emissive-probe method for electron temperature measurement in radio frequency plasmas. AB - A new method to measure electron temperature by an emissive probe has been proposed. The method is based on measurement of the functional relationship between the floating potential and the heating voltage of emissive probe. From the measured data of the floating potential change as a function of the heating voltage, the electron temperature could be determined by comparing with the theoretical curve obtained under the assumption of Maxwellian distribution. The overall characteristic of the floating potential change could be explained as a function of the heating voltage. The electron temperatures obtained by the present method were consistent with those measured by the rf-compensated Langmuir probe within the error. These experimental verifications were made in the electron density range of 2.6x10(11)-2.8x10(12) cm(-3). It was stressed that the present method is advantageous in that the probe is operated in a floating condition, hence applicable to plasmas produced in an insulated container. PMID- 18163729 TI - Neutron production from feedback controlled thermal cycling of a pyroelectric crystal. AB - The LLNL Crystal Driven Neutron Source is operational and has produced record ion currents of approximately 10 nA and neutron output of 1.9(+/-0.3)x10(5) per thermal cycle using a crystal heating rate of 0.2 degrees C/s from 10 to 110 degrees C. A 3 cm diameter by 1 cm thick LiTaO(3) crystal with a socket secured field emitter tip is thermally cycled with feedback control for ionization and acceleration of deuterons onto a deuterated target to produce D-D fusion neutrons. The entire crystal and temperature system is mounted on a bellows which allows movement of the crystal along the beam axis and is completely contained on a single small vacuum flange. The modular crystal assembly permitted experimental flexibility. Operationally, flashover breakdowns along the side of the crystal and poor emitter tip characteristics can limit the neutron source. The experimental neutron results extend earlier published work by increasing the ion current and pulse length significantly to achieve a factor-of-two higher neutron output per thermal cycle. These findings are reviewed along with details of the instrument. PMID- 18163730 TI - Compact cantilever force probe for plasma pressure measurements. AB - A simple, compact cantilever force probe (CFP) has been developed for plasma pressure measurements. It is based on the pull-in phenomenon well known in microelectromechanical-system electrostatic actuators. The probe consists of a thin (25 mum) titanium foil cantilever (38 mm of length and 14 mm of width) and a fixed electrode separated by a 0.75 mm gap. The probe is shielded by brass box and enclosed into boron nitride housing with a 9 mm diameter window for exposing part of cantilever surface to the plasma. When the voltage is applied between the cantilever and the electrode, an attractive electrostatic force is counterbalanced by cantilever restoring spring force. At some threshold (pull-in) voltage the system becomes unstable and the cantilever abruptly pulls toward the fixed electrode until breakdown occurs between them. The threshold voltage is sensitive to an additional externally applied force, while a simple detection of breakdown occurrence can be used to measure that threshold voltage value. The sensitivity to externally applied forces obtained during calibration is 0.28 V/microN (17.8 VPa for pressure). However, the resolution of the measurements is +/-0.014 mN (+/-0.22 Pa) due to the statistical scattering in measured pull-in voltages. The diagnostic temporal resolution is approximately 10 ms, being determined by the dynamics of pull-in process. The probe has been tested in the tokamak ISTTOK edge plasma, and a plasma force of approximately 0.07 mN (plasma pressure approximately 1.1 Pa) has been obtained near the leading edge of the limiter. This value is in a reasonable agreement with the estimations using local plasma parameters measured by electrical probes. The use of the described CFP is limited by a heat flux of Q approximately 10(6) W/m(2) due to uncontrollable rise of the cantilever temperature (DeltaT approximately 20 degrees C) during CFP response time. PMID- 18163731 TI - A cryogenic Quadraprobe scanning tunneling microscope system with fabrication capability for nanotransport research. AB - We describe the development and the capabilities of an advanced system for nanoscale electrical transport studies. This system consists of a low temperature four-probe scanning tunneling microscope (STM) and a high-resolution scanning electron microscope coupled to a molecular-beam epitaxy sample preparation chamber. The four STM probes can be manipulated independently with subnanometer precision, enabling atomic resolution STM imaging and four-point electrical transport study of surface electronic systems and nanostructured materials at temperatures down to 10 K. Additionally, an integrated energy analyzer allows for scanning Auger microscopy to probe chemical species of nanostructures. Some testing results are presented. PMID- 18163732 TI - Simultaneous correction of flat field and nonlinearity response of intensified charge-coupled devices. AB - Intensified charge-coupled devices (ICCDs) are used extensively in many scientific and engineering environments to image weak or temporally short optical events. Care has to be taken in interpreting the images from ICCDs if quantitative results are required. In particular, nonuniform gain (flat field) and nonlinear response effects must be properly accounted for. Traditional flat field corrections can only be applied in the linear regime of the ICCD camera, which limits the usable dynamic range. This paper reports a more general approach to image correction whereby the nonlinear gain response of each pixel of the ICCD is characterized over the full dynamic range of the camera. Image data can then be corrected for the combined effects of nonuniform gain and nonlinearity. The results from a two-color pyrometry measurement of soot field temperature are used to illustrate the capabilities of the new correction approach. PMID- 18163733 TI - Phase-referenced probe interferometer for biological surface profiling and displacement measurements. AB - We present a probe-based, phase-referenced low coherence interferometer in which the reference field is provided by a fiber end reflection. A gradient-index microlens focuses light onto a sample and collects reflected light. We use the probe interferometer to measure surface profiles of the compound eye of a housefly (Musca domestica) and measure nanometer-scale vibrations in a test sample. PMID- 18163734 TI - Adaptation of the Bridgman anvil cell to liquid pressure mediums. AB - The advantage of Bridgman anvil pressure cells is their wide pressure range and the large number of wires which can be introduced into the pressure chamber. In these pressure cells, soft solid pressure mediums such as steatite are used. We have succeeded in adapting the Bridgman cell to liquid pressure mediums. With this breakthrough, it is now possible to measure in very good hydrostatic pressure conditions up to 7 GPa, which is about twice the pressure attainable in piston-cylinder cells. The pressure gradient in the cell, estimated from the superconducting transition width of lead, is reduced by a factor of 5 in the liquid medium with respect to steatite. By using nonmagnetic materials for the anvils and the clamp and due to the small dimensions of the latter, our device is specially suitable for magnetotransport measurements in dilution fridges. This pressure cell has been developed to measure very fragile and brittle samples such as organic conductors. Resistivity measurements of (TMTTF)(2)BF(4) performed in a solid and a liquid pressure medium demonstrate the necessity of hydrostatic pressure conditions for the study of organic conductors at high pressures. PMID- 18163735 TI - Experimental requirements for measuring pneumatochemical impedances. AB - Hydrogen storage remains a bottleneck process on the way to the hydrogen economy. For practical applications, metal hydride systems offer interesting features, in particular, the possibility of reversibly storing large amounts of hydrogen at low or moderate pressure. However, they still suffer from unfavorable specific energy, with mass-fraction values ranging from 0 up to 5 wt % whereas transport applications require 6 wt % and more. Besides this, higher sorption/desorption kinetics and better chemical stability over long-term cycling are also needed. This is why many studies are carried out in the research community on hydride forming systems, to develop new materials meeting these requirements. Development and optimization of metal hydride reactors require coupled thermodynamic and kinetic characterization of metal-hydrogen systems. In particular, it is necessary to analyze the kinetics in terms of reaction mechanism, in order to identify the different steps of commonly observed multistep reaction paths, and to measure their individual rate parameters. By analyzing hydriding kinetics in the frequency (Fourier) domain, pneumatochemical impedance spectroscopy (PIS) now offers the possibility of measuring experimental impedances and identifying reaction steps. However, measurement of such impedances is indirect and nontrivial. The purpose of this paper is to detail the experimental requirements needed for correctly measuring gas-phase impedance diagrams. In particular, practical conditions of data sampling and data treatment are described. Experimental results obtained with the model LaNi(5)-H(2)(g) system are presented to illustrate the potentialities of PIS analysis. PMID- 18163736 TI - Chemical vapor synthesis of nanocrystalline perovskites using laser flash evaporation of low volatility solid precursors. AB - One key requirement for the production of multinary oxide films by chemical vapor deposition (CVD) or nanocrystalline multinary oxides particles by chemical vapor synthesis (CVS) is the availability of precursors with high vapor pressure. This is especially the case for CVS where much higher production rates are required compared to thin films prepared by CVD. However, elements, which form low valent cations such as alkaline earth metals, are typically only available as solid precursors of low volatility, e.g., in form of beta-diketonates. This study describes laser flash evaporation as precursor delivery method for CVS of nanocrystalline perovskites. Laser flash evaporation exploits the nonequilibrium evaporation of solid metal organic precursors of low vapor pressure by absorption of the infrared radiation of a CO(2) laser. It is shown that stoichiometric, nanocrystalline particles consisting of SrZrO(3) and SrTiO(3) can be formed from corresponding mixtures of beta-diketonates which are evaporated nonselectively and with high rates by laser flash evaporation. PMID- 18163737 TI - An apparatus for the study of high temperature water radiolysis in a nuclear reactor: calibration of dose in a mixed neutron/gamma radiation field. AB - The cooling water of nuclear reactors undergoes radiolytic decomposition induced by gamma, fast electron, and neutron radiation in the core. To model the process, recombination reaction rates and radiolytic yields for the water radical fragments need to be measured at high temperature and pressure. Yields for the action of neutron radiation are particularly hard to determine independently because of the beta/gamma field also present in any reactor. In this paper we report the design of an apparatus intended to measure neutron radiolysis yields as a function of temperature and pressure. A new methodology for separation of neutron and beta/gamma radiolysis yields in a mixed radiation field is proposed and demonstrated. PMID- 18163738 TI - A three-dimensional magnetometer for motion sensing of a balloon-carried atmospheric measurement package. AB - An instrument is described which carries three orthogonal geomagnetic field sensors on a standard meteorological balloon package, to sense rapid motion and position changes during ascent through the atmosphere. Because of the finite data bandwidth available over the UHF radio link, a burst sampling strategy is adopted. Bursts of 9 s of measurements at 3.6 Hz are interleaved with periods of slow data telemetry lasting 25 s. Calculation of the variability in each channel is used to determine position changes, a method robust to periods of poor radio signals. During three balloon ascents, variability was found repeatedly at similar altitudes, simultaneously in each of three orthogonal sensors carried. This variability is attributed to atmospheric motions. It is found that the vertical sensor is least prone to stray motions, and that the use of two horizontal sensors provides no additional information over a single horizontal sensor. PMID- 18163739 TI - The ion-optical prototype of the low energy neutral atom sensor of the Interstellar Boundary Explorer Mission (IBEX). AB - The direct measurement of the energetic neutral atoms originating from the heliospheric termination shock and beyond as well as neutral interstellar gas penetrating into the heliosphere requires a very sensitive neutral particle imaging instrument in the energy range of 10-1000 eV. We present the development of the prototype of the low energy sensor for the Interstellar Boundary Explorer (IBEX) mission: IBEX-Lo is a neutral particle mass spectrometer dedicated to the measurement of energetic neutral atoms in this energy range. The response of the sensor to incident neutral hydrogen, helium, and oxygen atoms is discussed as well as the properties of the sensor's ion optics, the neutral-to-negative conversion surfaces, and other instrumental parameters. PMID- 18163740 TI - A differential double-coil inductive transducer for measuring electrical conductivity. AB - A differential system of two double-coil inductive transducers for the contactless measurement of the electrical conductivity of conducting materials has been presented. The differential system can be employed in applications that require smaller measurement uncertainty than that provided by the single transducer system. A mathematical model of the differential system is based on the model of a single double-coil inductive transducer; in this case, a so-called processing function is defined as a ratio of voltages at the measurement coil terminals with and without the test sample. The relative differential voltage of the differential system is derived as a difference of processing functions of two single transducers and depends on a relative difference between conductivities of the test and reference samples. The conductivity of the test sample is obtained either using precalculated graphs or by numerically processing the equation of the differential voltage. In order to verify the obtained theoretical results, experimental investigations have been carried out using a computer-controlled measurement system with the differential system of the transducers. The conductivity measurements have been carried out using samples made of aluminum rods. During the measurements, the temperature of the reference sample was equal to room temperature (20 degrees C), whereas the temperature of the test sample was changed in the range of 0-20 degrees C to obtain the conductivity variation. The obtained experimental results confirmed the accuracy of the theoretical model of the differential transducer. PMID- 18163741 TI - Nanosecond electro-optical switching with a repetition rate above 20 MHz. AB - We describe an electro-optical switch based on a commercial electro-optic modulator (modified for high-speed operation) and a 340 V pulser having a rise time of 2.2 ns (at 250 V). It can produce arbitrary pulse patterns with an average repetition rate beyond 20 MHz. It uses a grounded-grid triode driven by transmitting power transistors. We discuss variations that enable analog operation, use the step-recovery effect in bipolar transistors, or offer other combinations of output voltage, size, and cost. PMID- 18163742 TI - Method for nonlinear characterization of radio frequency coils made of high temperature superconducting material in view of magnetic resonance imaging applications. AB - A contactless method based on reflectometry to accurately characterize an inductive radio frequency (rf) resonator even in the occurrence of a strong electrical nonlinearity is presented. Nonlinear extraction of the unloaded quality factor and resonance frequency is possible by combining an initial low level swept-frequency calibration with high-level single-frequency measurements. The extraction protocol relies on a simple intrinsic R, L, C model and does not involve a fitting procedure according to a particular nonlinearity model. It includes a correction for strong coupling conditions between the probe and the rf coil, which allows extending the analysis over a wide range of transmitted power. Electrical modeling based on the extracted intrinsic data allows predicting the coil behavior when loaded by any kind of matching network. The method will have implications in different domains such as Magnetic Resonance (MR) applications with superconducting probe heads or analysis of rf properties in nonlinear materials. The method is demonstrated here by characterizing a high temperature superconducting (HTS) coil dedicated to MR imaging at 64 MHz. The coil consists in a multiturn spiral design that is self-resonant close to the MR frequency of interest. The Q factor and the resonance frequency are determined as a function of the actual power dissipated in the HTS coil accounting for losses occurring in the measurement system. Further characteristics of the HTS coil are considered in the present paper. The relation between the transmitted power and the magnetic field generated by the coil, which is the most relevant characteristics for MR applications, is directly accessible. The equivalent impedance of the coil under test is also expressed as a function of the total current flowing in the windings. The method could be extended to assess the fundamental properties of the nonlinear material (e.g., the London penetration depth or the critical current density) by including any pertinent model. PMID- 18163743 TI - A high pressure cell for small angle neutron scattering up to 500 MPa in combination with light scattering to investigate liquid samples. AB - We report on a high pressure cell to use with small angle neutron scattering (SANS) in a pressure range up to 500 MPa. The cell offers the new possibility to investigate liquid samples by a specially designed sample chamber, which allows changing of samples relatively easily. Since the cell construction uses sapphire as window material, also light scattering investigations can be performed simultaneously to the SANS measurements. In this article we describe the construction of a high pressure cell and we demonstrate the applicability of the construction for SANS in combination with dynamic light scattering showing data on the biological molecule lysozyme. PMID- 18163744 TI - Design and implementation of an efficient acoustically levitated drop reactor for in stillo measurements. AB - We present the details necessary for building an efficient acoustic drop levitator with reduced electrical power consumption and greater drop stability compared to previous designs. The system is optimized so that the levitated drop may be used as a chemical reactor. By introducing a temperature, pressure, and relative humidity sensor for feedback control of a linear actuator for adjusting resonator length, we have built a completely automated system capable of continuous levitation for extended periods of time. The result is a system capable of portable operation and interfacing with a variety of detection instrumentation for in stillo (in drop) measurements. PMID- 18163745 TI - A study on the disk-shaped piezoelectric transformer with multiple outputs. AB - In this study, a modified disk-shaped multiple-output piezoelectric transformer operated at the fundamental radial vibration mode has been presented. A derived equivalent circuit for the multioutput piezoelectric transformer was used to analyze the performance. Two piezoelectric transformers, a symmetrically electroded piezoelectric transformer with dual outputs and an asymmetrically electroded piezoelectric transformer with triple outputs, were fabricated with lead zirconate titanate piezoelectric ceramics. The characteristics of the two piezoelectric transformers were investigated experimentally. The piezoelectric transformer with multiple outputs has potential to be used in power supply units and other electronic circuits. PMID- 18163746 TI - High-pressure phase equilibria with compressed gases. AB - An apparatus is described that is capable of determining high-pressure vapor liquid equilibrium, liquid-liquid equilibrium, solid-liquid-vapor equilibrium, vapor-liquid-liquid equilibrium, and mixture critical points and transitions. The device is capable of temperatures to 150 degrees C and pressures to 300 bars (higher with slight modifications). The construction and operation are described in detail and do not require the use of mercury. This method requires very low sample volumes and no analytical equipment nor system-specific calibration. The apparatus was verified by comparison with literature data for the decane-CO(2) mixture and CO(2)-ionic liquid [1-hexyl-3-methyl-imidazolium bis(trifyl)imide)] systems. The experimental data have excellent agreement with the literature data that used different experimental methods. A rigorous error analysis of the system is also presented. PMID- 18163747 TI - A probe for in situ, remote, detection of defects in buried plastic natural gas pipelines. AB - Several techniques are available to determine the integrity of in situ metal pipeline but very little is available in the literature to determine the integrity of plastic pipelines. Since the decade of the 1970s much of the newly installed gas distribution and transmission lines in the United States are fabricated from polyethylene or other plastic. A probe has been developed to determine the in situ integrity of plastic natural gas pipelines that can be installed on a traversing mechanism (pig) to detect abnormalities in the walls of the plastic natural gas pipeline from the interior. This probe has its own internal power source and can be deployed into existing natural gas supply lines. Utilizing the capacitance parameter, the probe inspects the pipe for flaws and records the data internally which can be retrieved later for analysis. PMID- 18163748 TI - Pressure measurements in laboratory-scale blast wave flow fields. AB - The present study examines the effects that temporal and spatial averagings due to finite size and finite response time of pressure transducers have on the pressure measurements in blast wave flow fields generated by milligram charges of silver azide. In such applications, the characteristic time and length scales of the physical process are of the same order of magnitude as the temporal and spatial characteristics of the transducer. The measured pressure values will then be spatially and temporally averaged, and important parameters for the assessment of blast effects may not be properly represented in the measured trace. In this study, face-on and side-on pressure transducer setups are considered. In the experiments, face-on and side-on readings at the same distance from the charge as well as time-resolved optical visualization of the whole flow field are obtained simultaneously for the same explosive event. The procedure of data extraction from the experimental pressure traces is revisited and discussed in detail. In the numerical modeling part of the study, numerical blast flow fields are generated using an Euler flow solver. A numerical pressure transducer model is developed to qualitatively simulate the averaging effects. The experimental and numerical data show that the results of pressure measurements in experiments with small charges must be used with great caution. The effective averaging of the pressure signal may lead to a significant underestimation of blast wave intensities. The side-on setup is especially prone to this effect. The face-on setup provides results close to those obtained from optical records only if the pressure transducer is sufficiently remote from the charge. PMID- 18163749 TI - The influence of preamplifiers on the piezoelectric sensor's dynamic property. AB - A charge amplifier or a voltage amplifier can be used as a signal conditioning circuit between a piezoelectric element and a meter or a data acquisition board. The outputs of the piezoelectric sensor are in an open-circuit state and a short circuit state if a voltage amplifier and a charge amplifier are used, respectively. When the electrodes are in different states, the piezoelectric element has rather different stiffness and thus different sensor resonant frequency. This phenomenon is theoretically analyzed in detail and validated by a carefully designed experiment. The results indicate that a much wider range of working frequency is achieved when a voltage amplifier is used. PMID- 18163750 TI - Scanning electron microscope for in situ study of crystallization of Ge2Sb2Te5 in phase-change memory. AB - By introducing electrical connections into the chamber of a scanning electron microscope (SEM) via its holder assembly, it has become feasible to in situ observe and electrically characterize electronic devices. The in situ SEM was applied to investigate electric-pulse-induced behavior of Ge(2)Sb(2)Te(5) in a lateral phase-change memory cell. Randomly distributed nuclei with sizes from 20 to 80 nm were initiated at a low voltage pulse. Initially, grain growth depended strongly on pulse amplitude at around 60.3 nm/V and then a weak pulse amplitude dependence was observed at around 13.5 nm/V. Device resistance during crystallization dropped by two to three orders of magnitude with two falling steps, which probably resulted from amorphous to face-centered-cubic and subsequently to hexagonal transitions, respectively. PMID- 18163751 TI - Microcantilever actuation via periodic internal heating. AB - This paper reports electrothermal actuation of silicon microcantilevers having integrated resistive heaters. Periodic electrical excitation induced periodic resistive heating in the cantilever, while the cantilever deflection was monitored with a photodetector. Excitation was either at the cantilever resonant frequency, f(0), f(0)/2, or f(0)/3. When the time averaged maximum cantilever temperature was 174 degrees C, the cantilever out-of-plane actuation amplitude was 484 nm near the cantilever resonance frequency of 24.9 kHz. This actuation was sufficiently large to operate the cantilever in intermittent contact mode and scan a calibration grating of height of 20 nm. PMID- 18163752 TI - Evaluation of a multimode fiber optic low coherence interferometer for path length resolved Doppler measurements of diffuse light. AB - The performance of a graded index multimode fiber optic low coherence Mach Zehnder interferometer with phase modulation is analyzed. Investigated aspects were its ability to measure path length distributions and to perform path length resolved Doppler measurements of multiple scattered photons in a turbid suspension of particles undergoing Brownian and translational motion. The path length resolution of this instrument is compared with a system using single mode fibers for illumination and detection. The optical path lengths are determined from the zero order moment of the phase modulation peak in the power spectrum. The weighted first moment, which is equal to the average Doppler shift, shows a linear response for different mean flow velocities within the physiological range. PMID- 18163753 TI - A new approach to the preionization of flash lamps in power lasers. AB - As long as neodymium amplifiers will be used in high-energy lasers, preionization problems will be present in firing the flash lamps. It is difficult to find papers describing technical solutions using preionization concept; only a few internal reports and proceedings from Pulsed Power conferences describe the usage of such circuits in power laser facilities. In all of them, a special negative high voltage power supply is used. A new simple solution, without any additional power supply, using only a small part of the energy gathered in the main capacitor bank for generating a preionization pulse is used. The firing order is split into two signals; the first one is triggering preionization circuit and the second one, after delaying, is triggering the main discharging circuit. PMID- 18163754 TI - Transient of scanning electron microscopic images for a buried microstructure in insulators. AB - We clarify the transient process and its mechanism of scanning electron microscope (SEM) images of a trench microstructure buried in insulators. First, interface charges of primary electrons trapped on the trench are derived from the charging model of a capacitor considering the electron beam induced current, and the surface potential is therefore assumed. The SEM signal current is then determined from its simplified relation with the surface potential. Calculated profiles of the secondary electron (SE) signal current and their time-evolution behaviors can well fit the transient of the experimental SEM images. Results show that the variation of the surface potential due to the transient interface charges and the effect of SE redistribution result in transients of the SEM imaging signal and the image width of the buried trench. PMID- 18163758 TI - Rayleigh-Benard convection with imposed heat flux. PMID- 18163757 TI - Polymer drop breakup in microchannels. PMID- 18163759 TI - Paraboloidal crystals. PMID- 18163760 TI - Shear faults in a model brittle solid. PMID- 18163761 TI - Avoided level crossings in the quantization of a mixed regular-chaotic system. PMID- 18163762 TI - Spatially heterogeneous dynamics in a granular system near jamming. PMID- 18163764 TI - Numerical study of the phase space of modulated exponentially coupled harmonic oscillators. PMID- 18163763 TI - Complex pore spaces create record-breaking methane storage system for natural-gas vehicles. PMID- 18163765 TI - Characterization of stickiness by means of recurrence. AB - We propose recurrence plots (RPs) to characterize the stickiness of a typical area-preserving map with coexisting chaotic and regular orbits. The difference of the recurrence properties between quasiperiodic and chaotic orbits is revisited, which helps to understand the complex patterns of the corresponding RPs. Moreover, several measures from the recurrence quantification analysis are used to quantify these patterns. Among these measures, the recurrence rate, quantifying the percentage of black points in the plot, is applied to characterize the stickiness of a typical chaotic orbit. The advantage of the recurrence based method in comparison to other standard techniques is that it is possible to distinguish between quasiperiodic and chaotic orbits that are temporarily trapped in a sticky domain, from very short trajectories. PMID- 18163766 TI - Random walks on the Comb model and its generalizations. AB - Microscopic models with anomalous diffusion, which include the Comb model and its generalization for the finite width of the backbone, have been considered in this paper. The physical mechanisms of the subdiffusion random walks have been established. The first comes from the permanent return of the diffusing particle to the initial point of the diffusion due to "effective reducing" of the dimensionality of the considered system to the quasi-one-dimensional system. This physical mechanism has been obtained in the Comb model and in the model with a strip. The second mechanism of the subdiffusion is connected with random capture on the traps of diffusing particles and their ensuing random release from the traps. It has been shown that these different mechanisms of subdiffusion have been described by the different generalized diffusion equations of fractional order. The solutions of these different equations have been obtained, and the physical sense of the fractional order generalized equations has been discussed. PMID- 18163767 TI - Optimal structure of complex networks for minimizing traffic congestion. AB - To design complex networks to minimize traffic congestion, it is necessary to understand how traffic flow depends on network structure. We study data packet flow on complex networks, where the packet delivery capacity of each node is not fixed. The optimal configuration of capacities to minimize traffic congestion is derived and the critical packet generating rate is determined, below which the network is at a free flow state but above which congestion occurs. Our analysis reveals a direct relation between network topology and traffic flow. Optimal network structure, free of traffic congestion, should have two features: uniform distribution of load over all nodes and small network diameter. This finding is confirmed by numerical simulations. Our analysis also makes it possible to theoretically compare the congestion conditions for different types of complex networks. In particular, we find that network with low critical generating rate is more susceptible to congestion. The comparison has been made on the following complex-network topologies: random, scale-free, and regular. PMID- 18163768 TI - On the absence of stable periodic orbits in domains of separatrix crossings in nonsymmetric slow-fast Hamiltonian systems. AB - We consider a two degree of freedom Hamiltonian system with one degree of freedom corresponding to fast motion and the other corresponding to slow motion. We assume that at frozen values of the slow variables there is a separatrix on the phase plane of the fast variables and there is a region in the phase space (the domain of separatrix crossings) where projections of phase points onto the plane of the fast variables repeatedly cross the separatrix in the process of evolution of the slow variables. Under rather general conditions, we prove that there are no stable periodic trajectories of any prescribed period inside the domain of separatrix crossings, except maybe for periodic trajectories passing anomalously close to the saddle point. PMID- 18163769 TI - Effect of dynamical traps on chaotic transport in a meandering jet flow. AB - We continue our study of chaotic mixing and transport of passive particles in a simple model of a meandering jet flow [Prants et al., Chaos 16, 033117 (2006)]. In the present paper we study and phenomenologically explain a connection between dynamical, topological, and statistical properties of chaotic mixing and transport in the model flow in terms of dynamical traps, singular zones in the phase space where particles may spend an arbitrarily long but finite time [Zaslavsky, Phys. D 168-169, 292 (2002)]. The transport of passive particles is described in terms of lengths and durations of zonal flights which are events between two successive changes of sign of zonal velocity. Some peculiarities of the respective probability density functions for short flights are proven to be caused by the so-called rotational-island traps connected with the boundaries of resonant islands (including the vortex cores) filled with the particles moving in the same frame and the saddle traps connected with periodic saddle trajectories. Whereas, the statistics of long flights can be explained by the influence of the so-called ballistic-islands traps filled with the particles moving from a frame to frame. PMID- 18163770 TI - Enhancing dominant modes in nonstationary time series by means of the symbolic resonance analysis. AB - We present the symbolic resonance analysis (SRA) as a viable method for addressing the problem of enhancing a weakly dominant mode in a mixture of impulse responses obtained from a nonlinear dynamical system. We demonstrate this using results from a numerical simulation with Duffing oscillators in different domains of their parameter space, and by analyzing event-related brain potentials (ERPs) from a language processing experiment in German as a representative application. In this paradigm, the averaged ERPs exhibit an N400 followed by a sentence final negativity. Contemporary sentence processing models predict a late positivity (P600) as well. We show that the SRA is able to unveil the P600 evoked by the critical stimuli as a weakly dominant mode from the covering sentence final negativity. PMID- 18163771 TI - Chaos suppression through asymmetric coupling. AB - We study pairs of identical coupled chaotic oscillators. In particular, we have used Roessler (in the funnel and no funnel regimes), Lorenz, and four-dimensional chaotic Lotka-Volterra models. In all four of these cases, a pair of identical oscillators is asymmetrically coupled. The main result of the numerical simulations is that in all cases, specific values of coupling strength and asymmetry exist that render the two oscillators periodic and synchronized. The values of the coupling strength for which this phenomenon occurs is well below the previously known value for complete synchronization. We have found that this behavior exists for all the chaotic oscillators that we have used in the analysis. We postulate that this behavior is presumably generic to all chaotic oscillators. In order to complete the study, we have tested the robustness of this phenomenon of chaos suppression versus the addition of some Gaussian noise. We found that chaos suppression is robust for the addition of finite noise level. Finally, we propose some extension to this research. PMID- 18163772 TI - On the complexity of periodic and nonperiodic behaviors of a hysteresis-based electronic oscillator. AB - We investigate the families of periodic and nonperiodic behaviors admitted by a hysteresis-based circuit oscillator. The analysis is carried out by combining brute-force simulations with continuation methods. As a result of the analysis, it is shown that the existence of many different periodic solutions and of the chaotic behaviors associated with them is organized by few codimension-2 bifurcation points. This implies the possibility of switching between different periodic solutions by controlling only two bifurcation parameters, which makes the oscillator a possible generator of nontrivial periodic solutions suitable, for instance for actual radiofrequency identification systems applications. PMID- 18163773 TI - Chaotic oscillations in a map-based model of neural activity. AB - We propose a discrete time dynamical system (a map) as a phenomenological model of excitable and spiking-bursting neurons. The model is a discontinuous two dimensional map. We find conditions under which this map has an invariant region on the phase plane, containing a chaotic attractor. This attractor creates chaotic spiking-bursting oscillations of the model. We also show various regimes of other neural activities (subthreshold oscillations, phasic spiking, etc.) derived from the proposed model. PMID- 18163774 TI - Multiscale vulnerability of complex networks. AB - We present a novel approach to quantify the vulnerability of a complex network, i.e., the capacity of a graph to maintain its functional performance under random damages or malicious attacks. The proposed measure represents a multiscale evaluation of vulnerability, and makes use of combined powers of the links' betweenness. We show that the proposed approach is able to properly describe some cases for which earlier measures of vulnerability fail. The relevant applications of our method for technological network design are outlined. PMID- 18163775 TI - Frequency dependence of phase-synchronization time in nonlinear dynamical systems. AB - It has been found recently that the averaged phase-synchronization time between the input and the output signals of a nonlinear dynamical system can exhibit an extremely high sensitivity to variations in the noise level. In real-world signal processing applications, sensitivity to frequency variations may be of considerable interest. Here we investigate the dependence of the averaged phase synchronization time on frequency of the input signal. Our finding is that, for typical nonlinear oscillator systems, there can be a frequency regime where the time exhibits significant sensitivity to frequency variations. We obtain an analytic formula to quantify the frequency dependence, provide numerical support, and present experimental evidence from a simple nonlinear circuit system. PMID- 18163776 TI - Deterministic single-file dynamics in collisional representation. AB - We re-examine numerically the diffusion of a deterministic, or ballistic single file with preassigned velocity distribution (Jepsen's gas) from a collisional viewpoint. For a two-modal velocity distribution, where half the particles have velocity +/-c, the collisional statistics is analytically proven to reproduce the continuous time representation. For a three-modal velocity distribution with equal fractions, where less than 12 of the particles have velocity +/-c, with the remaining particles at rest, the collisional process is shown to be inhomogeneous; its stationary properties are discussed here by combining exact and phenomenological arguments. Collisional memory effects are then related to the negative power-law tails in the velocity autocorrelation functions, predicted earlier in the continuous time formalism. Numerical and analytical results for Gaussian and four-modal Jepsen's gases are also reported for the sake of a comparison. PMID- 18163777 TI - Detecting and localizing the foci in human epileptic seizures. AB - We consider the electrical signals recorded from a subdural array of electrodes placed on the pial surface of the brain for chronic evaluation of epileptic patients before surgical resection. A simple and computationally fast method to analyze the interictal phase synchrony between such electrodes is introduced and developed with the aim of detecting and localizing the foci of the epileptic seizures. We evaluate the method by comparing the results of surgery to the localization predicted here. We find an indication of good correspondence between the success or failure in the surgery and the agreement between our identification and the regions actually operated on. PMID- 18163778 TI - Quantifying the complexity of excised larynx vibrations from high-speed imaging using spatiotemporal and nonlinear dynamic analyses. AB - In this paper, we investigate the biomechanical applications of spatiotemporal analysis and nonlinear dynamic analysis to quantitatively describe regular and irregular vibrations of twelve excised larynges from high-speed image recordings. Regular vibrations show simple spatial symmetry, temporal periodicity, and discrete frequency spectra, while irregular vibrations show complex spatiotemporal plots, aperiodic time series, and broadband spectra. Furthermore, the global entropy and correlation length from spatiotemporal analysis and the correlation dimension from nonlinear dynamic analysis reveal a statistical difference between regular and irregular vibrations. In comparison with regular vibrations, the global entropy and correlation dimension of irregular vibrations are statistically higher, while the correlation length is significantly lower. These findings show that spatiotemporal analysis and nonlinear dynamic analysis are capable of describing the complex dynamics of vocal fold vibrations from high speed imaging and may potentially be helpful for understanding disordered behaviors in biomedical laryngeal systems. PMID- 18163779 TI - Vulnerability to re-entry in simulated two-dimensional cardiac tissue: effects of electrical restitution and stimulation sequence. AB - Ventricular fibrillation is a lethal arrhythmia characterized by multiple wavelets usually starting from a single or figure-of-eight re-entrant circuit. Understanding the factors regulating vulnerability to the re-entry is essential for developing effective therapeutic strategies to prevent ventricular fibrillation. In this study, we investigated how pre-existing tissue heterogeneities and electrical restitution properties affect the initiation of re entry by premature extrastimuli in two-dimensional cardiac tissue models. We studied two pacing protocols for inducing re-entry following the "sinus" rhythm (S1) beat: (1) a single premature (S2) extrastimulus in heterogeneous tissue; (2) two premature extrastimuli (S2 and S3) in homogeneous tissue. In the first case, the vulnerable window of re-entry is determined by the spatial dimension and extent of the heterogeneity, and is also affected by electrical restitution properties and the location of the premature stimulus. The vulnerable window first increases as the action potential duration (APD) difference between the inside and outside of the heterogeneous region increases, but then decreases as this difference increases further. Steeper APD restitution reduces the vulnerable window of re-entry. In the second case, electrical restitution plays an essential role. When APD restitution is flat, no re-entry can be induced. When APD restitution is steep, re-entry can be induced by an S3 over a range of S1S2 intervals, which is also affected by conduction velocity restitution. When APD restitution is even steeper, the vulnerable window is reduced due to collision of the spiral tips. PMID- 18163780 TI - Exponential decay of correlations for randomly chosen hyperbolic toral automorphisms. AB - We consider pairs of toral automorphisms (A,B) satisfying an invariant cone property. At each iteration, A acts with probability p is in (0,1) and B with probability 1-p. We prove exponential decay of correlations for a class of Holder continuous observables. PMID- 18163781 TI - New eigenvalue based approach to synchronization in asymmetrically coupled networks. AB - Locally and globally exponential stability of synchronization in asymmetrically nonlinear coupled networks and linear coupled networks are investigated in this paper, respectively. Some new synchronization stability criteria based on eigenvalues are derived. In these criteria, both a term that is the second largest eigenvalue of a symmetrical matrix and a term that is the largest value of the sum of the column of the asymmetrical coupling matrix play a key role. Comparing with existing results, the advantage of our synchronization stability results is that they can be analytically applied to the asymmetrically coupled networks and can overcome the complexity of calculating eigenvalues of the coupling asymmetric matrix. Therefore, these conditions are very convenient to use. Moreover, a necessary condition of globally exponential synchronization stability criterion is also given by the elements of the coupling asymmetric matrix, which can conveniently be used in judging the synchronization stability condition without calculating the eigenvalues of the coupling matrix. PMID- 18163782 TI - Experimental synchronization of single-transistor-based chaotic circuits. AB - This work deals with nonautonomous chaotic circuits and, in particular, with the experimental characterization of the synchronization properties of two simple nonautonomous circuits. Two single-transistor chaotic circuits, which are among the simplest chaotic oscillators, are investigated. We studied synchronization of these circuits and found that the most appropriate technique to synchronize two single-transistor chaotic circuits is that based on the design of an inverse circuit. PMID- 18163783 TI - Regions of nonexistence of invariant tori for spin-orbit models. AB - The spin-orbit problem in celestial mechanics describes the motion of an oblate satellite moving on a Keplerian orbit around a primary body. We apply the conjugate points criterion for the nonexistence of rotational invariant tori. We treat both the conservative case and a case including a dissipative effect modeling a tidal torque generated by internal nonrigidity. As a by-product of the conjugate points criterion we obtain a global view of the dynamics, thanks to the introduction of a tangent orbit indicator, which allows us to discern the dynamical character of the motion. PMID- 18163784 TI - Multiple period-doubling bifurcation route to chaos in periodically pulsed Murali Lakshmanan-Chua circuit-controlling and synchronization of chaos. AB - We consider a simple nonautonomous dissipative nonlinear electronic circuit consisting of Chua's diode as the only nonlinear element, which exhibit a typical period doubling bifurcation route to chaotic oscillations. In this paper, we show that the effect of additional periodic pulses in this Murali-Lakshmanan-Chua (MLC) circuit results in novel multiple-period-doubling bifurcation behavior, prior to the onset of chaos, by using both numerical and some experimental simulations. In the chaotic regime, this circuit exhibits a rich variety of dynamical behavior including enlarged periodic windows, attractor crises, distinctly modified bifurcation structures, and so on. For certain types of periodic pulses, this circuit also admits transcritical bifurcations preceding the onset of multiple-period-doubling bifurcations. We have characterized our numerical simulation results by using Lyapunov exponents, correlation dimension, and power spectrum, which are found to be in good agreement with the experimental observations. Further controlling and synchronization of chaos in this periodically pulsed MLC circuit have been achieved by using suitable methods. We have also shown that the chaotic attractor becomes more complicated and their corresponding return maps are no longer simple for large n-periodic pulses. The above study also indicates that one can generate any desired n-period-doubling bifurcation behavior by applying n-periodic pulses to a chaotic system. PMID- 18163785 TI - Parametric investigation of nonlinear fluctuations in a dc glow discharge plasma. AB - Glow discharge plasmas exhibit various types of self-excited oscillations for different initial conditions like discharge voltages and filling pressures. The behavior of such oscillations associated with the anode glow has been investigated using nonlinear techniques like correlation dimension, largest Lyapunov exponent, etc. It is seen that these oscillations go to an ordered state from a chaotic state with an increase in input energy, i.e., with discharge voltages implying occurrence of inverse bifurcations. These results are different from the other observations wherein the fluctuations have been observed to go from ordered to chaotic state. PMID- 18163786 TI - The antisynchronization of a class of chaotic delayed neural networks. AB - In this paper, the antisynchronization problems of a class of chaotic delayed neural networks are investigated. Some criteria of the antisynchronization of the chaotic delayed neural networks are established by using the linear matrix inequality and Lyapunov stability theory. These criteria not only improve and generalize some known results, but are also some less conservative conditions. Finally, two numerical examples and the corresponding numerical simulations are used to illustrate the effectiveness of the obtained results. PMID- 18163787 TI - Stationary oscillation for chaotic shunting inhibitory cellular neural networks with impulses. AB - In this paper, we study stationary oscillation for general shunting inhibitory cellular neural networks with impulses which are complex nonlinear neural networks. In a recent paper [Z. J. Gui and W. G. Ge, Chaos 16, 033116 (2006)], the authors claimed that they obtained a criterion of existence, uniqueness, and global exponential stability of periodic solution (i.e., stationary oscillation) for shunting inhibitory cellular neural networks with impulses. We point out in this paper that the main result of their paper is incorrect, and presents a sufficient condition of ensuring existence, uniqueness, and global stability of periodic solution for general shunting inhibitory cellular neural networks with impulses. The result is derived by using a new method which is different from those of previous literature. An illustrative example is given to demonstrate the effectiveness. PMID- 18163788 TI - Dynamics of the chain of forced oscillators with long-range interaction: from synchronization to chaos. AB - We consider a chain of nonlinear oscillators with long-range interaction of the type 1l(1+alpha), where l is a distance between oscillators and 070%) among the European experts members of the project. Twelve items focus on Oral Health of children and adolescents: they are described in the present paper. CONCLUSION: Further developing phases aimed to establish standardised methodological criteria for the collection of oral health data are also described. PMID- 18163858 TI - Patient adherence: present state and future directions. PMID- 18163859 TI - Number needed to succeed in disease management. PMID- 18163860 TI - Healing the health care system. PMID- 18163861 TI - Cost and effects of performance feedback and nurse case management for medicare beneficiaries with diabetes: a randomized controlled trial. AB - Nurse case management has been shown to improve the quality of diabetes care in closed model health maintenance organizations and Veterans Affairs medical clinics. A randomized controlled trial of a similar intervention within HealthTexas Provider Network, a fee-for-service primary care network in North Texas, demonstrated no benefit in processes of care or clinical outcomes for Medicare diabetes patients. To investigate whether the case management model impacted the cost of diabetes care from the Medicare perspective, we compared the average payments and charges incurred between intervention arms: claims-based audit and feedback; claims- and medical-record-based audit and feedback; and claims- and medical-record-based audit and feedback plus a practice-based diabetes resource nurse. Following adjustment for baseline differences between groups, no significant differences were observed. Thus, within this setting, it appears the nurse case management model produced no improvement in either clinical quality or in costs associated with diabetes from a Medicare perspective. PMID- 18163862 TI - Iatrogenic disease management: moderating medication errors and risks in a pharmacy benefit management environment. AB - Disease Management (DM) programs have advanced to address costly chronic disease patterns in populations. This is in part due to the programs' significant clinical and economical value, coupled with interest by pharmaceutical manufacturers, managed care organizations, and pharmacy benefit management firms. While cost containment realizations for many such interventions have been less than anticipated, this article explores potentials in marrying Medication Error Risk Reduction into DM programs within managed care environments. Medication errors are an emergent serious problem now gaining attention in US health policy. They represent a failure within population-based health programs because they remain significant cost drivers. Therefore, medication errors should be addressed in an organized fashion, with DM being a worthy candidate for piggybacking such programs to achieve the best synergistic effects. PMID- 18163863 TI - Avoidable hospitalizations for diabetes: comorbidity risks. AB - This study examined the risk for avoidable diabetes hospitalizations associated with comorbid conditions and other risk variables. A retrospective analysis was conducted of hospitalizations with a primary diagnosis of diabetes in a 2004 sample of short stay general hospitals in the United States (N = 97,526.) Data were drawn from the Health Care Utilization Project National Inpatient Sample. Avoidable hospitalizations were defined using criteria from the Agency for Healthcare Research and Quality to analyze 2 types of ambulatory care sensitive conditions (ACSCs): short-term complications and uncontrolled diabetes. Maternal cases, patients younger than age 18, and transfers from other hospitals were excluded. Avoidable hospitalization was estimated using maximum likelihood logistic regression analysis, where independent variables included patient age, gender, comorbidities, uninsurance status, patient's rural-urban residence and income estimate, and hospital variables. Models were identified using multiple runs on 3 random quartiles and validated using the fourth quartile. Costs were estimated from charge data using cost-to-charge ratios. Results indicated that these 2 ACSCs accounted for 35,312 or 36% of all diabetes hospitalizations. Multiple types of comorbid conditions were related to risk for avoidable diabetes hospitalizations. Estimated costs and length of stay were lower among these types of avoidable hospitalizations compared to other diabetes hospitalizations; however, total estimated nationwide costs for 2004 short-term complications and uncontrolled diabetes hospitalizations totaled over $1.3 billion. Recommendations are made for how disease management programs for diabetes could incorporate treatment for comorbid conditions to reduce hospitalization risk. PMID- 18163865 TI - An energy balance concept for habitability. AB - Habitability can be formulated as a balance between the biological demand for energy and the corresponding potential for meeting that demand by transduction of energy from the environment into biological process. The biological demand for energy is manifest in two requirements, analogous to the voltage and power requirements of an electrical device, which must both be met if life is to be supported. These requirements exhibit discrete (non-zero) minima whose magnitude is set by the biochemistry in question, and they are increased in quantifiable fashion by (i) deviations from biochemically optimal physical and chemical conditions and (ii) energy-expending solutions to problems of resource limitation. The possible rate of energy transduction is constrained by (i) the availability of usable free energy sources in the environment, (ii) limitations on transport of those sources into the cell, (iii) upper limits on the rate at which energy can be stored, transported, and subsequently liberated by biochemical mechanisms (e.g., enzyme saturation effects), and (iv) upper limits imposed by an inability to use "power" and "voltage" at levels that cause material breakdown. A system is habitable when the realized rate of energy transduction equals or exceeds the biological demand for energy. For systems in which water availability is considered a key aspect of habitability (e.g., Mars), the energy balance construct imposes additional, quantitative constraints that may help to prioritize targets in search-for-life missions. Because the biological need for energy is universal, the energy balance construct also helps to constrain habitability in systems (e.g., those envisioned to use solvents other than water) for which little constraint currently exists. PMID- 18163866 TI - Quantitative habitability. AB - A framework is proposed for a quantitative approach to studying habitability. Considerations of environmental supply and organismal demand of energy lead to the conclusions that power units are most appropriate and that the units for habitability become watts per organism. Extreme and plush environments are revealed to be on a habitability continuum, and extreme environments can be quantified as those where power supply only barely exceeds demand. Strategies for laboratory and field experiments are outlined that would quantify power supplies, power demands, and habitability. An example involving a comparison of various metabolisms pursued by halophiles is shown to be well on the way to a quantitative habitability analysis. PMID- 18163867 TI - Actinides and Life's Origins. AB - There are growing indications that life began in a radioactive beach environment. A geologic framework for the origin or support of life in a Hadean heavy mineral placer beach has been developed, based on the unique chemical properties of the lower-electronic actinides, which act as nuclear fissile and fertile fuels, radiolytic energy sources, oligomer catalysts, and coordinating ions (along with mineralogically associated lanthanides) for prototypical prebiotic homonuclear and dinuclear metalloenzymes. A four-factor nuclear reactor model was constructed to estimate how much uranium would have been required to initiate a sustainable fission reaction within a placer beach sand 4.3 billion years ago. It was calculated that about 1-8 weight percent of the sand would have to have been uraninite, depending on the weight percent, uranium enrichment, and quantity of neutron poisons present within the remaining placer minerals. Radiolysis experiments were conducted with various solvents with the use of uraniumand thorium-rich minerals (metatorbernite and monazite, respectively) as proxies for radioactive beach sand in contact with different carbon, hydrogen, oxygen, and nitrogen reactants. Radiation bombardment ranged in duration of exposure from 3 weeks to 6 months. Low levels of acetonitrile (estimated to be on the order of parts per billion in concentration) were conclusively identified in 2 setups and tentatively indicated in a 3(rd) by gas chromatography/mass spectrometry. These low levels have been interpreted within the context of a Hadean placer beach prebiotic framework to demonstrate the promise of investigating natural nuclear reactors as power production sites that might have assisted the origins of life on young rocky planets with a sufficiently differentiated crust/mantle structure. Future investigations are recommended to better quantify the complex relationships between energy release, radioactive grain size, fissionability, reactant phase, phosphorus release, and possible abiotic production of sugars, amino acids, activated phosphorus, prototypical organometallic enzymes, and oligomer catalysts at a single putative beach site. PMID- 18163868 TI - Formate as an energy source for microbial metabolism in chemosynthetic zones of hydrothermal ecosystems. AB - Formate, a simple organic acid known to support chemotrophic hyperthermophiles, is found in hot springs of varying temperature and pH. However, it is not yet known how metabolic strategies that use formate could contribute to primary productivity in hydrothermal ecosystems. In an effort to provide a quantitative framework for assessing the role of formate metabolism, concentration data for dissolved formate and many other solutes in samples from Yellowstone hot springs were used, together with data for coexisting gas compositions, to evaluate the overall Gibbs energy for many reactions involving formate oxidation or reduction. The result is the first rigorous thermodynamic assessment of reactions involving formate oxidation to bicarbonate and reduction to methane coupled with various forms of iron, nitrogen, sulfur, hydrogen, and oxygen for hydrothermal ecosystems. We conclude that there are a limited number of reactions that can yield energy through formate reduction, in contrast to numerous formate oxidation reactions that can yield abundant energy for chemosynthetic microorganisms. Because the energy yields are so high, these results challenge the notion that hydrogen is the primary energy source of chemosynthetic microbes in hydrothermal ecosystems. PMID- 18163869 TI - A thermodynamic analysis of microbial growth experiments. AB - The common thread of energy release suggests that diverse microbial metabolic processes can be compared through thermodynamic analyses. The resulting energy and power requirements can provide quantitative constraints on habitability. Because previous thermodynamic analyses have focused on the minimum amount of energy needed for the growth of a microorganism or community, the focus of this study is to gain a fuller understanding of the microbial response to highly habitable conditions. This communication summarizes the results of a thermodynamic analysis of the energy and power consumed by microorganisms in experiments that were designed to optimize growth. Reports of microbial growth experiments taken from the literature were combined with speciation and standard state calculations to assess the overall Gibbs energy change during the experiments. Results show that similar numbers of cells (10(9) to 10(10) ) were produced in these experiments regardless of the duration of log phase growth (from <2 to >200 hours) or the total Gibbs energy change [from 1.3-29.6 kJ (mol electrons transferred)(1)]. As a result, optimal growth conditions appear to produce between 10(10) and 10(14) cells per watt of power consumed. PMID- 18163870 TI - Temporal changes in fluid chemistry and energy profiles in the vulcano island hydrothermal system. AB - In June 2003, the geochemical composition of geothermal fluids was determined at 9 sites in the Vulcano hydrothermal system, including sediment seeps, geothermal wells, and submarine vents. Compositional data were combined with standard state reaction properties to determine the overall Gibbs free energy (DeltaG(r) ) for 120 potential lithotrophic and heterotrophic reactions. Lithotrophic reactions in the H-O-N-S-C-Fe system were considered, and exergonic reactions yielded up to 120 kJ per mole of electrons transferred. The potential for heterotrophy was characterized by energy yields from the complete oxidation of 6 carboxylic acids- formic, acetic, propanoic, lactic, pyruvic, and succinic-with the following redox pairs: O(2)/H(2)O, SO(4) (2)/H(2)S, NO(3) ()/NH(4) (+), S(0)/H(2)S, and Fe(3)O(4)/Fe(2+). Heterotrophic reactions yielded 6-111 kJ/mol e(). Energy yields from both lithotrophic and heterotrophic reactions were highly dependent on the terminal electron acceptor (TEA); reactions with O(2) yielded the most energy, followed by those with NO(3) (), Fe(III), SO(4) (2), and S(0). When only reactions with complete TEA reduction were included, the exergonic lithotrophic reactions followed a similar electron tower. Spatial variability in DeltaG(r) was significant for iron redox reactions, owing largely to the wide range in Fe(2+) and H(+) concentrations. Energy yields were compared to those obtained for samples collected in June 2001. The temporal variations in geochemical composition and energy yields observed in the Vulcano hydrothermal system between 2001 and 2003 were moderate. The largest differences in DeltaG(r) over the 2 years were from iron redox reactions, due to temporal changes in the Fe(2+) and H(+) concentrations. The observed variations in fluid composition across the Vulcano hydrothermal system have the potential to influence not only microbial diversity but also the metabolic strategies of the resident microbial communities. PMID- 18163871 TI - Geochemical constraints on sources of metabolic energy for chemolithoautotrophy in ultramafic-hosted deep-sea hydrothermal systems. AB - Numerical models are employed to investigate sources of chemical energy for autotrophic microbial metabolism that develop during mixing of oxidized seawater with strongly reduced fluids discharged from ultramafic-hosted hydrothermal systems on the seafloor. Hydrothermal fluids in these systems are highly enriched in H(2) and CH(4) as a result of alteration of ultramafic rocks (serpentinization) in the subsurface. Based on the availability of chemical energy sources, inferences are made about the likely metabolic diversity, relative abundance, and spatial distribution of microorganisms within ultramafic hosted systems. Metabolic reactions involving H(2) and CH(4), particularly hydrogen oxidation, methanotrophy, sulfate reduction, and methanogenesis, represent the predominant sources of chemical energy during fluid mixing. Owing to chemical gradients that develop from fluid mixing, aerobic metabolisms are likely to predominate in low-temperature environments (<20-30 degrees C), while anaerobes will dominate higher-temperature environments. Overall, aerobic metabolic reactions can supply up to approximately 7 kJ of energy per kilogram of hydrothermal fluid, while anaerobic metabolic reactions can supply about 1 kJ, which is sufficient to support a maximum of approximately 120 mg (dry weight) of primary biomass production by aerobic organisms and approximately 20-30 mg biomass by anaerobes. The results indicate that ultramafic-hosted systems are capable of supplying about twice as much chemical energy as analogous deep-sea hydrothermal systems hosted in basaltic rocks. PMID- 18163872 TI - Radiolytic hydrogen and microbial respiration in subsurface sediments. AB - Radiolysis of water may provide a continuous flux of an electron donor (molecular hydrogen) to subsurface microbial communities. We assessed the significance of this process in anoxic marine sediments by comparing calculated radiolytic H(2) production rates to estimates of net (organic-fueled) respiration at several Ocean Drilling Program (ODP) Leg 201 sites. Radiolytic H(2) yield calculations are based on abundances of radioactive elements (uranium, thorium, and potassium), porosity, grain density, and a model of water radiolysis. Net respiration estimates are based on fluxes of dissolved electron acceptors and their products. Comparison of radiolytic H(2) yields and respiration at multiple sites suggests that radiolysis gains importance as an electron donor source as net respiration and organic carbon content decrease. Our results suggest that radiolytic production of H(2) may fuel 10% of the metabolic respiration at the Leg 201 site where organic-fueled respiration is lowest (ODP Site 1231). In sediments with even lower rates of organic-fueled respiration, water radiolysis may be the principal source of electron donors. Marine sedimentary ecosystems may be useful models for non-photosynthetic ecosystems on early Earth and on other planets and moons, such as Mars and Europa. PMID- 18163873 TI - Hydrogeologic controls on episodic H2 release from precambrian fractured rocks- energy for deep subsurface life on earth and mars. AB - Dissolved H(2) concentrations up to the mM range and H(2) levels up to 9-58% by volume in the free gas phase are reported for groundwaters at sites in the Precambrian shields of Canada and Finland. Along with previously reported dissolved H(2) concentrations up to 7.4 mM for groundwaters from the Witwatersrand Basin, South Africa, these findings indicate that deep Precambrian Shield fracture waters contain some of the highest levels of dissolved H(2) ever reported and represent a potentially important energy-rich environment for subsurface microbial life. The delta (2)H isotope signatures of H(2) gas from Canada, Finland, and South Africa are consistent with a range of H(2)-producing water-rock reactions, depending on the geologic setting, which include both serpentinization and radiolysis. In Canada and Finland, several of the sites are in Archean greenstone belts characterized by ultramafic rocks that have under gone serpentinization and may be ancient analogues for serpentinite-hosted gases recently reported at the Lost City Hydrothermal Field and other hydrothermal seafloor deposits. The hydrogeologically isolated nature of these fracture controlled groundwater systems provides a mechanism whereby the products of water rock interaction accumulate over geologic timescales, which produces correlations between high H(2) levels, abiogenic hydrocarbon signatures, and the high salinities and highly altered delta (18)O and delta (2)H values of these groundwaters. A conceptual model is presented that demonstrates how periodic opening of fractures and resultant mixing control the distribution and supply of H(2) and support a microbial community of H(2)-utilizing sulfate reducers and methanogens. PMID- 18163874 TI - Hydrothermal systems in small ocean planets. AB - We examine means for driving hydrothermal activity in extraterrestrial oceans on planets and satellites of less than one Earth mass, with implications for sustaining a low level of biological activity over geological timescales. Assuming ocean planets have olivine-dominated lithospheres, a model for cooling induced thermal cracking shows how variation in planet size and internal thermal energy may drive variation in the dominant type of hydrothermal system-for example, high or low temperature system or chemically driven system. As radiogenic heating diminishes over time, progressive exposure of new rock continues to the current epoch. Where fluid-rock interactions propagate slowly into a deep brittle layer, thermal energy from serpentinization may be the primary cause of hydrothermal activity in small ocean planets. We show that the time-varying hydrostatic head of a tidally forced ice shell may drive hydrothermal fluid flow through the seafloor, which can generate moderate but potentially important heat through viscous interaction with the matrix of porous seafloor rock. Considering all presently known potential ocean planets-Mars, a number of icy satellites, Pluto, and other trans-neptunian objects-and applying Earth-like material properties and cooling rates, we find depths of circulation are more than an order of magnitude greater than in Earth. In Europa and Enceladus, tidal flexing may drive hydrothermal circulation and, in Europa, may generate heat on the same order as present-day radiogenic heat flux at Earth's surface. In all objects, progressive serpentinization generates heat on a globally averaged basis at a fraction of a percent of present-day radiogenic heating and hydrogen is produced at rates between 10(9) and 10(10) molecules cm(2) s(1). PMID- 18163875 TI - Energy, chemical disequilibrium, and geological constraints on Europa. AB - Europa is a prime target for astrobiology. The presence of a global subsurface liquid water ocean and a composition likely to contain a suite of biogenic elements make it a compelling world in the search for a second origin of life. Critical to these factors, however, may be the availability of energy for biological processes on Europa. We have examined the production and availability of oxidants and carbon-containing reductants on Europa to better understand the habitability of the subsurface ocean. Data from the Galileo Near-Infrared Mapping Spectrometer were used to constrain the surface abundance of CO(2) to 0.036% by number relative to water. Laboratory results indicate that radiolytically processed CO(2)-rich ices yield CO and H(2)CO(3); the reductants H(2)CO, CH(3)OH, and CH(4) are at most minor species. We analyzed chemical sources and sinks and concluded that the radiolytically processed surface of Europa could serve to maintain an oxidized ocean even if the surface oxidants (O(2), H(2)O(2), CO(2), SO(2), and SO(4) (2)) are delivered only once every approximately 0.5 Gyr. If delivery periods are comparable to the observed surface age (30-70 Myr), then Europa's ocean could reach O(2) concentrations comparable to those found in terrestrial surface waters, even if approximately 10(9) moles yr(1) of hydrothermally delivered reductants consume most of the oxidant flux. Such an ocean would be energetically hospitable for terrestrial marine macrofauna. The availability of reductants could be the limiting factor for biologically useful chemical energy on Europa. PMID- 18163876 TI - Did earthquakes keep the early crust habitable? AB - The shallow habitable region of cratonal crust deforms with a strain rate on the order of approximately 10(19) s(1). This is rapid enough that small seismic events are expected on one-kilometer spatial scales and one-million-year timescales. Rock faulting has the potential to release batches of biological substrate, such as dissolved H(2), permitting transient blooms. In addition, the steady-state deformation of the brittle crust causes numerous small faults to be permeable enough (on the order of approximately 10(15) m(2)) for water to flow on a kilometer scale over relatively short geological times ( approximately 10(5) yr). Hence, active faults act as concentrated niches capable of episodically tapping resources in the bulk volume of the rock. Radiolysis and ferrous iron are potentially bases of sustainable hard-rock niches. PMID- 18163878 TI - Immune responses and protective efficacy of the gene vaccine expressing Ag85B and ESAT6 fusion protein from Mycobacterium tuberculosis. AB - Genetic immunity is a new promising approach for the development of novel tuberculosis vaccines. In this study, it is shown that DNA vaccines expressing the fusion protein of antigen 85B (Ag85B) and early secreted antigenic target 6 kDa antigen (ESAT6) can induce high levels of specific IgG2a antibody subtype in the mice. With the prolongation of postimmunization time, the levels of IgG2a antibody decrease gradually. Although a high-level specific IgG2a antibody subtype is also elicited by classical BCG, the ratio of antibody subtypes IgG2a to IgG1 changes 4 weeks after immunization, and IgG1 is gradually shifted to the main antibody subtype. DNA vaccines also elicit cellular immunity as shown by specific spleen lymphocytes proliferation to Ag85B or ESAT6 protein and the production of high levels of IFN-gamma and IL-2, which is similar to that elicited by BCG. Vaccination of mice with DNA vaccines expressing the fusion protein Ag85B-ESAT6 results in a significant level of protection against the subsequent high-dose challenge with virulent Mycobacterium tuberculosis (MTB) H37Rv. Dramatic reduction in the number of MTB colony-forming units in the spleens and lungs is observed. Pathological examination showed that recombinant plasmid and BCG groups have only minor damage and organizational structures that are kept relatively complete, while in the control group, spleens and lungs are damaged seriously. Therefore, although the reducing degree of mycobacterial loads in the organ of mice immunized with recombinant plasmid is not more than that of BCG, through the analysis of pathological changes, we may conclude that the protective effect provided by DNA vaccine expressing the Ag85B-ESAT6 fusion protein is equivalent to that afforded by the classical BCG. PMID- 18163879 TI - Heterogeneic dynamics of the structures of multiple gene clusters in two pathogenetically different lines originating from the same phytoplasma. AB - Phytoplasmas are phloem-limited plant pathogens that are transmitted by insect vectors and are associated with diseases in hundreds of plant species. Despite their small sizes, phytoplasma genomes have repeat-rich sequences, which are due to several genes that are encoded as multiple copies. These multiple genes exist in a gene cluster, the potential mobile unit (PMU). PMUs are present at several distinct regions in the phytoplasma genome. The multicopy genes encoded by PMUs (herein named mobile unit genes [MUGs]) and similar genes elsewhere in the genome (herein named fundamental genes [FUGs]) are likely to have the same function based on their annotations. In this manuscript we show evidence that MUGs and FUGs do not cluster together within the same clade. Each MUG is in a cluster with a short branch length, suggesting that MUGs are recently diverged paralogs, whereas the origin of FUGs is different from that of MUGs. We also compared the genome structures around the lplA gene in two derivative lines of the 'Candidatus Phytoplasma asteris' OY strain, the severe-symptom line W (OY-W) and the mild symptom line M (OY-M). The gene organizations of the nucleotide sequences upstream of the lplA genes of OY-W and OY-M were dramatically different. The tra5 insertion sequence, an element of PMUs, was found only in this region in OY-W. These results suggest that transposition of entire PMUs and PMU sections has occurred frequently in the OY phytoplasma genome. The difference in the pathogenicities of OY-W and OY-M might be caused by the duplication and transposition of PMUs, followed by genome rearrangement. PMID- 18163880 TI - Small-molecule inhibitors of Bcl-2 family proteins are able to induce tumor regression in a mouse model of pre-B-cell acute lymphocytic lymphoma. AB - The overexpression of prosurvival members of the Bcl-2 family is commonly associated with the enhanced malignancy of hematological tumors. There has been great interest in a novel set of agents that are able to mimic the function of the BH3 domain by binding to the groove of Bcl-2-like proteins and initiating the cell death sequence. We sought to examine the efficacy of BH3 mimetics in a spontaneous mouse model of B-cell neoplasia. We evaluated the ability of the BH3 mimetics to preferentially target tumor cells while sparing normal cells. In addition, we examined the contributions of Bim and Puma to the sensitivity of tumor cells to the BH3 mimetics. We report here that two BH3 mimetics (HA-14-1 and BH3-I-2') were able to induce apoptosis of murine B-cell lymphoma cells in vitro and in vivo. Tumors that arose from transplantation of primary lymphoma cells regressed following 7 days of treatment with BH3-mimetic drugs. The long term benefits of the transient treatment of tumor-bearing mice with the BH3 mimetics, however, could not be properly evaluated, due to the high levels of toxicity we observed in vivo with these drugs. Decreased expression of either Bim or Puma from B-cell tumor cells was able to protect these cells from the apoptosis induced by these BH3 mimetics, suggesting that they function through other means. We conclude that while the BH3-mimetic drugs are effective at inducing cell death of lymphoma cells in vitro and in vivo, their unclear molecular specificity and their ability to kill normal cells may limit their therapeutic uses in humans. PMID- 18163881 TI - Development of a human adipocyte model derived from human mesenchymal stem cells (hMSC) as a tool for toxicological studies on the action of TCDD. AB - Abstract Efforts were made to develop a human adipocyte model that is useful for toxicological studies in vitro. For this purpose, a stem cell line derived from human bone marrow cells, originally from an adult, was induced to differentiate towards adipocytes by treating them with insulin, dexamethasone, indomethacin and 3-isobutyl-1-methylxanthine for 3 d, followed by additional incubation for 3 d in Dulbecco's modified Eagle's medium supplemented with insulin only. In most cases, thus differentiated cells through such one cycle of differentiation treatment were further subjected to the second cycle of differentiation. The resulting 2 cycle differentiated cells were found to exhibit many characteristics of typical adipocytes. Dioxin (TCDD), when added at the beginning of their treatment with differentiation-inducing hormone cocktail, clearly prevented them from becoming adipocytes, as in the case of TCDD-treated 3T3-L1 cells. Furthermore, TCDD, even when administered to previously differentiated human mesenchymal stem cells (hMSC) adipocytes, consistently induced the sign of inflammatory responses during the early period of TCDD action (24 h), which was followed by gradual loss of adipocyte-specific markers during the 5-d incubation period. In conclusion, hMSC derived adipocytes appear to offer a promising human cell model suited for future toxicological studies. PMID- 18163882 TI - Secondary structures and functional requirements for thiM riboswitches from Desulfovibrio vulgaris, Erwinia carotovora and Rhodobacter spheroides. AB - Abstract Bacterial thiM riboswitches contain aptamer domains that bind the metabolite thiamine pyrophosphate (TPP). Binding of TPP to the aptamer domain induces structural rearrangements that are relayed to the expression domain, thereby interfering with gene expression. Here, we report identification of three putative thiM riboswitches from different bacteria and analysis of their secondary structures. Chemical probing revealed that the riboswitches share similar secondary structures in their aptamer domains that can communicate with the highly variant expression domains in a mechanism likely involving sequestration of the Shine-Dalgarno sequence. Remarkably, the aptamer domain of the thiM gene of Desulfovibrio vulgaris binds TPP with similar affinity and selectivity as that of Escherichia coli, although nucleotides previously shown to form direct contacts to the metabolite are mutated. We also designed small RNA hairpins for each riboswitch that bind the RNA only in the absence of the metabolite. Our study shows that aptamer domains in riboswitches with high similarity in their secondary structures can communicate with a broad variety of non-related expression domains by similar mechanisms. PMID- 18163883 TI - Evaluation of the 'side door' in carboxylesterase-mediated catalysis and inhibition. AB - Abstract Structures of mammalian carboxylesterases (CEs) reveal the presence of a 'side door' that is proposed to act as an alternative pore for the trafficking of substrates and products. p-Nitrobenzyl esterase (pnb CE) from Bacillus subtilis exhibits close structural homology and a similar side-door domain as mammalian CEs. We investigated the role of a specific 'gate' residue at the side door (i.e., Leu 362) during pnb CE-catalyzed hydrolysis of model esters, pesticides, and lipids. Recombinant pnb CE proteins containing mutations at position 362 demonstrated markedly lower kcat and kcat/Km values. The mutation with the most significant impact on catalysis was the L362R mutant (kcat/Km was 22-fold lower). Moreover, the ability of the L362R mutant to be inhibited by organophosphates (OP) was also lower. Investigation into the altered catalytic proficiency using pH-activity studies indicated that the catalytic triad of the mutant enzyme was preserved. Furthermore, viscosity variation and carbamate inhibition experiments indicated that rates of substrate association and acylation/deacylation were lower. Finally, recombinant CEs were found to possess lipolytic activity toward cholesteryl oleate and 2-arachidonylglycerol. In summary, the L362R mutant CE markedly slowed the rate of ester hydrolysis and was less sensitive to OP inhibition. The apparent causes of the diminished catalysis are discussed. PMID- 18163884 TI - Effects of gastric inhibitory polypeptide (GIP) and related analogues on glucagon release at normo- and hyperglycaemia in Wistar rats and isolated islets. AB - Glucose-dependent insulinotropic polypeptide (GIP) is an incretin hormone secreted by endocrine K-cells in response to nutrient absorption. This study has utilised numerous well-characterised dipeptidyl peptidase IV-resistant GIP analogues to evaluate the glucagonotropic actions of GIP in Wistar rats and isolated rat islets. Intraperitoneal administration of GIP analogues (25 nmol/kg body weight) in combination with glucose had no effect on circulating glucagon concentrations compared to controls in Wistar rats. However, plasma glucose concentrations were significantly (p<0.05 to p<0.001) lowered by the GIP-receptor agonists, N-AcGIP, GIP(Lys37)PAL and N-AcGIP(Lys37)PAL. The GIP antagonist, (Pro3)GIP, caused a significant (p<0.05) reduction in glucagon levels following concurrent administration with saline in Wistar rats. In isolated rat islets native GIP induced a significant (p<0.01) enhancement of glucagon release at basal glucose concentrations, which was completely annulled by (Pro3)GIP. Furthermore, glucagon release in the presence of GLP-1, GIP(Lys37)PAL, N AcGIP(Lys37)PAL and (Pro3)GIP was significantly (p<0.05 to p<0.001) decreased compared to native GIP in isolated rat islets. These data indicate a modest effect of GIP on glucagon secretion from isolated rat islets, which was not observed in vivo. However, the GIP agonists N-AcGIP, GIP(Lys37)PAL and N AcGIP(Lys37)PAL had no effect on glucagon release demonstrating an improved therapeutic potential for the treatment of type 2 diabetes. PMID- 18163885 TI - Functional tyrosine residue in the active center of human dipeptidyl peptidase III. AB - Abstract Human dipeptidyl peptidase III (DPP III) is a member of the metallopeptidase family M49 with an implied role in the pain-modulatory system and endogenous defense against oxidative stress. Here, we report the heterologous expression of human DPP III and the site-directed mutagenesis results which demonstrate a functional role for Tyr318 at the active site of this enzyme. The substitution of Tyr318 to Phe decreased kcat by two orders of magnitude without altering the binding affinity of substrate, or of a competitive hydroxamate inhibitor designed to interact with S1 and S2 subsites. The results indicate that the conserved tyrosine could be involved in transition state stabilization during the catalytic action of M49 peptidases. PMID- 18163886 TI - Transcriptional responses of Plasmodium falciparum to alpha difluoromethylornithine-induced polyamine depletion. AB - Abstract Polyamines are essential polycationic molecules involved in multiple cellular events, including cell differentiation, division and death. Inhibition of polyamine biosynthesis has been considered in diverse therapeutic strategies ranging from tumour suppressors to anti-parasitic agents. In the human malaria parasite, Plasmodium falciparum, inhibition of ornithine decarboxylase (ODC) results in the arrest of schizogony due to polyamine depletion. However, the exact physiological role of the polyamines in the parasite is unknown. Here, we present results of the depletion of polyamines in the malaria parasite by alpha difluoromethylornithine inhibition of ODC, as observed with differential transcriptome profiling. Upon depletion of their endogenous polyamines, the up- and downregulated parasite transcripts were selected with suppression subtractive hybridisation and differences were detected using blots or DNA microarrays. A direct linkage between polyamine depletion and the differential expression of two distinct transcripts was observed, indicating the existence of a transcriptional feedback response in the P. falciparum transcriptome upon drug challenge. The data presented provide input into the role of the polyamines in the cellular biology of P. falciparum and contribute towards the validation of polyamine biosynthesis as an antimalarial target. PMID- 18163887 TI - Tissue-specific promoter utilisation of the kallikrein-related peptidase genes, KLK5 and KLK7, and cellular localisation of the encoded proteins suggest roles in exocrine pancreatic function. AB - Abstract Tissue kallikrein (kallikrein 1) was first identified in pancreas and is the namesake of the kallikrein-related peptidase (KLK) family. KLK1 and the other 14 members of the human KLK family are encoded by 15 serine protease genes clustered at chromosome 19q13.4. Our Northern blot analysis of 19 normal human tissues for expression of KLK4 to KLK15 identified pancreas as a common expression site for the gene cluster spanning KLK5 to KLK13, as well as for KLK15 which is located adjacent to KLK1. Consistent with previous reports detailing the ability of KLK genes to generate organ- and disease-specific transcripts, detailed molecular and in silico analyses indicated that KLK5 and KLK7 generate transcripts in pancreas variant from those in skin or ovary. Consistently, we identified in the promoters of these KLK genes motifs which conform with consensus binding sites for transcription factors conferring pancreatic expression. In addition, immunohistochemical analysis revealed predominant localisation of KLK5 and KLK7 in acinar cells of the exocrine pancreas, suggesting roles for these enzymes in digestion. Our data also support expression patterns derived from gene duplication events in the human KLK cluster. These findings suggest that, in addition to KLK1, other related KLK enzymes will function in the exocrine pancreas. PMID- 18163888 TI - Secreted human apolipoprotein(a) kringle IV-10 and kringle V inhibit angiogenesis and xenografted tumor growth. AB - Abstract Angiogenesis plays an important role in normal physiology of blood vessel growth, but can contribute to the pathogenesis of diseases, such as cancer. A new anti-angiogenic recombinant kringle protein, composed of the fused domains of human apolipoprotein(a) carboxyl-terminal kringle IV-10 and kringle V, was expressed in Pichia pastoris and human colorectal carcinoma (HCT 116) cells to investigate its influence on angiogenesis and tumor growth. The mature recombinant protein exhibited the characteristic features of kringle-containing proteins (glycosylation and disulfide bond formation) and, when added to cultures of human umbilical vein endothelial cell, resulted in a 31% decrease in proliferation relative to untreated controls (p<0.05). The neo-angiogenesis was diminished by 63% in chick embryos treated with 10 mug recombinant protein compared with 7% for phosphate buffer solution-treated embryos (p<0.01). Transfection of a kringle IV-10-kringle V fusion protein construct into HCT 116 cells decreased tumorigenesis and inhibited tumor growth in vivo without affecting tumor cell proliferation. HCT 116 cells that expressed recombinant protein displayed a much lower relative growth ratio of 8% (p<0.01) against the control tumor cells. From these results, we conclude that human apolipoprotein(a) carboxyl-terminal kringle IV-10-kringle V fusion protein is an effective inhibitor of angiogenesis and angiogenesis-dependent tumor growth. PMID- 18163889 TI - Insulin-releasing properties of the frog skin peptide pseudin-2 and its [Lys18] substituted analogue. AB - Abstract Pseudin-2 is a cationic alpha-helical peptide that was first isolated from the skin of the paradoxical frog Pseudis paradoxa on the basis of its antimicrobial activity. We have investigated the insulin-releasing properties and cytotoxicity of the peptide, together with selected analogues with increased cationicity and hydrophobicity. At concentrations in the range 10(-9)-10(-6) m, pseudin-2, and its [Lys18], [Phe8], and [d-Lys3,d-Lys10,d-Lys14] derivatives, stimulated insulin release from the BRIN-BD11 clonal beta-cell line without increasing release of lactate dehydrogenase. The [Lys18] analogue was the most potent (46% increase in insulin release at 10(-9) m) and the most effective (215% increase in insulin release at 10(-6) m). The more cationic [Lys3,Lys10,Lys14] and [Lys3,Lys10,Lys14,Lys21] analogues lacked insulinotropic action and the more hydrophobic [Phe16] analogue was cytotoxic at concentrations > or =10(-7) m. Pseudin-2 and [Lys18]-pseudin-2 had no effect on intracellular calcium concentrations and stimulated insulin release in the absence of external calcium. [Lys18]-pseudin-2 (10(-8) m) stimulated insulin release in the presence of diazoxide and verapamil. Our results demonstrate that pseudin-2 stimulates insulin secretion from BRIN-BD11 cells by a mechanism involving Ca2+-independent pathways and identify [Lys18]-pseudin-2 as a peptide that may have potential for development as a therapeutically valuable insulinotropic agent for the treatment of type 2 diabetes. PMID- 18163890 TI - HNF4 alpha orchestrates a set of 14 genes to down-regulate cell proliferation in kidney cells. AB - Abstract Few genes are known to be involved in renal cell carcinoma (RCC) development and progression. The cell-specific transcription factor hepatocyte nuclear factor 4 alpha (HNF4 alpha) is down-regulated in RCC and we have shown that HNF4 alpha inhibits cell proliferation in the embryonic kidney cell line HEK293. To clarify the possible tumor suppressor activity of HNF4 alpha we analyzed the whole human expression profile in HEK293 cells upon HNF4 alpha induction. By comparing induced and uninduced cells, we identified 1411 differentially expressed genes. Using RNA interference, we screened 56 HNF4 alpha regulated genes for their possible role in mediating inhibition of cell proliferation triggered by HNF4 alpha. We demonstrate that 14 of these regulated genes are able to contribute to the inhibitory effect of HNF4 alpha on cell proliferation, including well-known cancer genes, such as CDKN1A (p21), TGFA, MME (NEP) and ADAMTS1. In addition, the genes SEPP1, THEM2, BPHL, DSC2, ANK3, ALDH6A1, EPHX2, NELL2, EFHD1 and PROS1 are also part of the network of HNF4 alpha target genes that regulate proliferation in HEK293 cells. Therefore, we postulate that HNF4 alpha orchestrates, at least, these 14 genes to regulate cell proliferation in HEK293 cells and that down-regulation of HNF4 alpha could contribute to the progression of kidney cancer. PMID- 18163891 TI - Cathepsin V, but not cathepsins L, B and K, may release angiostatin-like fragments from plasminogen. AB - Abstract Cathepsin V is a lysosomal cysteine peptidase highly expressed in corneal epithelium; however, its function in the eye is still unknown. Here, we describe the capability of cathepsin V to hydrolyze plasminogen, which is also expressed in human cornea at levels high enough to produce physiologically relevant amounts of angiostatin-related molecules. The co-localization of these two proteins suggests an important role for the enzyme in the maintenance of corneal avascularity, essential for optimal visual performance. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of plasminogen digestion by cathepsin V revealed the generation of three major products of 60, 50 and 40 kDa, which were electrotransferred to polyvinylidene difluoride membranes and excised for characterization. NH(2)-terminal amino acid sequencing of these fragments revealed the sequences EKKVYL, TEQLAP and LLPNVE, respectively. These data are compatible with cleavage sites at plasminogen F94-E95, S358-T359 and V468-L469 peptide bonds generating fragments of the five-kringle domains. In contrast, we did not detect any plasminogen degradation by cathepsins B, K and L. Using a Matrigel assay, we confirmed the angiogenesis inhibition activity on endothelial cells caused by plasminogen processing by cathepsin V. Our results suggest a novel physiological role for cathepsin V related to the control of neovascularization in cornea. PMID- 18163892 TI - Refractive surgery in adults with binocular vision "abnormalities.". PMID- 18163893 TI - The eyes had it first. PMID- 18163895 TI - Sixth nerve palsy post intravitreal bevacizumab for AMD: a new possibly causal relationship and complication? PMID- 18163896 TI - Safety stitch: a modification to postoperatively adjustable suture strabismus surgery of the inferior rectus muscle. AB - OBJECTIVE: To introduce a variation of adjustable suture recession surgery of the inferior rectus muscle by adding a non- absorbable "safety stitch" to reduce post surgery overcorrection. METHODS: Eleven patients with vertical strabismus who needed inferior rectus recession were the subjects of this study. The vertical deviation was measured preoperatively, 24 hours after the adjustment, and after a minimum of a year followup. An adjustable suture technique through a limbal incision with a silicon sheet was used. We added a non-absorbable suture in the medial edge of the tendon of the inferior rectus muscle and fixed it at the scleral insertion of the muscle. This area of the inferior rectus tendon was exposed for the adjustment, which was performed 24-48 hours after the surgery. The safety suture was then fastened with a knot and 4-6 prism diopters (pd) of undercorrection in down gaze was intentionally left. RESULTS: The average preoperative vertical deviation was 17 prism diopters (pd) in primary position, and 21.6 pd in down gaze. Six of the eleven patients were adjusted postop' leaving an average residual vertical deviation of 2 pd in primary position and 4.7 pd in down gaze. After a year of followup, the average vertical deviation was 0.4 pd in primary position and 2 pd in down gaze. Ten of the eleven patients were considered to have "successful" primary surgery using this technique. The eleventh required a second operation for an undercorrection which resulted from inadequate original placement of the safety stitch. CONCLUSIONS: The non absorbable safety stitch technique provided satisfactory results, superior to previously reported techniques for postop' adjustable recession strabismus surgery of the inferior rectus muscle. PMID- 18163897 TI - Radio-opaque modification/substitute for the wright superior oblique tendon extender for superior oblique muscle overaction strabismus. AB - Kenneth Wright developed a technique for graded weakening of the superior oblique by increasing the effective length of this extraocular muscle's long tendon with a piece of silicone rubber retinal encircling band commonly used by eye surgery for retinal detachment repairs. In the absence of any specific retinal bands in our Children's Hospital, the following technique was developed affording a non invasive ability to monitor, which was less intricate than the technique so well described by Demer. We substituted the "Mini Vessel Loop" (by Maaxxim Medical or Henley International). It is an elastic smooth silicone rubber cord that is radio opaque, and can easily be seen on X-rays and CT scans. It is not an ophthalmologic medical device but it rather is designed to loop around and identify and gently retract blood vessels and nerves in any form of surgery where needed. We demonstrated success similar to that achieved by Wright in 43 patients using these radio-opaque, silicon Mini Vessel loops. PMID- 18163898 TI - Essential infantile esotropia in neurologically impaired pediatric patients: is botulinum toxin better primary treatment than surgery? AB - A prospective study was performed over a 10 year period on 25 children with infantile esotropia and neurological problems to answer this question. From November 1996 to March 2006 they were treated with injections of botulinum toxin (Botox) of both medial rectus extraocular muscles. Mean age was 26.4 months, (range 9-76 months) and mean initial angle was 35 prism diopters (PD)(range 20-60 PD). RESULTS: 18 patients (72%) remained orthotropic+/-10 PD at 29 months (range 6-59 months). Average number of injection treatments was 1.5 per patient. We compared our success rate data with those obtained with primary conventional strabismus surgical procedures in 2 previously published series. Treatment with botulinum toxin seemed to produce better results than one surgical series and at least equally similar results to the other one. Because there are, as well, so many other advantages to the injection procedure including superior safety and economy, we now use botulinum injections as our primary treatment in these patients. PMID- 18163899 TI - Surgical correction of synergistic divergence strabismus. A report of three cases. AB - BACKGROUND AND PURPOSE: To review and explain some of the characteristics of synergistic divergence and compare the surgical effect on each of those characteristics. METHODS OF STUDY: Three patients demonstrating findings characteristic of synergistic divergence, two bilateral and one unilateral, are reported. Surgery consisted of denervation extirpation of the lateral rectus muscle along with resection of the medial rectus muscle of the affected eye or supramaximal recession of the lateral rectus muscle and resection of the antagonist medial rectus muscle of the affected eye. RESULTS: The simultaneous abduction and the abnormal head posture responded well to surgery. The exotropia was reduced. The deficient adduction, the total absence of active abduction and the infraduction of the synkinetically abducting eye remained unchanged. CONCLUSIONS: Synergistic divergence is a severe exotropic form of Duane's Syndrome where the feature of simultaneous abduction is the most striking. To date, no surgical procedure leads to satisfactory results on all the characteristics of synergistic divergence. Although some important features can be satisfactorily improved, some others do not respond to current surgical techniques. PMID- 18163900 TI - Outcome study of two standard and graduated augmented modified Kestenbaum surgery protocols for abnormal head postures in infantile nystagmus. AB - BACKGROUND AND PURPOSE: Since Kestenbaum and Anderson, several ophthalmologists have reported the results of different surgical procedures, for abnormal head posture in infantile nystagmus. In this study, we tried to evaluate the surgical results of Parks' original 5-6-7-8 mm modified Kestenbaum procedure and our own 6 7-6-7 mm modified Kestenbaum procedure, designed to reduce some of the problems encountered with other variations of these techniques. METHODS: Medical records of 92 patients, who had modified Kestenbaum surgery (5-6-7-8 mm or 6-7-6-7 mm) at The Yonsei Medical Center, from March 1991 to September 2001 with a follow-up period of more than 6 months, were reviewed retrospectively. We compared Parks' modified Kestenbaum surgery (5- 6-7-8 mm) performed on 51 patients with our own modified Kestenbaum surgery (6-7-6-7 mm) on 41 patients. Each procedure was done with graded augmentation according to the amount of the face turn and the null point in electro-oculography. RESULTS: In the follow-up of an average 33 months, 45 out of 51 patients (88.2%) who underwent Parks' modified procedures showed face turn less than 10 degrees. In the follow-up of an average 29 months, 36 out of 41 patients (87.8%) with 6-7-6-7 mm procedure had face turn less than 10 degrees. CONCLUSIONS: We suggest that 6-7-6-7 mm modified Kestenbaum procedures with a graded augmentation may be a safe and efficient procedure to correct abnormal head posture in infantile nystagmus with a minimum decrease in ocular motility. PMID- 18163901 TI - Cocaine treatment admissions at three sentinel sites in South Africa (1997-2006): findings and implications for policy, practice and research. AB - BACKGROUND: Accurate prevalence data on cocaine use, that points to where problems exist and the extent of these problems, is necessary to guide the formulation of effective substance abuse policy and practice. The purpose of this study was to provide surveillance information about the nature and extent of problematic cocaine use in South Africa. METHODS: Data were collected between January 1997 and December 2006 on admissions for drug abuse treatment through a regular monitoring system involving 56 drug treatment centres and programmes in Cape Town, Gauteng Province (Johannesburg and Pretoria) and the Eastern Cape every six months as part of the South African Community Epidemiology Network on Drug Use (SACENDU). A one-page form was completed by treatment centre personnel to obtain demographic data, the patients' primary and secondary substances of abuse, the mode, frequency and age of first use of substance, and information on prior treatment. RESULTS: Treatment indicators point to a significant increase in cocaine related admissions over time in all sites, but with substantial inter site variation, particularly in recent years. The data indicate high levels of crack cocaine use and high levels of daily usage among patients, most of whom were first time admissions. Patients with cocaine related problems continue to be predominantly male, with a mean age of around 30 years. Substantial changes in the racial profile of patients have occurred over time. Poly drug use is high with cocaine often used with alcohol, cannabis and other drugs. CONCLUSION: These trends point to the possibility of cocaine use becoming a serious health and social issue in South Africa and demonstrate the utility of continued monitoring of cocaine treatment admissions in the future. They also highlight the need to address cocaine use in national and provincial policy planning and intervention efforts. In terms of treatment, the findings highlight the need to ensure that treatment practitioners are adequately trained to address stimulant problems, poly drug use, and HIV and other risk behaviour related to crack cocaine use. Possible gaps in access to treatment by certain sectors of the population should be addessed as a matter of urgency. PMID- 18163902 TI - A three-arm randomised controlled trial comparing Gonadotrophin Releasing Hormone (GnRH) agonist long regimen versus GnRH agonist short regimen versus GnRH antagonist regimen in women with a history of poor ovarian response undergoing in vitro fertilisation (IVF) treatment: Poor responders intervention trial (PRINT). AB - BACKGROUND: Poor response to ovarian stimulation with exogenous gonadotrophins occurs in 9-24% of women undergoing in vitro fertilisation (IVF) treatment, which represents an estimated 4000-10,000 women per year in the UK. Poor responders often have their treatment cycle cancelled because of expected poor outcome.One treatment strategy that may influence outcome is the choice of pituitary suppression regimen prior to the initiation of ovarian stimulation. The three commonly used pituitary suppression regimens in IVF treatment are:(1) the GnRH agonist long regimen,(2) the GnRH agonist short regimen and(3) the GnRH antagonist regimen.A systematic review of randomised controlled trials of these pituitary suppression regimens has shown the evidence to be either inconclusive or inconsistent. We therefore designed a three arm randomised trial to evaluate the effectiveness of these regimens in women who had poor ovarian response in a previous IVF treatment cycle. METHODS/DESIGN: Consenting, eligible women will be randomised to one of the three regimens using an internet-based trial management programme that ensures allocation concealment and employs block randomisation and minimisation for prognostic variables. The primary outcome is the number of oocytes retrieved. Other outcomes include total dose of follicle stimulating hormone (FSH) used for ovarian stimulation, mature oocytes retrieved, embryos available for transfer, implantation rate and clinical pregnancy rate.The sample size for this trial has been estimated as 102 participants with 34 participants in each of the three arms. Appropriate interim analysis will be conducted by a Data Monitoring and Ethics Committee (DMEC), and the final analysis will be by intention to treat. TRIAL REGISTRATION: ISRCTN27044628. PMID- 18163903 TI - BGLAP is expressed in pancreatic cancer cells and increases their growth and invasion. AB - BACKGROUND: Bone gamma-carboxyglutamate protein (BGLAP; osteocalcin) is a small, highly conserved molecule first identified in the mineralized matrix of bone. It has been implicated in the pathophysiology of various malignancies. In this study, we analyzed the expression and role of BGLAP in the normal human pancreas, chronic pancreatitis (CP), and pancreatic ductal adenocarcinoma (PDAC) using quantitative RT-PCR, immunohistochemistry, immunocytochemistry and enzyme immunoassays, as well as cell proliferation and invasion assays. Gene silencing was carried out using specific siRNA molecules. RESULTS: Compared to the normal pancreas, BGLAP mRNA and protein levels were not significantly different in CP and PDAC tissues. BGLAP was faintly present in the cytoplasm of normal acinar cells but was strongly expressed in the cytoplasm and nuclei of tubular complexes and PanIN lesions of CP and PDAC tissues. Furthermore, BGLAP expression was found in the cancer cells in PDAC tissues as well as in 4 cultured pancreatic cancer cell lines. TNFalpha reduced BGLAP mRNA and protein expression levels in pancreatic cancer cell lines. In addition, BGLAP silencing led to reduction of both cell growth and invasion in those cells. CONCLUSION: BGLAP is expressed in pancreatic cancer cells, where it potentially increases pancreatic cancer cell growth and invasion through autocrine and/or paracrine mechanisms. PMID- 18163904 TI - Posterior mediastinal hematoma - a rare case following a fall from standing height: a case report. AB - INTRODUCTION: We present a previously unreported complication of a fall from standing height. CASE PRESENTATION: A 76-year-old woman sustained blunt chest trauma resulting from a fall from standing height. She was diagnosed with a mediastinal hematoma, and did well with supportive care. Follow up CT angiograms on days 2 and 4 of hospital stay revealed a stable hematoma and she did not require any intervention. CONCLUSION: Mediastinal hematoma has been reported secondary to trauma, coagulation abnormalities and hematologic malignancies, but it not been reported secondary to a fall from standing height. Factors predisposing to a hematoma in this case were aspirin therapy and a modest elevation of INR secondary to chronic hepatitis C. PMID- 18163905 TI - Health-related quality of life of patients following selected types of lumbar spinal surgery: a pilot study. AB - BACKGROUND: Over 500,000 spinal surgeries are performed annually in the United States. Although pain relief and improved health-related quality of life (HRQOL) are expectations following lumbar spinal surgery, there is limited research regarding this experience from the individual's perspective. In addition, no studies have examined the HRQOL of persons who have had this surgery using a comprehensive approach. The intent of this study was to address this deficiency by an assessment of both the individual and environmental factors that impact perceived HRQOL using the Wilson and Cleary Model for Health-Related Quality of Life in persons who have undergone lumbar spinal surgery. METHODS: This was a pilot study of 57 adult patients undergoing elective lumbar spinal surgery for either herniated disk and/or degenerative changes. Individuals completed questionnaires measuring perceived pain, mood, functional status, general health perceptions, social support and HRQOL preoperatively and three months following surgery. Descriptive statistics, dependent t-tests, and MANOVAs were used to describe and compare the differences of the study variables over time. RESULTS: Preliminary results indicate overall perceived physical HRQOL was significantly improved postoperatively (t [56] = 6.45, p < .01), however, it was lower than the published norms for patients with low back pain. Both functional disability (t [56] = 10.47, p < .001) and pain (t [56] = 10.99, p < .001) were significantly improved after surgery. Although levels of fatigue and vigor were also significantly improved after surgery, both were less than the published norms. There was no change in the level of social support over time; however, level of support was consistent with that reported by patients with chronic illness. CONCLUSION: Although perceived physical HRQOL was significantly improved three months postoperatively, fatigue and lack of vigor were issues for subjects postoperatively. Excessive fatigue and low vigor may have implications for successful rehabilitation and return to work for patients following lumbar spinal surgery. Further research is needed with a larger sample size and subgroup analyses to confirm these results. PMID- 18163906 TI - Epigenetic hereditary transcription profiles II, aging revisited. AB - BACKGROUND: Previously, we have shown that deviations from the average transcription profile of a group of functionally related genes can be epigenetically transmitted to daughter cells, thereby implicating nuclear programming as the cause. As a first step in further characterizing this phenomenon it was necessary to determine to what extent such deviations occur in non-tumorigenic tissues derived from normal individuals. To this end, a microarray database derived from 90 human donors aged between 22 to 87 years was used to study deviations from the average transcription profile of the proteasome genes. RESULTS: Increase in donor age was found to correlate with a decrease in deviations from the general transcription profile with this decline being gender specific. The age-related index declined at a faster rate for males although it started from a higher level. Additionally, transcription profiles from similar tissues were more alike than those from different tissues, indicating that deviations arise during differentiation. CONCLUSION: These findings suggest that aging and differentiation are related to epigenetic changes that alter the transcription profile of proteasomal genes. Since alterations in the structure and function of the proteasome are unlikely, such changes appear to occur without concomitant change in gene function. These findings, if confirmed, may have a significant impact on our understanding of the aging process. PMID- 18163907 TI - Design of a trans protease lentiviral packaging system that produces high titer virus. AB - BACKGROUND: The structural and enzymatic proteins of the human immunodeficiency virus (HIV) are initially generated as two long polyproteins encoded from overlapping reading frames, one producing the structural proteins (Gag) and the second producing both structural and enzymatic proteins (Gag-Pol). The Gag to Gag Pol ratio is critical for the proper assembly and maturation of viral particles. To minimize the risk of producing a replication competent lentivirus (RCL), we developed a "super-split" lentiviral packaging system in which Gag was separated from Pol with minimal loss of transducibility by supplying protease (PR) in trans independently of both Gag and Pol. RESULTS: In developing this "super-split" packaging system, we incorporated several new safety features that include removing the Gag/Gag-Pol frameshift, splitting the Gag, PR, and reverse transcriptase/integrase (RT/IN) functions onto separate plasmids, and greatly reducing the nucleotide sequence overlap between vector and Gag and between Gag and Pol. As part of the construction of this novel system, we used a truncated form of the accessory protein Vpr, which binds the P6 region of Gag, as a vehicle to deliver both PR and RT/IN as fusion proteins to the site of viral assembly and budding. We also replaced wt PR with a slightly less active T26S PR mutant in an effort to prevent premature processing and cytoxicity associated with wt PR. This novel "super-split" packaging system yielded lentiviral titers comparable to those generated by conventional lentiviral packaging where Gag-Pol is supplied intact (1.0 x 106 TU/ml, unconcentrated). CONCLUSION: Here, we were able to create a true "split-function" lentiviral packaging system that has the potential to be used for gene therapy applications. This novel system incorporates many new safety features while maintaining high titers. In addition, because PR is supplied in trans, this unique system may also provide opportunities to examine viral protein processing and maturation. PMID- 18163908 TI - Changeovers of vasoactive drug infusion pumps: impact of a quality improvement program. AB - BACKGROUND: Hemodynamic instability following the changeover of vasoactive infusion pump (CVIP) is a common problem in the intensive care unit. Several empiric methods are used to achieve CVIP. We hypothesized that the variation in these procedures could generate some morbidity. We sought to assess the effects of the standardization of practice, as a quality improvement program, on the CVIP induced incidents. MATERIALS AND METHOD: We performed a prospective before-and after intervention study including all adult patients with a diagnosis of cardiovascular failure who received a continuous infusion of vasoactive drugs or inotropic drugs. After a baseline preimplementation period (phase 1), a standardized 'quick change method' of CVIP using two syringe drivers was implemented in our intensive care unit (phase 2). Endpoints (rate and distribution of incidents: variations of systolic blood pressure >20 mmHg or heart rate >20 beats/min, and arrhythmias) were registered in both 3-month phases. RESULTS: We studied a total of 913 CVIP events (phase 1, 435 events; phase 2, 478 events) from 43 patients. Patient characteristics were not significantly different among phases, with a majority of the patients having septic shock. The frequency of incidents was significantly (P < 0.0001) reduced in phase 2 (5.9%, n = 28) versus phase 1 (17.8%, n = 78). This effect was observed whichever catecholamine was used. More than 98% of incidents were blood pressure variations, with a similar distribution of the nature of incidents in both phases. CONCLUSION: The present study illustrates that adverse events are common following CVIP, and illustrates the positive impact of a quality improvement program to enhance inpatient safety related to this current process of care. PMID- 18163909 TI - Marathon related death due to brainstem herniation in rehydration-related hyponatraemia: a case report. AB - INTRODUCTION: Identifying marathon runners at risk of neurological deterioration at the end of the race (within a large cohort complaining of exhaustion, dehydration, nausea, headache, dizziness, etc.) is challenging. Here we report a case of rehydration-related hyponatraemia with ensuing brain herniation. CASE PRESENTATION: We report the death of runner in his 30's who collapsed in the recovery area following a marathon. Following rehydration he developed a respiratory arrest in the emergency room. He was found to be hyponatraemic (130 mM). A CT brain scan showed severe hydrocephalus and brain stem herniation. Despite emergency insertion of an extraventricular drain, he was tested for brainstem death the following morning. Funduscopy demonstrated an acute-on chronic papilledema; CSF spectrophotometry did not reveal any trace of oxyhemoglobin or bilirubin, but ferritin levels were considerably raised (530 ng/mL, upper reference value 12 ng/mL), consistent with a previous bleed. Retrospectively it emerged that the patient had suffered from a thunderclap headache some months earlier. Subsequently he developed morning headaches and nausea. This suggests that he may have suffered from a subarachnoid haemorrhage complicated by secondary hydrocephalus. This would explain why in this case the relatively mild rehydration-related hyponatremia may have caused brain swelling sufficient for herniation. CONCLUSION: Given the frequency of hyponatraemia in marathon runners (serum Na <135 mM in about 13%), and the non-specific symptoms, we discuss how a simple screening test such as funduscopy may help to identify those who require urgent neuroimaging. PMID- 18163910 TI - Late aortic lymphocele and residual ovary syndrome after gynecological surgery. AB - BACKGROUND: Gynecological surgery, as radical hysterectomy or pelvic and aortic lymphadenectomy, accounts for more than 50% of iatrogenic injuries. In premenopausal women, an hysterectomy with ovarian sparing and concomitant lateral ovarian transposition is frequently performed. However, the fate of the retained ovary is complicated by the residual ovarian syndrome (ROS) and one of the most common postoperative complications of the lymphadenectomy procedure is the lymphocele, with an average incidence of 22-48.5%. The differential diagnosis of a postoperative fluid collection includes, in addition to a lymphocele, urinoma, hematoma, seroma or abscess and the computed tomography (CT) findings alone is not enough. CASE PRESENTATION: We describe a patient, affected by ROS concomitant with a asymptomatic lymphocele, initially confused with an aortic lymph nodes relapse, after abdominal radical hysterectomy. The patient was subjected to a surgical approach, included a diagnostic open laparoscopy and laparotomy with sovraombelico-pubic incision, wide opening of the pelvic peritoneum and retroperitoneum. Examination of the mass revealed, macroscopically, a ovary with multiloculated cystic masses filled with clear or yellow serous fluid and the layers were composed by flat or cuboidal mesothelial cells. CONCLUSION: The tribute of this case illustrates the atypical appearance with uncertain aetiology after complex imaging. Gynecologist and radiologist should acquaint with the appearance of fluid collection (urinoma, lymphocele, seroma, hematoma, abscess) in gynecologic oncology follow-up to properly differentiated from tumor recurrence. PMID- 18163911 TI - Neighborhood consistency in mental arithmetic: Behavioral and ERP evidence. AB - BACKGROUND: Recent cognitive and computational models (e.g. the Interacting Neighbors Model) state that in simple multiplication decade and unit digits of the candidate answers (including the correct result) are represented separately. Thus, these models challenge holistic views of number representation as well as traditional accounts of the classical problem size effect in simple arithmetic (i.e. the finding that large problems are answered slower and less accurate than small problems). Empirical data supporting this view are still scarce. METHODS: Data of 24 participants who performed a multiplication verification task with Arabic digits (e.g. 8 x 4 = 36 - true or false?) are reported. Behavioral (i.e. RT and errors) and EEG (i.e. ERP) measures were recorded in parallel. RESULTS: We provide evidence for neighborhood-consistency effects in the verification of simple multiplication problems (e.g. 8 x 4). Behaviorally, we find that decade consistent lures, which share their decade digit with the correct result (e.g. 36), are harder to reject than matched inconsistent lures, which differ in both digits from the correct result (e.g. 28). This neighborhood consistency effect in product verification is similar to recent observations in the production of multiplication results. With respect to event-related potentials we find significant differences for consistent compared to inconsistent lures in the N400 (increased negativity) and Late Positive Component (reduced positivity). In this respect consistency effects in our paradigm resemble lexico-semantic effects earlier found in simple arithmetic and in orthographic input processing. CONCLUSION: Our data suggest that neighborhood consistency effects in simple multiplication stem at least partly from central (lexico-semantic') stages of processing. These results are compatible with current models on the representation of simple multiplication facts - in particular with the Interacting Neighbors Model - and with the notion of decomposed representations of two-digit numbers in general. PMID- 18163912 TI - Light chain deposition disease presenting as paroxysmal atrial fibrillation: a case report. AB - INTRODUCTION: Light chain deposition disease (LCDD) can involve the heart and cause severe heart failure. Cardiac involvement is usually described in the advanced stages of the disease. We report the case of a woman in whom restrictive cardiomyopathy due to LCDD presented with paroxysmal atrial fibrillation. CASE PRESENTATION: A 55-year-old woman was admitted to our emergency department because of palpitations. In a recent blood test, serum creatinine was 1.4 mg/dl. She was found to have high blood pressure, left ventricular hypertrophy and paroxysmal atrial fibrillation. An ACE-inhibitor was prescribed but her renal function rapidly worsened and she was admitted to our nephrology unit. On admission serum creatinine was 9.4 mg/dl, potassium 6.8 mmol/l, haemoglobin 7.7 g/dl, N-terminal pro-brain natriuretic peptide 29894 pg/ml. A central venous catheter was inserted and haemodialysis was started. She underwent a renal biopsy which showed kappa LCDD. Bone marrow aspiration and bone biopsy demonstrated kappa light chain multiple myeloma. Echocardiographic findings were consistent with restrictive cardiomyopathy. Thalidomide and dexamethasone were prescribed, and a peritoneal catheter was inserted. Peritoneal dialysis has now been performed for 15 months without complications. DISCUSSION: Despite the predominant tubular deposition of kappa light chain, in our patient the first clinical manifestation of LCDD was cardiac disease manifesting as atrial fibrillation and the correct diagnosis was delayed. The clinical management initially addressed the cardiovascular symptoms without paying sufficient attention to the pre-existing slight increase in our patient's serum creatinine. However cardiac involvement is a quite uncommon presentation of LCDD, and this unusual case suggests that the onset of acute arrhythmias associated with restrictive cardiomyopathy and impaired renal function might be related to LCDD. PMID- 18163913 TI - Serum diagnosis of diffuse large B-cell lymphomas and further identification of response to therapy using SELDI-TOF-MS and tree analysis patterning. AB - BACKGROUND: Currently, there are no satisfactory biomarkers available to screen for diffuse large B cell lymphoma (DLBCL) or to identify patients who do not benefit from standard anti-cancer therapies. In this study, we used serum proteomic mass spectra to identify potential serum biomarkers and biomarker patterns for detecting DLBCL and patient responses to therapy. METHODS: The proteomic spectra of crude sera from 132 patients with DLBCL and 75 controls were performed by SELDI-TOF-MS and analyzed by Biomarker Patterns Software. RESULTS: Nine peaks were considered as potential DLBCL discriminatory biomarkers. Four peaks were considered as biomarkers for predicting the patient response to standard therapy. The proteomic patterns achieved a sensitivity of 94% and a specificity of 94% for detecting DLBCL samples in the test set of 85 samples, and achieved a sensitivity of 94% and a specificity of 92% for detecting poor prognosis patients in the test set of 66 samples. CONCLUSION: These proteomic patterns and potential biomarkers are hoped to be useful in clinical applications for detecting DLBCL patients and predicting the response to therapy. PMID- 18163914 TI - Regional body composition in college-aged Caucasians from anthropometric measures. AB - BACKGROUND: Quantitating fat and lean tissue in isolated body regions may be helpful or required in obesity and health-outcomes research. However, current methods of regional body composition measurement require specialized, expensive equipment such as that used in computed tomography or dual energy x-ray absorptiometry (DEXA). Simple body size or circumference measurement relationships to body composition have been developed but are limited to whole body applications. We investigated relationships between body size measurements and regional body composition. METHODS: Using DEXA technology we determined the fat and lean tissue composition for six regions of the body in predominantly Caucasian, college-aged men (n = 32) and women (n = 67). Circumference measurements as well as body weight and height were taken for each individual. Equations relating body measurements to a respective regional fat and lean mass were developed using multiple regression analysis. RESULTS: Multiple regression R2 values ranged from 0.4451 to 0.8953 and 0.1697 to 0.7039 for regional fat and lean mass relationships to body measurements, respectively. CONCLUSION: The equations developed in this study offer a simple way of estimating regional body composition in a college-aged adult population. The parameters used in the equations are common body measurements that can be obtained with the use of a measuring tape and weight scale. PMID- 18163915 TI - The effects of atorvastatin therapy on endothelial function in patients with coronary artery disease. AB - BACKGROUND: Statins improve the endothelial function in patients with coronary artery disease (CAD). However, they contribute to the substantial decrease in coronary heart disease by reducing plasma cholesterol levels. They also, reduce oxidative stress, stabilize the atherosclerotic plaque and inhibit inflammatory response. These functions of statins have been briefly described as pleiotropic effects. The aim of our study was to evaluate the effect of atorvastatin therapy on endothelial functions in patients with CAD. METHODS: Fourty-nine patients (40 men, 9 women, mean age 59 +/- 11 years) with diagnosed CAD were selected as the study group. The patients were given 10 mg/day atorvastatin for 12 weeks. If the target cholesterol levels has not been achieved 6 weeks after the treatment, then the daily atorvastatin dosage has been increased. The endothelial function was evaluated by flow mediated dilatation (FMD) of the brachial artery. RESULTS: It has been figured out that 12 weeks later, atorvastatin caused a statistically significant decrease in the plasma levels of LDL-cholesterol and total cholesterol (p < 0,0001). Meanwhile, it was determined that the FMD got statistically significant improved 12 weeks after the atorvastatin therapy (8,1% 4,2%, p < 0,001). However there was no statistically significant change in non endothelium dependent dilatation (NID). CONCLUSION: Endothelium derived vasodilatation (EBD), which was non-invasively detected via brachial artery ultrasonography, had statistically significant improvment within 12 weeks of atorvastatin therapy whereas non-endothelium dependent dilatation (NID) had no change. PMID- 18163916 TI - Nanobiotechnology today: focus on nanoparticles. AB - In the recent years the nanobiotechnology field and the Journal of Nanobiotechnology readership have witnessed an increase in interest towards the nanoparticles and their biological effects and applications. These include bottom up and molecular self-assembly, biological effects of naked nanoparticles and nano-safety, drug encapsulation and nanotherapeutics, and novel nanoparticles for use in microscopy, imaging and diagnostics. This review highlights recent Journal of Nanobiotechnology publications in some of these areas http://www.jnanobiotechnology.com. PMID- 18163917 TI - Is there a body of evidence for the treatment of patients with Adolescent Idiopathic Scoliosis (AIS)? AB - Historically, the treatment options for AIS, the most common form of scoliosis are; exercises; in-patient rehabilitation; braces and surgery. While there is evidence in the form of prospective controlled studies that Scoliosis Intensive Rehabilitation (SIR) and braces can alter the natural history of the condition, there is no prospective controlled study comparing the natural history with surgical treatment.One aim of the Scoliosis Society (SOSORT) should be; to help develop a body of research regarding the outcomes of conservative and operative treatment as well, and to highlight the problems of treatment indications in patients with AIS and other spinal deformities. Another aim is to help to improve the safety of patients who have surgery. By producing evidence-based information that can be used to develop guidelines that could aid both professionals and patients in making decisions about surgical and conservative options.Although 'Scoliosis' is the official journal of the SOSORT and is the main forum for experts in the field of conservative management of patients with spinal deformities, there needs to be more wide spread attempt to develop a fuller body of evidence focussing on spine surgery as well. PMID- 18163918 TI - The Malawi National Tuberculosis Programme: an equity analysis. AB - BACKGROUND: Until 2005, the Malawi National Tuberculosis Control Programme had been implemented as a vertical programme. Working within the Sector Wide Approach (SWAp) provides a new environment and new opportunities for monitoring the equity performance of the programme. This paper synthesizes what is known on equity and TB in Malawi and highlights areas for further action and advocacy. METHODS: A synthesis of a wide range of published and unpublished reports and studies using a variety of methodological approaches was undertaken and complemented by additional analysis of routine data on access to TB services. The analysis and recommendations were developed, through consultation with key stakeholders in Malawi and a review of the international literature. RESULTS: The lack of a prevalence survey severely limits the epidemiological knowledge base on TB and vulnerability. TB cases have increased rapidly from 5,334 in 1985 to 28,000 in 2006. This increase has been attributed to HIV/AIDS; 77% of TB patients are HIV positive. The age/gender breakdown of TB notification cases mirrors the HIV epidemic with higher rates amongst younger women and older men. The WHO estimates that only 48% of TB cases are detected in Malawi. The complexity of TB diagnosis requires repeated visits, long queues, and delays in sending results. This reduces poor women and men's ability to access and adhere to services. The costs of seeking TB care are high for poor women and men - up to 240% of monthly income as compared to 126% of monthly income for the non-poor. The TB Control Programme has attempted to increase access to TB services for vulnerable groups through community outreach activities, decentralising DOT and linking with HIV services. CONCLUSION: The Programme of Work which is being delivered through the SWAp is a good opportunity to enhance equity and pro-poor health services. The major challenge is to increase case detection, especially amongst the poor, where we assume most 'missing cases' are to be found. In addition, the Programme needs a prevalence survey which will enable thorough equity monitoring and the development of responsive interventions to promote service access amongst 'missing' women, men, boys and girls. PMID- 18163919 TI - Effectiveness of low-dose doxycycline (LDD) on clinical symptoms of Sjogren's syndrome: a randomized, double-blind, placebo controlled cross-over study. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are proteolytic enzymes that may contribute to tissue destruction in Sjogren's syndrome (SS). Low-dose doxycycline (LDD) inhibits MMPs. We evaluated the efficacy of LDD for the subjective symptoms in primary SS patients. This was a randomized, double blind, placebo controlled cross-over study. 22 patients were randomly assigned to receive either 20 mg LDD or matching placebo twice a day for 10 weeks. The first medication period was followed by 10-week washout period, after which the patient received either LDD or placebo, depending on the first drug received, followed by the second washout period. Stimulated saliva flow rates and pH were measured before and after one and ten weeks of each medication and after washout periods. VAS scale was used to assess the effect of LDD and placebo on following six subjective symptoms: xerostomia; xerophtalmia; difficulty of swallowing; myalgia; arthralgia; and fatigue. The effect was evaluated for each medication and washout period separately. RESULTS: Overall, the effects of medications on subjective symptoms were minor. Wilcoxon test demonstrated increased fatigue with LDD during medication (p < 0.05). The differences may, however, reflect normal fluctuation of symptoms in SS patients. CONCLUSION: LDD may not be useful in reducing the primary SS symptoms. PMID- 18163922 TI - Re: Mechanical failure rate of da Vinci robotic system. PMID- 18163923 TI - Screening men for prostate cancer and colorectal cancer: is practice evidence based? AB - INTRODUCTION: Controversy persists about whether men should be screened for prostate cancer. On the other hand, the benefit of colorectal cancer screening has been proven for men starting at age 50. We aimed to examine the rate of exposure to previous screening tests for prostate cancer and colorectal cancer in a cohort of men living in Quebec. MATERIALS AND METHODS: As part of an event promoting early prostate cancer detection, 347 men aged 50 to 69 without an established diagnosis of prostate cancer agreed to reply to questions in a previously validated questionnaire. The self-administered questionnaire, which asked about previous screening tests for prostate cancer and colorectal cancer, was completed on-site. RESULTS: Among men aged 50 to 69, previous exposure to a digital rectal examination (DRE), a prostate-specific antigen (PSA) test, a fecal occult blood test (FOBT), and sigmoidoscopy were reported by 132 men (62.9%), 73 men (34.8%), 37 men (17.6%), and 39 men (18.6%) , respectively. Across all age strata (< 50, 50-69, > or = 70 years), PSA and DRE testing were highest in men aged 50 to 69 and were 2- to 3-fold higher than screening tests for colorectal cancer. CONCLUSIONS: In this cohort of asymptomatic Canadian men, overall and age stratified exposure to tests to detect colon cancer early is far from ideal. Conversely, far more men have been subjected to PSA testing and DRE. Patients should be informed of the benefits and risks of colorectal cancer screening and PSA testing. PMID- 18163924 TI - A new suture technique for anastomosis in radical retropubic prostatectomy and early removal of urethral catheter. AB - INTRODUCTION: A modified suture technique for urethro-vesical anastomosis during radical retropubic prostatectomy was developed and utilized in a cohort of patients. Comparative analysis of postoperative outcome was performed with a previous group of patients who had an anastomosis with the conventional technique. MATERIAL AND METHODS: A consecutive group of patients who underwent radical retropubic prostatectomy in our department with the diagnosis of localized prostate cancer was included in this retrospective study. Urethro vesical anastomosis was performed either with the new or conventional method (CM). Outcome data of these two different patient sets were compared. RESULTS: One-hundred and one consecutive patients (mean age of 61.9 years) who were operated by either one of the two anastomotic suture techniques composed our study group. The mean follow-up period was 18 months (min: 12-max: 24). Urethro vesical "U" (UVU) suture was performed in 51 patients, and CM in 50 patients. Foley catheter was removed at postoperative fourth day in 33 (64.7%) and 18 (36%) patients in UVU and CM groups, respectively (p < 0.01). Incontinence rate was significantly lower in the UVU group at postoperative first year (p < 0.0005). Anastomotic strictures were observed in only 1.9% of the cases in UVU group, compared to 4% in CM group. CONCLUSION: Outcome data from our patient group indicate that UVU suture may allow a high quality urethro-vesical anastomosis with a very favorable outcome in terms of early catheter removal, high continence and low stricture formation rates in patients undergoing radical retropubic prostatectomy. Further validation of these results requires a prospective randomized trial. PMID- 18163925 TI - Pediatric renal cell carcinoma as second malignancy: reports of two cases and a review of the literature. AB - Pediatric renal cell carcinoma (RCC) is relatively rare and appears to comprise a group of tumors distinct from RCCs typically seen in adults. Recently described tumors show an association with neuroblastoma or specific chromosomal translocations. Only rarely have other childhood cancers been associated with pediatric RCC. We present two cases of pediatric RCC following treatment of other childhood malignancies not previously described, supratentorial primitive neuroectodermal tumor and acute lymphoblastic leukemia, and review the literature on pediatric RCC. As the RCCs were discovered as incidental radiologic findings, we emphasize the importance of close follow-up (including imaging) at routine intervals in survivors of childhood malignancies, not only to monitor for recurrence or metastasis, but also for development of a second malignancy. PMID- 18163926 TI - Bladder contracture: review for intravesical bacillus Calmette-Guerin complication. AB - INTRODUCTION: We are reporting a case of bladder contracture post intravesical bacillus Calmette-Guerin (BCG) therapy; to our knowledge only two cases were reported. We present the clinical history/presentation investigation and the outcome of the treatment. Approximately 75%-85% of patients with bladder cancer present with disease confined to the mucosa (stage Ta-CIS) or submucosa (stage T1). The management of non-muscle invasive bladder cancer has become more complex with regard to initial investigation, treatment and follow-up. In high-grade tumors, BCG therapy has proven to be superior to intravesical chemotherapy. BCG therapy prevents, or at least delays, tumor progression. METHODS AND RESULTS: A case of high grade superficial bladder caner treated with intravesical BCG which has successfully cleared her bladder cancer nevertheless has lead to bladder contracture for which case she may need bladder reconstruction/augmentation surgery if she remained disease free added to her psychological and social effects on her life. CONCLUSION: Although BCG is considered a very effective treatment; consensus exists that not every patient with superficial bladder cancer should be treated with BCG due to its increased risk of toxicity. Ultimately, the choice of treatment will depend upon the patient's risk of recurrence and progression. Assuming that maintenance therapy is necessary for optimal efficacy, the issue of BCG toxicity becomes more relevant. Due to the more pronounced side effects of BCG compared to intravesical chemotherapy, reluctance still exists about BCG use. However, with increased experience in applying BCG, the side effects now appear to be less prominent and few. Serious side effects are encountered in less than 5% of patients and this case carries one of the rarest, yet drastic, side effects of intravesical BCG. PMID- 18163927 TI - Unusual presentation of seminal vesiculitis in an infertile man. AB - Seminal vesiculitis is generally not investigated in infertile men, since it is uncommon and often nonspecific in signs and symptoms. In this article, the author reports an unusual presentation of seminal vesiculitis, incidentally diagnosed in a man referred for semen analysis to investigate infertility. Analyses of physical properties of the ejaculate (coagulation, liquefaction, volume, viscosity and pH) and of biochemical markers of the prostate (total calcium and zinc) and seminal vesicle (fructose and inorganic phosphorus) detected a dysfunction of the seminal vesicles that was subsequently diagnosed as vesiculitis using transrectal ultrasound. After treatment with a single 500-mg dose of oral ciprofloxacin, the patient was referred again for semen analysis. The analysis showed considerable improvement of the seminal vesicle function. The clinical significance of these findings is discussed. PMID- 18163928 TI - Robot-assisted radical cystectomy versus open radical cystectomy: assessment of postoperative pain. AB - INTRODUCTION: To date, no study has compared postoperative pain and requirement for pain medications in open versus robot-assisted radical cystectomy. Patient reported pain and opiate use were reviewed retrospectively using prospectively collected data from postoperative day one to day of discharge. MATERIALS AND METHODS: Twenty consecutive robot-assisted radical cystectomy patients were compared to the prior 20 patients who underwent open radical cystectomy. Data was collected prospectively to determine opiate requirements and pain scores in each group. Daily opiate use was converted to morphine sulfate equivalents (MSE) to facilitate comparison. A Likert pain perception scale was used to assess perceived pain. Statistical models were used to test for differences in opiate usage and pain perception between groups of patients who underwent open versus robot-assisted surgery. RESULTS: Seven patients were excluded from the study (three from the open group, and four from the robotic group): five due to preoperative opiate usage, one due to missing pain data, and one whose procedure was aborted due to unresectable disease. All patients were similar with respect to age, body mass index and pathological parameters. Average MSE usage differed significantly between the two groups on all postoperative days (p < 0.007) whereas average pain scores were similar in the two groups. CONCLUSION: Patients who underwent robot-assisted radical cystectomy achieved similar pain control but required less opiates than those who underwent open radical cystectomy. PMID- 18163929 TI - Pheochromocytoma of the urinary bladder. AB - OBJECTIVE: Pheochromocytoma of the urinary bladder is rare. Herein, we report our experience with pheochromocytoma of the urinary bladder in three patients. MATERIALS AND METHODS: Records of three consecutive patients diagnosed with bladder pheochromocytoma were reviewed. Patients' age, sex, presentation, associated conditions, diagnostic and imaging modalities utilized, management and follow up were recorded. RESULTS: The three patients included one child and two adults. An 11-year-old female presented with hematuria and bladder mass. Transurethral biopsy was non-diagnostic and she underwent partial cystectomy with eventual diagnosis of pheochromocytoma. Of the two adults, one was a 35-year-old female with history of gestational tumor who was being followed with computed tomography (CT) scan. A bladder mass was incidentally discovered. Transurethral resection of bladder tumor revealed pheochromocytoma and she underwent partial cystectomy. In retrospect, she has had symptoms related to micturition. The third patient is a 32-year-old male, who presented with fainting on voiding which suggested pheochromocytoma. He was also managed with partial cystectomy. There were no perioperative complications in any of the three patients. CONCLUSIONS: Pheochromocytoma of the urinary bladder has unique characteristics. A high index of suspicion should be present in patients who present with suggestive symptoms associated with voiding. In this series, all patients were successfully managed with partial cystectomy. PMID- 18163930 TI - Obstructive uropathy secondary to inguinal bladder hernia: case report and review of the literature. AB - We report a case of a large inguino-scrotal bladder hernia presenting with obstructive lower urinary symptoms and causing massive unilateral uretero hydronephrosis with ipsilateral renal function deterioration. Careful anamnesis and computer tomography (CT) scan of the abdomen and pelvis were important for the diagnosis. Open surgery with mesh hernia repair was successful. The obstructive lower tract symptoms subsided following surgery while the deteriorated renal unit regained only part of its function. PMID- 18163931 TI - Primary signet-ring cell carcinoma of the prostate. AB - We report a rare case of primary signet-ring cell carcinoma of the prostate in an advanced stage. A 62-year-old man with serum level of prostate-specific antigen at 364.70 ng/ml was diagnosed as having cT4N1M1c prostatic signet-ring cell carcinoma of Gleason score 5 + 4 = 9. Immunohistochemical examination demonstrated cytoplasmic immunoreactivity to prostate-specific antigen in signet ring cancer cells. The intracytoplasmic vacuoles in the signet-ring cells showed mucin production with a positive staining with periodic acid-Schiff. Although the patient received hormonal therapy, the disease progressed and lead to death 15 months after the diagnosis. The clinical and immunohistochemical characteristics of this malignancy are also reviewed. PMID- 18163932 TI - Mixed gonadal dysgenesis and Denys-Drash syndrome: urologists should screen for nephrotic syndrome. AB - OBJECTIVE: We present a child with Denys-Drash syndrome recognized after surgery for mixed gonadal dysgenesis, and discuss screening procedures the urologist should consider in similar circumstances. CASE REPORT: A 1-year-old child with XY gonadal dysgenesis underwent genital reconstruction. The postoperative period was complicated by incisional drainage, which led to the recognition of a nephrotic syndrome. Molecular analysis of the WT-1 gene confirmed a mutation associated with the Denys-Drash syndrome. CONCLUSION: The Denys-Drash syndrome should be suspected in children with XY gonadal dysgenesis. The presence of urine protein should be sought in such children, and if present, consultation with genetic and nephrology specialists is warranted. PMID- 18163933 TI - Embolization with Tornado coils to control bleeding from an arterioureteral fistula. AB - Arterioureteral fistulae are rare, but potentially life-threatening causes of bleeding. We present a case of an 82 year-old woman with refractory, transfusion dependent bleeding from an arterial fistula to her right ureteral stump, following right radical nephrectomy for advanced renal cell carcinoma. Cystoscopy with retrograde ureteral stump embolization using Tornado (Cook Medical, Bloomington, Indiana, USA) coils plus a slurry of thrombin-soaked Gelfoam (Pfizer Inc., New York, New York, USA) was performed, which led to prompt resolution of the patient's hematuria requiring no further hospitalizations or transfusions. Retrograde insertion of coils and injection of thrombin-soaked Gelfoam can be a minimally invasive, safe, and durable alternative for controlling hemorrhage from an arterioureteral fistula to a ureteral stump. PMID- 18163934 TI - Laparoscopic pyeloplasty for ureteropelvic junction obstruction in a horseshoe kidney. AB - With a continued movement toward minimally invasive surgical interventions, the range of applications treated with laparoscopic surgery will continue to grow. Laparoscopy is a preferred method for various reasons, including decreased postoperative pain, shorter inpatient hospital stays, and decreased convalescence. Ureteropelvic junction obstruction (UPJO) has traditionally been treated by open pyeloplasty. In patients with horseshoe kidneys, the blood supply is aberrant, which adds complexity to the procedure. We present the second reported case of a pediatric patient with a horseshoe kidney found to have UPJO who was successfully treated with transperitoneal laparoscopic pyeloplasty. PMID- 18163936 TI - Open clinical uro-oncology trials in Canada. PMID- 18163938 TI - Salvage options for biochemical recurrence after primary therapy for prostate cancer. AB - Despite excellent success rates with radical prostatectomy and radiotherapy for the treatment of prostate cancer, a significant number of patients will experience a rise in their serum prostate specific antigen (PSA) level. A variety of salvage options in this scenario have been investigated and the choice to pursue surveillance, single therapy or combination therapy depends on clinical assessment of risk and location of tumor recurrence. After radical prostatectomy, for example, patients with low risk local disease may not require secondary therapy or may benefit from salvage radiotherapy. Those with higher risk disease, based on PSA kinetics and tumor pathology may require systemic androgen deprivation therapy (ADT) with or without radiotherapy. Local recurrence after radiotherapy has the options of cryotherapy, brachytherapy or salvage surgery. ADT can also be applied in these patients at high risk of disease progression and cancer-specific mortality. Risk assessment in these settings is paramount as all secondary therapy options for prostate cancer have potential side effects that may significantly affect quality of life. We review the literature and discuss the current methods of risk assessment and the treatment options in prostate cancer once primary therapy fails. PMID- 18163939 TI - Managing prostate cancer: the role of hormone therapy. AB - Androgen deprivation therapy has been the mainstay of treatment for men with metastatic prostate cancer and now plays a more active role in the management of less advanced cancers as neoadjuvant and adjuvant treatment. Investigative uses include primary therapy for patients unsuitable for definitive therapy and as a complement to ablative procedures, brachytherapy, and chemotherapy. Intermittent androgen deprivation therapy is being considered as an alternative to continuous therapy and further evaluated as triple androgen blockade in conjunction with finasteride. Many accepted and potential management schemes incorporating hormonal therapy are increasingly employed despite indeterminate indications for use. Here, we review currently available data on the efficacy of hormonal therapy with regard to complete androgen ablation, primary, neoadjuvant, and adjuvant therapy. Additionally, we examine the usefulness of delayed versus immediate administration, intermittent androgen deprivation, and other prospective applications for hormonal therapy. PMID- 18163940 TI - Radiotherapy for localized prostate cancer. AB - Over 200,000 cases of prostate cancer will be diagnosed in the United States in 2007. Management of this common malignancy is controversial with essentially equal long-term survival and local control with either surgery or radiation therapy stage for stage in the setting of localized disease. Factors that can affect treatment recommendations include stage and grade of disease, the pre treatment PSA, physician bias and patient choice. This paper examines several of the radiotherapeutic options for the treatment of prostate cancer, and will also discuss evolving modalities that may offer additional treatment choices in the future. PMID- 18163941 TI - Salvage cryosurgical ablation of the prostate for local recurrence after radiation therapy: improved outcomes utilizing a capromab pendetide scan and biopsy algorithm. AB - PURPOSE: We assessed the efficacy, complications and technical advancements in salvage cryosurgical ablation of the prostate for recurrent prostate cancer after radiation therapy. METHODS: A total of 58 patients were evaluated for salvage cryosurgery using an algorithm of capromab pendetide scan and prostate biopsy from January 2003-July 2007. Forty-seven patients underwent salvage cryosurgery and biochemical recurrence free survival and complications were retrospectively reviewed. Mean follow-up was 24 months. RESULTS: Seventy percent of patients achieved a nadir PSA < 0.5 ng/ml. Overall, 51% of patients achieved a durable PSA response with a pre-salvage serum PSA < 10 predictive of success. There were no major complications and minor complications were few. CONCLUSION: Salvage cryotherapy in experienced hands utilizing third-generation technology provides for excellent biochemical control with minimal morbidity. PMID- 18163942 TI - Options and recent advances in permanent brachytherapy for prostate cancer. AB - Permanent interstitial brachytherapy with I-125 or Pd-103 seeds is a well established approach as single modality for low-risk prostate cancer patients and as part of a multi-modality program for intermediate- and high-risk patients. There are multiple approaches that have been developed to deliver high-quality implants, including pre-planned and real-time intra-operative techniques. In the hands of experienced users, either approach can provide consistently excellent outcomes. We believe that the combination of real-time intra-operative dosimetry and connected seeds may provide for improved consistency due to decreased seed migration. PMID- 18163943 TI - The emergence of imaging technology in advanced prostate cancer. AB - Rapid advances in imaging technology have whetted our collective appetites for practical clinical applications to assist the physician and patient in therapeutic decisions. Current limitations of imaging technology are being addressed by the convergence of technology in materials science, the computer industry, and biology which have led to improvements of diagnostic imaging. Refinements in image acquisition, fusion of images, and outcomes data now suggest use for image-guided therapy. Novel imaging agents and technologies appear to provide improved capabilities to detect malignant lymph nodes. Future applications of optical coherence tomography, electron paramagnetic resonance imaging, nanotechnology, and other forms of molecular imaging promise further refinements to enhance our diagnostic armamentarium. PMID- 18163944 TI - Current management of small renal masses. AB - The incidence of small renal masses (< 4 cm) is increasing due to the widespread use of imaging studies. Many of these incidental lesions may remain asymptomatic or in fact be benign, and recent insight into their natural course has contributed to modifications in management. With improvements in biopsy technique and minimally invasive technologies, appropriate diagnosis and treatment of these masses are further being evaluated. Other contemporary approaches, including surveillance, laparoscopic partial nephrectomy, enucleation, ablative procedures, and high-intensity focused ultrasound, are weighed against open nephron-sparing surgery, the current gold standard for treatment. Here, we review currently available data on the efficacy of these treatment options. Additionally, we examine the natural history of small renal masses, the role of diagnostic biopsy, and follow-up strategies for proper management. PMID- 18163945 TI - Molecular targeted therapies for renal cell carcinoma. AB - New advances in technology to directly target specific molecular events in the proliferation of cancer have led to promising results in renal cell carcinoma. Response rates in excess of 70% and complete responses in advanced (metastatic) renal cell carcinoma have caused a change in the paradigm of treatment from immunotherapy. Toxicities are significant, but manageable and pushing the toxicity to tolerability may increase the response rate. PMID- 18163946 TI - Medical management of benign prostatic hypertrophy. AB - Benign prostatic hyperplasia (BPH) is a common condition of the aging male. The bladder outlet obstruction caused by this condition occurs despite variations in prostate size. Symptoms of BPH include the irritative and obstructive voiding symptoms termed lower urinary tract symptoms (LUTS). While transurethral surgery has long been the gold standard for treatment of LUTS, medical treatment has emerged as the first line of treatment for those men who fail expectant or watchful waiting treatment. Medical options include: alpha blockers, 5alpha reductase inhibitors and newly identified PDE 5 inhibitors, drugs for erectile dysfunction that have a relieving effect on the symptoms of LUTS. Newer prostate selective alpha blockers have replaced older nonselective agents as first choice in treatment of most men, especially those with smaller prostates and in whom preservation of sexual function is important. While tamsulosin has the effect of an ejaculation, alfuzosin preserves ejaculatory function. 5alpha-reductase inhibitors may decrease ejaculate volume, libido and sexual function. While this effect is frequently a self limited, it can be a compliance issue for many men. PDE 5 inhibitors, while effective in relieving LUTS symptoms, have not shown effectiveness in reducing post void residual volumes or increasing urinary flow rates. PMID- 18163947 TI - Evaluation of the patient with incontinence. AB - The incontinent patient is evaluated in order to make a presumptive diagnosis so that treatment can be offered. The evaluation begins with a history and a physical examination. The history focuses on the description of the patient's incontinence. Assessing the patient's bother and determining their expectations of treatment may further guide how aggressive one needs to be both with the evaluation and the presentation of treatment options. The important parts of the physical exam are an examination of the abdomen and pelvis including a provocative stress test. A urinalysis and a post-void residual (PVR) should be performed in all incontinent patients. Incontinence questionnaires, voiding diaries, and pad weight tests can provide more objective data than the history alone. Upper tract imaging is indicated in the patient with a history of hematuria and in patients with suspected hydroureteronephrosis. Other imaging may be useful to further evaluate other suspected pelvic pathology. Urodynamics are performed to determine if the incontinence is due to bladder or urethral dysfunction or both, to assess if the patient has a storage or emptying problem and lastly in an effort to identify patients whose upper tracts are at risk due to high bladder storage pressures. Cystoscopy is indicated in the work up of some incontinent patients. The evaluation of the incontinent patient consists of a history, a physical, urinalysis and a post-void residual. Optional evaluative tests consist of a variety of urodynamic tests, imaging studies and cystoscopy. PMID- 18163948 TI - Androgen deficiency in the aging male: a guide to diagnosis and testosterone replacement therapy. AB - A steady decline in androgen levels occurs in males as they age. Evidence suggests that this decline may be at least partially responsible for a variety of physical and mental changes associated with the aging process. For instance, abnormally low levels of androgens can lead to profound changes in bone density, body composition, as well as sexual and cognitive function. Testosterone replacement has been shown to produce improvements in many of these areas. However, this practice is not without risks, both proven and theoretic. Also, the diagnosis of androgen deficiency and the decision to treat is not always straightforward. The purpose of this article is to familiarize the clinician with issues associated with androgen deficiency in the aging male. The clinical symptoms of androgen deficiency as well as the risks and benefits of androgen replacement will be discussed. This should help clinicians better identify those patients in whom testosterone replacement therapy should be considered. PMID- 18163949 TI - Peyronie's disease: update on medical management and surgical tips. AB - Peyronie's disease is a scarring phenomenon of the penis causing various deformities; initially pain with erection, and in most patients is associated with some element of erectile dysfunction. Studies of the natural history of the disease show that Peyronie's disease is a self-limited condition. In its stable and quiescent phase, patients have stable deformity, and in some cases that deformity then requires surgery. For the most part, pharmacologic therapy is confined to the immature or active phase of the disease. Pharmacotherapy is aimed at trying to adjust or interfere with the scarring process, so that the resultant scar causes as little disability as possible to the patient. Most pharmacotherapy is thus useful only in the active/immature phase of disease. In the mature or quiescent phase of the disease, therapy is aimed at undoing the effects of the scarring lesion. Those therapies for the most part can be considered "scar revisions". There is no best surgical therapy, and unfortunately because the disease process generally evolves with the background of erectile dysfunction, often times with surgery there is progression of the erectile dysfunction. All patients should be counseled with regards to the option of continued watchful waiting. Patients who are operated on must be counseled with regards to realistic outcomes. PMID- 18163950 TI - Do as we say and as we do: the interplay of descriptive and injunctive group norms in the attitude-behaviour relationship. AB - Past research on the social identity approach to attitude-behaviour relations has operationalized group norms as a mixture of both descriptive information (i.e. what most people do themselves) and injunctive information (i.e. what most people approve of). Two experiments (Study 1=185 participants; Study 2=238 participants) were conducted to tease apart the relative effects of descriptive and injunctive group norms. In both studies, university students' attitudes towards current campus issues were obtained, the descriptive and injunctive group norms were manipulated, and participants' post-manipulation attitudes, behavioural willingness, and behaviour were assessed. Study 2 also examined the role of norm source (i.e. in-group vs. out-group injunctive and descriptive norms). In both studies, the injunctive and descriptive in-group norms interacted significantly to influence attitudes, behavioural willingness, and behaviour. Study 2 revealed that out-group norms were largely ineffective. The research illustrates that in groups interactively influence decisions, not only by what they say, but also by what they do, and asserts the value of considering the interaction of descriptive and injunctive norms in accounts of normative influence. PMID- 18163951 TI - Origin of CD8+ effector and memory T cell subsets. AB - It is well accepted that CD8+ T cells play a pivotal role in providing protection against infection with intracellular pathogens and some tumors. In many cases protective immunity is maintained for long periods of time (immunological memory). Over the past years, it has become evident that in order to fulfill these multiple tasks, distinct subsets of effector and memory T cells have to be generated. Until today, however, little is known about the underlying mechanisms of subset differentiation and the timing of lineage fate decisions. In this context, it is of special importance to determine at which level of clonal expansion functional and phenotypical heterogeneity is achieved. Different models for T cell subset diversification have been proposed; these differ mainly in the time point during priming and clonal expansion (prior, during, or beyond the first cell division) when differentiation programs are induced. Recently developed single-cell adoptive transfer technology has allowed us to demonstrate that individual precursor cell still bears the full plasticity to develop into a plethora different T cell subsets. This observation targets the shaping of T cell subset differentiation towards factors that are still operative beyond the first cell division. These findings have important implications for vaccine development, as the modulation of differentiation patterns towards distinct subsets could become a powerful strategy to enhance the efficacy and quality of vaccines. PMID- 18163953 TI - Epithelial cell apoptosis and lung remodeling. AB - Lung epithelium is the primary site of lung damage in various lung diseases. Epithelial cell apoptosis has been considered to be initial event in various lung diseases. Apoptosis signaling is classically composed of two principle pathways. One is a direct pathway from death receptor ligation to caspase cascade activation and cell death. The other pathway triggered by stresses such as drugs, radiation, infectious agents and reactive oxygen species is mediated by mitochondria. Endoplasmic reticulum has also been shown to be the organelle to mediate apoptosis. Epithelial cell death is followed by remodeling processes, which consist of epithelial and fibroblast activation, cytokine production, activation of coagulation pathway, neoangiogenesis, re-epithelialization and fibrosis. Epithelial and mesenchymal interaction plays important roles in these processes. Further understanding of apoptosis signaling and its regulation by novel strategies may lead to effective treatments against various lung diseases. We review the recent advances in the understanding of apoptosis signaling and discuss the involvement of apoptosis in lung remodeling. PMID- 18163952 TI - The interferon signaling network and transcription factor C/EBP-beta. AB - Cytokines like interferons (IFNs) play a central role in regulating innate and specific immunities against the pathogens and neoplastic cells. A number of signaling pathways are induced in response to IFN in various cells. One classic mechanism employed by IFNs is the JAK-STAT signaling pathway for inducing cellular responses. Here we describe the non-STAT pathways that participate in IFN-induced responses. In particular, we will focus on the role played by transcription factor C/EBP-beta in mediating these responses. PMID- 18163954 TI - Phenotypic and functional analysis of LCMV gp33-41-specific CD8 T cells elicited by multiple peptide immunization in mice revealed the up-regulation of PD-1 expression on antigen-specific CD8 T cells. AB - The phenotype and function of antigen-specific CD8 T cells are closely associated with the efficacy of a therapeutic vaccination. Here we showed that multiple immunizations with LCMV gp33-41 peptide (KAV) in Freund's adjuvant could induce KAV-specific CD8 T cells with low expression of CD127 and CD62L molecules. The inhibitory receptor PD-1 was also expressed on a substantial part of KAV-specific CD8 T cells, and its expression level on KAV-specific CD8 T cells in spleen and lymph nodes was much higher when compared to those in peripheral blood. Furthermore, KAV-specific CD8 T cells could specifically kill KAV-pulsed target cells in vivo but the efficiency was low. These data suggest that prime-boost vaccination schedule with peptide in Freund's adjuvant can elicit antigen specific CD8 T cells of effector-like phenotype with partial functional exhaustion, which may only provide short-term protection against the pathogen. PMID- 18163955 TI - Protective effects of overexpression TCR Vbeta5.2-HSP70 and TCR Vbeta8.2-HSP70 against collagen-induced arthritis in rats. AB - Collagen-induced arthritis (CIA) is an animal model, which closely resembles human rheumatoid arthritis (RA) in pathogenesis and pathology. Evidence suggests that the inhibition of T lymphocytes or their functions can alleviate the progression of arthritis. So the administration of arthritogenic T cell receptor (TCR) variable region peptide or DNA vaccines encoding pathogenic TCR Vbeta variable region may provide useful information for designing specific immunotherapies against autoimmune diseases. Heat shock proteins (HSPs) have the function of raising antigenic immunogenicity and HSP70 has a protective effect against arthritis. We previously demonstrated the presence of pathogenic predominant T cell receptor Vbeta5.2 and Vbeta8.2 clonotypes in the joints of CIA rats. In this study, we constructed the recombinant eukaryotic expression vectors pTARGET-TCR Vbeta5.2/8.2-HSP70, and evaluated their protective effects on CIA rats. Protective effects were observed in CIA rats by injecting these recombinant DNA vaccines, which could alleviate arthritis index, decrease the levels of IFN gamma and anti-CII antibody in serum, and increase the levels of IL-4. Pathological changes were not as serious as those observed in control CIA rats. The rat injected with two combined vaccines showed better protective effects than CIA rats administered with individual vaccine. These results showed that recombinant DNA vaccines pTARGET-TCR Vbeta5.2-HSP70 and pTARGET-TCR Vbeta8.2 HSP70 could significantly alleviate the arthritic symptoms of CIA rats, and better protective effects could be achieved if these two vaccines were used in combination. PMID- 18163956 TI - Mitochondria play a role in the development of non-apoptotic programmed cell death of neutrophils induced by ONO-AE-248. AB - We previously reported that ONO-AE-248, a selective EP3 receptor agonist, has been shown to cause neutrophil death without the typical features of apoptosis and necrosis. However, the mechanism of the neutrophil death is unclear. By using Western blotting, flow cytometry (FACS) and confocal laser scanning microscopy (CLSM), we investigated the cellular signal transduction pathways of the neutrophil death. The research results showed that the neutrophil death induced by ONO-AE-248 did not show the morphologic changes of apoptosis and was not associated with the activity of caspase-3, caspase-8, and phosphorylation of p38 MAPK. However, impairment of mitochondria transmembrane potential has been found during the process of cell death. These findings suggested that ONO-AE-248 induced a non-apoptotic programmed cell death of neutrophils through partially mitochondria signaling transduction pathway. PMID- 18163957 TI - Correlation between expression of DcR3 on tumor cells and sensitivity to FasL. AB - To investigate the correlation between sensitivity to Fas ligand (FasL) and expression level of decoy receptor 3 (DcR3) on tumor cell surface, Fas/DcR3 mRNA expression was detected by RT-PCR. Anti-DcR3 mAb was used to detect expression level of DcR3 on surface of tumor cells by flow cytometry. Caspase-8, caspase-9, caspase-3, Bcl-2 expressions were analyzed by Western blot, respectively. Sensitivity to apoptosis induced by FasL was determined by Annexin V apoptosis kit. The expressions of DcR3 on the surface of tumor cells from high to low were approximately 35.3% in BGC823 cells, and 21.6% in MCF-7 cells, respectively. The apoptotic rates induced by FasL from low to high were 15.6% in BGC823 cells, and 58.2% in MCF-7 cells, respectively. There was a significant correlation between the expression levels of DcR3 with FasL-inducing apoptosis. PMID- 18163958 TI - Functional identification of the stable transfection C5aR cell line Molt-4. AB - The complement C5 anaphylatoxin receptor is a member of the seven transmembrane spanning G protein-coupled receptor superfamily that signals through Galphai and Galpha16. C5aR is mostly expressed on neutrophils, macrophages and endothelial cells. C5a and C5aR interaction plays an important role in numerous biological effects such as in vivo cytokine storm which results in inflammatory damage. Considering the limitation of collection of human peripheral blood neutrophils and their short half life, the stably transfected cell line for studying the biological effects of C5aR is needed. In this study, we transfected C5aR gene into Molt-4 cell line and examined the function of ectopic C5aR. Our results showed stable expression of the C5aR in Molt-4 cell line and their interaction with human C5a induced ERK1/2 phosphorylation, Ca++ influx. This stable transfected cell line may provide a useful tool for studying signal pathways related to C5a and C5aR interplay and antibody development specific for C5aR. PMID- 18163959 TI - Isolation and identification of cancer stem-like cells from murine melanoma cell lines. AB - In current study, cancer stem-like cells in the murine melanoma B16F10 cells were investigated. CD phenotypes of the B16F10 cells were analyzed by flow cytometry, and the specific CD phenotype cells from the B16F10 cells were isolated by MACS. Then we used colony formation assay in soft agar media, the cell growth assay in serum-free culture media as well as the tumorigenicity investigation of the specific CD phenotype cells in C57BL/6 mice, respectively, to identify cancer stem-like cells in the B16F10 cells. The results showed that the B16F10 cells could form spherical clones in serum-free culture media, and the rate of clonegenesis of CD133+, CD44+ and CD44+CD133+ cells was higher than that of CD133 , CD44- and CD44+CD133- cells in soft agar media, respectively. The tumorigenic potential of CD133+, CD44+, CD44+CD133+ cells and CD44+CD133+CD24+ cells was stronger than that of CD133-, CD44-, CD44+CD133- cells and CD44+CD133+CD24- cells in mice, respectively. In conclusion, the CD44+CD133+CD24+ cells have some biological properties of cancer stem-like cells or are highly similar to the characteristics of cancer stem cells (CSC). These results provide an important method for identifying cancer stem-like cells in B16F10 cells and for further cancer target therapy. PMID- 18163960 TI - Construction and identification of a human liver specific microRNA eukaryotic expression vector. AB - MiR-122 is one of the non-coding RNAs which showed its effects on the lipo metablism, virus infection and HCC forming through regulation of liver gene expression. Its eukaryotic expression vector was constructed by using pSuper which was widely applied in the siRNA expression. The precursor of human miR-122 gene was amplified by polymerase chain reaction (PCR) from the human genomic DNA. The positive clones were screened by PCR and restriction enzyme digestion. The new expression vector of miR-122 was named pHsa-m122. PHsa-m122 and its controls were transfected to HepG2 cells. The miR-122 expression activity was evaluated by GFP122i sensor reporter plasmid through fluorescence detection and Western blot. It was shown that the fluorescence intensity of GFP122si and pHsa-m122 co transfection group was weaker than that of the controls, so the functional activity of expressed miR-122 was detected. When HepG2 cells were co-transfected with HBV1.3 and pHsa-m122 plasmids, the results showed miR-122 may down-regulate the gene expression of HBV. The human liver specific microRNA eukaryotic expression vector of miR-122 was constructed successfully, which may facilitate further study of its function in the development of liver virus infection diseases and HCC. PMID- 18163964 TI - An overlooked source of physician-scientists. AB - A shortage of physician-scientists in the United States is an ongoing problem. Various recommendations have been made to address this issue; however, none of them have ameliorated the situation. Foreign medical school graduates with postdoctoral training in the United States are an overlooked and untapped resource for combating the dearth of physician-scientists. Evaluation of the scientific staff at the University of Texas Southwestern Medical Center revealed that 11% of all postdoctoral fellows were international medical graduates. Interestingly, a survey taken by these individuals revealed a lack of institutional and/or mentor support for career development and preparation for becoming physician-scientists. Foreign postdoctoral fellows with medical degrees are not even eligible for physician-scientist grants and awards since they are not US citizens. Although physicians educated in the United States usually matriculate from medical school with high educational debt that prevents most of them from entering into scientific careers, doctors trained outside the United States generally have minimal, if any, debt. Furthermore, many of them have a keen interest in remaining in the United States once they complete their postdoctoral training. Thus, foreign-trained medical professionals who have pursued scientific training in the United States can be one of the solutions for the current dearth of physician-scientists. PMID- 18163965 TI - Integrating international medical graduates into the physician-scientist pool: solution to the problem of decreasing physician-scientists in the United States. AB - There is an increasing shortage of physician-scientists in the United States, threatening future medical research. There are several factors that dissuade US medical graduates from entering into physician scientists careers. This article proposes that international medical graduates (IMGs) who have contributed to the physician work force in the under serviced rural health system could also be a great source to meet the increasing physician-scientist demand. Mechanisms to allow IMGs to enter into the physician-scientist career track in the United States are suggested. PMID- 18163966 TI - Contributions of international medical graduates to US biomedical research: The experience of US medical schools. AB - International medical graduates (IMGs) constitute an appreciable fraction of full time faculty at US medical schools and of principal investigators (PIs) on National Institutes of Health (NIH) research project grants. Information from the Faculty Roster of the Association of American Medical Colleges (AAMC) and from the NIH Consolidated Grant Applicant File (CGAF) was examined to assess IMGs' contribution to US medical school faculty and research. The study found that the number of IMG full-time faculty more than doubled over two decades-from 7,866 individuals in 1984 to 17,085 individuals in 2004, but that IMGs remained relatively stable as a share of physician full-time faculty (from 18.8 to 19.4%); the share is somewhat higher (20.0% of full-time physician faculty in 1984 to 23.7% in 2004) if faculty with degrees of unknown provenance are included. From 1984 to 2004, IMGs increased as a share of full-time physician faculty who are principal investigators on NIH research grants from 16.5% (540) to 21.3% (1,143). Including faculty with incomplete data on degree provenance, the corresponding IMG share increases to 18.0 and 24.0% respectively. Thus, IMGs comprise at least one-fifth and more likely one-fourth of all full-time faculty physicians who are PIs on NIH research project grants. The proportion of IMG full-time physician faculty who are in basic science departments is about twice that of their US/Canadian counterparts, as is the proportion of IMG physician PIs. Slightly fewer than half (48%) of full-time IMG faculty PIs pursue human subjects research (as coded by the NIH), while the majority of US/Canadian counterparts pursue human subjects research. PMID- 18163967 TI - Disparities in cardiovascular disease risk and treatment: demographic comparison. AB - BACKGROUND: Cardiovascular morbidity and mortality can be prevented by identification and modification of specific risk factors. Ethnic minorities have a higher incidence of cardiovascular risk factors. Additionally, ethnic minorities often reside in medically underserved areas and are subject to health care disparities. We hypothesized that ethnic minorities residing in medically underserved areas would experience greater health care disparities related to cardiovascular disease (CVD) prevention and treatment compared with those residing near an urban academic medical center. METHODS: We performed a retrospective chart review (N = 200) comparing an urban academic medical center with a rural community center. We evaluated the effects of ethnicity, demographics, and the absence or presence of CVD on cardiovascular risk factor prevalence, risk factor reduction, and CVD prevention and treatment. RESULTS: We found that Hispanics had more cardiovascular risk factors, including diabetes mellitus and low high-density lipoprotein cholesterol, compared with non-Hispanic whites. However, there were no ethnically based differences in risk factor prevalence by location. Additionally, ethnicity had no impact on the management of cardiovascular risk factors. However, patients with CVD residing in the rural location, regardless of ethnicity, received significantly fewer secondary prevention treatments compared with patients residing near the urban academic medical center, including aspirin or antiplatelets (p < .0001); beta-blockers or calcium channel blockers (p < or = .0001); diuretics, angiotensin-converting enzyme inhibitors, or angiotensin receptor blockers (p = .014); and statins (p < or = .0001). CONCLUSIONS: Hispanics have more CVD risk factors than non-Hispanic whites but receive equivalent prevention initiatives. Residing in a rural, medically underserved area, regardless of ethnicity, was associated with the largest CVD treatment and health care disparity. PMID- 18163968 TI - Toward early identification of acute lung injury in the emergency department. AB - BACKGROUND: There are no studies evaluating the epidemiology of pediatric acute lung injury (ALI) in the emergency department (ED), where early identification and interventions are most likely to be helpful. The purpose of this study was to describe the epidemiology of the ALI precursor acute hypoxemic respiratory failure (AHRF) in the ED. METHODS: We analyzed 11,664 pediatric patient records from 16 EDs. Records were selected if oxygen saturation (SpO(2)) was recorded during the visit. Virtual partial pressure of oxygen (pO(2)) was calculated from SpO(2), thus allowing calculation of ratios of pO(2) to fraction of inspired oxygen (FiO(2)) (PFRs). Patients with a PFR < 300 were classified as having AHRF. Univariate analyses and logistic regression were used to test the association of clinical factors with the presence of AHRF and intubation. RESULTS: AHRF criteria (ie, PFR < 300) were met in 121 (2.9%) of the 4,184 patients with an oxygenation measurement. The following variables were independently associated with ALI: higher Pediatric Risk of Admission II score (adjusted odds ratio [95% confidence interval (CI)] = 1.12 [1.08-1.16]; p < .001), higher heart rate (1.02 [1.01 1.03]; p = .009), a positive chest radiograph (2.35 [1.02-5.43]; p = .045), and lower temperature (0.49 [0.36-0.68]; p < .001).The final model had an R(2) = .20. CONCLUSION: We found nonintubated AHRF to be prevalent in the ED. The low R(2) for the regression model for AHRF underscores the lack of criteria for early identification of patients with respiratory compromise. Our findings represent an important first step toward establishing the true incidence of ALI in the pediatric ED. PMID- 18163969 TI - Evaluation of C-reactive protein in primary and secondary prevention. AB - Inflammation is pivotal in atherosclerosis, and C-reactive protein (CRP) is an inflammatory marker that predicts cardiovascular events. Several population-based studies have demonstrated that baseline CRP levels predict future cardiovascular events. CRP testing may thus have a major adjunctive role in the global assessment of cardiovascular risk. Recently, the National Cholesterol Education Program, through the Adult Treatment Panel III guidelines, identified CRP and another marker of inflammation, the fibrinogen, as "emerging risk factors," suggesting that their measurement may improve the estimations of absolute risk obtained using the traditional cardiovascular risk factors. In terms of clinical application, CRP seems to be a strong predictor of cardiovascular events such as low-density lipoprotein cholesterol and adds prognostic information to all levels of calculated Framingham Risk Score and to the cluster of risk factors of metabolic syndrome. Several pharmacologic agents proven to reduce vascular risk also reduce CRP levels. Among these, all statins lower CRP, with more potent statins having greater effects; on average, median CRP levels decline from 15 to 25% after 6 weeks of starting therapy. Although limitations concerning the inflammatory screening remain, available data suggest that CRP has the potential to play an important role in adding more information for the evaluation of global risk assessment in primary prevention of cardiovascular disease. PMID- 18163970 TI - Mutation analysis of WASF2 and GALE genes in one Chinese family with benign familial infantile convulsions with a novel locus. AB - Benign familial infantile convulsions (BFIC) are an autosomal dominant form of idiopathic epilepsy in which partial and generalized seizures commence in the first 3 months of life and spontaneously remit by age 1 year. As it is still unknown whether WASF2 and GALE genes are responsible for pure BFIC syndromes, in this article, mutations of the WASF2 and GALE genes in the proband of one Chinese family with pure BFICs were studied. Mutation analysis was carried out by polymerse chain reaction and leoxyribonueleic acid direct sequencing. One exonic variant (1047A-->G) and one intronic variant (IVS10+13A-->G), neither causing a modification of the physiologic messenger ribonucleic acid maturation, were found. The WASF2 and GALE genes do not appear to be involved in the ethiopathogenesis of pure BFIC syndromes, at least in the Chinese family we studied. PMID- 18163971 TI - Anti-Saccharomyces cerevisiae antibodies in acute myocardial infarction. AB - BACKGROUND: Elevated anti-Saccharomyces cerevisiae antibody (ASCA) immunoglobulin (IgG) and IgA levels were first described in the serum of Crohn disease patients and have increasingly been reported in other inflammatory diseases. The role of in situ and remote inflammation in atherosclerosis is a major area of interest. In this study, we compared ASCA IgG and IgA levels in acute myocardial infarction (AMI) and controls to investigate the possible role of ASCA in AMI. METHODS: Serum samples were obtained from 140 consecutive patients who presented to the emergency department with acute chest pain. AMI was diagnosed by electrocardiography and serial enzymes. Patients ruled out for acute coronary event were grouped as controls. ASCA IgA and IgG levels were determined using enzyme-linked immunosorbent assay. Groups were compared for statistically significant difference. RESULTS: ASCA IgG titers ranged between 0.1 and 31.0 RIU/mL (mean 4.92) in the AMI group and 0.1 and 6.0 (mean 0.84) in the controls. The groups were found to differ very significantly (p = .001). ASCA IgA titers ranged between 2.0 and 200.0 RIU/mL (mean 13.73) in the AMI group and 2.0 and 11.5 RIU/mL, (mean 4.25) in controls. The groups differed significantly (p = .32). AMI and controls were also analyzed for ASCA IgA and IgG positivity. Both groups differed significantly from controls (p = .013). CONCLUSION: Elevated ASCA IgA and IgG levels as well as ASCA positivity in the AMI might suggest use of ASCA as a marker for atherosclerotic plaque instability. It might also provide a link between inflammatory processes and increased cardiovascular risk. Further studies are needed on a Saccharomyces cerevisiae-based diet, related intestinal colonization, and associated inflammation, autoimmune disorders, and cardiovascular events. PMID- 18163974 TI - Cementoblast response to low- and high-intensity ultrasound. AB - OBJECTIVE: It has been shown that ultrasound stimulation accelerates repair of orthodontically induced root resorption. However, the mechanism of such adaptive change is still unclear. The purpose of this study was to evaluate pulsed ultrasound on the differentiation-potential and cementoblast-mediated osteoclastogenesis using a cementoblastic cell line. DESIGN: Cultured cementoblasts (murine cementoblastic cell line, OCCM-30) were subjected to ultrasound exposure (frequency = 1 MHz; pulsed 1:4; spatial average temporal average intensities=30 or 150 mW/cm(2)) or sham exposure for 15 min per day. Expression levels of alkaline phosphatase (ALP), type I collagen (COL-I), osteoprotegerin (OPG) and receptor activator of nuclear factor small ka, CyrillicB ligand (RANKL) mRNAs were analysed by a real-time PCR analysis. Furthermore, ALP activity, collagen synthesis, and protein levels of OPG and RANKL were examined after 6-day ultrasound exposure. RESULTS: Real-time PCR analysis indicated that, irrespective of the intensity, single ultrasound exposure increased the expression of transcripts for COL-I and ALP after 24h; the expression of OPG and RANKL also increased after 1 and 4h, respectively. Cultured cementoblasts receiving ultrasound stimulation for 6 days showed a significant (p<0.05 or 0.01) increase in cell number and collagen synthesis. ALP activity and OPG synthesis were also significantly (p<0.05) upregulated by ultrasound stimulation with 150 mW/cm(2). CONCLUSIONS: These results demonstrated that ultrasound stimulation especially with 150 mW/cm(2) might be a better candidate as a medical remedy to protect against root resorption and/or accelerate its repair. PMID- 18163975 TI - "Just looking at food makes me gain weight": experimental induction of thought shape fusion in eating-disordered and non-eating-disordered women. AB - Thought-shape fusion (TSF) is a cognitive distortion that can be induced experimentally and is associated with eating pathology. The current study was conducted in order to elucidate the effects of TSF induction in females with eating disorders (n=35), as well as in restrained eaters (n=38) and unrestrained eaters (n=39). It was hypothesized that TSF induction would result in anxiety, guilt, increased feelings of fatness, perceived weight gain and feelings of moral wrong-doing relative to an anxiety and control induction. It was further hypothesized that restrained eaters and individuals with eating disorders would exhibit a stronger reaction to a TSF induction than would unrestrained eaters. The results indicated that, as predicted, TSF can be induced in individuals both with and without eating disorders, and individuals with eating disorders reported the highest levels of "state" TSF after the induction relative to the non clinical controls. However, contrary to expectations, restrained eaters reported higher levels of perceived weight gain and moral wrong-doing after the anxiety induction (but not the TSF induction) relative to the control induction. Potential mechanisms for this pattern of results are discussed, and the clinical implications of research on TSF are also considered. PMID- 18163976 TI - Small-molecule inhibition of Aurora kinases triggers spindle checkpoint independent apoptosis in cancer cells. AB - Aurora kinases are key regulators of mitotic progression and have also been implicated in tumorigenesis. Small molecules that inhibit Aurora kinases have shown impressive anticancer activity in preclinical studies and are currently under clinical evaluation. In this study, our data show that suppression of Aurora activity with a specific inhibitor prevents the proliferation of breast cancer cells. Molecular modeling studies indicate that the Aurora inhibitor suppresses Aurora activity by competitive displacement of ATP. Mechanistically, the Aurora inhibitor causes the accumulation of multinucleated cells, leading to profound apoptosis in the absence of caspase-3 activity. Further studies show that the sensitivity of cancer cells to the Aurora inhibitor is independent of the spindle checkpoint. In addition, the Aurora inhibitor acts synergistically with the vinca alkaloids but not with the taxanes in inhibiting cell proliferation and inducing apoptosis. These results suggest that Aurora inhibitors might be effective in spindle checkpoint-defective cancer cells and a combination of Aurora inhibitors with the vinca alkaloids is a promising approach for cancer chemotherapy. PMID- 18163977 TI - A Systematic single nucleotide polymorphism screen to fine-map alcohol dependence genes on chromosome 7 identifies association with a novel susceptibility gene ACN9. AB - BACKGROUND: Chromosome 7 has shown consistent evidence of linkage with a variety of phenotypes related to alcohol dependence in the Collaborative Study on the Genetics of Alcoholism (COGA) project. With a sample of 262 densely affected families, a peak logarithm of odds (LOD) score for alcohol dependence of 2.9 was observed at D7S1799. The LOD score in the region increased to 4.1 when a subset of the sample was genotyped with the Illumina Linkage III panel for the Genetic Analysis Workshop 14 (GAW14). To follow up on this linkage region, we systematically screened single nucleotide polymorphisms (SNPs) across a 2 LOD support interval surrounding the alcohol dependence peak. METHODS: The SNPs were selected from the HapMap Phase I CEPH data to tag linkage disequilibrium bins across the region. Across the 18-Mb region, genotyped by the Center for Inherited Disease Research (CIDR), 1340 SNPs were analyzed. Family-based association analyses were performed on a sample of 1172 individuals from 217 Caucasian families. RESULTS: Eight SNPs showed association with alcohol dependence at p < .01. Four of the eight most significant SNPs were located in or very near the ACN9 gene. We conducted additional genotyping across ACN9 and identified multiple variants with significant evidence of association with alcohol dependence. CONCLUSIONS: These analyses suggest that ACN9 is involved in the predisposition to alcohol dependence. Data from yeast suggest that ACN9 is involved in gluconeogenesis and the assimilation of ethanol or acetate into carbohydrate. PMID- 18163979 TI - Effects of a polyacetylene from Panax ginseng on Na+ currents in rat dorsal root ganglion neurons. AB - The root of ginseng (Panax ginseng) has been used as a traditional medicine in the far east countries since ancient times. Ginseng extracts produce analgesia among other various biologically beneficial effects. A polyacetylenic compound, (9R,10S)-epoxyheptadecan-4,6-diyn-3-one (EHD), has been isolated from ginseng extract, whose biological activity is largely unknown. Voltage-gated Na(+) channels in primary sensory neurons play important roles in pain perception. We investigated the effects of EHD on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) Na(+) currents in acutely dissociated rat dorsal root ganglion neurons. EHD inhibited both Na(+) currents in a concentration dependent manner with an equal potency (K(d) values were both 14.3 microM). The activation voltage was not affected by EHD in either type of Na(+) current. However, EHD accelerated the inactivation of both Na(+) currents and produced a hyperpolarizing shift of the steady-state inactivation curve. In addition EHD suppressed the maximal Na(+) current at negative holding potentials at which the channels are relieved from inactivation. Thus EHD appears to bind both resting and inactivated channels. The recovery from inactivation of both Na(+) currents was also slowed by EHD. EHD inhibition of TTX-S Na(+) current but not TTX-R Na(+) current was frequency-dependent. This is the first report that a polyacetylene from ginseng inhibits Na(+) currents in primary sensory neurons. EHD by inhibiting Na(+) currents may contribute to the ginseng analgesia. PMID- 18163978 TI - The CHRNA5/A3/B4 gene cluster variability as an important determinant of early alcohol and tobacco initiation in young adults. AB - BACKGROUND: One potential site of convergence of the nicotine and alcohol actions is the family of the neuronal nicotinic acetylcholine receptors. Our study examines the genetic association between variations in the genomic region containing the CHRNA5, A3, and B4 gene cluster (A5A3B4) and several phenotypes of alcohol and tobacco use in an ethnically diverse young adult sample. Significant results were then replicated in a separate adult population-representative sample. METHODS: In a selected sample, nine single nucleotide polymorphisms (SNPs) were tested for association with various nicotine and alcohol phenotypes, including age of initiation and measures of frequency, quantity, and subjective responses to the substances. Analysis was conducted with the statistical genetics program WHAP in the full sample (1075 subjects) including ethnicities as covariates and within each ethnic group sub-sample. Replication of the significant results in a separate population-based sample was carried out with the PBAT statistical genetics program. RESULTS: Two linked SNPs (rs8023462 and rs1948) located in a conserved region of the A5A3B4 gene cluster significantly predicted early age of initiation for tobacco with a hazard ratio (HR) of 1.35 (95% confidence interval [CI]1.08-1.70) for the CC genotype of rs8023462 and a HR of 1.29 (95% CI 1.01-1.63) for the TT genotype of rs1948 [corrected]. These findings were then replicated in a separate population-representative sample, showing rs1948 and rs8023462 to be associated with age of initiation for both tobacco and alcohol use (p < .01 and p < .001). CONCLUSIONS: Variations in A5A3B4 genes might influence behaviors that promote early age of experimentation with drugs. PMID- 18163980 TI - Genomic and proteomic analysis of the effects of cannabinoids on normal human astrocytes. AB - Delta-9-tetrahydrocannabinol (Delta(9)-THC), the main psychoactive component of marijuana, is known to dysregulate various immune responses. Cannabinoid (CB)-1 and -2 receptors are expressed mainly on cells of the central nervous system (CNS) and the immune system. The CNS is the primary target of cannabinoids and astrocytes are known to play a role in various immune responses. Thus we undertook this investigation to determine the global molecular effects of cannabinoids on normal human astrocytes (NHA) using genomic and proteomic analyses. NHA were treated with Delta(9)-THC and assayed using gene microarrays and two-dimensional (2D) difference gel electrophoresis (DIGE) coupled with mass spectrometry (MS) to elucidate their genomic and proteomic profiles respectively. Our results show that the expression of more than 20 translated protein gene products from NHA was differentially dysregulated by treatment with Delta(9)-THC compared to untreated, control NHA. PMID- 18163981 TI - Repetitive stimulation of adenosine A1 receptors in vivo: changes in receptor numbers, G-proteins and A1 receptor agonist-induced hypothermia. AB - Adenosine is an important neuromodulator and neuroprotective molecule, which is produced in the brain as a function of neuronal activity, coupling energy expenditure to energy supply. Under conditions of increased need and reduced availability of energy, including hypoxia and prolonged wakefulness, there is an increase in adenosine turnover and adenosine receptor stimulation. The aim of the present study was to examine how repetitive adenosine receptor stimulation affects receptor function and adenosinergic signaling in the brain. Adult male Wistar rats received daily intraperitoneal injections of the adenosine A1 receptor agonist N(6)-cyclopentyladenosine (CPA; 0.25 mg/kg; once per day) and effects on adenosine signaling were established with receptor and G-protein autoradiography. Injections of CPA for 5 consecutive days caused a significant decrease in adenosine A1 receptor numbers in the hippocampus and somatosensory cortex. In contrast, while the amount of adenosine A1 receptor-activated G proteins was not affected in most regions, a significant increase was found in the somatosensory cortex. On the level of physiological output, CPA-induced hypothermia was significantly attenuated, suggesting a functional desensitization of the A1 receptor system. Taken together, the present findings suggest that repetitive stimulation of the A1 receptors can affect elements of the adenosinergic signaling cascade in the rat brain in a region-specific manner. PMID- 18163982 TI - Osteosarcoma cell line growth inhibition by zoledronate-stimulated effector cells. AB - Bisphosphonates have a profound effect on bone resorption and are widely used in the treatment of osteoclast-mediated bone diseases. Zoledronic acid (ZA), a third generation biphosphonate, has a potent antitumor activity and expands gammadelta (gammadelta) T cells endowed of major histocompatibility complex-unrestricted lytic activity. Many solid tumors express tumor-specific antigens on their surface, representing targets for immune effector T cells. Nevertheless, the immune surveillance against clinically manifested tumors is relatively inefficient. Therefore, we investigated the hitherto unknown effects of ZA activated gammadelta T cells of normal donors on osteosarcoma cell lines. gammadelta T cells were stimulated with ZA and low doses of interleukin-2, and then analyzed for proliferation and generation of effector activity against osteosarcoma cell lines. Our results show the potent anti-tumor activity of ZA stimulated gammadelta T cells and the enhanced immunosensitivity of osteosarcoma cell lines to gammadelta T cells suggesting that osteosarcoma is another gammadelta T cell susceptible tumor type. PMID- 18163984 TI - Inhibition of porcine liver carboxylesterase by a new flavone glucoside isolated from Deverra scoparia. AB - An endemic North African Saharan plant from of the Apiaceae family, Deverra scoparia, used locally for medicinal preparations, showed a strong inhibitory effect on porcine liver carboxylesterase. The active compound from the aerial part of the plant was purified by semi-preparative HPLC and photodiode array detection, and structurally determined by (1)H, (13)C NMR and mass spectroscopy methods. This compound was identified as flavone-3,4',7-trihydroxy-3'-methoxy-7 glucoside and it was found to be a powerful competitive inhibitor of porcine liver carboxylesterase with a inhibition constant value of 16 microM. Based on the structural features of the inhibitor and the enzyme active site region, it seems that the flavonoside binds to the surface of the enzyme. The low K(i) value suggests some physiological significance of such inhibitory activity, especially concerning the bio-transformation of xenobiotics. PMID- 18163983 TI - Dithiocarbamates and viral IL-10 collaborate in the immortalization and evasion of immune response in EBV-infected human B lymphocytes. AB - Epstein-Barr virus (EBV) is implicated in the development of a number of human malignancies including several subtypes of non-Hodgkin lymphoma (NHL) [G. Pallesen, S.J. Hamilton-Dutoit, X. Zhou, The association of Epstein-Barr virus (EBV) with T cell lymphoproliferations and Hodgkin's disease: two new developments in the EBV Field, Adv. Cancer Res. 62 (1993) 179-239]. Lymphoproliferative disease and NHL occurring in severely immunosuppressed individuals almost always involve EBV and have been extensively studied and modeled in vitro. EBV has also been causally associated with some cases of NHL occurring in otherwise immunocompetent individuals. However, a direct role for EBV in the pathogenesis of neoplasms developing in the presence of an otherwise competent immune system has not been established. We investigated potential interactions between dithiocarbamates (DTC), an important class of thiono-sulfur compounds, and EBV leading to immortalization of human B lymphocytes and evasion of cell-mediated immune response in culture. Primary lymphocyte cultures employing wild-type and recombinant EBV mutants were used to assess the respective roles of DTC and viral genes in lymphocyte transformation and survival. Pretreatment of EBV-infected human B lymphocytes with DTC directly enhanced transformation in the absence of T cells (5 nM) and independently increased survival of transformed cells in the presence of competent autologous T cells (10 nM). Both DTC-induced transformation and immortalization of EBV infected B lymphocytes were dependent on the expression of viral IL-10. These results provide a biological basis for studying collaborations between chemical and virus that alter lymphocyte biology, and provide a rationale for further molecular epidemiology studies to better understand the potential influence of these interactions on the development of NHL and perhaps other viral-associated malignancies. PMID- 18163985 TI - The proportionator: unbiased stereological estimation using biased automatic image analysis and non-uniform probability proportional to size sampling. AB - The proportionator is a novel and radically different approach to sampling with microscopes based on the well-known statistical theory (probability proportional to size-PPS sampling). It uses automatic image analysis, with a large range of options, to assign to every field of view in the section a weight proportional to some characteristic of the structure under study. A typical and very simple example, examined here, is the amount of color characteristic for the structure, marked with a stain with known properties. The color may be specific or not. In the recorded list of weights in all fields, the desired number of fields is sampled automatically with probability proportional to the weight and presented to the expert observer. Using any known stereological probe and estimator, the correct count in these fields leads to a simple, unbiased estimate of the total amount of structure in the sections examined, which in turn leads to any of the known stereological estimates including size distributions and spatial distributions. The unbiasedness is not a function of the assumed relation between the weight and the structure, which is in practice always a biased relation from a stereological (integral geometric) point of view. The efficiency of the proportionator depends, however, directly on this relation to be positive. The sampling and estimation procedure is simulated in sections with characteristics and various kinds of noises in possibly realistic ranges. In all cases examined, the proportionator is 2-15-fold more efficient than the common systematic, uniformly random sampling. The simulations also indicate that the lack of a simple predictor of the coefficient of error (CE) due to field-to-field variation is a more severe problem for uniform sampling strategies than anticipated. Because of its entirely different sampling strategy, based on known but non uniform sampling probabilities, the proportionator for the first time allows the real CE at the section level to be automatically estimated (not just predicted), unbiased-for all estimators and at no extra cost to the user. PMID- 18163986 TI - Boveri's long experiment: sea urchin merogones and the establishment of the role of nuclear chromosomes in development. AB - Theodor Boveri's major intellectual contribution was his focus on the causality of nuclear chromosomal determinants for embryological development. His initial experimental attempt to demonstrate that the character of the developing embryo is determined by nuclear rather than cytoplasmic factors was launched in 1889. The experimental design was to fertilize enucleate sea urchin eggs with sperm of another species that produces a distinguishably different embryonic morphology. Boveri's "hybrid merogone" experiment provided what he initially thought was empirical evidence for the nuclear control of development. However, for subtle reasons, the data were not interpretable and the experiment was repeated and contested. At the end of his life, Boveri was finally able to explain the technical difficulties that had beset the original experiment. However, by 1902 Boveri had carried out his famous polyspermy experiments, which provided decisive evidence for the role of nuclear chromosomal determinants in embryogenesis. Here we present the history of the hybrid merogone experiment as an important case of conceptual reasoning paired with (often difficult) experimental approaches. We then trace the further history of the merogone and normal species hybrid approaches that this experiment had set in train, and review their results from the standpoint of current insights. The history of Boveri's hybrid merogone experiment suggests important lessons about the interplay between what we call "models", the specific intellectual statements we conceive about how biology works, and the sometimes difficult task of generating experimental proof for these concepts. PMID- 18163987 TI - Drosophila ELMO/CED-12 interacts with Myoblast city to direct myoblast fusion and ommatidial organization. AB - Members of the CDM (CED-5, Dock180, Myoblast city) superfamily of guanine nucleotide exchange factors function in diverse processes that include cell migration and myoblast fusion. Previous studies have shown that the SH3, DHR1 and DHR2 domains of Myoblast city (MBC) are essential for it to direct myoblast fusion in the Drosophila embryo, while the conserved DCrk-binding proline rich region is expendable. Herein, we describe the isolation of Drosophila ELMO/CED 12, an approximately 82 kDa protein with a pleckstrin homology (PH) and proline rich domain, by interaction with the MBC SH3 domain. Mass spectrometry confirms the presence of an MBC/ELMO complex within the embryonic musculature at the time of myoblast fusion and embryos maternally and/or zygotically mutant for elmo exhibit defects in myoblast fusion. Overexpression of MBC and ELMO in the embryonic mesoderm causes defects in myoblast fusion reminiscent of those seen with constitutively-activated Rac1, supporting the previous finding that both the absence of and an excess of Rac activity are deleterious to myoblast fusion. Overexpression of MBC and ELMO/CED-12 in the eye causes perturbations in ommatidial organization that are suppressed by mutations in Rac1 and Rac2, demonstrating genetically that MBC and ELMO/CED-12 cooperate to activate these small GTPases in Drosophila. PMID- 18163988 TI - The actin-related protein hArp8 accumulates on the mitotic chromosomes and functions in chromosome alignment. AB - The actin family consists of conventional actin and various actin-related proteins (Arps). Some of these Arps are localized in the nucleus, and a fraction of each of these nuclear Arps is functionally involved in chromatin remodeling and histone acetyltransferase complexes. On the other hand, in mitotic cells, the localization and function of the nuclear Arps are largely unknown. Human Arp8 (hArp8), an ortholog of yeast nuclear Arp8, was recently found to be associated with the hINO80-chromatin remodeling complex along with hArp5. Here we report that hArp8, but not hArp5, accumulates on mitotic chromosomes. This is the first example where a member of the actin family is found to be associated with mitotic chromosomes. Expression of truncated hArp8 proteins and depletion of endogenous hArp8 by RNA interference caused misalignment of mitotic chromosomes, suggesting that chromosome-associated hArp8 has a role in chromosome behavior. In contrast, depletion of hIno80 and hArp5 did not cause misalignment of chromosomes, suggesting that the role of hArp8 at mitotic chromosomes is independent of the activity of hINO80 complexes. These findings provide the first insight into a novel function of actin family members in mitosis. PMID- 18163989 TI - Retinoid processing in cone and Muller cell lines. AB - To determine whether cones and Muller cells in the rod dominated retina cooperate to regenerate the 11-cis retinal chromophore via the retinoid cycle, two cell lines from the rod dominated retinas of Murine were used for this study: 661W, a mouse cell line derived from cones, and rMC-1, a rat Muller cell line. Retinoid cycle enzymes were analyzed by RT-PCR, and their catalytic activity was detected by incubation with retinoids and analyzed by HPLC. We found that 661W cells are capable of reducing all-trans retinal to all-trans retinol due to the presence of multiple dehydrogenases and to generate minor amounts of retinyl-ester. The rMC-1 cells take up all-trans retinol and oxidize it to all-trans retinal or esterify it to retinyl-ester, but are incapable of isomerizing all-trans retinoids to 11 cis retinoids. This could be a reflection of lack of necessary activities in Muller cells in vivo, which suggests that Muller cells do not contribute to retinoid cycling by regenerating 11-cis retinoids. Alternatively, this could be due to the potential that rMC-1, as a transformed cell line, has stopped expressing the proteins needed for the regeneration of 11-cis retinoids. PMID- 18163990 TI - Trypanosoma cruzi: the role of PGE2 in immune response during the acute phase of experimental infection. AB - Chagas disease is characterized by cardiac lesions and a high level of PGE2. Our objective was to investigate the role of PGE2 in cardiac lesions. BALB/c mice were infected with Trypanosoma cruzi (1x10(3) trypomastigote forms) and were treated daily with PBS, meloxicam (0.5 mg/kg) or etoricoxib (0.6 mg/kg). The animals were sacrificed on the 21st day of infection and we collected the cardiac tissue and spleen cells for tissue culture. We observed that treatment with COX-2 inhibitors was able to decrease synthesis of PGE2 by spleen cells. This reduction was accompanied by reduction of the inflammatory infiltrate, parasite nets, cardiac fibrosis and fewer COX-2 positive cells in cardiac tissue obtained from these animals. In conclusion, treatment with COX-2 inhibitors, and consequent inhibition of PGE2 synthesis, was able to reduce the cardiac damage observed during the acute phase of experimental Chagas disease, thus demonstrating the involvement of this mediator in the cardiac lesion. PMID- 18163991 TI - Gene expression in women conceiving spontaneously over the age of 45 years. AB - OBJECTIVE: To examine gene expression profiles of women conceiving spontaneously after the age of 45. DESIGN: Expression profiling by complementary DNA microarray analysis. SETTING: University departments. PATIENT(S): Eight women 45 years or older (study group) who conceived spontaneously and were 6 months after their last delivery were compared with six women aged 45 years old (control group) who had their last delivery before the age of 30. INTERVENTION(S): Blood samples were collected from each woman for RNA isolation from peripheral blood mononuclear cells (PBMC). MAIN OUTCOME MEASURE(S): Expression profiles of PBMC isolated from each woman were determined by using Affymetrix DNA microarray analysis covering about 15,000 identified genes. RESULT(S): Microarray of global gene expression revealed 671 genes that showed statistically significant differential expression between the study and control groups: 383 genes were overexpressed and 288 were underexpressed. The most significant functional groups defining these genes were: apoptosis, ubiquitination, and energy production. As many as 60 genes also participated in ovarian physiology. CONCLUSION(S): These observations suggest that extended fertility is associated with a unique ability to enrich cellular processes, leading to delayed ovarian senescence. PMID- 18163992 TI - A new laparoscopic-transvaginal technique for rectosigmoid resection in patients with endometriosis. AB - OBJECTIVE: To present our experience with a new technique for laparoscopic rectosigmoid resection in patients with endometriosis. DESIGN: Prospective collaborative cohort study. SETTING: Gynecologic departments of two university hospitals. PATIENT(S): Thirty-three women with rectosigmoid endometriotic lesions requiring segmental bowel resection. INTERVENTION(S): Laparoscopic intracorporeal division of the distal bowel and exteriorization of the affected segment via a colpotomy incision to complete the resection. MAIN OUTCOME MEASURE(S): Intraoperative and postoperative complications, and relief from symptoms. RESULT(S): The only intraoperative complication was bleeding from the inferior mesenteric artery that required conversion to laparotomy to obtain hemostasis. No patient required a temporary colostomy. No anastomotic complications were identified. Postoperative complications included a symptomatic pelvic seroma that required operative drainage in 1 patient and urinary retention that required intermittent self-catheterization in 3 women. The median follow-up duration was 13 months (range, 3-27 mo). Twenty-seven women were symptom free at the time of last follow-up evaluation. No patient had recurrent cyclic rectal bleeding, rectal pain on defecation, or tenesmus. Postoperatively, 4 of 13 patients who tried to conceive were successful. CONCLUSION(S): Segmental colorectal resection with a combined laparoscopic-transvaginal approach, avoiding the extension of port-site incisions, represents a viable option for the treatment of bowel endometriosis. PMID- 18163993 TI - Obesity differentially affects serum levels of androstenedione and testosterone in polycystic ovary syndrome. AB - OBJECTIVE: To assess androstenedione (A) and T levels in obese and nonobese patients with polycystic ovary syndrome (PCOS) after GnRH and oral glucose tolerance tests (OGTT). DESIGN: Cross sectional study. SETTING: Clinical research center. PATIENT(S): Thirty patients with PCOS, of whom 15 were obese and 15 were nonobese, and 7 women without PCOS were included in the study. INTERVENTION(S): The GnRH test and OGTT were performed in all subjects. MAIN OUTCOME MEASURE(S): Basal and stimulated levels of LH, FSH, insulin, A, and total T were measured. Areas under the curve (AUCs) and AUC change after stimulation for these hormones were calculated. RESULT(S): The basal T levels were significantly higher in obese than in nonobese patients with PCOS. In contrast, the basal levels of A were similar in obese and nonobese patients with PCOS. The T(AUC) after GnRH was significantly greater in obese than in nonobese patients with PCOS but was not significantly different after OGTT. The A(AUC) after GnRH and OGTT was significantly greater in nonobese than in obese patients with PCOS. However, there were no significant differences in T(AUC) and A(AUC) changes after GnRH and OGTT. CONCLUSION(S): A different pattern in the levels of T and A with respect to obesity in PCOS was observed, suggesting a shift in ovarian enzymatic function. PMID- 18163994 TI - Influence of polymorphism of glutathione S-transferase T1 on Chinese infertile patients with varicocele. AB - To investigate the influence of glutathione S-transferase T1 (GSTT1) gene polymorphism on Chinese infertile patients with varicocele, 63 infertile patients with varicocele and 54 healthy fertile controls were recruited in this case control study. Our results show that oxidative damage may be the cause of infertility in patients with varicocele, and GSTT1 null genotype predisposes to over oxidative damage to spermatocytes of infertile patients with varicocele. PMID- 18163995 TI - Modeling the early endometriotic lesion: mesothelium-endometrial cell co-culture increases endometrial invasion and alters mesothelial and endometrial gene transcription. AB - OBJECTIVE: To determine the role of peritoneal mesothelial cells (PMCs) in the process of endometrial invasion into the peritoneum and to evaluate gene expression after endometrial-PMC co-culture. DESIGN: In vitro study. SETTING: University laboratory. PATIENT(S): Reproductive-age women without endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The rate of endometrial invasion through modeled peritoneum in the presence and absence of PMCs was evaluated. The influence of endometrial-PMC attachment on the expression of target genes, implicated in the pathogenesis of endometriosis, was examined by using reverse transcription polymerase chain reaction. RESULT(S): Endometrial stromal cell (ESC) invasion through invasion chambers coated with Matrigel (MTGL) and with growth factor-reduced Matrigel (GFR-MTGL) was increased 10-fold when a PMC monolayer was present. Endometrial epithelioid cell (EM42) invasion increased greater than threefold through the MTGL and GFR-MTGL-coated membranes when a PMC monolayer was present. Endometrial stromal cell, EM42, and PMC transcription of extracellular signal-related kinase, colony stimulating factor-1, c-fms, and c Met was increased after endometrial-PMC attachment. Similar changes were not seen when endometrial cells were exposed to PMC-conditioned media and when PMCs were exposed to endometrial cell conditioned media. CONCLUSION(S): Peritoneal mesothelial cells increased invasion of ESCs and EM42s through modeled peritoneum. Endometrial-PMC co-culture led to alterations in gene transcription by endometrial cells and PMCs. This study suggests that PMCs contribute to the process of endometrial invasion into the peritoneum. PMID- 18163996 TI - The destiny of myomas: should we treat small submucous myomas in women of reproductive age? AB - OBJECTIVE: To carry out a thorough analysis aimed at demonstrating that a "wait and-see" approach is no longer acceptable in women of reproductive age with small submucous myomas, even if they are asymptomatic. DESIGN: Review article. SETTING: University hospitals. PATIENT(S): Women of reproductive age with small (<1.5 cm) submucous myomas. INTERVENTION(S): "See-and-treat" hysteroscopy performed in an outpatient setting. MAIN OUTCOME MEASURES(S): a) The real endometrial surface and volume occupied by a submucous myoma; b) the high potential of a small myoma to grow during the reproductive age; c) its negative impact on reproduction through normal or assisted conception; d) the inability to perform a reliable and "safe" diagnosis, with respect to malignancy, without an eye-guided biopsy; and e) the effectiveness of "see-and-treat" hysteroscopy in removing small submucous myomas. RESULT(S): Small myomas, as hormone-dependent benign tumors, have a high potential to grow and either to become symptomatic or to cause complications during natural or assisted conception and pregnancy. Furthermore, not withstanding the risk of malignancy is rare, even the most experienced operator cannot replace the histological analysis to exclude malignancy or premalignant lesions. "See-and-treat" hysteroscopy has been demonstrated to be safe and effective in removing such small submucous myomas. CONCLUSION(S): A "wait-and see" approach is no longer acceptable in women of reproductive age with small submucous myomas, especially if the lesion could be easily and safely removed in an outpatient setting with minimal patient's discomfort. PMID- 18163997 TI - Curcumin ameliorates aflatoxin-induced toxicity in mice spermatozoa. AB - OBJECTIVE: To investigate the ameliorative effect of curcumin on aflatoxin induced toxicity in mice spermatozoa. DESIGN: Prospective controlled animal study. SETTING: University research laboratory. PATIENT(S): Young, inbred, Swiss strain male albino mice (Mus musculus), weighing approximately 37 to 40 g, were obtained from Cadila Health Care, Ahmedabad, India. INTERVENTION(S): Aflatoxin was orally administered in 25 (low dose) and 50 (high dose) microg/0.2 mL olive oil to each animal, each day (750 and 1500 microg/kg body weight), respectively, with and without curcumin for 45 days. On 46th day the animals were killed by cervical dislocation. The cauda epididymis were removed and weighed, then was teased in normal saline to obtain spermatozoa. MAIN OUTCOME MEASURE(S): Sperm count, viability, and motility were assessed immediately. Sperm semidried smears were prepared on slides and stained by Papanicolaou stain to study sperm morphologic features. RESULT(S): There was a dose-dependent spermicidal effect of aflatoxin. Sperm count, viability, and motility were statistically significantly reduced. Different morphologic abnormalities were encountered. Treatment with curcumin along with aflatoxin ameliorated aflatoxin-induced sperm count, immobilization, and viability, and improved the morphologic characteristics of the sperm. CONCLUSION(S): Curcumin has an ameliorative effect on sperm parameters and improves morphologic features of sperm in mice. PMID- 18163998 TI - Growth and development of children born after in vitro fertilization. AB - OBJECTIVE: To evaluate growth and development of children born after IVF treatment. DESIGN: Literature review. CONCLUSION(S): At present there is substantial evidence that children born after IVF are at increased risk for adverse perinatal outcome, congenital malformations, and rare epigenetic defects. It is still unclear whether observed health problems originate from the IVF procedure itself or the underlying subfertility problems of the parents. Current follow-up studies regarding postnatal growth and morbidity rates are scarce with conflicting results and other areas of long-term research in children born after IVF are still in its infancy. The importance of the worldwide continuing monitoring of children born after IVF to investigate potential long-term consequences including the development of cardiovascular diseases is therefore highlighted. PMID- 18163999 TI - Coasting vs. cryopreservation of all embryos for prevention of ovarian hyperstimulation syndrome in in vitro fertilization. AB - In the patient at risk for ovarian hyperstimulation syndrome, coasting will result in fewer eggs retrieved and embryos produced when compared with cryopreservation of all embryos. However, both strategies are associated with a similar incidence of ovarian hyperstimulation syndrome, with achievement of comparable cumulative pregnancy rates. PMID- 18164000 TI - First trimester screening for Down's syndrome after assisted reproductive technology: non-male factor infertility is associated with elevated free beta human chorionic gonadotropin levels at 10-14 weeks of gestation. AB - We retrospectively compared the first trimester Down's syndrome serum screening markers free beta-hCG (fbetahCG) and pregnancy-associated plasma protein-A (PAPP A) at 11-14 weeks of gestation in 4,088 women with naturally conceived pregnancies and in women pregnant after ICSI (n = 163), IVF (n = 59) and frozen thawed embryo transfer (n = 31), and we searched for a potential relationship between infertility cause and marker levels. We found lower serum PAPP-A levels in pregnancies after IVF and ICSI compared with spontaneously conceived pregnancies and non-male factor infertility was associated with elevated serum fbetahCG levels at 11-14 weeks of gestation. PMID- 18164001 TI - Low-dose oral contraceptive pill for dysmenorrhea associated with endometriosis: a placebo-controlled, double-blind, randomized trial. AB - OBJECTIVE: To evaluate the efficacy of a low-dose oral contraceptive pill (OCP) for patients with dysmenorrhea associated with endometriosis. DESIGN: A double blind, randomized, placebo-controlled trial. SETTINGS: Clinical trial sites in Japan. PATIENT(S): One hundred patients with dysmenorrhea associated with endometriosis. Most enrolled patients had radiologic evidence of endometriosis rather than surgical diagnosis. INTERVENTION(S): Patients were randomly assigned to receive either monophasic OCP (ethinylestradiol plus norethisterone) or placebo. Participants used their usual pain medications as needed during the trial. MAIN OUTCOME MEASURE(S): After four cyclic treatments, we used a zero- to three-point verbal rating scale and a visual analogue scale to measure the severity of disability because of dysmenorrhea in daily life, and the patients' use of analgesics. RESULT(S): Total dysmenorrhea scores assessed by the verbal rating scale were significantly decreased at the end of treatment in both groups. From the first cycle through the end of treatment, dysmenorrhea in the OCP group was significantly milder than in the placebo group. Nonmenstrual pelvic pain was present at baseline in 24.5% (12 of 49) of the OCP group and 34.0% (16 of 47) of the placebo group. The volume of endometrioma (larger than 3 cm in diameter) was significantly decreased in the OCP group, but not in the placebo group. No serious adverse events related to using OCPs occurred. CONCLUSION(S): The present study clearly demonstrated for the first time that OCPs could be used to effectively and safely treat pain associated with endometriosis. PMID- 18164003 TI - Fracture angulation-what about modern technology? PMID- 18164002 TI - Gonadectomy reveals sex differences in circadian rhythms and suprachiasmatic nucleus androgen receptors in mice. AB - In mammals, it is well established that circadian rhythms in physiology and behavior, including the rhythmic secretion of hormones, are regulated by a brain clock located in the suprachiasmatic nucleus (SCN) of the hypothalamus. While SCN regulation of gonadal hormone secretion has been amply studied, the mechanisms whereby steroid hormones affect circadian functions are less well known. This is surprising considering substantial evidence that sex hormones affect many aspects of circadian responses, and that there are significant sex differences in rhythmicity. Our previous finding that "core" and "shell" regions of the SCN differ in their expression of clock genes prompted us to examine the possibility that steroid receptors are localized to a specific compartment of the brain clock, with the discovery that the androgen receptor (AR) is concentrated in the SCN core in male mice. In the present study, we compare AR expression in female and male mice using Western blots and immunochemistry. Both of these methods indicate that ARs are more highly expressed in males than in females; gonadectomy eliminates and androgen treatment restores these sex differences. At the behavioral level, gonadectomy produces a dramatic loss of the evening activity onset bout in males, but has no such effect in females. Treatment with testosterone, or with the non-aromatizable androgen dihydrotestosterone, restores male locomotor activity and eliminates sex differences in the behavioral response. The results indicate that androgenic hormones regulate circadian responses, and suggest an SCN site of action. PMID- 18164004 TI - Ilizarov treatment for femoral mal-union or non-union associated with fatigue fracture of an intramedullary nail. PMID- 18164005 TI - Male external genitalia injury caused by 'elephant stampede'. PMID- 18164006 TI - Fixation of long bone segmental defects: a biomechanical study. AB - OBJECTIVES: Obtaining stable fixation in cases of long bone non-union with segmental bone defects can be challenging. Bone quality is often sub-optimal. Locking plates and structural allografts have both been used clinically in these cases. The objective of this study was to determine the biomechanical characteristics of three constructs that have been employed in this context. METHODS: A biomechanical study was performed using 3rd Generation Composite Femurs as specimens. A diaphyseal segmental defect was created and fixed with one of three constructs: (1) lateral locking plate (LP); (2) lateral non-locking plate and medial allograft strut (S); (3) lateral non-locking plate and intramedullary fibula allograft (F). The "allografts" were fashioned from 3rd generation composite bones. Axial, torsional and bending stiffness as well as load to failure were determined using a materials testing machine. RESULTS: Overall, construct S was the stiffest and construct LP was the least stiff. Construct F had intermediate characteristics. Axial load to failure for construct S (6108N) and for construct F (5344N) was significantly greater than for construct LP (2855N). CONCLUSION: When maximal stiffness is desired, a construct with a structural allograft should be chosen over a locking plate. However, biological and anatomic factors must also be taken into account when using these constructs clinically. PMID- 18164007 TI - Missed injury and the tertiary trauma survey. AB - Missed injury in the context of major trauma remains a persistent problem, both from a clinical and medico-legal point-of-view. Estimates of the incidence vary widely, dependent on the precise parameters of the studied population, the definition of missed injury and the extent of follow-up, but may be as high as 38%. The tertiary survey, in which formal repeated examination of the patient is undertaken after initial resuscitation and treatment have taken place, has been suggested as a way of identifying injuries not found at presentation. This paper appraises the concept of the tertiary survey, and also reviews the literature on missed injury in order to identify the risk factors, the types of injury and the reasons for error. PMID- 18164009 TI - We must never lose our capacity for wonder. PMID- 18164010 TI - Comparison of four regimens of misoprostol after methotrexate for early abortion. PMID- 18164011 TI - Short-term antibiotic treatment of pelvic actinomycosis. PMID- 18164012 TI - Uterine inversion associated with uterine sarcoma. PMID- 18164013 TI - High HDL cholesterol does not protect against coronary artery disease when associated with combined cholesteryl ester transfer protein and hepatic lipase gene variants. AB - Cholesteryl ester transfer protein (CETP) and hepatic lipase (HL) are two HDL modifying proteins that have both pro- and anti-atherogenic properties. We hypothesized that CETP and HL synergistically affect HDL cholesterol and atherosclerotic risk. To test our hypothesis, we analysed the genotype frequencies of CETP Taq1B (rs708272) and LIPC-514C/T (rs1800588) polymorphisms in male coronary artery disease patients (CAD; n=792) and non-symptomatic controls (n=539). Cases and controls had similar allele frequencies, but the occurrence of the combined genotypes differed (p=0.027). In CAD patients, 1.3% had the CETP B2B2/LIPC-TT genotype, with only 0.2% in controls (p=0.033). The presence of the CETP lowering B2 allele and the HL lowering LIPC-T allele synergistically increased HDL cholesterol from 0.87+/-0.19 mmol/L in the B1B1/CC (n=183) to 1.21+/-0.25 mmol/L in the B2B2/TT carriers (n=10). The B1B1/CC carriers had an increased CAD risk (OR 1.4; p=0.025). Despite their high HDL cholesterol, the B2B2/TT individuals also had an increased CAD risk (OR 3.7; p=0.033). In a 2-year follow up, the loss of coronary artery lumen diameter in these patients was higher than in all other patients combined (0.34+/-0.70 versus 0.10+/-0.29 mm; p=0.044). We conclude that a high HDL cholesterol does not protect against coronary artery disease when associated with combined CETP- and HL-lowering gene variants. PMID- 18164014 TI - Paraoxonase (PON1) and the risk for coronary heart disease and myocardial infarction in a general population of Dutch women. AB - There is strong evidence from both animal- and in vitro-models that paraoxonase (PON1) is involved in the onset of cardiovascular disease. In humans there is no consensus on this issue and therefore we investigated the effect of PON1 genotype and activity on the incidence of coronary heart disease (CHD) and acute myocardial infarction (AMI) in a large prospective cohort of 17,357 middle-aged women. We applied a case-cohort design using the CHD (n=211) and AMI cases (n=71) and a random sample from the baseline cohort (n=1527). A weighted Cox proportional hazards model was used to estimate age- and multivariate-adjusted hazard ratios (HR) for the PON1 genetic variants (192Q > R and -107C > T) and tertiles of the PON1 arylesterase- and paraoxonase activities. Neither the PON1 genetic variants, nor the PON1 activities affected the incidence of CHD in general, but, an increased paraoxonase activity was associated with a higher risk of AMI: the second and third tertile HR were 1.31 and 2.07, respectively (P trend=0.029, multivariate model). In the subgroup of never-smokers, paraoxonase activity was associated with an increased risk for AMI: the second and third tertile HR were 4.1 and 4.7, respectively (P-trend=0.009, multivariate model). Additionally, when compared to the lowest paraoxonase tertile in never-smokers, the highest paraoxonase tertile in current-smokers showed a 19.2-fold higher risk for AMI (95%CI: 5.3-69.5, P < 0.0001, multivariate model). In conclusion, this study shows that in middle-aged women paraoxonase activity was associated with an increased risk for AMI and that the risk was modified by the effects of smoking. PMID- 18164015 TI - Genetic modulation of anti-inflammatory effects of atorvastatin; probably a multi gene condition. PMID- 18164016 TI - Glycated LDL increases monocyte CC chemokine receptor 2 expression and monocyte chemoattractant protein-1-mediated chemotaxis. AB - BACKGROUND: Previous reports have suggested that levels of advanced glycation end product-modified LDL (AGE-LDL) increase in patients with diabetes due to elevated plasma glucose. However, understanding of the mechanisms by which AGE-LDL may accelerate atherogenesis remains incomplete. METHODS AND RESULTS: Microarray and reverse transcription real-time PCR (RT-PCR) analyses revealed that AGE-LDL significantly increased levels of CC chemokine receptor 2 (CCR2) mRNA in human macrophages compared with LDL, an effect accompanied by increased levels of CCR2 protein. Flow cytometry also showed that AGE-LDL increases CCR2 expression on the cell surface following stimulation (48h) (P<0.05). This effect appeared to depend on the receptor for AGE (RAGE), since an anti-RAGE antibody significantly blocked increased CCR2 mRNA. Functional studies demonstrated that exposure of THP-1 monocytoid cells to AGE-LDL increases chemotaxis mediated by monocyte chemoattractant protein-1 (MCP-1) up to 3-fold compared to LDL treatment (P<0.05). CONCLUSIONS: These data show that AGE-LDL can increase CCR2 expression in macrophages and stimulate the chemotactic response elicited by MCP-1. This novel mechanism may contribute to accelerated atherogenesis in diabetic patients. PMID- 18164017 TI - Sex differences in the associations between birthweight and lipid levels in middle-age: findings from the 1958 British birth cohort. AB - OBJECTIVE: To examine sex differences in birthweight-lipid associations. METHODS AND RESULTS: Using prospectively collected data on birthweight and non-fasting lipid levels at age 44-45 y from the 1958 British birth cohort (3603 men and 3583 women), sex differences in birthweight-lipid associations were examined. There were inverse associations between birthweight and total and low-density lipoprotein (LDL)-cholesterol among women (a 1 kg increase in birthweight was associated with a 0.13 mmol/L reduction in total cholesterol (p<0.001) and a 0.07 mmol/L reduction in LDL-cholesterol (p=0.02)) but no associations among men (p=0.005 and p=0.01, respectively, for birthweight x sex interactions). There was an inverse association between birthweight and triglycerides of a similar magnitude in both sexes (a 1 kg increase in birthweight was associated with a 7% reduction in triglyceride levels in sex-adjusted models (p<0.001)). There was no association between birthweight and high-density-lipoprotein-cholesterol. Associations were largely unaltered after adjustment for covariates. Of birthweight, current height and BMI, the latter was the strongest predictor of lipid levels. CONCLUSIONS: The finding of an inverse association between birthweight and triglycerides in both sexes and of inverse associations between birthweight and total and LDL-cholesterol only in women suggests that the mechanisms underlying the associations with birthweight may vary for different lipids. PMID- 18164018 TI - Prevalence and determinants of coronary artery disease in males and females with familial combined hyperlipidaemia. AB - BACKGROUND: Familial combined hyperlipidaemia (FCH) is an inherited dyslipidaemia that is related to a high risk of coronary artery disease (CAD). We evaluated the prevalence of CAD in a large FCH population and the association of risk factors with CAD according to gender. METHODS: In this single-center, observational study, lipid and lipoprotein variables were measured in untreated patients with FCH (565 males and 302 females). CAD was defined as a documented history of myocardial infarction or coronary revascularization, or an abnormal coronary angiogram (stenosis of >50% in an epicardial coronary artery), or angina plus abnormal imaging stress test. RESULTS: Males had higher triglyceride level (P<0.001) but lower total cholesterol (P<0.001) and HDL-cholesterol level (P<0.001) compared to women. The prevalence of CAD was 22.2% in men and 4.6% in women (P<0.001). In logistic regression analysis, male gender was associated with a higher risk of CAD independent of lipid parameters and other risk factors (adjusted ORs for CAD 9.4, P<0.001). In gender-specific analysis, age (OR=1.06 per 1-year increase, P<0.001), diabetes (OR=2.42, P<0.01) and Lp(a) (OR=1.09 per 1-mg/dL increase, P<0.01) were independent predictors of CAD in men. In women, age (OR=1.24, P<0.01), total cholesterol (OR=1.022 per 1-mg/dL increase, P<0.05) and fasting glucose (OR=1.031 per 1-mg/dL increase, P<0.05) were independently associated with CAD. CONCLUSIONS: In FCH patients, the prevalence of CAD is higher in males than in females, independent of lipidaemic profile and other risk factors. Among lipid variables, Lp(a) and cholesterol level are predictors of CAD in males and females respectively. PMID- 18164019 TI - Increased adiponectin synthesis in the visceral adipose tissue in men with coronary artery disease treated with pravastatin: a role of the attenuation of oxidative stress. AB - Pravastatin is reported to increase the adiponectin level in humans, but the mechanism remains unclear. We examined plasma and gene expressions of adiponectin, tumor necrosis factor (TNF)-*, interleukin (IL)-6 and protein carbonyl level, an indicator of oxidative stress, in visceral and subcutaneous adipose tissue from 32 patients with coronary artery disease undergoing coronary artery bypass grafting (CABG). Fourteen patients with serum LDL-cholesterol level >100mg/dl were treated with pravastatin at 10mg/day for 2 months before CABG (Statin), and the other 18 with LDL-cholesterol 0.05). However, the aircrew sampled population was almost two times more sensitive to BLM G0 clastogenic effects than controls (p<0.05). Therefore, our data suggest that chronic exposure of aircrew to cosmic radiation increases the in vitro chromosomal sensitivity of their peripheral lymphocytes to BLM (at least in the G0 stage of the cell cycle), and that occupational exposure of flight personnel to cosmic radiation does not induce an adaptive response to this radiomimetic compound. Our results justify further studies aimed at determine if those aircrew members hypersensitive to BLM are more prone to develop environmental cancer than BLM-insensitive individuals. PMID- 18164040 TI - Polymorphisms in genes involved in the inflammatory response and interaction with NSAID use or smoking in relation to lung cancer risk in a prospective study. AB - Lung cancer risk was investigated in relation to single nucleotide polymorphisms in genes involved in the inflammatory response. The aim was to see if polymorphisms modifying the inflammatory response are associated with risk of lung cancer and if there were interactions between the same polymorphism and factors, which modify an inflammatory response, such as smoking status, duration, and intensity, and use of NSAID. The functional SNPs IL-1B T-31C, IL6 G-174C, IL8 T-251A, IL10 C-592T, COX2 C8473T, COX2 A-1195G and PPARgamma2 Pro(12)Ala were included. A case-cohort study including 428 lung cancer cases and a sub-cohort of 800 persons was nested within a population-based prospective study of 57,053 individuals. Variant allele carriers of IL-1B T-31C were at increased risk of lung cancer (IRR=1.51, 95% CI=1.08-2.12). There was interaction between the polymorphism COX-2 T8473C and smoking status. Thus, non-smoking variant allele carriers were at 5.75-fold (95% CI=1.25-26.43) higher risk of lung cancer than for homozygous wild type allele carriers. Lung cancer risk was similar for all genotype carriers among past and current smokers. There were, however, very few non-smoking lung cancer cases. There was interaction between IL-1B T-31C, COX-2 A 1195G and PPARgamma2 Pro(12)Ala and NSAID use in relation to lung cancer risk. For the two latter, NSAID use was only associated with a lower cancer risk among homozygous wild type allele carriers. p for interaction was 3 x 10(-6) for COX-2 A-1195G and 9 x 10(-5) for PPARgamma2 Pro(12)Ala. The results suggest that NSAID use may modify risk of lung cancer differently depending on the genotype. PMID- 18164041 TI - Amphetamine-induced decreases in dopamine transporter surface expression are protein kinase C-independent. AB - Amphetamine (AMPH) is a potent dopamine (DA) transporter (DAT) inhibitor that markedly increases extracellular DA levels. In addition to its actions as a DAT antagonist, acute AMPH exposure induces DAT losses from the plasma membrane, implicating transporter-specific membrane trafficking in amphetamine's actions. Despite reports that AMPH modulates DAT surface expression, the trafficking mechanisms leading to this effect are currently not defined. We recently reported that DAT residues 587-596 play an integral role in constitutive and protein kinase C (PKC)-accelerated DAT internalization. In the current study, we tested whether the structural determinants required for PKC-stimulated DAT internalization are necessary for AMPH-induced DAT sequestration. Acute amphetamine exposure increased DAT endocytic rates, but DAT carboxy terminal residues 587-590, which are required for PKC-stimulated internalization, were not required for AMPH-accelerated DAT endocytosis. AMPH decreased DAT endocytic recycling, but did not modulate transferrin receptor recycling, suggesting that AMPH does not globally diminish endocytic recycling. Finally, treatment with a PKC inhibitor demonstrated that AMPH-induced DAT losses from the plasma membrane were not dependent upon PKC activity. These results suggest that the mechanisms responsible for AMPH-mediated DAT internalization are independent from those governing PKC-sensitive DAT endocytosis. PMID- 18164042 TI - Consciousness and its function. AB - It is plain that an individual's being conscious and an individual's being conscious of various things are both crucial for successful functioning. But it is far less clear how, if at all, it is also useful for a person's psychological states to occur consciously, as against those states occurring but without being conscious. Restricting attention to cognitive and desiderative states, a number of suggestions are current about how the consciousness of those states may be useful. It has been held that such consciousness enhances processes of rational thought and planning, intentional action, executive function, and the correction of complex reasoning. I examine these and related proposals in the light of various empirical findings and theoretical considerations and conclude that the consciousness of cognitive and desiderative states is unlikely to be useful in these or related ways. This undermines a reliance on evolutionary selection pressures in explaining why such states so often occur consciously in humans. I propose an alternative explanation, on which cognitive and desiderative states come to be conscious as a result of other highly useful psychological developments, some involving language. But on this explanation the consciousness of these states itself adds no significant function to that of those other developments. PMID- 18164043 TI - Association of cognitive restraint with ghrelin, leptin, and insulin levels in subjects who are not weight-reduced. AB - Despite widespread efforts at weight loss, the prevalence of obesity continues to rise. Restrained eating is a pattern of attempted weight control characterized by cognitive restriction of food intake that has paradoxically been linked with overeating and/or weight gain. It is not known whether restrained eating is associated with abnormalities in appetite-regulating hormones, independent of its effects on body weight. To address this question, we assessed cognitive restraint using the Three-Factor Eating Questionnaire and obtained fasting measurements of ghrelin, leptin and insulin from 24 healthy, non-obese (body mass index (BMI) 19.7 to 29.6 kg/m(2)) adult subjects who were at a stable, lifetime maximum weight. We chose to study subjects at stable maximum weight to avoid the secondary effects of weight reduction on body-weight regulating hormones. Subjects were classified by cognitive restraint scale score into Low, Indeterminate, and High Restraint groups. Higher ghrelin levels were significantly associated with restraint in an unadjusted model (P=0.004) and after adjustment for BMI (P=0.007). No relationships were found between restraint scores and either leptin (P=0.75) or insulin (P=0.36). These findings show an orexigenic hormonal profile in restrained eaters, independent of changes in body weight. PMID- 18164044 TI - 7-O-Methylated anthocyanidin glycosides from Catharanthus roseus. AB - Anthocyanins were isolated from orange-red flowers of Catharanthus roseus cv 'Equator Deep Apricot', and identified as rosinidin 3-O-[6-O-(alpha rhamnopyranosyl)-beta-galactopyranoside] (1), and also 7-O-methylcyanidin 3-O-[6 O-(alpha-rhamnopyranosyl)-beta-galactopyranoside] (2) by chemical and spectroscopic methods. Pigment 1 was found to be a major anthocyanin in the flowers of this cultivar. By contrast, the distribution of rosinidin glycosides is very limited in plants, and reported only in the flowers of Primula. Pigment 2 was found in smaller concentrations, but its aglycone, 7-O-methylcyanidin, has been reported only once before, from the fruit of mango. PMID- 18164045 TI - [Cleft palate and Williams syndrome]. AB - INTRODUCTION: Cleft palate is not one of the malformations in the Williams-Beuren syndrome. OBSERVATION: We report the case of a female patient affected by Williams syndrome, presenting with a cleft palate. The diagnosis of Williams Beuren syndrome was confirmed by genetic analysis. DISCUSSION: Cleft palate with a Williams syndrome can be fortuitous given these two affections' frequency. The prevalence of Williams syndrome is 1/7500 to 1/10,000 births and palatine division 1/2000 to 1/2500. Cleft palate might also be a rare entity of Williams syndrome. We cannot be sure that the two are linked. PMID- 18164046 TI - Uncertainties in stormwater E. coli levels. AB - Although water-quality monitoring programs have been widely used to identify and understand the level of pollution in urban stormwater systems, these data are often used without due consideration of the inherent uncertainties contained within these measurements. This study focuses on the uncertainties associated with the monitored levels of Escherichia coli, a common microbial indicator, in urban stormwater. Four sites located in Melbourne, Australia, were used to assess the uncertainty of six stormwater flow and E. coli variables: (1) discrete E. coli concentration, (2) stormwater flow rate, (3) stormwater event volume, (4) event mean concentration (EMC) of E. coli (i.e. a flow-weighted average of an event's E. coli concentrations), (5) E. coli load for each measured event, and (6) site mean E. coli concentration (SMC) (i.e. a volume-weighted average of the E. coli EMCs). Uncertainties of discrete E. coli samples were greater than 30%, while the uncertainty in stormwater flow measurements averaged greater than 97%, mainly due to the high uncertainties in measurements of very low flows. Propagation of these uncertainties, through their respective formulas, found that E. coli EMC uncertainties varied between 10% and 52% and that uncertainties relating to SMC estimates ranged from 35% to 55%. These results show the importance of considering uncertainty when using monitored data sets for any application, including those relating to stormwater management decisions. Suggestions are made about how to increase the accuracies of E. coli monitoring in urban stormwater and how to balance the different sources of uncertainties so that the overall combined uncertainties are minimised while keeping costs at a minimum. PMID- 18164047 TI - Effectiveness of soil washing, nanofiltration and electrochemical treatment for the recovery of metal ions coming from a contaminated soil. AB - This research was conducted to integrate soil washing, nanofiltration (NF) membranes and electrochemical treatment as feasible methods for the remediation of contaminated soils. For this investigation, two acidic leachates (pH=2) were prepared using HCl and H(2)SO(4)-NaCl as soil-washing agents. The results of the soil washing indicated that HCl and the combined H(2)SO(4)-NaCl were effective for the extraction of ions resulting from a contaminated soil. It was observed that both leachates presented similar chemical compositions. Following this procedure, the leachate solutions were pre-filtered by microfiltration, followed by NF using the Desal-5 (DK) membrane. The experiment results showed that NF membranes presented a high ion-retention rate for the two leachates. In general, better retentions were observed with the leachate prepared with HCl than H(2)SO(4)-NaCl. In order to treat the concentrate resulting from the NF treatment, it underwent an electrochemical procedure (electrochemical deposition) as an alternative method for safe disposal. The results showed a high reduction of toxic ions, such as Pb and Cu, from the solution. These three processes applied in conjunction not only indicated that the treatment of solutions heavily contaminated with inorganic pollutants resulting from contaminated soils were feasible but also suggested the possibility of treating different types of heavy industrial effluents. PMID- 18164048 TI - Source apportionment of pollutants and flows of combined sewer wastewater. AB - To reduce the combined sewer overflows (CSOs) and improve surface water quality in the Greater Milwaukee area, construction of a 19.5-mile (31.4 km) inline storage system (ISS) was completed in 1994 to capture and convey sanitary sewage and stormwater for treatment. However, one to six overflows per year still occur. Chemical mass balance (CMB) and positive matrix factorization (PMF) modeling was done in order to find origins of flows and pollutants in CSOs. Based on overflow events from 2000 to 2006 (CMB) and 2004 to 2006 (PMF), we found that between 27% and 56% of the total overflow is from sanitary sewage and most of the remaining from stormwater with possible minor contribution (< or = 8%) from groundwater. Most total suspended solids and metals (Cd, Cr, Cu, Pb, Ni, Hg, and Zn) are from stormwater, while sanitary sewage carries large contributions (> or = 28%) of BOD5, NH3, and total phosphorus. The fraction of NH3 is especially high, i.e., > or = 58%. Implications for a possible future sewer separation in combined sewer areas into separate sanitary and storm sewers are discussed. PMID- 18164049 TI - The degradation of dissolved organic nitrogen associated with melanoidin using a UV/H2O2 AOP. AB - The aim of this study was to examine the simultaneous degradation of dissolved organic nitrogen (DON) and associated colour from wastewater containing melanoidins by an advanced oxidation process (AOP). UV irradiation of H2O2 was used as the mechanism to create the hydroxyl radical for oxidation. Melanoidins are large nitrogenous organic compounds that are refractory during biological wastewater treatment processes. The simultaneous degradation of DON and colour, present as a result of these compounds, was investigated using an AOP. The oxidation process was much more capable of removing colour (99% degradation), dissolved organic carbon (DOC) (50% degradation) and DON (25% degradation) at the optimal applied dose of hydrogen peroxide for the system (3300 mg l(-1)). This indicated that colour and DON removal were decoupled problems for the purpose of treating melanoidin by an AOP and thus colour removal can not be used as an indication of DON removal Colour was caused by organic molecules with molecular weight greater than 10 kDa. Oxidation caused a partial reduction of the DON (41 15% of the total dissolved nitrogen) and DOC (29-14% of the DOC) associated with the large molecular weight fraction (>10 kDa) and almost complete colour removal (87-3% of the total colour). The degraded DON was mostly accounted for by the formation of ammonia (31% of the nitrogen removed from the large fraction) and small molecular weight compounds (66% of the nitrogen removed from the large fraction). The degraded DOC appeared to be mostly mineralised (to CO2) with only 20% of the degraded compounds appearing as small molecular weight DOC. PMID- 18164050 TI - Biochemical characterization of a multiple heavy metal, pesticides and phenol resistant Pseudomonas fluorescens strain. AB - Pseudomonas fluorescens SM1 isolate was found to be resistant to some major water pollutants namely Cd2+, Cr6+, Cu2+, Ni2+, Pb2+, BHC, 2,4-D, mancozeb and phenols up to a concentration four times to the normal levels occurring in the highly pollulated regions. Curing experiment brought about the loss of one or more resistance markers indicating the plasmid born resistance. Plasmid profile of SM1 strain showed the presence of one DNA band of 43.6 kb. This Plasmid was isolated from SM1 strain and introduced into Escherichia coli DH5 alpha with a transformation frequency of 6.7 x 10(-4)transformants/recipient cell. The test SM1 strain was also capable of biotransforming Cr(VI) to Cr(III) which is less toxic compounds. Present studies further indicated that the test SM1 strain was not only resistant to 2,4-D, phenols and catechol but also capable of bioremediating these toxicants quite efficiently. Moreover, studies with inhibitors like sodium azide, 2,4-DNP and chloramphenicol suggested that the major mechanism for the bioremediation of the heavy metals other than Cr6+ would be the biosorption process. PMID- 18164051 TI - Surface detail reproduction of elastomeric impression materials related to rheological properties. AB - OBJECTIVES: The purpose of this work was to discern, for elastomeric impression materials, the important rheological properties and importance of hydrophilicity for detail reproduction. METHODS: Viscosity, modulus and tan delta were measured using a controlled-stress rheometer in cone/plate configuration. The flow of the materials, immediately after mixing and at the manufacturer's stated working time, was measured using a shark fin test and the interaction with moist surfaces was determined by taking impressions from two different sized grooves in moist gypsum casts. RESULTS: Tan delta was found to be the parameter most indicative of the accuracy of the impression and the flow of the material. Impregum samples, a polyether material, exhibited the highest initial tan delta (7.4), the largest shark fins at both time periods and the most accurate impressions from both grooves. Aquasil, a polyvinylsiloxane material, had similar initial tan delta values (6.9) and impressions taken on the deep groove with this material closely matched the groove. The other two polyvinylsiloxane materials (Affinis and Flexitime) had significantly lower initial tan delta values (3.1 and 2.9, respectively), exhibited much smaller shark fins and a worse ability to accurately reproduce the deep groove. SIGNIFICANCE: For large features, it is clear that the higher the initial tan delta of the impression material the better the ability to replicate larger features. However, with smaller features the relative hydrophobicity of the material becomes an important factor, with more hydrophilic materials better able to reproduce fine detail. PMID- 18164052 TI - The effect of load cycling on nanoleakage of deproteinized resin/dentin interfaces as a function of time. AB - OBJECTIVES: To evaluate the effects of cyclic loading on nanoleakage as a function of time at resin-dentin interfaces with and without removal of collagen. METHODS: Bovine flat mid-coronal dentin received one of the following surface treatments: (1) acid-etch or (2) acid etched+5% NaOCl for 2m. The teeth were then bonded with Single Bond (3M ESPE), Scotchbond Multi-Purpose (3M ESPE), One-Step Plus (Bisco) or All-Bond 2 (Bisco) adhesive systems and restored with Z250 composite. Half of the bonded teeth were randomly assigned to receive 200,000 cycles of loading at 50 N. Teeth were sectioned into 1 mm x 1 mm thick slices and stored in distilled water for 24h and 6 months. After water storage, beams were prepared for nanoleakage evaluation and observed under the SEM. Data were statistically analyzed using three-way ANOVA and Fisher's PLSD test (p<0.05). RESULTS: Use of NaOCl did not affect nanoleakage at the interface of all adhesive systems (p>0.05) when compared to their respective controls. After 6 months of immersion in water, except for One Step Plus, collagen-depleted groups and control groups presented similar leakage values. After cyclic loading, the deproteinized group revealed a higher degree of silver nitrate deposits when compared to the control group (p<0.05). All four adhesive systems presented a high degree of silver nitrate deposits after 6 months of water storage. SIGNIFICANCE: The hybrid layer is important as a stress-absorbing layer and it may not represent the weak link for initiation of the nanoleakage phenomenon. PMID- 18164053 TI - Interaction and relationship development in stable young couples: effects of positive engagement, psychological aggression, and withdrawal. AB - This study tested associations among observed interaction patterns and relationship satisfaction in a subsample of young at-risk couples (n=47) from the Oregon Youth Study who remained stable over 7 years; each partner's positive engagement, psychological aggression, and withdrawal within a particular conflict structure (his vs. her topic) was used to predict satisfaction over time using multilevel growth curve modeling. Women's positive engagement during both topics predicted higher satisfaction for both partners at within-couple and between couple levels. Women's psychological aggression showed topic-specific associations with lower satisfaction for each partner, and increases in both men's and women's psychological aggression during their partner's topic related to lower satisfaction over time for women. Both partners' withdrawal during men's topics predicted less decline in satisfaction for men. PMID- 18164054 TI - A new experimental method assessing attitudes toward adolescent dating and sibling violence using observations of violent interactions. AB - The present study provides experimental data comparing emerging adults' attitudes toward dating and sibling violence in adolescence using a new methodology in which participants observe a violent interaction between adolescents. The reported amount of violence experienced in dating and sibling relationships among emerging adults is also compared. The participants included 148 emerging adults (111 females, 37 males) who were in the dating violence condition and 134 emerging adults (93 females, 41 males) in the sibling violence condition. The results provide initial psychometric data on a new measure to assess attitudes toward interpersonal violence in adolescence-the Attitudes toward Interpersonal Violence Assessment (AIVA). Using this new observational measure in an experimental design, empirical evidence was found that (1) male initiated violence was less acceptable than female initiated violence, (2) females were less accepting of violence than males, (3) individuals were more complacent toward sibling violence than dating violence in adolescence, (4) males reported perpetrating more injuries than females, and (5) sibling violence was reported at higher frequencies than dating violence. Implications of the findings are discussed. PMID- 18164055 TI - Differential gene expression involved in oxidative stress response caused by triethylene glycol dimethacrylate. AB - Triethylene glycol dimethacrylate (TEGDMA) is a comonomer that is released from dental resin-based materials into hydrophilic solvents. The compound reduces cell vitality, and causes genotoxicity in mammalian cells in vitro. Here, we used gene expression profiling, combined with pathway analysis tools, to identify the molecular events associated with TEGDMA cytotoxicity in human fibroblasts using Affymetrix HG-U133A 2.0 GeneChip arrays. Increased ROS production and a cell cycle delay caused by 3mm TEGDMA after a 6h exposure were related to a cell response at the transcriptional level. The predominant biological processes associated with the genes that were differentially expressed in untreated and treated cell cultures included oxidative stress, cellular growth, proliferation and morphology, cell death, gene expression as well as DNA replication and repair. The most significantly upregulated genes were GEM (17-fold), KLHL24, DDIT4, TGIF, DUSP5 and ATF3, which are all related to the regulation of the cell structure, stress response, and cell proliferation. TXNIP was the most downregulated transcript (five-fold), whose gene product regulates the cellular redox balance. The downregulation of NRG1, ASPM, FBXO5, and PLK2 is linked to the regulation of cell proliferation and cell structure. The underlying mechanisms of the up- and downregulation of genes seem to be activated by the production of ROS, and the related regulation of the cellular redox balance disturbed in the presence of TEGDMA appears to be of the utmost importance. The coordinated induction of genes coding for oxidative stress response and antioxidant proteins is a critical mechanism of protection against TEGDMA-induced cell damage. PMID- 18164056 TI - Small-diameter biodegradable scaffolds for functional vascular tissue engineering in the mouse model. AB - The development of neotissue in tissue engineered vascular grafts remains poorly understood. Advances in mouse genetic models have been highly informative in the study of vascular biology, but have been inaccessible to vascular tissue engineers due to technical limitations on the use of mouse recipients. To this end, we have developed a method for constructing sub-1mm internal diameter (ID) biodegradable scaffolds utilizing a dual cylinder chamber molding system and a hybrid polyester sealant scaled for use in a mouse model. Scaffolds constructed from either polyglycolic acid or poly-l-lactic acid nonwoven felts demonstrated sufficient porosity, biomechanical profile, and biocompatibility to function as vascular grafts. The scaffolds implanted as either inferior vena cava or aortic interposition grafts in SCID/bg mice demonstrated excellent patency without evidence of thromboembolic complications or aneurysm formation. A foreign body immune response was observed with marked macrophage infiltration and giant cell formation by post-operative week 3. Organized vascular neotissue, consisting of endothelialization, medial generation, and collagen deposition, was evident within the internal lumen of the scaffolds by post-operative week 6. These results present the ability to create sub-1mm ID biodegradable tubular scaffolds that are functional as vascular grafts, and provide an experimental approach for the study of vascular tissue engineering using mouse models. PMID- 18164057 TI - PYY transgenic mice are protected against diet-induced and genetic obesity. AB - The gut-derived hormone, peptide YY (PYY) reduces food intake and enhances satiety in both humans and animals. Obese individuals also have a deficiency in circulating peptide YY, although whether this is a cause or a consequence of obesity is unclear. Our aims were to determine whether peptide YY (PYY) over expression may have therapeutic effects for the treatment of obesity by altering energy balance and glucose homeostasis. We generated PYY transgenic mice and measured body weight, food intake, temperature, adiposity, glucose tolerance, circulating hormone and lipid concentrations and hypothalamic neuropeptide levels (neuropeptide Y; proopiomelanocortin, and thyrotropin-releasing hormone) under chow and high-fat feeding and after crossing these mice onto the genetically obese leptin-deficient ob/ob mouse background. PYY transgenic mice were protected against diet-induced obesity in association with increased body temperature (indicative of increased thermogenesis) and sustained expression of thyrotropin releasing hormone in the paraventricular nucleus of the hypothalamus. Moreover, PYY transgenic mice crossed onto the genetically obese ob/ob background had significantly decreased weight gain and adiposity, reduced serum triglyceride levels and improved glucose tolerance compared to ob/ob controls. There was no effect of PYY transgenic over expression on basal or fasting-induced food intake measured at 11-12 weeks of age. Together, these findings suggest that long-term administration of PYY, PYY-like compounds or agents that stimulate PYY synthesis in vivo can reduce excess adiposity and improve glucose tolerance, possibly via effects on the hypothalamo-pituitary-thyroid axis and thermogenesis. PMID- 18164059 TI - Using a classical weight-of-evidence approach for 4-years' monitoring of the impact of an accidental oil spill on sediment quality. AB - In the present report, the successful application of a Weight of evidence approach (WOE) to sediment quality assessment during a four year impact period following an oil spill is discussed. The study assesses the sediment quality on the Galician Coast (NW Spain) which was impacted by an accidental spill associated with the sinking of the tanker Prestige (2002). The assessment is based on three lines of evidence: physicochemical characterization of the sediments; determination of acute toxicity by conducting sediment toxicity tests and benthic alteration including taxonomic identifications along with community descriptive statistics. The data obtained were integrated using a WOE approach by means of two different methodologies: multivariate analysis and ANOVA-based pie charts. Results confirm that PAHs related to the Prestige oil spill are the main contaminant associated with biological effects in the area which has since recovered from the initial acute impact. Also, the WOE allowed the identification of metal contamination not previously described in the area responsible for toxicity in sediments analyzed. In addition, the methodology proposed to link the 3 lines of evidence results shows the use for the first time of an objective indice based on factor analysis which allows pollution of the sediments studied to be qualitatively and quantitatively evaluated while demonstrating the WOE approach to be recommendable in monitoring environmental quality. PMID- 18164060 TI - Persistent organic pollutants (PCBs and DDTs) in small size specimens of bluefin tuna (Thunnus thynnus) from the Mediterranean Sea (Ionian Sea). AB - The concentration levels and pattern distribution of polychlorinated biphenyls (PCBs) and organochlorine pesticides (DDTs), were investigated in liver of small size specimens of bluefin tuna (Thunnus thynnus) from Mediterranean Sea (Ionian Sea). PCB concentrations (average: 526 ng g(-1) lipid wt) were comparable with DDT levels (average: 435 ng g(-1) lipid wt). The specific analysis of PCB congeners revealed a profile dominated by hexa-, penta- and heptachlorinated congeners, while among DDTs, the compounds with the higher concentration was p,p' DDE, constituting 91.9% of the total DDT. The total 2,3,7,8-TCDD toxic equivalent (TEQs) of coplanar PCBs, including mono- and non-ortho congeners, was 0.55 pg g( 1) wet weight with. The congeners with highest TEQs values were non-ortho congeners followed by mono-ortho ones. PMID- 18164061 TI - Bacterial DNA indicated as an important inducer of fish cathelicidins. AB - Cathelicidins are antimicrobial peptides indicated as important in the control of the natural microflora as well as in the fight against bacterial invasion in mammals. Little is known about cathelicidins in fish and here the Chinook salmon (Oncorhynchus tshawytscha) embryo cell line (CHSE-214) was used as a model system to study the expression of cathelicidins due to fish pathogenic bacteria. The cDNA of cathelicidin from CHSE-214 cells (csCath) was cloned and shown to be closely related to gene 2 of both rainbow trout and Atlantic salmon. The deducted amino acid sequence showed highest sequence identity to rtCath2 with 95% and 72% for the cathelin and the antibacterial part, respectively. Cathelicidin gene expression was studied and various Gram positive and Gram negative bacteria caused the upregulation of the gene (csCath). Bacterial DNA and protein were shown important for the induction of cathelicidin expression in these cells. LPS (Escherichia coli) also causes the upregulation of cathelicidins, but digestion of the LPS with DNase I before incubation of the cells, totally abolished the upregulation of cathelicidin and suggests DNA contamination in the LPS to be the trigger for this effect. These results could explain the limited responsiveness of fish cells towards pure LPS and confirm previous suggestions that fish cells are less sensitive to LPS than mammalian cells. PMID- 18164058 TI - Inhibitory control and emotional stress regulation: neuroimaging evidence for frontal-limbic dysfunction in psycho-stimulant addiction. AB - This review focuses on neuroimaging studies that examined stress processing and regulation and cognitive inhibitory control in patients with psycho-stimulant addiction. We provide an overview of these studies, summarizing converging evidence and discrepancies as they occur in the literature. We also adopt an analytic perspective and dissect these psychological processes into their sub components, to identify the neural pathways specific to each component process and those that are more specifically involved in psycho-stimulant addiction. To this aim we refer frequently to studies conducted in healthy individuals. Despite the separate treatment of stress/affect regulation, stress-related craving or compulsive drug seeking, and inhibitory control, neural underpinnings of these processes overlap significantly. In particular, the ventromedial prefrontal regions including the anterior cingulate cortex, amygdala and the striatum are implicated in psychostimulant dependence. Our overarching thesis is that prefrontal activity ensures intact emotional stress regulation and inhibitory control. Altered prefrontal activity along with heightened striatal responses to addicted drug and drug-related salient stimuli perpetuates habitual drug seeking. Further studies that examine the functional relationships of these neural systems will likely provide the key to understanding the mechanisms underlying compulsive drug use behaviors in psycho-stimulant dependence. PMID- 18164062 TI - The antimicrobial peptide hepcidin exerts an important role in the innate immunity against bacteria in the bony fish gilthead seabream. AB - Antimicrobial peptides (AMPs) are important mediators of the immune response against bacteria and hepcidin is a 20-25 residues member with known functions in iron regulation and the innate immune response. Most studies have focused on mammalian organisms but very little is known about other vertebrate groups including teleost fish. Thus, based on the sequence of an EST database, we have characterized hepcidin gene organization, gene expression, distribution and in vitro and in vivo regulation, as well as the biological activity of a synthetic peptide in the teleost fish gilthead seabream (Sparus aurata L.). First, it was found that the seabream hep gene genomic organization is formed by 3 exons and 2 introns, while the mRNA transcript is constitutively detected in most of the fish tissues but mainly in peritoneal leucocytes, head-kidney, liver and skin. Moreover, we have identified for the first time that hep is much more highly expressed in acidophilic granulocytes than in monocyte-macrophages and lymphocytes. In vitro, hep expression is up-regulated by several mitogens, pathogen-associated molecular patterns (PAMPs) and particulated antigens. Not surprisingly, intraperitoneal injection of bacteria or virus led to a significant gene up-regulation in the liver, head-kidney, peritoneal exudate or spleen. These observations suggest a major role for seabream hepcidin in the immune response to bacteria and viruses. Furthermore, the synthetic seabream Hep exerted an important antimicrobial activity against several bacterial strains in vitro reducing their viability. To conclude, seabream hep gene expression, up regulation after in vitro or in vivo treatment with mitogens, PAMPs or particulated antigens and the direct in vitro biological activity against bacteria demonstrate that it is an important antimicrobial peptide and probably plays an important role in the innate immune response of fish. PMID- 18164063 TI - Endothelial cell loss after Descemet's stripping endothelial keratoplasty in a large prospective series. AB - PURPOSE: To report the donor endothelial cell loss in the first year after Descemet's stripping endothelial keratoplasty (DSEK) for the treatment of endothelial dysfunction. DESIGN: Prospective noncomparative interventional case series. PARTICIPANTS: Eighty eyes of 78 patients with corneal edema. METHODS: Eighty eyes with endothelial failure were entered into a prospective study of endothelial keratoplasty (EK). The donor central endothelial cell density (ECD) was recorded postoperatively at 6 months (n = 80) and 12 months (n = 80) and then compared with the preoperative eye bank measurements. The subsets of eyes with the donor prepared manually (DSEK; n = 19) and the donor prepared with a microkeratome (Descemet's stripping automated EK [DSAEK]; n = 61) were also evaluated and compared. MAIN OUTCOME MEASURES: Preoperative and postoperative central ECDs were prospectively evaluated and the cell loss calculated for each postoperative time point. RESULTS: The average and standard deviation ECD at 6 months was 1908+/-354 cells/mm(2), representing a mean cell loss from preoperative donor cell measurements of 34+/-12%. At 12 months, ECD was 1856+/ 371 cells/mm(2) (35+/-13% cell loss). The 1% additional cell loss from 6 to 12 months was not significant (P = 0.233). In the subset of DSEK eyes (n = 19), the cell loss from preoperatively to 6 months was 34%, and at 12 months it was 39%. In the subset of DSAEK eyes (n = 61), the cell loss from preoperatively to 6 months was 34%, and at 12 months it was 34%. There was no statistical difference between the cell loss from DSEK and that from DSAEK at 6 months (P = 0.884) or at 12 months (P = 0.224). CONCLUSIONS: Descemet's stripping EK using our surgical technique has a mean donor endothelial cell loss of 34% at the 6-month postoperative examination, and this average cell loss remains relatively stable up to at least 1 year. We found no difference in cell loss between the DSEK and DSAEK techniques over this 1-year postoperative period. PMID- 18164064 TI - Randomized trial of early phacoemulsification versus peripheral iridotomy to prevent intraocular pressure rise after acute primary angle closure. AB - PURPOSE: To compare the efficacy of early phacoemulsification versus laser peripheral iridotomy (LPI) in the prevention of intraocular pressure (IOP) rise in patients after acute primary angle closure (APAC). DESIGN: Prospective randomized controlled trial. PARTICIPANTS: Sixty-two eyes of 62 Chinese subjects, with 31 eyes in each arm. METHODS: Subjects were randomized to receive either early phacoemulsification or LPI after aborting APAC by medications. Patients were followed up on day 1; week 1; and months 1, 3, 6, 12, and 18. Predictors for IOP rise were studied. MAIN OUTCOME MEASURES: Prevalence of IOP rise above 21 mmHg (primary) and number of glaucoma medications, IOP, and Shaffer gonioscopy grading (secondary). RESULTS: Prevalences of IOP rise for the LPI group were 16.1%, 32.3%, 41.9%, and 46.7% for the follow-ups at 3, 6, 12, and 18 months, respectively. There was only one eye (3.2%) in the phacoemulsification group that had IOP rise at all follow-up time points (P<0.0001). Treatment by LPI was associated with significantly increased hazard of IOP rise (hazard ratio [HR], 14.9; 95% confidence interval [CI], 1.9-114.2; P = 0.009). In addition, a maximum IOP at presentation > 55 mmHg was associated with IOP rise (HR, 4.1; 95% CI, 1.3 13.0; P = 0.017). At 18 months, the mean number of medications required to maintain IOP 55 mmHg is an added risk factor for subsequent IOP rise. For patients with coexisting cataract and presenting IOP of >55 mmHg, early phacoemulsification can be considered as a definitive treatment to prevent IOP rise. PMID- 18164065 TI - Incidence of and factors associated with glaucoma after surgery for congenital cataract: findings from the British Congenital Cataract Study. AB - PURPOSE: To report the incidence of and factors associated with postoperative open-angle glaucoma in a nationally representative group of children undergoing surgery for congenital or infantile cataract. DESIGN: Noncomparative interventional cohort study. PARTICIPANTS: All children in the United Kingdom who were newly diagnosed with congenital or infantile cataract in a 12-month period in 1995 and 1996 (the British Congenital/Infantile Cataract Study) were eligible for this study. One hundred sixty-five children with congenital or infantile cataract underwent cataract surgery. METHODS: All the children were traced through their managing ophthalmologists. Standardized outcome data were collected at least 6 years after diagnosis. For children undergoing cataract extraction, Cox regression analysis was performed to determine incidence of postoperative open-angle glaucoma and the effect of key factors considered, a priori, potentially to be associated with it (i.e., age at detection and surgery, type of cataract surgery, primary intraocular lens implantation, severe postoperative uveitis, and microphthalmia). MAIN OUTCOME MEASURES: Development of open-angle glaucoma after cataract surgery. RESULTS: Postoperative glaucoma developed in 27 of 275 eyes of 165 children who underwent cataract surgery. The overall annual incidence of postoperative glaucoma was 5.25 per 100 cataract operations. The median time to development of postoperative glaucoma was 1.34 years (range, 0.39 months-6.73 years). Younger age at detection of cataract was the only factor independently associated with the development of glaucoma when all other factors of interest (which were all statistically associated with age at detection) were accounted for. A 10-fold increase in the age at detection (for example, 30 days compared with 3 days) was associated with a 64% decrease in the hazard ratio (95% confidence interval, 41%-79%; P<0.001). CONCLUSIONS: Median time to development of postoperative open-angle glaucoma in the present study was lower than that reported previously, emphasizing the need for vigilance from the early postoperative period. Earlier detection of cataract was the only significant factor associated with the development of glaucoma after surgery for congenital cataract. PMID- 18164066 TI - Alleles in the HtrA serine peptidase 1 gene alter the risk of neovascular age related macular degeneration. AB - OBJECTIVE: To examine if the genes encoding the pleckstrin homology domain containing protein gene (PLEKHA1), hypothetical LOC387715/ARMS2 gene, and HtrA serine peptidase 1 gene (HTRA1) located on the long arm of chromosome 10 (10q26 region) confer risk for neovascular age-related macular degeneration (AMD) in an independent or interactive manner when controlling for complement factor H gene (CFH) genotype and smoking exposure. DESIGN: Retrospective matched-pair case control study. PARTICIPANTS: Hospital clinic-based sample of 134 unrelated patients with neovascular AMD who have a sibling with normal maculae (268 subjects). METHODS: Disease status was ascertained by at least 2 investigators by review of fundus photographs and/or fluorescein angiography according to the Age Related Eye Disease Study grading scale. If necessary, a home retinal examination was performed (n = 6). A combination of direct sequencing and analysis of 8 highly polymorphic microsatellite markers was used to genotype 33 megabases of the 10q26 region on leukocyte DNA. Smoking history was obtained via a standardized questionnaire and measured in pack-years. The family-based association test, haplotype analysis, multiple conditional logistic regression, and linkage analysis were used to determine significant associations. MAIN OUTCOME MEASURE: Neovascular AMD status. RESULTS: Of the 23 variants we identified in the 10q26 region, 6 were significant. Four of the 6 were novel and included 2 genotypes that reduced risk of AMD. Many single-nucleotide polymorphisms (SNPs), including the previously reported variants rs10490924 (hypothetical LOC387715/ARMS2) and rs11200638 (HTRA1), defined 2 significant haplotypes associated with increased risk of neovascular AMD. The coding HTRA1 SNP rs2293870, not part of the significant haplotypes containing rs10490924 and rs11200638, showed as strong an association with increased susceptibility to neovascular AMD. Linkage analysis supported our findings of SNP association (P<10(-15)). No significant interactions were found between any of the SNPs in the 10q26 and smoking or between these SNPs and CFH genotype. CONCLUSIONS: Independent of CFH genotype or smoking history, an individual's risk of AMD could be increased or decreased, depending on their genotype or haplotype in the 10q26 region. PMID- 18164067 TI - Testability of the Retinomax autorefractor and IOLMaster in preschool children: the Multi-ethnic Pediatric Eye Disease Study. AB - PURPOSE: To determine the testability of Retinomax and IOLMaster ocular biometry in preschool children. DESIGN: Population-based study of inner city preschool children in Los Angeles County. PARTICIPANTS: Two thousand five hundred forty five Hispanic and 2178 African American children 6 to 72 months old. METHODS: Subjects were identified by door-to-door screening within previously identified contiguous census tracts. Pediatric ophthalmologists or optometrists performed comprehensive eye examinations on all subjects. Refractive error and keratometry measurements were attempted on all subjects with the Retinomax autorefractor after cycloplegia. Axial length measurements with the IOLMaster partial coherence interferometer were attempted on those subjects ages 30 to 72 months. MAIN OUTCOME MEASURES: Ability to obtain high confidence autorefraction readings or axial length measurements on both eyes. RESULTS: Overall, 89% were testable in both eyes with the Retinomax device, and 91% of the children were testable with the IOLMaster. Testability rose sharply with age, so that by age 36 months 98% of children were testable with the Retinomax device and 90% were testable with IOLMaster. There were no consistent gender- or ethnicity-related differences in testability overall or when stratified by age for either device. CONCLUSIONS: Young children can be reliably tested for ocular biometry with the Retinomax and IOLMaster devices. This may impact strategies for management of cataracts and refractive errors in preschool children. PMID- 18164068 TI - Efficacy of the Ahmed S2 glaucoma valve compared with the Baerveldt 250-mm2 glaucoma implant. AB - OBJECTIVE: To compare the efficacy of the Ahmed S2 Glaucoma Valve with the Baerveldt 250-mm(2) Glaucoma Implant in the treatment of adult glaucoma. DESIGN: Comparative case series. PARTICIPANTS: Fifty-nine eyes of 59 patients who received the Ahmed S2 Glaucoma Valve and 133 eyes of 133 patients who received the Baerveldt 250-mm(2) Glaucoma Implant by the Indiana University Glaucoma Service from 1996 to 2003. METHODS: Eyes that had previous drainage implant procedures were excluded from both groups. If both eyes of a single patient received an implant, the second eye to undergo implantation was excluded from the study. MAIN OUTCOME MEASURES: Kaplan-Meier survival with success defined as intraocular pressure (IOP) > 5 mmHg and < 22 mmHg and at least 20% reduction from preoperative IOP (with or without antiglaucoma medications) and without loss of light perception. Secondary outcome measures included intraocular pressure, visual acuity, number of glaucoma medications, and surgical complications. RESULTS: The 2 groups were similar with regards to age, gender, race, neovascular glaucoma diagnosis, number of prior ocular surgeries, preoperative IOP, and number of preoperative glaucoma medications. Mean durations of follow-up were 20.0 months for Ahmed eyes and 22.9 months for Baerveldt eyes. Cumulative successes in the Ahmed group were 0.73 at 1 year and 0.62 at 2 years, whereas cumulative successes in the Baerveldt group were 0.92 at 1 year and 0.85 at 2 years (Kaplan-Meier survival functions: P = 0.03, log rank test). Male gender, African descent, neovascular glaucoma, and Ahmed implantation were found to be significant predictors of failure. At last follow-up visit, eyes in the Ahmed group had a significantly higher mean IOP (19.8+/-9.5 vs. 15.8+/-7.9 mmHg, P = 0.003, t test) and more antiglaucoma medications (1.4+/-1.2 vs. 0.9+/-1.1 medications, P = 0.008, Mann-Whitney test) than eyes in the Baerveldt group. Two methods for avoiding hypotony after Baerveldt 250-mm(2) implantation had similar outcomes. CONCLUSIONS: Our study suggests that the Ahmed S2 Glaucoma Valve may be less effective at lowering IOP than the Baerveldt 250-mm(2) Glaucoma Implant. PMID- 18164069 TI - Rainbow glare as an optical side effect of IntraLASIK. AB - PURPOSE: To report a new optical side effect associated with the IntraLASIK procedure. DESIGN: Retrospective chart review and questionnaire of all patients treated with IntraLASIK from June 2004 to August 2005. PARTICIPANTS: A cohort of 585 eyes (312 patients) was divided sequentially into those treated with the older-model IntraLase laser (group 1: 399 eyes of 215 patients) and those treated with the newer model (group 2: 186 eyes of 97 patients). METHODS: Preoperative and postoperative refraction, visual acuity, pupillometry, pachymetry, intraocular pressure, mean topographic power and cylinder, wavefront aberrations, and answers to a questionnaire of symptoms were recorded. The precise spatial extent of the rainbow spectrum was drawn by select patients for calculation of the diffractive grating size responsible for the light scatter. A glass slide was also irradiated and photographed using the older model to simulate the rainbow spectrum. MAIN OUTCOME MEASURES: The symptom of rainbow glare was correlated with preoperative and postoperative factors and time between the most recent service call and surgery. RESULTS: In group 1, 90.2% of patients were successfully contacted, and 37 (69 eyes) reported symptoms of rainbow glare around lights, for an incidence of 19.07%. In group 2, 88.6% of patients were contacted, and 2 (4 eyes) reported similar symptoms, for an incidence of 2.32%. The odds ratio (OR) between the 2 groups is 9.4 (P<0.001). For every 10-mum increase in ablation depth, the OR is 1.2 (P = 0.038), and for every 30 days from the last service call, it is 1.5 (P = 0.007). The spatial extent of the color pattern was used to calculate the spacing array at approximately 9 mum, corresponding to the laser spot and line separation. CONCLUSIONS: Light scattering from the back surface of the IntraLASIK flap creates a spectral pattern whose visual impact is clinically inconsequential in the majority of patients. The spectral pattern and visual angle correspond to a grating size that matches the raster spot separation of IntraLase pulsing. Eyes treated with newer focusing optics of higher numerical aperture reduced the rainbow symptom. Variance in optical quality and numerical aperture can impact the spot size and uniformity of IntraLase flap creation. PMID- 18164070 TI - The role of protein binding of trivalent arsenicals in arsenic carcinogenesis and toxicity. AB - Three of the most plausible biological theories of arsenic carcinogenesis are protein binding, oxidative stress and altered DNA methylation. This review presents the role of trivalent arsenicals binding to proteins in arsenic carcinogenesis. Using vacuum filtration based receptor dissociation binding techniques, the lifetimes of unidentate (<1s), bidentate (1-2min) and tridentate (1-2h) arsenite containing peptide binding complexes were estimated. According to our experimental data some of the protein targets to which arsenite may bind in vivo include tubulin, poly(ADP-ribose)polymerase (PARP-1), thioredoxin reductase, estrogen receptor-alpha, arsenic(+3)methyltransferase and Keap-1. Arsenite binding to tubulin can lead to several of the genetic effects observed after arsenic exposures (aneuploidy, polyploidy and mitotic arrests). Among many other possible arsenite binding sites are rat hemoglobin, the DNA repair enzyme xeroderma pigmentosum protein A (XPA), and other C2H2, C3H and C4 zinc finger proteins including members of the steroid receptor superfamily (e.g. glucocorticoid receptor). Macromolecules to which arsenite does not bind to include calf thymus DNA, mixed Type II-A histones and bovine H3/H4 histone. Although all six tested arsenicals released iron from ferritin, radioactive arsenite did not bind to the protein horse ferritin. PMID- 18164071 TI - Synthesis and properties of a highly soluble dihydoxo(tetra-tert butylphthalocyaninato)antimony(V) complex as a precursor toward water-soluble phthalocyanines. AB - The title complex cation, [Sb(tbpc)(OH)(2)](+) (where tbpc denotes tetra(tert butyl)phthalocyaninate, C(48)H(48)N(8)(2-)), has been prepared by oxidizing [Sb(tbpc)]I(3) with tert-butyl perbenzoate in CH(2)Cl(2), CHCl(3), o dichlorobenzene and also without solvent in the range of 20-80 degrees C. This species has been isolated as I(3)(-) salt and characterized by elemental analysis, ESI-MS, FT-IR, optical absorption and emission, and magnetic circular dichroism spectroscopy. This compound is quite well soluble in common polar organic solvents (e.g., CH(2)Cl(2), acetonitrile, acetone) without detectable aggregation at least up to ca. 10(-4)M while much less soluble (e.g., benzene, chloronaphthalene) or insoluble (hexane) in non-polar solvents. Although this compound is insoluble in water, it makes hydrophilic colloids in acetone-water mixtures. The most intense absorption band (Q-band) in a specific solvent red shifts with an increase in the refractive index of the solvent. However, considerable deviation of the Q-band positions in donor-solvents from linear correlation between the positions and Onsager's solvent polarity function suggests that there are significant specific chemical interactions between the axial hydroxyl groups and the surrounding donor molecules. The low fluorescence quantum yield (ca. 0.01) for [Sb(tbpc)(OH)(2)](+) suggests that the singlet excited state of this species is considerably quenched by the presence of antimony ion in the chromophore. PMID- 18164072 TI - In vitro and ex vivo storage phosphor imaging of short-living radioisotopes. AB - Storage phosphor imaging may be of value for biodistribution studies of short living radiotracers in small animals. Efficiency, sensitivity and resolution of imaging plates for short-living radioisotopes vary considerably but linear response to many radioisotopes was shown previously. However, these properties have not been compared directly for larger series of short-living radioisotopes, and only few studies have directly compared data obtained from phosphor images of tissue slices with results from dissection biodistribution studies. Therefore, we evaluated the properties of imaging plates for 11 short-living radioisotopes (18F, 32P, 67Ga, 89Sr, (99m)Tc, 90Y, 111In, 123I, 125I, 131I and 201Tl). We also evaluated the biodistribution of [123I]FP-CIT in rat brain using both the phosphor technique and conventional dissection methods. The imaging system showed a linear response for all tested radioisotopes over a wide range of radioactive concentrations and the efficiency, sensitivity and resolution varied greatly for the tested radioisotopes. Shielding experiments revealed the contribution of the various emission products of radioisotopes to these properties. However, quantitative biodistribution studies with radiotracers that are labeled with all tested radioisotopes, even 123I, are feasible. The results from the ex vivo biodistribution study, using [123I]FP-CIT as a radiotracer were similar for the phosphor imaging technique as compared to the dissection technique. Advantages of phosphor imaging in radiotracer distribution studies in rat brain as compared to dissection experiments may be more precise measurements, possibility to reanalyze imaging data and 3D-reconstruction. In conclusion, phosphor imaging is an attractive alternative for biodistribution studies of short-living radiotracers in small animals. PMID- 18164073 TI - A simple, efficient tool for assessment of mice after unilateral cortex injury. AB - A refined battery of neurological tests, SNAP (Simple Neuroassessment of Asymmetric Impairment), was developed and validated to efficiently assess neurological deficits induced in a mouse model of traumatic brain injury. Four to 7-month old mice were subjected to unilateral controlled cortical impact or sham injury (craniectomy only). Several behavioral tests (SNAP, beam walk, foot fault, and water maze) were used to assess functional deficits. SNAP was unique among these in that it required no expensive equipment and was performed in less than 5 min per mouse. SNAP demonstrated a high level of sensitivity and specificity as determined by receiver-operator characteristics curve analysis. Interrater reliability was good, as determined by Cohen's Kappa method and by comparing the sensitivity and specificity across various raters. SNAP detected deficits in proprioception, visual fields, and motor strength in brain-injured mice at 3 days, and was sensitive enough to detect magnitude and recovery of injury. The contribution of individual battery components changed as a function of time after injury, however, each was important to the overall SNAP score. SNAP provided a sensitive, reliable, time-efficient and cost-effective means of assessing neurological deficits in mice after unilateral brain injury. PMID- 18164074 TI - Hypofrontality in subjects at high genetic risk of schizophrenia with depressive symptoms. AB - BACKGROUND: Subjects at high risk of schizophrenia for genetic reasons were found to demonstrate increased levels of depressive symptoms compared to controls. The current study sought to investigate the neural correlates of depression in these subjects. We hypothesised abnormal activation of dorsolateral prefrontal regions in those at high risk with depression. METHODS: Depression was rated according to DSM-IV criteria. FMRI data was available from 90 high risk subjects, comprising 78 not depressed (HRD-) and 12 depressed (HRD+) subjects. Activation during the Hayling Sentence Completion Task was compared to 25 healthy control subjects without depression. RESULTS: The HRD+ group demonstrated reduced activation of the right middle/superior frontal gyrus compared to both healthy controls and the HRD- group. Increased left superior temporal gyrus activation was also found in the HRD+ group versus the HRD- group. These results survived controlling for the presence of positive psychotic symptoms at the time of the scan. CONCLUSION: Reduced activation of dorsolateral prefrontal regions, widely reported in established schizophrenia and seen here in people at high familial risk with depressive features, may be related to the presence of affective symptoms of the disorder rather than to the presence of positive psychotic symptoms. Since studies have indicated that depressive symptoms antecede illness, these findings may be relevant to the early features of developing psychosis. PMID- 18164075 TI - Comorbidity of depressive disorders among adolescents in community and clinical settings. AB - The aim of this article was to examine the comorbidity rates and clinical characteristics of depressive disorders among adolescents in community and clinical settings, using the same methodology. One thousand and thirty-five adolescents from the community setting and 200 adolescents from the clinical setting were interviewed using the computerized Munich version of the Composite International Diagnostic Interview. In both settings, the comorbidity of depressive disorders with other psychiatric disorders was high. About 58% of the depressed adolescents in the community setting had at least one additional disorder, compared with 63.5% in the clinical setting. The most common comorbid pattern was that of depressive and anxiety disorders. Among adolescents with anxiety and depression, 72% and 62% of those in the community and clinical settings, respectively, reported the occurrence of anxiety before that of depression. Depressed adolescents with comorbid disorders used significantly more mental health services than adolescents with depression only. The findings suggest the need to design assessment and intervention strategies to deal with adolescents with multiple disorders. PMID- 18164076 TI - Immunosuppressive mechanisms in human tumors: why we still cannot cure cancer. AB - Tumor cells often evoke specific immune responses that, however, fail to eliminate all the tumor cells. The development of cancer immunotherapies is, therefore, mostly focused on the generation of large numbers of activated anti tumor effector cells by vaccination or adoptive T cell transfer. These developments are built on an ever-extended list of identified tumor-associated antigens and corresponding T cell epitopes, and a steady flow of reports from proof-of-principle animal model experiments demonstrating cure from disease by immune interventions. However, the promises have not translated into clinical successes for cancer patients. Even where tumor regression or complete responses were achieved there is usually relapse of the disease. Increasing numbers of reports over recent years highlight potential immunosuppressive mechanisms that act in tumors and systemically in cancer patients to block effective anti-tumor immune responses. They account in large parts for the failures of cancer immunotherapy and need to be overcome before progress can be expected. We review here the current state of the research on immunosuppressive networks in human cancer. PMID- 18164077 TI - The interleukin 23 receptor gene in multiple sclerosis: a case-control study. AB - Variants of the gene coding for interleukin 23 receptor (IL23R) have been confirmed to be associated with inflammatory bowel disease (IBD). Based on similarities in the autoimmune features of IBD and multiple sclerosis (MS), we selected this gene for analysis as a candidate gene in MS. We tested 32 single nucleotide polymorphisms tagging all parts of the gene including 10 markers previously studied in IBD for association with MS. The study population consisted of 1114 MS patients and 1235 controls of Scandinavian origin. None of the tests showed more than very modest differences between patients and controls. Haplotypes were constructed and were also similarly distributed in patients and controls. We conclude that it is unlikely that the IL23R gene confers any significant risk for MS. PMID- 18164078 TI - The selective 5-HT(6) receptor antagonist Ro 04-6790 attenuates psychotomimetic effects of the NMDA receptor antagonist MK-801. AB - There are experimental evidences indicating that the non-competitive NMDA receptor antagonist MK-801 impairs cognition and produces a series of schizophrenia-like symptoms in rodents (hypermotility, stereotypies and ataxia). The present study was designed to investigate the efficacy of the selective 5 HT(6) receptor antagonist Ro 04-6790 in counteracting these MK-801-induced behavioural effects in the rat. The effects of Ro 04-6790 in antagonizing MK-801 induced memory deficits were assessed using the object recognition task. The ability of this 5-HT(6) receptor antagonist in counteracting hypermotility, stereotypies and ataxia produced by MK-801 were evaluated in a motor activity cage. Post-training administration of Ro 04-6790 (10 and to some extent also 3mg/kg) antagonized MK-801-induced performance deficits in a recognition memory test. In a subsequent study, Ro 04-6790 (3 and 10 mg/kg) reversed hypermotility and ataxia produced by MK-801. This 5-HT(6) receptor antagonist also alleviated MK-801-induced certain stereotypies. Our findings indicate that Ro 04-6790 attenuates behavioural effects related to the hypofunction of the NMDA receptor suggesting that this compound might be involved in the psychotomimetic effects of non-competitive NMDA receptor antagonists. PMID- 18164079 TI - Transcriptionally active PCR for antigen identification and vaccine development: in vitro genome-wide screening and in vivo immunogenicity. AB - We have evaluated a technology called transcriptionally active PCR (TAP) for high throughput identification and prioritization of novel target antigens from genomic sequence data using the Plasmodium parasite, the causative agent of malaria, as a model. First, we adapted the TAP technology for the highly AT-rich Plasmodium genome, using well-characterized P. falciparum and P. yoelii antigens and a small panel of uncharacterized open reading frames from the P. falciparum genome sequence database. We demonstrated that TAP fragments encoding six well characterized P. falciparum antigens and five well-characterized P. yoelii antigens could be amplified in an equivalent manner from both plasmid DNA and genomic DNA templates, and that uncharacterized open reading frames could also be amplified from genomic DNA template. Second, we showed that the in vitro expression of the TAP fragments was equivalent or superior to that of supercoiled plasmid DNA encoding the same antigen. Third, we evaluated the in vivo immunogenicity of TAP fragments encoding a subset of the model P. falciparum and P. yoelii antigens. We found that antigen-specific antibody and cellular immune responses induced by the TAP fragments in mice were equivalent or superior to those induced by the corresponding plasmid DNA vaccines. Finally, we developed and demonstrated proof-of-principle for an in vitro humoral immunoscreening assay for down-selection of novel target antigens. These data support the potential of a TAP approach for rapid high throughput functional screening and identification of potential candidate vaccine antigens from genomic sequence data. PMID- 18164080 TI - Identification of flotillin-1 as an interacting protein for antisecretory factor. AB - Antisecretory factor (AF) also named S5a/Rpn10 was originally identified through its capacity to inhibit intestinal hypersecretion and was later shown to be a component in the proteasome complex. AF is also a potent anti-inflammatory agent and can act as a neuromodulator. In this study we used yeast two-hybrid screens, with yeast strain PJ692A transformed with the bait vector pGBKT7 (AF aa 1-105) against yeast strain Y187 pretransformed with human brain or placenta cDNA libraries, to identify AF-binding proteins. Flotillin-1 was identified as a specific interacting factor with AF. Immunohistochemistry showed co-localization of AF and flotillin-1 in nervous tissue. Flotillin-1 is an integral membrane protein and a component of lipid rafts, a membrane specialization involved in transport processes. Intracellular AF may affect secretory processes by regulating the localization of signal proteins to lipid rafts. PMID- 18164081 TI - Wrong theories on the origin of blood vessels: Polybus and De Natura Hominis. AB - Polybus of Cos (approximately 400 B.C.) was the son-in-law and the successor of Hippocrates. He is credited with founding the school of Dogmatism, and writing "The Nature of Man" which was important in advancing the theory of the four body humors (humoralism). Some earlier scholars negated Polybus' role as an independent medical figure. However, Corpus Aristotelicum quoted him as having a unique theory regarding the body vasculature which stated that this system was composed of four pairs of blood vessels originating from the head and that these supplied the whole body. In an interpretation of this theory, we opined that numerological mysticism might have been the common motive for both Hippocrates' humoralism and Polybus' theory of the vasculature. A discussion on this issue is presented. PMID- 18164082 TI - An unusual case of partial anomalous pulmonary venous drainage: utility of the cardiac MRI. PMID- 18164083 TI - Effect of atherosclerosis on thermo-mechanical properties of arterial wall and its repercussion on plaque instability. AB - Data from the literature report febrile reactions prior to myocardial infarction in patients with normal coronary arteries and that coronary syndromes seem to be triggered by bacterial and viral infections, being fever the common symptom. The thermo-mechanical behavior of thoracic aortas of New Zealand White rabbits with different degrees of atherosclerosis was measured by means of pressure-diameter tests at different temperatures. Specific measurements of the thermal dilatation coefficient of atheroma plaques were performed by means of tensile tests. Results show a different thermo-mechanical behavior, the dilatation coefficient of atheroma plaque being at least twice that of the arterial wall. Temperature induced mechanical stress at the plaque-vessel interface could be enough to promote plaque rupture. Therefore, increases of corporal temperature, either local or systemic, can play a role in increasing the risk of acute coronary syndromes and deserve a more comprehensive study. PMID- 18164084 TI - Vanishing multiple thrombi in left ventricle of a patient with dilated cardiomyopathy. PMID- 18164085 TI - The chakragati mouse shows deficits in prepulse inhibition of acoustic startle and latent inhibition. AB - The chakragati (ckr) mouse, which was serendipitously created as a result of a transgenic insertional mutation, has been proposed as a model of aspects of schizophrenia. The mice exhibit circling, hyperactivity, reduced social interactions, and enlarged lateral ventricles, which parallel aspects of the pathophysiology of schizophrenia. Deficits in sensorimotor gating and processing of the relevance of stimuli are core features of schizophrenia, which underlie many of the symptoms presented. Measures of prepulse inhibition (PPI) and latent inhibition (LI) can assess sensorimotor gating and processing of relevance in both humans and animal models. We investigated PPI of acoustic startle and LI of aversive conditioning in wild-type, heterozygous, and ckr mice. The ckr mice, which are homozygous for the transgene insertion, but not heterozygous littermates, showed impaired PPI in the absence of any difference in acoustic startle amplitude and showed deficits in LI of conditioning of a light stimulus to footshock, measured as suppression of licking for water in water-restricted mice. Together with the previous evidence for hyperactivity, reduced social interactions, and enlarged lateral ventricles, these data lend further support to the suggestion that the ckr mouse has utility as an animal model of aspects of schizophrenia. PMID- 18164086 TI - Neural transdifferentiation of bone marrow stromal cells obtained by chemical agents is a short-time reversible phenomenon. AB - Bone marrow stromal cells (BMSC) can acquire morphological and immunohistochemical features of neural cells when they are treated with diverse chemical agents, a finding interpreted as result of cell transdifferentiation. With the purpose of a better knowledge of the possible utility of BMSC for strategies of Nervous System (NS) repair, we have studied the morphological and immunohistochemical changes induced in BMSC by chemical agents, in comparison with those that happen when BMSC are co-cultured with Schwann cells. While chemical BMSC transdifferentiation is a short-time reversible phenomenon, BMSC transdifferentiation obtained by Schwann cell-derived neurotrophic factors remains stable after it has been reached. These findings question the possible clinical utility of BMSC trandifferentiation using chemical agents, and support that neural transdifferentiation of BMSC is a biological phenomenon that can be obtained in vivo because of the presence of environmental factors. PMID- 18164087 TI - Role of primate cerebellar lobulus petrosus of paraflocculus in smooth pursuit eye movement control revealed by chemical lesion. AB - The primate lobulus petrosus (LP) of the cerebellar paraflocculus receives inputs from visual system-related pontine nuclei, and projects to eye movement-related cerebellar nuclei. To reveal a potential involvement of LP in oculomotor control, we lesioned LP unilaterally by local injections of ibotenic acid in three Macaca fuscata. We examined the effects of lesion on eye movements evoked by step (3 degrees )-ramp (5-15 degrees/s) moving target. To step-ramp moving target, the monkeys showed an initial slow eye movement and later a small catch-up saccade, which was followed by the post-saccadic pursuit nearly matching to the velocity of the ramp target motion. After LP lesioning, the velocity of post-saccadic pursuits in the ipsiversive and down-ward directions decreased by 20-40% in all three monkeys. These deficits lasted for at least 1 month, and some recovery was observed. In the amplitudes of catch-up saccades, no consistent changes were seen among the three monkeys after LP lesioning. These results suggest an involvement of LP in the primate smooth pursuit eye movement control. PMID- 18164088 TI - Screening of Argentine plant extracts: impact on growth parameters and aflatoxin B1 accumulation by Aspergillus section Flavi. AB - The effect of essential oils, ethanolic and aqueous extract of 41 vegetable species on Aspergillus section Flavi growth was evaluated. The in vitro screen was a two-stage process. A wide-spectrum initial screen which identified promising antifungal plant extracts was carried out first. After that, identified extracts were studied in more detail by in vitro assays. A total of 96 plant extracts were screened. Essential oils were found to be the most effective extract controlling aflatoxigenic strains. Clove, mountain thyme, poleo and eucalyptus essential oils were selected to study their antifungal effect. Studies on percentage of germination, germ-tube elongation rate, growth rate, and aflatoxin B1 accumulation were carried out. Clove, mountain thyme and poleo essential oils showed the most antifungal effect under all growth parameters analyzed as well as aflatoxin B1 accumulation. Our results suggest that mountain thyme and poleo, which are native vegetal species of Argentina, and clove essential oils could be used alone or in conjunction with other substances to control the presence of aflatoxigenic fungi in stored maize. PMID- 18164089 TI - Foodborne disease outbreaks caused by sucrose-nonfermenting and beta galactosidase-deficient variants of Vibrio cholerae. AB - We reported four foodborne disease outbreaks in Taiwan caused by sucrose nonfermenting and by beta-galactosidase-deficient variants of non-O1, non-O139 Vibrio cholerae. The sucrose-nonfermenting vibrios collected from three outbreaks were biochemically identified to be V. mimicus and the beta-galactosidase deficient vibrios from an outbreak to be V. alginolyticus. However, molecular methods including DNA-DNA hybridization, fatty acid profile analysis, and sequence analysis of 16S rRNA, oriC, pyrH, recA, and rpoA indicated that these vibrios should be V. cholerae. These V. cholerae variants carried two hemolysin genes, hlyA and hlx, but contained neither cholera toxin gene, ctx, V. mimicus hemolysin gene, vmh, nor thermo-directed hemolysin, tdh. The sucrose nonfermenting variants of V. cholerae shared a high level of genetic relatedness; they could derive from a common clone. In our record from 1995 to date, this was the first time that V. cholerae variants were discovered as etiologic agents for foodborne disease outbreaks in Taiwan. PMID- 18164090 TI - [Exercise training for lower limb amputees]. AB - Lower limb amputee have lower exercise capacities, proportionally to the delay necessary to use their well-fitted prosthesis. Exercise training is a valid therapeutic to improve local factors (residual limb), muscle strength and endurance, locomotor performance and to decrease the cardiovascular risk factors. The programs for exercise training used for amputees are derivate from the vascular diseases and adapted (upper limb ergometer, cycloergometer with intact limb, pharmacological stress). Exercise training must be personalised because the population with lower limb amputation is very heterogeneous for deficiency and capacity (orthopaedic, vascular and cardiac) and for their socioprofessional project. PMID- 18164092 TI - [Early muscular strengthening after total hip arthroplasty: association of two models of rehabilitation]. AB - OBJECTIVE: To assess the effect of combining two rehabilitation models during early-stage muscle reinforcement in patients having undergone total hip arthroplasty (THA). METHODOLOGY: Our study was carried out on a total of 81 unilateral THA patients aged between 61 and 84. Forty-one control group (CG) patients participated in a standard rehabilitation (SR) program, whereas the 40 patients in the test group (TG) performed both SR and isometric exercises with electromyographic feedback. We performed three tests in each group, on the day before surgery (T(1)) and then three days (T(2)) and seven days (T(3)) after surgery. These tests enabled measurement of the integrated electromyographic peak (EMG(ipeak)) for the gluteus medius, the peak voluntary isometric strength (VIS(peak)) of each leg (i.e., both the operated and nonoperated sides) and the operated gluteus medius' maintenance time at 50% of the EMG(ipeak) during isometric exercises with electromyographic feedback. RESULTS: For the operated gluteus medius, the TG's EMG(ipeak) values were similar to those measured in the CG (P>0.05) at T(1) and T(2). Likewise, the VIS(peak) values for the corresponding legs at T(1) and T(2) did not differ significantly. However, at T(3), the EMG(ipeak) of the gluteus medius and the leg VIS(peak) on the operated side increased in both groups and were higher in the TG than in the CG (P<0.001). Compared with the CG, the TG also displayed a longer maintenance time at 50% of the EMG(ipeak) for the operated gluteus medius during isometric exercises with electromyographic feedback. CONCLUSION: This preliminary study showed that a combination of two rehabilitation models leads to faster post-THA recovery than the application of SR alone. PMID- 18164093 TI - [Thoracic aorta aneurysm as a complication after transpedicular stabilization of the thoracic vertebral column--a case study]. AB - INTRODUCTION: Incorrectly positioned transpedicular stabilization accounts for 4 to 30% of the complications occurring after vertebral column stabilization operations. OBSERVATIONS: We describe a specific complication following transpedicular stabilization of TH5-TH7, performed due to fracture of the TH6 vertebra. Given that the patient complained of pain in the thoracic vertebrae, a CT scan of the TH5-TH7 region was performed and revealed that the transpedicular stabilization was incorrectly located; it was immediately adjacent to the thoracic aorta and had led to the formation of a false aneurysm. The stabilization was removed and the patient is under the constant supervision of a vascular surgeon. CONCLUSIONS: Thoracic aorta false aneurysm is a rare complication after vertebral column stabilization. However, there is a high risk of the aorta wall damage during stabilization removal. PMID- 18164094 TI - A comparison of preprandial insulin glulisine versus insulin lispro in people with Type 2 diabetes over a 12-h period. AB - A comparison of the plasma glucose and insulin day profiles between two prandial rapid-acting insulin analogues, insulin glulisine (glulisine) and insulin lispro (lispro), in 18 obese subjects with Type 2 diabetes. Subjects (body mass index: males, 36.7 [33.2-43.8] kg/m(2); females, 40.0 [35.7-46.5] kg/m(2)) received subcutaneous glulisine or lispro (0.15 U/kg) at 4-h intervals immediately (within 2 min) before three standard test meals during each of two 12-h, randomised, open label, crossover studies (7+/-2-day interval between each). Overall, preprandial subtracted glucose concentrations (area under the curve) were similar on the glulisine and lispro study days. However, the mean of the three maximal preprandial subtracted plasma glucose concentrations (DeltaGLU(max)) were lower with glulisine versus lispro (12%; p<0.01). Mean concentrations of insulin analogue were significantly higher post-meal with glulisine (p<0.01 for all). Post hoc analysis showed a significantly faster absorption rate for glulisine versus lispro in the first 30 min post-meal (estimated difference 0.48 microU/min; p<0.0001). Only two cases of hypoglycaemia were reported; both from one subject during the lispro day. When glulisine is injected immediately before a meal in obese patients with Type 2 diabetes, glulisine achieves significantly lower glucose excursions over lispro. Significantly faster absorption with higher and sustained post-meal levels of insulin analogue was achieved at every meal with glulisine versus lispro. PMID- 18164095 TI - Automatic covariate selection in logistic models for chest pain diagnosis: a new approach. AB - A newly established method for optimizing logistic models via a minorization majorization procedure is applied to the problem of diagnosing acute coronary syndromes (ACS). The method provides a principled approach to the selection of covariates which would otherwise require the use of a suboptimal method owing to the size of the covariate set. A strategy for building models is proposed and two models optimized for performance and for simplicity are derived via 10-fold cross validation. These models confirm that a relatively small set of covariates including clinical and electrocardiographic features can be used successfully in this task. The performance of the models is comparable with previously published models using less principled selection methods. The models prove to be portable when tested on data gathered from three other sites. Whilst diagnostic accuracy and calibration diminishes slightly for these new settings, it remains satisfactory overall. The prospect of building predictive models that are as simple as possible for a required level of performance is valuable if data-driven decision aids are to gain wide acceptance in the clinical situation owing to the need to minimize the time taken to gather and enter data at the bedside. PMID- 18164096 TI - Digital-analog hybrid control model for eukaryotic heat shock response illustrating the dynamics of heat shock protein 70 on exposure to thermal stress. AB - We are introducing in this paper a digital-analog hybrid model approach for the study of a complete gene regulatory network; the heat shock response (HSR) network of eukaryotes. HSR is a crucial and widely studied cellular phenomenon occurring due to various stresses on the cell, and is characterised by the induction of heat shock genes resulting in the production of heat shock proteins (HSPs) which restores cellular homeostasis by maintaining protein integrity. We are proposing a model which incorporates simple digital and analog components which mimic the functioning of biological molecules involved in HSR and model their dynamics and behaviour. The simulation result of the circuit for the production of HSP70 has been found to be consistent with published experimental results. The qualitative behaviour of the HSR is expressed through a truth table. Through this novel approach, the authors have tried to develop a level of understanding of the interactions of the parts of the HSR system and of this system as a whole. PMID- 18164097 TI - A novel electromyographic signal simulator for muscle contraction studies. AB - Mathematical simulation has been widely used in biomedical and biological sciences. In the case of the surface electromyographic (SEMG) activity, some models have been proposed aiming to study muscle contraction strategies that are used during different tasks and conditions. Most of SEMG simulators are based on energy modulation of a Gaussian noise. This work proposes a novel simulator in which the user-defined parameters are associated with the motor units (MUs) recruitment and their firing rate. Comparison between the mean spectrum of real SEMG signals collected in isometric contraction of the muscle biceps brachii and the mean spectrum obtained from simulated SEMG signals showed a good agreement, pointing the proposed simulator seems to be capable to generate consistent electromyographic signals in time and frequency domains and that can be used in many studies, in particular in the evaluation of automatic methods aimed to detect muscular contraction. PMID- 18164098 TI - Wavelet-based ECG compression by bit-field preserving and running length encoding. AB - Efficient electrocardiogram (ECG) compression can reduce the payload of real-time ECG transmission as well as reduce the amount of data storage in long-term ECG recording. In this paper an ECG compression/decompression architecture based on the bit-field preserving (BFP) and running length encoding (RLE)/decoding schemes incorporated with the discrete wavelet transform (DWT) is proposed. Compared to complex and repetitive manipulations in the set partitioning in hierarchical tree (SPIHT) coding and the vector quantization (VQ), the proposed algorithm has advantages of simple manipulations and a feedforward structure that would be suitable to implement on very-large-scale integrated circuits and general microcontrollers. PMID- 18164099 TI - The influence of provider characteristics on resection rates and survival in patients with localized non-small cell lung cancer. AB - Surgery remains the mainstay of treatment for localized non-small cell lung cancer (NSCLC). However, wide variations have been reported regarding rates of operative therapy. We examined the influence of characteristics of the hospital of diagnosis on the likelihood of receiving surgical treatment and on survival. We evaluated patients with primary, first-time, localized NSCLC diagnosed from 1998 to 2003 in the region of the Amsterdam Cancer Registry. Treatment and survival data were extracted from the registry database. We investigated which provider characteristics (hospital category, mean annual lung cancer caseload, presence of a cardiothoracic surgery unit) were predictive of receiving surgical treatment and of survival. 1591 patients were diagnosed with clinically localized NSCLC, of which 1097 (69%) had surgery. Resection rates varied significantly between the various hospitals (48-90%, chi(2), P<0.001). Patients diagnosed at specialized centers or higher volume hospitals were more likely to receive surgical therapy, especially for patients over 80 years of age. In addition, there was a trend that octogenarians had higher odds of undergoing surgery when diagnosed in a center with a cardiothoracic surgery unit. Patients had a better survival after resection than without surgery (P<0.001). Survival after surgery did not differ between the various hospital categories. In conclusion, there is wide institutional variability in rates of surgical treatment in lung cancer patients. In addition to patient characteristics, attributes of the hospital of diagnosis also have significant influence on the likelihood of receiving surgical therapy. Future studies should examine the underlying mechanisms for this phenomenon. PMID- 18164100 TI - Role of carnosine in preventing thioacetamide-induced liver injury in the rat. AB - Carnosine (beta-alanyl-L-histidine) is a dipeptide with antioxidant properties. Free radicals are involved in the pathogenesis of acute liver injury induced by thioacetamide (TAA). In this study, we investigated the effect of carnosine treatment on TAA-induced oxidative stress and hepatotoxicity. Rats were injected intraperitoneally with TAA (500 mg/kg) and carnosine (250 mg/kg, intraperitoneal) was co-administered with TAA. All animals were killed 24 h after injections. TAA administration resulted in hepatic necrosis, significant increases in plasma transaminase activities as well as hepatic lipid peroxide levels. In addition, hepatic antioxidant system was found to be depressed following TAA administration. When carnosine was co-administered with TAA in rats, plasma transaminase activities were found to approach to normal values in rats. Histological findings also suggested that carnosine has preventive effect on TAA induced hepatic necrosis. Carnosine treatment caused significant decreases in lipid peroxide levels in TAA-treated rats without any changes in enzymatic and non-enzymatic antioxidants except vitamin E in the liver of rats. Our findings indicate that carnosine, in vivo may have a preventive effect on TAA-induced oxidative stress and hepatotoxicity by acting as an non-enzymatic antioxidant itself. PMID- 18164101 TI - Cyclo-glycyl-glutamine inhibits ethanol intake in P and Sprague-Dawley rats. AB - Peptide inhibitors of ethanol consumption have shown promise. The purpose of this study was to test the cyclized form of the opioid-derived dipeptide, glycyl-L glutamine to reduce ethanol consumption after either peripheral injections or site-specific injections into the nucleus accumbens (NAC) of high drinking and low drinking rats. Following I.P. cyclo-glycyl-glutamine (c-GQ), the data show a mean decrease in ethanol intake of 34.4% in P rats, and 39.4% in Sprague-Dawley rats at doses between 5 and 25mg/kg. The data show that peripherally administered c-GQ is effective in reducing ethanol consumption in both high (P) and low (SD) drinking strains of rats and suggests a therapeutic potential. PMID- 18164102 TI - Apoptosis induced by domoic acid in mouse cerebellar granule neurons involves activation of p38 and JNK MAP kinases. AB - In mouse cerebellar granule neurons (CGNs) the marine neurotoxin domoic acid (DomA) induces neuronal cell death, either by apoptosis or by necrosis, depending on its concentration, with apoptotic damage predominating in response to low concentrations (100 nM). DomA-induced apoptosis is due to selective activation of AMPA/kainate receptors, and is mediated by DomA-induced oxidative stress, leading to mitochondrial dysfunction and activation of caspase-3. The p38 MAP kinase and the c-Jun NH2-terminal protein kinase (JNK) have been shown to be preferentially activated by oxidative stress. Here we report that DomA increases p38 MAP kinase and JNK phosphorylation, and that this effect is more pronounced in CGNs from Gclm (-/-) mice, which lack the modifier subunit of glutamate-cysteine ligase, have very low glutathione (GSH) levels, and are more sensitive to DomA-induced apoptosis than CGNs from wild-type mice. The increased phosphorylation of JNK and p38 kinase was paralleled by a decreased phosphorylation of Erk 1/2. The AMPA/kainate receptor antagonist NBQX, but not the NMDA receptor antagonist MK 801, prevents DomA-induced activation of p38 and JNK kinases. Several antioxidants (GSH ethyl ester, catalase and phenylbutylnitrone) also prevent DomA induced phosphorylation of JNK and p38 MAP kinases. Inhibitors of p38 (SB203580) and of JNK (SP600125) antagonize DomA-induced apoptosis. These results indicate the importance of oxidative stress-activated JNK and p38 MAP kinase pathways in DomA-induced apoptosis in CGNs. PMID- 18164103 TI - Interactions between the products of the Herpes simplex genome and Alzheimer's disease susceptibility genes: relevance to pathological-signalling cascades. AB - The products of the Herpes simplex (HSV-1) genome interact with many Alzheimer's disease susceptibility genes or proteins. These in turn affect those of the virus. For example, HSV-1 binds to heparan sulphate proteoglycans (HSPG2), or alpha-2-macroglobulin (A2M), and enters cells via nectin receptors, which are cleaved by gamma-secretase (APH1B, PSEN1, PSEN2, PEN2, NCSTN). The virus also binds to blood-borne lipoproteins and apolipoprotein E (APOE) is able to modify its infectivity. Viral uptake is cholesterol- and lipid raft-dependent (DHCR24, HMGCR, FDPS, RAFTLIN, SREBF1). The virus is transported to the nucleus via the dynein and kinesin (KNS2) motors associated with the microtubule network (MAPT). Amyloid precursor protein (APP) plays a role in this transport. Nuclear export is mediated via disruption of the nuclear lamina and binding to LMNA. Herpes simplex activates kinases (CDC2 and casein kinase 2) whose substrates include APOE, APP, MAPT, PSEN2, and SREBF1. A viral protein is also able to delete mitochondrial DNA, a situation prevalent in Alzheimer's disease. The virus binds to the host transcription factors transcription factor CP2 (TFCP2) and POU2F1 that control many other genes associated with Alzheimer's disease. Viral latency is controlled by IL6 and IL1B and at different stages of its life cycle the virus can either promote or attenuate apoptosis via Fas and tumor necrosis factor pathways (FAS, TNF, DAPK1, PARP1). Viral evasion strategies include inhibition of the antigen processor TAP2, the production of an Fc immunoglobulin receptor mimic (FCER1G) and inhibition of the viral-activated kinase EIF2AK2. These and other host/viral interactions, targeted to certain Alzheimer's disease susceptibility genes, support the idea that some form of synergy between the pathogen and genetic factors may play a role in the pathology of late-onset Alzheimer's disease. PMID- 18164104 TI - Nursing students' learning of managing cross-infections--scenario-based simulation training versus study groups. AB - Up to 33% of all health care-associated infections are preventable [Alexander, M., 2006. Nursing Practice. Hospital and Home. Elsevier, Churchill Livingstone]. Nurses must consider how they can address infection prevention, control and management within health care settings. An outbreak of any infectious disease can occasionally occur, even when protocols have been followed. A well-coordinated, multidiscipline response will minimize the impact of the outbreak by controlling and managing the impact and ongoing risks. In this study nursing students were challenged to manage cross-infections based on two hypothetical cases, MRSA and Norovirus. The purpose of the study was to determine the most efficient teaching strategies for managing cross-infections and determining the teachers' role by comparing three different teaching methods; scenario-based study groups with and without teacher and simulation training. A class of 141 2nd year nursing students participated as part of their learning programme on infection control. Out of these 21 students were randomly selected to take part in three focus groups assigned to evaluate the program. Overall, the findings indicated that scenario based simulation training made the students more aware of how complex each scenario was. Events occurred that they had not expected, and this lead to a better recollection of details. By asking appropriate questions, giving feedback and hypothetical examples, the teacher's role was crucial in both teaching strategies. PMID- 18164105 TI - The impact of assessment methods on the learning of nursing students. AB - Assessment methods can influence the learning approaches of nursing students. Assessments designed to assess understanding rather than rote learning can contribute to a deep learning approach. Multiple-choice questions can be formulated at high cognitive levels for nursing assessments. This study aimed to examine the effect of high quality multiple-choice tests on the learning approaches of students enrolled in a mental health nursing course as measured by the revised two-factor study process questionnaire (R-SPQ-2F). Data were collected from 136 students in a higher diploma (HD) in nursing programme and 142 baccalaureate (BSN) nursing students. Two-step cluster analysis yielded three clusters represented by students who were predominantly deep, predominantly surface or intermediate in their orientation to learning. There was a general decrease in the deep learning score and an increase in the surface learning score of the students on the course, possibly related to the excessive workload in nursing studies. However, there was no direct association with multiple-choice assessment. The academic achievement of students correlated negatively with their surface learning approach. Students (n=31) in four focus group interviews believed that assessments constructed using scenario-based questions, simulated role-play situations and case studies could direct desired learning in order to facilitate their understanding of knowledge, their use of critical thinking, and their application of knowledge. PMID- 18164106 TI - Safety and immunogenicity of co-administered quadrivalent human papillomavirus (HPV)-6/11/16/18 L1 virus-like particle (VLP) and hepatitis B (HBV) vaccines. AB - Adolescents and young adults are at high risk for human papillomavirus (HPV) and hepatitis B virus (HBV) infections, which are preventable by currently available, safe and effective, prophylactic vaccines. However, development of a combined immunization strategy may lead to better compliance for these vaccines, thereby contributing to the overall goal of protection against these diseases. This study assessed the safety and immunogenicity of co-administered quadrivalent HPV 6/11/16/18 L1 VLP and HBV vaccines in women (n=1877) aged 16-23 years. Co administration of HPV and HBV vaccines induced robust anti-HPV-6, HPV-11, HPV-16, HPV-18 geometric mean titers (GMTs) and > or =99% seroconversion rates (Month 7) that were both non-inferior (p<0.001) to those induced by HPV vaccine alone. High Month 7 anti-HBs GMTs were also observed following concomitant vaccination. These GMTs were lower compared to those induced by the HBV vaccine itself; however, >96% of subjects achieved an anti-HBs seroprotection level of > or =10 mIU/mL that was non-inferior (p<0.001) to that of HBV vaccine alone. Overall, co administered and individual vaccines were generally well-tolerated and did not interfere with the immune response of either vaccine (ClinicalTrials.gov number, NCT00092521). PMID- 18164107 TI - Dynamic model for the assessment of radiological exposure to marine biota. AB - A generic approach has been developed to simulate dynamically the uptake and turnover of radionuclides by marine biota. The approach incorporates a three compartment biokinetic model based on first order linear kinetics, with interchange rates between the organism and its surrounding environment. Model rate constants are deduced as a function of known parameters: biological half lives of elimination, concentration factors and a sample point of the retention curve, allowing for the representation of multi-component release. The new methodology has been tested and validated in respect of non-dynamic assessment models developed for regulatory purposes. The approach has also been successfully tested against research dynamic models developed to represent the uptake of technetium and radioiodine by lobsters and winkles. Assessments conducted on two realistic test scenarios demonstrated the importance of simulating time dependency for ecosystems in which environmental levels of radionuclides are not in equilibrium. PMID- 18164108 TI - Plant induced changes in concentrations of caesium, strontium and uranium in soil solution with reference to major ions and dissolved organic matter. AB - For a better understanding of the soil-to-plant transfer of radionuclides, their behavior in the soil solution should be elucidated, especially at the interface between plant roots and soil particles, where conditions differ greatly from the bulk soil because of plant activity. This study determined the concentration of stable Cs and Sr, and U in the soil solution, under plant growing conditions. The leafy vegetable komatsuna (Brassica rapa L.) was cultivated for 26 days in pots, where the rhizosphere soil was separated from the non-rhizosphere soil by a nylon net screen. The concentrations of Cs and Sr in the rhizosphere soil solution decreased with time, and were controlled by K+NH(4)(+) and Ca, respectively. On the other hand, the concentration of U in the rhizosphere soil solution increased with time, and was related to the changes of DOC; however, this relationship was different between the rhizosphere and non-rhizosphere soil. PMID- 18164109 TI - Availability and immobilization of 137Cs in subtropical high mountain forest and grassland soils. AB - To understand the behavior of (137)Cs in undisturbed soils after nuclear fallout deposition between the 1940s and 1980s, we investigated the speciation of (137)Cs in soils in forest and its adjacent grassland from a volcano and subalpine area in Taiwan. We performed sequential extraction of (137)Cs (i.e., fractions readily exchangeable, bound to microbial biomass, bound to Fe-Mn oxides, bound to organic matter, persistently bound and residual). For both the forest and grassland soils, (137)Cs was mainly present in the persistently bound (31-41%) and residual (22-62%) fractions. The proportions of (137)Cs labile fractions--bound to exchangeable sites, microbial biomass, Mn-Fe oxides, and organic matter--were lower than those of the recalcitrant fractions. The labile fractions in the forest soils were also higher than those in the grassland soils, especially in the volcanic soil. The results suggest that the labile form of (137)Cs was mostly transferred to the persistently bound and resistant fractions after long-term deposition of fallout. The readily exchangeable (137)Cs fraction was higher in soils with higher organic matter content or minor amounts of 2:1 silicate clay minerals. PMID- 18164110 TI - Base cation depletion, eutrophication and acidification of species-rich grasslands in response to long-term simulated nitrogen deposition. AB - Pollutant nitrogen deposition effects on soil and foliar element concentrations were investigated in acidic and limestone grasslands, located in one of the most nitrogen and acid rain polluted regions of the UK, using plots treated for 8-10 years with 35-140 kg N ha(-2)y(-1) as NH(4)NO(3). Historic data suggests both grasslands have acidified over the past 50 years. Nitrogen deposition treatments caused the grassland soils to lose 23-35% of their total available bases (Ca, Mg, K, and Na) and they became acidified by 0.2-0.4 pH units. Aluminium, iron and manganese were mobilised and taken up by limestone grassland forbs and were translocated down the acid grassland soil. Mineral nitrogen availability increased in both grasslands and many species showed foliar N enrichment. This study provides the first definitive evidence that nitrogen deposition depletes base cations from grassland soils. The resulting acidification, metal mobilisation and eutrophication are implicated in driving floristic changes. PMID- 18164111 TI - Women and prostate cancer support groups: the gender connect? AB - There are more than 100 prostate cancer support groups (PCSGs) in Canada, most of which meet on a monthly basis-yet little attention has been paid to the role of women at these groups. As part of an ongoing ethnographic study of PCSGs, we examined women's motivations for attending the groups, their ways of functioning in PCSGs and the benefits they accrued. Participant observations conducted at 13 British Columbian-based PCSGs and individual interview data from 20 women who regularly attended PCSG meetings were analyzed. Although the groups did not overtly limit women's attendance, the women's decisions to attend and their participation at group meetings were subject to much self-reflection, uncertainty and tension. Motivations to access a PCSG included a desire to support their partners, develop understandings about the illness and disease, and to manage their own experience of prostate cancer. Our analyses revealed that women assume three roles in PCSGs: social facilitator, background supporter and cancer co survivor. The women reported many interrelated benefits as a result of attending, including information, hope and reassurance, and connecting with other women in similar circumstances. The results from this study reveal how traditional feminine ideals, such as nurturing and caring for the men in their lives, facilitating social connections and the desire to share emotional experiences guided the behaviors. Based on the study findings, we suggest that efforts to support women's involvement in PCSGs are critical to enhancing the effectiveness of the groups for both men and women. PMID- 18164112 TI - Family context of mental health risk in Tsunami affected mothers: findings from a pilot study in Sri Lanka. AB - This study investigated direct and indirect influences of Tsunami exposure on mothers' PTSD and depressive symptoms using survey data from 325 Tsunami-affected families living in two villages in southern Sri Lanka. Findings generally support the hypothesized model in that life and property destruction contributed to the PTSD and depressive symptoms of mothers. Detrimental influences of Tsunami exposure also operated through the generation of more proximal secondary Tsunami risks such as post-Tsunami family problems. In addition, religious participation, familism, number of children, intact family status, and community support reduced mothers' depressive symptoms. Finally, intact family status and high religious participation moderated the detrimental influence of Tsunami exposure on mothers' PTSD symptoms. These findings can contribute directly to improving ongoing recovery and reconstruction programs and help to formulate future programs for families affected by the Tsunami and other natural disasters. PMID- 18164113 TI - Relationship of race and poverty to lower extremity function and decline: findings from the Women's Health and Aging Study. AB - Race- and poverty-related disparities in physical function are well documented, though little is known about effects of race and poverty on functional decline and the progression of disability. We examined cross-sectional and longitudinal relationships between race, poverty and lower extremity function using data from moderately to severely disabled women in the U.S. Women's Health and Aging Study. Severity of lower extremity functional limitation was determined from scaled responses of reported difficulty walking (1/4) mile, walking across a room, climbing stairs, and stooping, crouching or kneeling. Usual walking speed assessed over 4m was our objective measure of function. Of the 996 women who described themselves as black or white, 284 (29%) were black and 367 (37%) were living at or below 100% of the federal poverty level. Independent of demographic and health-related factors, among white women, the poor exhibited consistently worse lower extremity function than the non-poor; this association, however, was not observed in black women. Among the non-poor, black women had slower walking speeds, and reported more limitation in lower extremity function than their non poor white counterparts, even after adjusting for demographic variables and health-related characteristics. After 3 years, accounting for baseline function, demographic and health-related factors, race and poverty status were unrelated to functional decline. Thus, while race and poverty status were associated with functional deficits in old age, they do not appear to impact the rate of functional decline or progression of disability over 3 years. PMID- 18164114 TI - Provider and clinic cultural competence in a primary care setting. AB - A multilevel approach that enhances the cultural competence of clinicians and healthcare systems is suggested as one solution to reducing racial/ethnic disparities in healthcare. The primary objective of this cross-sectional study was to determine if there is a relationship between the cultural competence of primary care providers and the clinics where they work. Forty-nine providers from 23 clinics in Baltimore, Maryland and Wilmington, Delaware, USA completed an on line survey which included items assessing provider and clinic cultural competence. Using simple linear regression, it was found that providers with attitudes reflecting greater cultural motivation to learn were more likely to work in clinics with a higher percent of nonwhite staff, and those offering cultural diversity training and culturally adapted patient education materials. More culturally appropriate provider behavior was associated with a higher percent of nonwhite staff in the clinic, and culturally adapted patient education materials. Enhancing provider and clinic cultural competence may be synergistic strategies for reducing healthcare disparities. PMID- 18164115 TI - Protective effect of minocycline, a semi-synthetic second-generation tetracycline against 3-nitropropionic acid (3-NP)-induced neurotoxicity. AB - 3-Nitropropionic acid (3-NP) is an irreversible inhibitor of the electron transport enzyme succinate dehydrogenase, a mitochondrial Complex II enzyme. Minocycline is a semi-synthetic second-generation tetracycline with neuroprotective activity and has the capability to effectively cross the blood brain barrier. We investigated the effects of minocycline on behavioral, biochemical, inflammation related and neurochemical alterations induced by the sub-chronic administration of 3-nitropropionic acid to rats. Chronic pre administration of minocycline (50 and 100mg/kg) dose dependently prevented 3-NP induced dysfunction behavioral (hypoactivity, memory retention, locomotor and rota-rod activity). In addition, 3-NP produced a marked increase in lipid peroxidation levels whereas decreased the activities of catalase and succinate dehydrogenase. In contrast, pretreatment of 3-NP injected rats with minocycline resulted in the attenuation of all these alterations. A marked increase in an inflammatory cytokine TNF-alpha by 3-NP was also decreased by minocycline treatment. Neurochemically, the administration of 3-NP significantly decreased the levels of catecholamines in the brain homogenates (dopamine, norepinephrine and serotonin) which were reversed by pretreatment of minocycline. The present finding explains the neuroprotective effect of minocycline against 3-NP toxicity by virtue of its antioxidant and anti-inflammatory activity. PMID- 18164116 TI - beta-Naphthoflavone enhances oxidative stress responses and the induction of preneoplastic lesions in a diethylnitrosamine-initiated hepatocarcinogenesis model in partially hepatectomized rats. AB - The tumour-promoting effects of beta-naphthoflavone (BNF), a novel aryl hydrocarbon receptor (AhR) agonist, were investigated using a medium-term hepatocarcinogenesis model in rats. Six-week-old male F344 rats received an intraperitoneal injection of N-diethylnitrosamine (DEN) at a dose of 200mg/kg body weight and were fed a diet containing 0% (basal diet), 0.5% or 1% BNF for 6 weeks from 2 weeks after DEN treatment. All animals were subjected to two-thirds partial hepatectomy 1 week after the BNF treatment. The number and area of glutathione S-transferase placental form (GST-P) positive foci significantly increased in the livers of rats treated with BNF with concomitantly increased cell proliferation compared to those in the livers of the DEN alone group. Global gene expression analysis and subsequent quantitative real-time reverse transcription-polymerase chain reaction revealed that BNF induced not only the 'AhR gene battery'Cyp1a1, Cyp1a2, Cyp1b1, Nqo1, Aldh3a1 and Ugt1a6 but also the transcription factor NF-E2-related factor 2 (Nrf2)-regulated genes such as Gstm1, Gpx2, Akr7a3 and Yc2 (and also Nqo1), presumably due to the adaptive response against BNF-triggered oxidative stress responses. Reactive oxygen species production increased in microsomes isolated from the livers of BNF-treated rats, and this enhancement was suppressed by the P450 inhibitor SKF-525A. Furthermore, BNF enhanced oxidative DNA damage and lipid peroxidation, estimated by the levels of 8-hydroxydeoxyguanosine (8-OHdG) and thiobarbituric acid-reactive substances. These results suggest that the administration of BNF at a high dose and over a long-term enhance oxidative stress responses which may contribute to its hepatocarcinogenic potential in rats. PMID- 18164117 TI - Goal-directed hemodynamic optimization in the post-cardiac arrest syndrome: a systematic review. AB - AIMS: The treatment recommendations from the 2005 International Consensus Conference on Cardiopulmonary Resuscitation and Emergency Cardiovascular Care Science (hosted by the American Heart Association) advocate a goal-directed treatment strategy for hemodynamic optimization after return of spontaneous circulation (ROSC) in post-cardiac arrest care. We performed a systematic review to (1) examine the available evidence for goal-directed hemodynamic support in the post-cardiac arrest syndrome, (2) determine the effect of such a treatment strategy on survival, and (3) define the specific hemodynamic goals, if any, that have been tested in clinical trials of post-cardiac arrest patients. METHODS: We conducted a systematic review of the Cochrane Library, MEDLINE, CINAHL, conference proceedings, clinical practice guidelines, and other sources using a comprehensive strategy to identify randomized controlled trials and quasi experimental studies of goal-directed hemodynamic optimization in patients with ROSC after cardiac arrest. RESULTS: The comprehensive search yielded a total of 1184 potential publications and after a relevance screen, five studies were eligible for full article review. None of the studies were eligible for inclusion in the final analysis. CONCLUSIONS: To date, no clinical trials have examined hemodynamic optimization in post-cardiac arrest patients. Although clinical acumen may support the concept that hemodynamic derangements after ROSC should be normalized, there is currently no evidence available to indicate the best strategy for goal-directed hemodynamic support. The current study indicates the need for future clinical investigations designed to determine both the efficacy of hemodynamic optimization in post-cardiac arrest patients and the best endpoints to target as part of a goal-directed strategy. PMID- 18164118 TI - A case of Commotio cordis in a young child caused by a fall. AB - Commotio cordis or ventricular fibrillation caused by a blow to the chest is a rare cause of cardiac arrest in a well child. We report a case of a young child falling from a low height landing chest first with rapid onset of unconsciousness, apnoea and cyanosis. Cardiopulmonary resuscitation was given by parents under telephone instruction from an ambulance dispatch centre. On arrival of officers, 7 min after the fall, ventricular fibrillation was present but responded to defibrillation (biphasic 3 J/kg). No clinical or CT evidence of chest or brain trauma was present and investigations (ECGs, cardiac MRI, echocardiography, viral tests, metabolic tests, drug tests, serum electrolytes) did not reveal any cardiac illness or abnormal cardiac anatomy. Specifically, a long QT was absent and a Flecainide challenge for Brugada syndrome was negative. There was no family history of sudden death. No further dysrrhythmia occurred and the child recovered neurologically well after 3 days of therapeutic hypothermia (for cerebral ischaemia) and 7 days of mechanical ventilation. PMID- 18164119 TI - The effectiveness of cardiopulmonary resuscitation instruction: animation versus dispatcher through a cellular phone. AB - INTRODUCTION: We developed a cardiopulmonary resuscitation (CPR) instruction programme using motion capture animation integrated into cellular phones. We compared the effectiveness of animation-assisted CPR instruction with dispatcher assisted instruction in participants with no previous CPR training. METHODS: This study was a single blind cluster randomized trial. Participants were allocated to either animation-assisted CPR (AA-CPR; 8 clusters, 44 participants) group or dispatcher-assisted CPR (DA-CPR; 8 clusters, 41 participants). The overall performance and time of each step of CPR cycle were recorded on a checklist by 3 assessors. The objective performances were evaluated using the Resusci Anne SkillReporter Manikin. Differences between the groups were compared using an independent t-test adjusted for the effect of clustering. RESULTS: The AA-CPR group had a significantly better checklist score (p<0.001) and time to completion of 1 CPR cycle (p<0.001) than the DA-CPR group. In an objective assessment of psychomotor skill, the AA-CPR group demonstrated more accurate hand positioning (68.8+/-3.6%, p=0.033) and compression rate (72.4+/-3.7%, p=0.015) than DA-CPR group. However, the accuracy of compression depth (p=0.400), ventilation volume (p=0.977) and flow rate (p=0.627) were below 30% in both groups. CONCLUSION: Audiovisual animated CPR instruction through a cellular phone resulted in better scores in checklist assessment and time interval compliance in participants without CPR skill compared to those who received CPR instructions from a dispatcher; however, the accuracy of important psychomotor skill measures was unsatisfactory in both groups. PMID- 18164120 TI - Bilateral ovarian mucinous cystadenoma in an adolescent presenting as abdominal compartment syndrome. PMID- 18164121 TI - The missing ink. PMID- 18164122 TI - Prioritisation of farm scale remediation efforts for reducing losses of nutrients and faecal indicator organisms to waterways: a case study of New Zealand dairy farming. AB - The international competitiveness of the New Zealand (NZ) dairy industry is built on low cost clover-based systems and a favourable temperate climate that enables cows to graze pastures mostly all year round. Whilst this grazed pasture farming system is very efficient at producing milk, it has also been identified as a significant source of nutrients (N and P) and faecal bacteria which have contributed to water quality degradation in some rivers and lakes. In response to these concerns, a tool-box of mitigation measures that farmers can apply on farm to reduce environmental emissions has been developed. Here we report the potential reduction in nutrient losses and costs to farm businesses arising from the implementation of individual best management practices (BMPs) within this tool-box. Modelling analysis was carried out for a range of BMPs targeting pollutant source reduction on case-study dairy farms, located in four contrasting catchments. Due to the contrasting physical resources and management systems present in the four dairy catchments evaluated, the effectiveness and costs of BMPs varied. Farm managements that optimised soil Olsen P levels or used nitrification inhibitors were observed to result in win-win outcomes whereby nutrient losses were consistently reduced and farm profitability was increased in three of the four case study farming systems. Other BMPs generally reduced nutrient and faecal bacteria losses but at a small cost to the farm business. Our analysis indicates that there are a range of technological measures that can deliver substantial reductions in nutrient losses to waterways from dairy farms, whilst not increasing or even reducing other environmental impacts (e.g. greenhouse gas emissions and energy use). Their implementation will first require clearly defined environmental goals for the catchment/water body that is to be protected. Secondly, given that the major sources of water pollutants often differed between catchments, it is important that BMPs are matched to the physical resources and management systems of the existing farm businesses. PMID- 18164123 TI - 1-Bromopropane induces macrophage activation via extracellular signal-regulated kinase 1/2 MAPK and NF-kappaB pathways. AB - 1-Bromopropane (1-BP) has been used in the workplace as an alternative to ozone depleting solvents. This study examined the effects of 1-BP on the production of nitric oxide (NO) and on proinflammatory cytokines, and analyzed the mechanisms involved in macrophages. 1-BP dose-dependently induced the production of NO and proinflammatory cytokines, such as IL-1beta, IL-6, and TNF-alpha, and expression levels of these genes also increased in a dose-dependent manner. The NF-kappaB sites were identified in the promoter of the iNOS and proinflammatory cytokine genes. Transient transfection and electrophoretic mobility shift assays revealed that NF-kappaB-mediated the 1-BP-induced increase in the iNOS and proinflammatory cytokine expression levels. Pretreating the macrophages with the NF-kappaB inhibitor, BAY 11-7082, and the ERK inhibitor, PD98059, inhibited NO production and iNOS expression induced by 1-BP. This demonstrates that 1-BP stimulates macrophage activation via NF-kappaB transactivation and ERK1/2 MAP kinase phosphorylation. These results suggest that 1-BP has the potential to be inflammatory and that it has previously unrecognized immunomodulating activity. PMID- 18164124 TI - Adipocyte culture medium stimulates production of macrophage inhibitory cytokine 1 in MDA-MB-231 cells. AB - Obesity is one of the potential risk factors in causing breast cancer. As a result, adipose tissue surrounding breast ductal cells may play an important role in the breast cancer development or progression. To identify the genes that are regulated by factors secreted from adipocytes in breast cancer cells, MDA-MB-231 cells were treated with the culture medium of adipocytes. Most of induced genes were related to immune function and wound healing, which share a common gene expression signature with cancer progression. In present study macrophage inhibitory cytokine 1 (MIC-1) gene was studied among the induced genes. It was found that both MIC-1 mRNA and protein were dramatically increased by the culture medium of adipocytes. Furthermore, proteinase K-treated adipocyte culture supernatants also induced MIC-1 expression. These findings indicate that proteins are not major MIC-1 inducing factors in adipocyte culture medium. Consequently, we examined the effect of free fatty acids such as palmitate and oleate on MIC-1 induction and found that palmitate markedly induced MIC-1 gene expression, whereas oleate did not. Adipocyte culture medium- and palmitate-induced MIC-1 gene expression was mediated by the activation of p38 MAPK, but not by the activation of JNK, ERK, and NF-kappaB pathway. In addition, adipocyte-CM-induced MIC-1 also increased invasiveness of MDA-MB-231 cells. PMID- 18164125 TI - Optimized and validated spectrophotometric method for the determination of uranium(VI) via complexation with meloxicam. AB - An optimized and validated spectrophotometric method has been developed for the determination of uranyl ion in the presence of other metal ions. The method is based on the chelation of uranyl ion with meloxicam via beta-diketone moiety to produce a yellow colored complex, which absorbs maximally at 398 nm. Beer's law is obeyed in the concentration range of 5-60 microg/mL with apparent molar absorptivity and Sandell's sensitivity of 5.02 x 10(4)L/mol/cm and 0.1 microg/cm2/0.001 absorbance unit, respectively. The method has been successfully applied for the determination of uranyl ion in synthetic mixture and soil samples. Results of analysis were statistically compared with those obtained by Currah's spectrophotometric method showing acceptable recovery and precision. PMID- 18164126 TI - Photocatalyzed degradation of flumequine by doped TiO2 and simulated solar light. AB - Titanium dioxide was obtained in its pure form (TiO2) and in the presence of urea (u-TiO2) and thiourea (t-TiO2) using the sol-gel technique. The obtained powders were characterized by BET surface area analysis, Infrared Spectroscopy, Diffuse Reflectance Spectroscopy and the Rietveld refinement of XRD measurements. All the prepared catalysts show high anatase content (>99%). The a and b-cell parameters of anatase increase in the order TiO2u TiO2>TiO2. The photocatalytic activities of the samples were determined on flumequine under solar-simulated irradiation. The most active catalysts were u TiO2 and t-TiO2, reaching values over 90% of flumequine degradation after 15 min irradiation, compared with values of 55% for the pure TiO2 catalyst. Changing simultaneously the catalyst amount (t-TiO2) and pH, multivariate analysis using the response surface methodology was used to determine the roughly optimal conditions for flumequine degradation. The optimized conditions found were pH below 7 and a catalyst amount of 1.6 g L(-1). PMID- 18164127 TI - Poly(vinyl pyridine-poly ethylene glycol methacrylate-ethylene glycol dimethacrylate) beads for heavy metal removal. AB - Poly(vinyl pyridine-poly ethylene glycol methacrylate-ethylene glycol dimethacrylate) [poly(VP-PEGMA-EGDMA)] beads with an average size of 30-100 microm were prepared by suspension polymerization. Poly(VP-PEGMA-EGDMA) beads were characterized by swelling studies, scanning electron microscopy (SEM), elemental analysis, Fourier Transform Infrared Spectroscopy (FTIR). The beads with a swelling ratio of 65% were used for the heavy metal removal studies. Chelation capacity of the beads for the selected metal ions, i.e., Pb(II), Cd(II), Cr(III) and Cu(II) were investigated in aqueous media containing different amounts of these ions (5-80 mg/l) and at different pH values (2.0 10.0). The maximum chelation capacities of the poly(VP-PEGMA-EGDMA) beads were 18.23 mg/g for Pb(II), 16.50 mg/g for Cd(II), 17.38 mg/g for Cr(III) and 18.25 mg/g for Cu(II). The affinity order on mass basis was observed as follows: Cu(II)>Pb(II)>Cr(III)>Cd(II). pH significantly affected the chelation capacity of VP incorporated beads. Heavy metal adsorption on the poly(PEGMA-EGDMA) control microspheres was negligible. Regeneration of the chelating beads was easily performed with 0.1 M HNO3. It was shown that these beads can be used effectively for heavy metal removal from aqueous solutions with repeatedly adsorption desorption operations. These features show that poly(VP-PEGMA-EGDMA) beads are potential candidate sorbent for heavy metal removal. PMID- 18164128 TI - Removal of Fe(II) from tap water by electrocoagulation technique. AB - Electrocoagulation (EC) is a promising electrochemical technique for water treatment. In this work electrocoagulation (with aluminum as electrodes) was studied for iron Fe(II) removal from aqueous medium. Different concentration of Fe(II) solution in tap water was considered for the experiment. During EC process, various amorphous aluminum hydroxides complexes with high sorption capacity were formed. The removal of Fe(II) was consisted of two principal steps; (a) oxidation of Fe(II) to Fe(III) and (b) subsequent removal of Fe(III) by the freshly formed aluminum hydroxides complexes by adsorption/surface complexation followed by precipitation. Experiments were carried out with different current densities ranging from 0.01 to 0.04 A/m2. It was observed that the removal of Fe(II) increases with current densities. Inter electrode distance was varied from 0.005 to 0.02 m and was found that least inter electrode distance is suitable in order to achieve higher Fe(II) removal. Other parameters such as conductivity, pH and salt concentration were kept constant as per tap water quality. Satisfactory iron removal of around 99.2% was obtained at the end of 35 min of operation from the initial concentration of 25 mg/L Fe(II). Iron concentration in the solution was determined using Atomic absorption spectrophotometer. By products obtained from the electrocoagulation bath were analyzed by SEM image and corresponding elemental analysis (EDAX). Cost estimation for the electrocoagulation was adopted and explained well. Up to 15 mg/L of initial Fe(II) concentration, the optimum total cost was 6.05 US$/m3. The EC process for removing Fe(II) from tap water is expected to be adaptable for household use. PMID- 18164129 TI - Distribution of c-Fos immunoreactive neurons in the brain after intraperitoneal injection of apelin-12 in Wistar rats. AB - The present study surveyed activation of central neurons following peripheral administration of apelin-12 (AP12), an apelin peptide homologue, by examining the distribution of neurons expressing c-Fos protein. AP12 is known to induce gastric acid secretion among other physiological functions such as regulation of circulation. It was recently reported that apelin counteracted the effect of arginine vasopressin (AVP) in the maintenance of body fluid homeostasis. We attempted to clarify which neurons in the central nervous system express c-Fos protein after intraperitoneal injection of AP12. Intraperitoneally administered AP12 induced expression of c-Fos protein in several nuclei throughout the brain. In the paraventricular nucleus of the hypothalamus (PAH), lateral hypothalamic area (LH), paraventricular nucleus of the thalamus (PVT), periaqueductal gray matter (PAG), bed nucleus of the stria terminalis (BNST), locus coeruleus (LC), lateral parabrachial nucleus (Pbl), the complex of the solitary tract nucleus (NTS) and dorsal motor nucleus of the vagus nerve (DMX), numbers of neurons expressing c-Fos protein were much higher in test than in control experiments. These findings suggest that AP12 stimulates central neurons that may play roles in the regulation of gastric acid, and hypothalamic neurons that may play roles in the maintenance of body fluid homeostasis as well as other physiological functions. PMID- 18164130 TI - Toll-like receptor 2 contributes to glial cell activation and heme oxygenase-1 expression in traumatic brain injury. AB - Traumatic brain injury is accompanied by glial cell activation around the site of the injury. In this study, we investigated the role of toll-like receptor 2 (TLR2) in glial cell activation using a stab-wound injury (SWI) model with TLR2 knock-out mice. Penetration of a normal mouse brain with a 26-G needle using a stereotaxic instrument resulted in an 18- and 4-fold upregulation of GFAP and CD11b mRNA, respectively, along the needle track in the injury area. However, in the TLR2 knock-out mice, the induced expression of these genes was reduced by 70% and 40%, respectively. Likewise, there was a reduction in the area of activated glial cells detected by immunohistochemistry and the glial cells had a less activated morphology in the TLR2 knock-out mice. In addition, the expression of the heme oxygenase-1 (HO-1) gene, a glia-expressing wound-responsive gene, was reduced after SWI in TLR2 knock-out mice. Taken together, these data argue that TLR2 contributes to the glial cell activation and HO-1 gene expression associated with traumatic brain injury. PMID- 18164131 TI - Auditory neuropathy in streptozotocin-induced diabetic mouse. AB - An investigation of the mechanism of damage to the peripheral nervous system and central nervous system in diabetes mellitus (DM) is highly important in current neurological research. Auditory neuropathy is a hearing disorder in which the auditory brainstem evoked potential is absent or severely abnormal. This study investigated auditory neuropathy caused by streptozotocin in mouse model. In order to assess diabetic auditory neuropathy, we evaluated auditory brainstem response (ABR) for the evaluation of sensorineural function in peripheral auditory nerve. Auditory middle latency response (AMLR) was employed to assess the middle response in the midbrain. STZ groups significantly increased the absolute latencies IV and the interpeak latencies I-III and I-IV of ABR compared with STZ 0 group. Pa latency of AMLR also significantly increased in proportion to STZ dosage. Taken together, our results demonstrate that STZ-induced DM may impair the auditory pathway from peripheral auditory nerve to midbrain in the mouse model. We suggest that the STZ-induced diabetic mouse model may be useful for the evaluation of auditory pathway impairment by using ABR and AMLR tests. PMID- 18164132 TI - Investigation of the dopamine D5 receptor gene (DRD5) in adult attention deficit hyperactivity disorder. AB - Several lines of evidence from neuroimaging, pharmacology and genetics support the involvement of the dopaminergic system in the etiology of Attention Deficit Hyperactivity Disorder (ADHD). Previous candidate gene studies have investigated the association between a dinucleotide (CA)(n) repeat polymorphism, located 18.5 kb from the start codon of the DRD5 gene, and ADHD. Association between the 148 bp allele and ADHD has been reported in some studies, however replication of the finding has not been consistent. We tested for an association between the (CA)(n) repeat and adult ADHD in a sample comprised of 119 families with adult ADHD probands and 88 unrelated adult ADHD cases with a corresponding number of controls matched for age, ethnicity and sex. In the family sample we found a non significant trend for association between the 148 bp allele and ADHD (Z=1.91, p=0.055). An excess of non-transmissions was detected for the 150 and 152bp alleles (Z=-2.26, p=0.023; Z=-2.20, p=0.028). Quantitative analysis performed using the Brown Attention Deficit Disorder Scale (BADDS) showed association between the 150 bp allele and lower total score (p=0.011), and lower effort (p=0.008), activation (p=0.008) and attention (p=0.01) cluster scores. We did not replicate association findings in the case-control group, likely due to the lack of statistical power of this sample. Our findings add to the literature suggesting DRD5 (CA)(n) repeat has a modest effect in modulating susceptibility to adult ADHD but further studies are required. PMID- 18164133 TI - Dorsal horn neurons having input from low back structures in rats. AB - The mechanisms of nociception in the low back are poorly understood, partly because systematic recordings from dorsal horn neurons with input from the low back are largely missing. The purpose of this investigation was to (1) identify spinal segments and dorsal horn neurons receiving input from the low back, (2) test the effect of nerve growth factor (NGF) injected into the multifidus muscle (MF) on the neurons' responsiveness, and (3) study the influence of a chronic MF inflammation on the responses. In rats, microelectrode recordings were made in the segments L2, L3, and L5 to find dorsal horn neurons having input from the low back (LB neurons). In control animals, the proportion of LB neurons in L2 was larger than in L3 and L5. Most LB neurons had a convergent input from several tissues. Injections of NGF into MF increased the proportion of LB neurons significantly. A chronic MF inflammation likewise increased the proportion of LB neurons and the input convergence. The centers of the neurons' receptive fields (RFs) were consistently located 2-3 segments caudally relative to their recording site. The results show that (1) input convergence from various tissues is common for LB neurons, (2) the input from structures of the low back is processed 2-3 segments cranially relative to the vertebral level of the RFs, and (3) the responsiveness of LB neurons is increased during a pathologic alteration of the MF. The above findings may be relevant for some cases of chronic low back pain in patients. PMID- 18164134 TI - Relapse prevention: still neglected after all these years. PMID- 18164135 TI - Clinical symptoms and diagnosis of encephalitozoonosis in pet rabbits. AB - Infections with Encephalitozoon cuniculi in rabbits are observed at increasing frequency and are known as opportunistic infections in immunocompromised humans. 191 pet rabbits with suspected encephalitozoonosis, presented at the Animal Hospital of the Veterinary University of Vienna (Austria), were included in this study. Rabbits were serologically examined for antibodies against E. cuniculi (144 positive out of 184 rabbits with suspected encephalitozoonosis compared to 14 positive out of 40 clinically healthy rabbits tested as part of a standard health check) and Toxoplasma gondii (8 positive out of 157). Of the 144 seropositive rabbits with clinical signs, 75% showed neurological symptoms, 14.6% demonstrated phacoclastic uveitis and 3.5% suffered from renal failure. 6.9% of the animals had combined symptoms. Vestibular disease dominated within the rabbits that showed neurological symptoms. Polymerase chain reaction (PCR) could not detect parasite DNA in urine or cerebrospinal fluid (CSF), but did so in 4 out of 5 samples of liquefied lens material in cases with phacoclastic uveitis due to lens capsule rupture. Additionally further diagnostic procedures, such as inspection of the external ear canal (N=69), radiography of the tympanic bullae (N=65) were performed to rule out differential diagnosis. 54.2% of the patients exhibiting neurological symptoms recovered within a few days, while 87.5% of the rabbits suffering from renal failure died or had to be euthanized. PMID- 18164136 TI - Effects of l-arginine and N(G)-nitro-l-arginine methyl ester treatments on expression of neuronal nitric oxide synthase in the guinea-pig bladder after partial bladder outlet obstruction. AB - This study was aimed to examine the effects of pharmacological intervention on partial bladder outlet obstruction (PBOO) on expression of neuronal nitric oxide synthase (nNOS) and nitric oxide (NO) production and NO-related free radical damage using nitrotyrosine as a marker in the guinea-pig bladder. Partial urethral ligation was performed in young male guinea pigs which were then intraperitoneally administered l-arginine, N(G)-nitro-l-arginine methyl ester (l NAME) or vehicle (saline) for 2 or 4 weeks. At the respective time points, the bladder was removed for nNOS immunohistochemistry, Western blot analysis, nitrotyrosine enzyme-linked immunosorbent assay test and NO colorimetric assay. In l-arginine-treated animals killed at 2 and 4 weeks, the total number of nNOS positive intramural neurons was significantly increased when compared with the corresponding control. Some neurons projected long extending fibers that were closely associated with the blood vessels. Furthermore, at 4 weeks, the nNOS protein content and NO production as reflected by the concentration of nitrite and nitrate were drastically elevated as measured by Western blot analysis and NO colorimetric assay, respectively. In l-NAME-treated group killed at 2 weeks, the number of nNOS positive neurons was markedly reduced when compared with the controls, but the change was not significant at 4 weeks. In the latter, however, the NO production as reflected by the concentration of nitrite and nitrate was markedly reduced; in addition, the nitrotyrosine concentration was significantly lower than the control. The present results support the role of NO in the pathophysiological changes following PBOO. We suggest the potential therapeutic application of l-arginine and l-NAME in PBOO; however, ultimately balancing the bidirectional effects of NO would be essential. PMID- 18164137 TI - Alpha calcium/calmodulin dependent protein kinase II in learning-dependent plasticity of mouse somatosensory cortex. AB - Calcium/calmodulin dependent protein kinase II (CaMKII), and more specifically its alpha subunit, is widely believed to be fundamental for hippocampal synaptic plasticity. In the cerebral cortex, deprivation-evoked plasticity was shown to depend on alphaCaMKII autophosphorylation abilities. Here we analyzed how learning-induced functional reorganization of cortical representations affected alphaCaMKII in adult Swiss mice. Mice were subjected to short-lasting sensory training in which stimulation of whiskers was paired with tail shock. The pairing results in enlargement of functional representation of vibrissae activated during the training. alphaCaMKII protein and its autophosphorylation level were determined by Western-blotting in somatosensory cortex crude synaptosomal fraction (P2) and postsynaptic protein-enriched, Triton X-100 insoluble fraction (TIF). The first training session resulted in an increase in alphaCaMKII autophosphorylation at autonomy site observed in TIF. A similar increase was also observed after the first session of just whiskers stimulation, which alone does not induce rearrangement of cortical representations. These data indicate that increased autophosphorylation of postsynaptic alphaCaMKII is not a correlate of induction phase of plasticity related reorganization of cortical representation of vibrissae. The increase observed in both experimental groups was transient and did not persist in the maintenance phase of the plastic change. Furthermore, we found that the training caused a delayed upregulation of alphaCaMKII protein level in crude synaptosomal fraction, but not in TIF, and the upregulation was not accompanied by an increase in autophosphorylation level of the kinase. The result indicates alphaCaMKII involvement in the late phase of plastic change and suggests the participation of a presynaptic pool of kinase rather than postsynaptic at this point. PMID- 18164138 TI - Urocortin 1 microinjection into the mouse lateral septum regulates the acquisition and expression of alcohol consumption. AB - Previous studies using genetic and lesion approaches have shown that the neuropeptide urocortin 1 (Ucn1) is involved in regulating alcohol consumption. Ucn1 is a corticotropin releasing factor (CRF) -like peptide that binds CRF1 and CRF2 receptors. Perioculomotor urocortin-containing neurons (pIIIu), also known as the non-preganglionic Edinger-Westphal nucleus, are the major source of Ucn1 in the brain and are known to innervate the lateral septum. Thus, the present study tested whether Ucn1 could regulate alcohol consumption through the lateral septum. In a series of experiments Ucn1 or CRF was bilaterally injected at various doses into the lateral septum of male C57BL/6J mice. Consumption of 20% volume/volume ethanol or water was tested immediately after the injections using a modification of a 2-h limited access sweetener-free "drinking-in-the-dark" procedure. Ucn1 significantly suppressed ethanol consumption when administered prior to the third ethanol drinking session (the expression phase of ethanol drinking) at doses as low as 6 pmol. Ethanol intake was differentially sensitive to Ucn1, as equivalent doses of this peptide did not suppress water consumption. In contrast, CRF suppressed both ethanol and water intake at 40 and 60 pmol, but not at lower doses. Repeated administration of Ucn1 during the acquisition of alcohol consumption showed that 40 pmol (but not 2 or 0.1 pmol) significantly attenuated ethanol intake. Repeated administration of Ucn1 also resulted in a decrease of ethanol intake in sham-injected animals, a finding suggesting that the suppressive effect of Ucn1 on ethanol intake can be conditioned. Taken together, these studies confirm the importance of lateral septum innervation by Ucn1 in the regulation of alcohol consumption. PMID- 18164139 TI - Enhanced neural activation in brain regions mediating sexual responses following exposure to a conditioned stimulus that predicts copulation. AB - Stimuli associated with sexual behavior increase reproductive success if presented prior to copulation. In Japanese quail, inseminations that take place in a context that predicts the arrival of a female are more likely to result in fertilized eggs. We demonstrate here that in male Japanese quail a sexual conditioned stimulus (CS) also enhances activity in two brain regions that mediate sexual behavior, the medial preoptic area and the medial part of the bed nucleus of the stria terminalis. C-fos expression, a marker of neural activation, was higher in these areas in subjects exposed sequentially to a sexual CS and copulation than in subjects exposed to copulation or the CS alone or in subjects exposed to no sexual stimulus, either an identical, untrained CS or an empty arena. These results suggest a link between a proximate result of sexual CS presentation, male brain activation, and a known ultimate outcome, increased fertilizations. PMID- 18164140 TI - Attention-deficit/hyperactivity disorder, Tourette's syndrome, and restless legs syndrome: the iron hypothesis. AB - Preliminary but increasing evidence suggests that attention-deficit/hyperactivity disorder (ADHD), Tourette's syndrome (TS), and restless legs syndrome (RLS) may be comorbid. In the present article, we hypothesize that ADHD, TS, and RLS may be part of a spectrum, and that iron deficiency contributes to the pathophysiology underlying this spectrum. Iron deficiency might lead to ADHD, RLS and TS symptoms via its impact on the metabolism of dopamine and other catecholamines, which have been involved into the pathophysiology of ADHD, TS, and RLS. We speculate that the catecholaminergic systems are differently impacted in each of the three disorders, contributing to a different specific phenotypic expression of iron deficiency. MRI studies assessing brain iron levels in ADHD, TS, and childhood RLS, as well as genetic studies on the specific molecular pathways involved in iron deficiency, are greatly needed to confirm the iron hypothesis underlying ADHD, TS, and RLS. This body of research may set the basis for controlled trials assessing the effectiveness and tolerability, as well as the most appropriate dose, duration and type (oral vs. intravenous) of iron supplementation. In conclusion, the iron hypothesis may help us progress in the understanding of pathophysiological links between ADHD, RLS, and TS, suggesting that iron supplementation might be effective for all these three impairing conditions. PMID- 18164141 TI - Is vascular endothelial growth factor (VEGF) the main mediator in ovarian hyperstimulation syndrome (OHSS)? AB - Ovarian hyperstimulation syndrome (OHSS) remains one of the most life-threatening and potentially fatal complications of assisted reproduction treatments (ART), due to excessive stimulation of the ovaries by exogenously administrated gonadotropins. The pathogenesis of OHSS remains obscure and patient treatments are empirical. However, vascular endothelial growth factor (VEGF) has been suggested to be responsible for OHSS. The present hypothesis aims to discuss the possible role of epithelial ion channels particularly cystic fibrosis transmembrane conductance regulator (CFTR) involvement in the pathogenesis of OHSS. This may provide grounds for the development of a better treatment strategy to reduce the risk of OHSS and improve in vitro fertilization (IVF) outcome. PMID- 18164142 TI - Configural and elemental processing in associative learning: commentary on Melchers, Shanks and Lachnit. AB - The paper by Melchers, Shanks, and Lachnit (2007) reviews the available evidence suggesting that there is flexible processing, such that some associative learning tasks can be solved either configurally or elementally. We find the evidence provocative but limited in its demonstrated generality and silent with respect to the theoretical mechanisms that might regulate the alleged flexibility of processing. Further research is invited to determine the scope of the variation involved and how best to account for it. At present, theories, either elemental or configural, that include mechanisms to shift the weight assigned to component stimuli appear to be potential candidates for embracing the data. PMID- 18164143 TI - Species differences in the identification of acoustic stimuli by birds. AB - The perceptual organization of auditory stimuli can reveal a great deal about how the brain naturally groups events. The current study uses identification techniques to investigate the abilities of two species of birds in identifying zebra finch song as well as synthetically generated speech stimuli. Budgerigars (Melopsittacus undulatus) and zebra finches (Taeniopygia guttata) were trained to differentially peck keys in response to the presentation of various complex stimuli. Although there were no clear differences in performance during the training paradigm between the two species, budgerigars were far more adept at learning to identify both sets of complex stimuli than were zebra finches, requiring far less trials to reach criterion. The non-singing but vocally plastic budgerigars vastly outperformed zebra finches at identifying both zebra finch song and synthetically designed human speech despite known similarities in auditory sensitivities between the two species and seemingly equivalent learning capacity. The flexibility that budgerigars seem to have at identifying various stimuli is highlighted by their enhanced performance in these tasks. These results are discussed in the context of what is known about both general and specialized processes which may contribute to any differences or similarities in performance. PMID- 18164144 TI - Levodopa pharmacotherapy for cocaine dependence: choosing the optimal behavioral therapy platform. AB - BACKGROUND: The dopamine precursor levodopa has shown some, albeit relatively weak, promise in treating cocaine dependence. This study sought to identify the most appropriate behavioral therapy platform for levodopa pharmacotherapy by evaluating its effect when administered in combination with behavioral platforms of varying intensities. METHOD: A total of 161 treatment-seeking cocaine dependent subjects received sustained release levodopa/carbidopa (400/100mg bid, Sinemet) or placebo delivered in combination with Clinical Management (ClinMan); ClinMan+cognitive behavioral therapy (CBT); or ClinMan+CBT+voucher-based reinforcement therapy (VBRT) in a 12-week randomized, placebo-controlled, double blind (for medication condition) trial. Medication compliance was monitored with riboflavin (100mg/capsule) and the Medication Event Monitoring System. Protocol compliance was addressed in weekly, 10-min nurse-delivered ClinMan sessions. Weekly, 1-h CBT sessions focused on coping skills training. VBRT (with escalating reinforcer value) provided cash-valued vouchers contingent on cocaine-negative urine toxicology results. Urine benzoylecgonine assays collected thrice-weekly were analyzed by intention-to-treat criteria using generalized linear mixed models. RESULTS: Levodopa main effects were found on all outcome measures of cocaine use. Contrasts testing the levodopa-placebo difference within each behavioral platform found reliable effects, favoring levodopa, only in the VBRT platform. Levodopa treatment with vouchers produced higher proportions of cocaine negative urines and longer periods of consecutive abstinence compared to other treatment combinations. CONCLUSION: This is the first study to find a significant treatment effect for levodopa and, in doing so, to demonstrate that the magnitude of this effect is dependent upon conditions of the behavioral therapy platform. The data support use of levodopa with abstinence-based reinforcement therapy as one efficacious combination in cocaine dependence disorder treatment. PMID- 18164145 TI - Behavioral drug and HIV risk reduction counseling (BDRC) with abstinence contingent take-home buprenorphine: a pilot randomized clinical trial. AB - This pilot randomized clinical trial evaluated whether the efficacy of office based buprenorphine maintenance treatment (BMT), provided with limited counseling or oversight of medication adherence is improved by the addition of individual drug counseling and abstinence-contingent take-home doses of buprenorphine. After a 2-week buprenorphine and stabilization period, heroin dependent individuals (n=24) in Muar, Malaysia were randomly assigned to Standard Services BMT (physician administered advice and support, and weekly, non-contingent medication pick-up) or Enhanced Services (nurse-delivered manual-guided behavioral drug and HIV risk reduction counseling (BDRC) and abstinence-contingent take-home buprenorphine (ACB), 7 day supply maximum). Outcomes included retention, proportion of opioid-negative urine tests, self-reported drug use, and self reported HIV risk behaviors. 12/12 (100%) of Enhanced Services and 11/12 (92%) of Standard Services participants completed the entire protocol. The proportion of opioid-negative urine tests increased significantly over time for both groups (p<0.001), and the reductions were significantly greater in the Enhanced Services group (p<0.05); Enhanced Services group achieved higher overall proportions of opiate negative urine toxicology tests (87% vs. 69%, p=0.04) and longer periods of consecutive abstinence from opiates (10.3 weeks vs. 7.8 weeks, p=0.154). Both groups significantly reduced HIV risk behaviors during treatment (p<0.05), but the difference between Enhanced and Standard Services (26% vs. 17% reductions from the baseline levels, respectively) was not statistically significant (p=0.9). Manual-guided behavioral drug and HIV risk reduction counseling and abstinence-contingent take-home buprenorphine appear promising for adding to the efficacy of office-based BMT provided with limited drug counseling and medication oversight. PMID- 18164146 TI - Socio-demographic and clinical characteristics of 3129 heroin users in the first methadone maintenance treatment clinic in China. AB - BACKGROUND: This study aimed to determine the characteristics of heroin users in the first methadone maintenance treatment (MMT) clinic in China. METHODS: In a retrospective chart review, the notes of 3127 heroin users who received both detoxification and MMT at the clinic were analyzed. Their socio-demographic and clinical data were collected, and the frequency of human immunodeficiency virus (HIV), Hepatitis C virus (HCV), and syphilis infections was investigated. RESULTS: The main findings are as follows: (1) 66.5% of the patients were younger than 35 years; (2) 55.1% were married at admission; (3) 32% were non-local residents; (4) the majority had high school level education, a history of smoking and alcohol consumption prior to the initial heroin use, and did not have stable jobs; (5) 28.4% were self-employed; (6) 5.4% of the sample had total hearing loss coupled with loss of speech; (7) 83.4% sniffed heroin or injected it intravenously at the time of admission, but 87.2% had smoked the drug when they first began abusing it; (8) a significant proportion of the patients were infected with HIV, HCV, and syphilis. CONCLUSIONS: Some of the above findings are not consistent with the results of previous studies conducted in Western countries and China. The unique socio-cultural and clinical characteristics of heroin abusers in different regions of China should be considered when MMT services are planned. PMID- 18164147 TI - Transmission of methicillin-resistant Staphylococcus intermedius between humans and animals. PMID- 18164148 TI - Performance of clinical signs to detect bluetongue virus serotype 8 outbreaks in cattle and sheep during the 2006-epidemic in The Netherlands. AB - The performance of clinical signs as a diagnostic test for the detection of BTV-8 outbreaks during the 2006-epidemic in The Netherlands was evaluated by constructing and analysing receiver operating characteristic (ROC) curves. The area under the ROC curve of the BT-associated clinical signs in cattle was 0.77. An optimal efficient test (maximising both sensitivity and specificity) in cattle herds combined a sensitivity (Se) of 67% with a specificity (Sp) of 72%, comprising the following clinical signs: ulcerations and/or erosions of oral mucosa or erosions of lips/crusts in or around nostrils or oedema of the nose or hyperaemic/purple coloration of tongue, tongue protrusion or coronitis or apathy/tiredness or muscle necrosis, stiffness of limbs or loathing or refusal to move, prostration or torticollis or anoestrus. The area under the ROC curve of the BT-associated clinical signs in sheep was 0.81. The optimal efficient test in sheep flocks combined a Se of 76% with a Sp of 72%, comprising the following clinical signs: ulcerations of oral mucosa or serous nasal discharge or erosions/ulceration of tongue mucosa or hypersensitivity of the skin or muscle necrosis, stiffness of limbs or coronitis or grinding of teeth or salivation or weakness/paresis. PMID- 18164149 TI - Feeding with supplemental squalene enhances the productive performance in boars. AB - In the present study, we investigated whether squalene could enhance the reproductive performance of boars. Boars at 12 months of age (weighting at 130+/ 5 kg) were randomly allocated into four groups and treated with basal diet (control) or with basal diet supplemental squalene at 10, 20 or 40 mg squalene/kg/day for 60 days. The impact of squalene feeding on the reproductive performance and the levels of serum leptin and testosterone were evaluated before and after feeding. Feeding with squalene at 10 mg/kg/day neither significantly increase the reproductive performance and serum testosterone levels, nor reduced the levels of leptin in boars. Surprisingly, feeding with supplemental squalene at 20 or 40 mg/kg/day significantly improved the reproductive performance as evidenced by dramatically reduced the time for mating, increased semens' volume and motility, and increased the size of litter as compared with that in controls. Furthermore, feeding with a higher dose of squalene significantly reduced the levels of serum leptin, accompanied by elevated levels of testosterone, as compared with that in controls. These data clearly indicate that feeding with squalene can improve the reproductive performance in boars. Therefore, dietary supplementation with squalene may increase the productivity of pig industry and potentially benefit for other animal production. PMID- 18164150 TI - Characterization and aerodynamic evaluation of spray dried recombinant human growth hormone using protein stabilizing agents. AB - The effect of the protein stabilizers on the stability and aerosol performance of spray dried recombinant human growth hormone (SD rhGH) was investigated. rhGH solution was spray dried alone, with polysorbate 20 (at three concentrations of 0.05%, 0.01%, and 0.005%), Zn(2+) (by Zn(2+):rhGH molar ratio of 2:1 and 4:1), and/or lactose (by lactose:rhGH weight ratio of 2:1). Size exclusion chromatography (SEC) analysis of spray dried powders demonstrated that of all the potential protein stabilizers, the combination of polysorbate 20 (0.05%), Zn(2+) (Zn(2+):rhGH molar ratio of 2:1) and lactose (lactose:rhGH weight ratio of 2:1) was the most effective at protecting rhGH against aggregation during spray drying. The results of circular dichroism (CD) analysis revealed that using of polysorbate 20 (in all concentrations) and Zn(2+) (by Zn(2+):rhGH molar ratio of 2:1) together in the formulations would preserve rhGH conformational stability during the process. The particle size distribution data obtained by laser diffraction method showed all SD rhGH formulations had volume median diameter and mean diameter below 5mum. The characterization of the aerosol performance of the spray dried powders by Andersen cascade impactor (ACI) showed that by increasing the concentration of polysorbate 20 in the formulations the aerodynamic efficiency of the resultant particles was reduced. In conclusion, the optimum amounts of polysorbate 20, Zn(2+) and lactose satisfied both physical stability during spray drying process (2.37% aggregation) and good aerosol performance (fine particle fraction; FPF=38.52%). PMID- 18164151 TI - Antifungal effect of high- and low-molecular-weight chitosan hydrochloride, carboxymethyl chitosan, chitosan oligosaccharide and N-acetyl-D-glucosamine against Candida albicans, Candida krusei and Candida glabrata. AB - OBJECTIVES: Generally, chitosan is a water-insoluble polyaminosaccharide with antimicrobial activity. The antifungal activity of water-soluble low- and high molecular-weight chitosan hydrochloride, carboxymethyl chitosan, chitosan oligosaccharide and N-acetyl-d-glucosamine against Candida albicans, Candida krusei and Candida glabrata was investigated. METHODS: Solutions of the tested substances in different concentrations (1, 0.5, 0.25, 0.1, 0.05, 0.025, 0.01, 0.005, and 0.0025%) were prepared and the influence on C. albicans DSM 11225, C. krusei ATCC 6258 and C. glabrata DSM 11226 was investigated. Yeasts (3 x 10(5) cells/mL) were incubated with Sabouraud liquid medium at 30 degrees C. Measurements were done with a microplate nephelometer (NEPHELOstar Galaxy, BMG LABTECH Ltd.) for 24 h. High values of light scattering correlate with strong cultural growth. Results were shown as growth curves and histograms displaying 24 h end points. These were compared with control by Mann-Whitney test. Furthermore, MIC(50%), MIC(80%) and Spearman correlation coefficients were calculated. RESULTS: C. albicans and C. krusei were the most sensitive species. C. glabrata was also inhibited, whereas 1% of tested substances could not prevent its growth completely. However, only both chitosan hydrochlorides showed a definite antifungal effect with high correlation between inhibition and test concentration. Carboxymethyl chitosan, chitosan oligosaccharide and N-acetyl-D glucosamine showed only a weak or no antifungal activity, respectively. CONCLUSIONS: Antifungal activity decreases with declining molecular mass (chitosan oligosaccharide and N-acetyl-D-glucosamine) and increasing masking of the protonated amino groups with functional groups (carboxymethyl chitosan). PMID- 18164152 TI - Valuation of anti-inflammatory and antinociceptive activities of Erica species native to Turkey. AB - Erica L. species (Ericaceae) have been popularly used as antirheumatic, diuretic, astringent and treatment of urinary infections. In order to evaluate this information, anti-inflammatory and antinociceptive activities of different extracts prepared with methanol, chloroform, ethyl acetate, n-butanol and water from the aerial parts of Erica arborea L., Erica manipuliflora Salisb., Erica bocquetii (Pesmen) P.F. Stevens and Erica sicula Guss. subsp. libanotica (C.&W. Barbey) P.F. Stevens (Ericaceae) of Turkish origin were investigated by using in vivo methods. For the anti-inflammatory activity, carrageenan-induced hind paw edema model, PGE(2)-induced hind paw edema model, and 12-O-tetradecanoyl-13 acetate (TPA)-induced mouse ear edema model and for the antinociceptive activity p-benzoquinone-induced writhing test in mice were employed. The ethyl acetate extracts of Erica arborea (EAE), Erica bocquetii (EBE) and Erica manipuliflora (EME) exhibited notable inhibition against carrageenan-induced (24.1-32.3%, 23.8 36.1%, 29.2-35.1%, respectively) and PGE(2)-induced (21.2-37.7%, 6.8-29.7%, and 6.2-34.1%, respectively) hind paw edema as well as TPA-induced mouse ear edema models in mice, while the ethyl acetate extract of Erica sicula subsp. libanotica (ESE) (10.7-29.7%) displayed potent anti-inflammatory activity only on the PGE(2) induced hind paw edema model. However, the remaining extracts were found to be inactive against inflammatory models. Same extracts, i.e., EAE, EBE and EME were also found to exhibit remarkable antinociceptive activity in p-benzoquinone induced abdominal constriction test at a dose of 100mg/kg (46.5%, 27.7% and 36.3%, respectively). PMID- 18164153 TI - Investigation of antimicrobial model of Hemsleya pengxianensis W.J. Chang and its main active component by metabolomics technique. AB - AIM OF THE STUDY: H.pengxianensi is a native plant in China. Dihydrocucurbitacin F-25-O-acetate is one of the major chemical components in H.pengxianensis. In this investigation, we attempted to quantify the content of dihydrocucurbitacin F 25-O-acetate in H.pengxianensis and find the possible antibacterial mode of dihydrocucurbitacin F-25-O-acetate and H.pengxianensis. MATERIAL AND METHODS: First, the content of dihydrocucurbitacin F-25-O-acetate was quantified by HPLC. Then, metabolic profiles of Staphylococcus aureus treated by dihydrocucurbitacin F-25-O-acetate, extract of H.pengxianensis and nine antibacterial substances with the known modes of action were acquired by HPLC/DAD/ESI-MS. Principal component analysis was carried out upon all metabolic profiles to classify those drugs according to their mechanisms. RESULTS AND CONCLUSION: The HPLC result show the content of dihydrocucurbitacin F-25-O-acetate in H.pengxianensis (5.3+/-0.4 mg/g). From the result obtained by principal component analysis, dihydrocucurbitacin F-25-O-acetate was found to be the component playing the main antimicrobial roles on Staphylococcus aureus in H.pengxianensis and the possible antibacterial mode of dihydrocucurbitacin F-25-O-acetate and H.pengxianensis may be to inhibit cell wall synthesis. PMID- 18164154 TI - Toll-like receptor ligand-induced activation of murine DC2.4 cells is attenuated by Panax notoginseng. AB - The medicinal herb, Panax notoginseng, has been used for thousands of years in traditional Chinese medicine and possesses anti-inflammatory properties. Dendritic cells (DCs) play a central role in the regulation of both inflammation and adaptive immunity. The aim of this study was to investigate the potential for notoginseng extracts to modulate Toll-like receptor (TLR) ligand-induced activation of cultured DC2.4 cells. Following stimulation with LPS, CpG or poly(I:C) and treatment with 0-50micorg/ml notoginseng extract for 24 h, DCs were evaluated for various phenotypic and functional readouts. Notoginseng reduced the LPS-, CpG- and poly(I:C)-induced production of TNF-alpha by DC2.4 cells. Also, IL 6 production by notoginseng-treated cells stimulated with LPS and CpG but not poly(I:C) was reduced when compared to controls. TLR ligand-induced CD40 expression was attenuated by notoginseng. In contrast, notoginseng decreased CD86 levels on DCs activated with LPS and poly(I:C) but not CpG. Inhibition of TNF alpha production was time-dependent in LPS-stimulated cells, occurring only with pretreatment or concurrent treatment of notoginseng but not after delayed addition of the herbal extract. Additionally, ginsenoside Rg1 more effectively inhibited LPS-stimulated cytokine production by DC2.4 cells than ginsenoside Rb1. Taken together, these results demonstrate that notoginseng inhibits the production of specific inflammatory molecules and innate immune responsiveness by DCs following TLR activation. PMID- 18164155 TI - Diagnosis of congenital hydrocephalus and delivery of its patients in Japan. AB - The study population included 193 patients with prenatally diagnosed (fetal) hydrocephalus and 181 with postnatally (12 or less than 12 months after birth) diagnosed (infantile) hydrocephalus identified by a nationwide questionnaire survey of congenital hydrocephalus performed in 2000. Of 180 patients with fetal, 101 (56.1%) were diagnosed before week 32 of gestation and 18 (10%) were diagnosed week 37 and later of gestation. In patients with fetal hydrocephalus, US was used in more than 80% of the cases, whereas in patients with infantile hydrocephalus, CT was used in more than half of the cases. For diagnosis of fetal hydrocephalus, either US or MRI had become dominantly utilized and CT had gone out of use in 1996-2000. The adoption ratio of cesarean delivery to transvaginal delivery was around 7 to 3 in patients with fetal hydrocephalus, and 2 to 7 in patients with infantile hydrocephalus, respectively, with significant difference between fetal hydrocephalus and infantile hydrocephalus groups (p < 0.001). Clinical outcomes in patients with fetal hydrocephalus was better in those delivered transvaginally than in those by cesarean delivery, although without no statistical significance (p = 0.124) and those in patients with infantile hydrocephalus showed almost no difference between transvaginal and cesarean delivery groups. There was a tendency for the Apgar score at 5 min to be lower in smaller birth weight infants with a body weight of less than 2000g. This score could be useful as an index for predicting immediate postnatal death in patients with fetal hydrocephalus. PMID- 18164156 TI - Preoperative 3T MR imaging of rectal cancer: local staging accuracy using a two dimensional and three-dimensional T2-weighted turbo spin echo sequence. AB - PURPOSE: The purpose of our study was to evaluate the image quality and diagnostic performance of two-dimensional (D) turbo spin echo (TSE) and 3D T2 weighted TSE MR imaging in local staging of rectal cancer at 3T. MATERIALS AND METHODS: 3T phased-array MR imaging was performed in 36 consecutive patients with biopsy-proven rectal cancer. High-resolution 2D TSE images in three planes and 3D TSE images of the rectum were obtained. Two independent observers performed an image quality assessment using eight image quality characteristics. All 2D and 3D datasets were evaluated separately. MR images were prospectively evaluated by two experienced radiologists in consensus with regard to local disease. Total mesorectal excision was used as the standard of reference. The sensitivity, specificity, positive and negative predictive value, and overall accuracy were calculated. Areas under the receiver operating characteristic (ROC) curve (AUC) were determined. RESULTS: Twenty-two patients who underwent a total mesorectal excision were enrolled in this study. Significantly more motion artifacts were present with 3D TSE imaging (P=0.04). The overall sensitivity, specificity, and accuracy of muscularis propria invasion in rectal cancer using 2D T2-weighted images were 100%, 66%, and 95%, respectively. There was a statistical significant greater AUC using 2D T2-weighted images compared to 3D T2-weighted MR images (P=0.04). The ROC curves describing the results of the interpretation of 2D and 3D T2-weighted datasets regarding perirectal tissue invasion showed no statistical significant difference (P=0.41). CONCLUSIONS: In this study, high local staging accuracies with 3T 2D T2-weighted MR imaging were demonstrated. 3D T2-weighted MR imaging cannot replace 2D MR imaging for local staging of rectal cancer. However, 3D MR imaging can be used for visualization of the complex pelvic anatomy for treatment planning purposes. PMID- 18164157 TI - Usefulness of 64-slice MDCT for follow-up of young children with coronary artery aneurysm due to Kawasaki disease: initial experience. AB - To evaluate the initial application and value of 64-slice multidetector computed tomography as an alternative diagnostic modality in the follow-up of young children with coronary artery aneurysm due to Kawasaki disease. Twelve boys (mean age 5.1 years, range 1.8-7.8 years) for follow-up (time range from 1.1 to 5.1 years) of known Kawasaki disease and coronary artery aneurysm underwent 64-slice MDCT ECG-gated coronary angiography. All data were acquired without breath holding. Two pediatric radiologists independently assessed image quality and the diameter of all coronary segments were measured for each patient. The number, position, shape and size of each coronary artery aneurysm were observed and compared with those of ECHO performed previously. A total of 118/156 segments permitted visualization with diagnostic image quality, the CT measurements showed good inter-observer and intra-observer reliability, coefficients were 0.93 and 0.88, respectively. A total of 30 coronary artery aneurysms were identified with measured mean of 7.5+/-3.8 mm in diameter, and of 12.4+/-9.1 mm in longitudinal lengths.10 tumors were small, 8 tumors were medium and 12 tumors were giant aneurysm. The affected segments included LM7/12(58.3%), 9/12(75%) of LAD1, 4/12(33.3%) of LAD2, 2/12(16.7%) of LCX1; 6/12(50%) of RCA1, 9/12(75%) of RCA2 and 4/12(33.3%) of RCA3, including affected two segments in 9 tumors and three segments in 1 tumor. Calcifications were found in 5 aneurysms and 3/5 with thrombosis; six stenotic segments were found. ECHO failed to detect 8 tumors with 2/8 in LAD, 1/8 in LCX and 5/8 in RCA, and those included 4 small aneurysms. The use of 64-slice MDCT angiography proved valuable for monitoring young children with Kawasaki disease. However, further study is necessary to specify the sensitivity and specificity of MDCT in the follow-up. PMID- 18164158 TI - The phase shift index for marking functional asynchrony in Alzheimer's disease patients using fMRI. AB - Our previous study suggested that the functional magnetic resonance imaging MRI (fMRI) COSLOF Index (CI) could be used as a quantitative biomarker for Alzheimer's disease (AD). The fMRI CI was lowest in the AD group (0.13+/-0.10), followed by the mild cognitive impairment (MCI) group (0.20+/-0.05) and the control group (0.34+/-0.09). The current study continues an investigation into which of the following two factors has a dominant role in determining the CI: the signal-to-noise ratio (SNR) or the phase shift of spontaneous low-frequency (SLF) components. By using a theoretical model for SLF components, we demonstrated that the normalized CI does not depend on the SNR of the SLF components. Further analysis shows that by taking the ratio of the cross-correlation coefficient to the maximum-shifted cross-correlation coefficient, the SNR factor can be canceled. Therefore, the determination of the phase shift index (PSI) method is independent of the SNR, and the PSI provides an accurate measure of the phase shift between SLF components. By applying this PSI method to the control, MCI and AD groups of subjects, experimental results demonstrated that the PSI was highest in the AD group (72.6+/-11.3 degrees ), followed by the MCI group (58.6+/-5.7 degrees ) and, finally, the control group (40.6+/-8.4 degrees ). These results suggest that the larger is the PSI value, the more asynchrony exists between SLF components. PMID- 18164159 TI - On-site screening and GC-MS analysis of cocaine and heroin metabolites in body packers urine. AB - The illicit transportation of cocaine and heroin either swallowed or inserted into the rectum and/or vagina of individuals, defined as "body-packers", is becoming increasingly common. Assessment of smuggling by urinalysis from body packers has been sparsely reported and on-site rapid screening methods are essentially lacking. We screened the presence of cocaine and heroin metabolites in urine from suspected body-packers by an on-site immunochromatographic test and confirmed the obtained results by gas chromatography-mass spectrometry and X-ray examination. Samples were collected from 64 individuals (45 men, 19 women) stopped at Fiumicino and Ciampino airports of Rome (Italy) for suspicion of internal concealment of cocaine and heroin between October 2006 and July 2007. Urine was immediately screened on-site by Cozart rapid urine test. Irrespective of test results, individuals underwent X-ray examination and urine samples were analyzed by gas chromatography-mass spectrometry (GC-MS). In 48 out of 64 cases (24 positives and 24 negatives) screening results were confirmed by GC-MS assay and X-ray examination. In 5 cases, positive to the on-site test and GC-MS analysis, abdominal radiography was negative and individuals resulted to be drug users. In 11 cases, negative to the on-site test and radiological investigation, GC-MS analysis found benzoylecgonine in 10 cases and morphine in one case. Concentration of both substances was in all cases lower than 50ng/ml and compatible with personal drug use. From obtained results, on-site detection of cocaine and heroin metabolites in the urine of suspected body-packers appears to be a reliable screening test to disclose internally concealed drugs and justify subsequent radiological investigations. PMID- 18164160 TI - Application of a self-modeling curve resolution method for studying the photodegradation kinetics of nitrendipine and felodipine. AB - Dihydropyridine (DHP) derivatives, as calcium channel blockers with cardiovascular activity, are highly photosensitive and converted in the presence of light to compounds that are inactive. In this work, a self-modeling curve resolution method was applied to study the photodegradation kinetics of nitrendipine and felodipine by spectrophotometric method. The methanolic solutions of drugs were separately exposed to UV and daylight, respectively. A fully soft-modeling multivariate curve resolution method based on the combination of iterative target transformation and Kubista methods were used to analyze the recorded absorbance data, extracting the concentration profiles and pure spectra of the drugs and their photodegradation products. By fitting the concentration profiles of the studied DHP drugs to different kinetic equations, it was found that at the beginning of lighting, the reaction is zero-order and in the case of nitrendipine it changes to a first-order kinetic when the concentration of products exceeded than that of the initial compounds. PMID- 18164161 TI - Novel omics technologies in nutrition research. AB - A key scientific objective of nutrition research is to determine the role of diet in metabolic regulation and to improve health. There are many new opportunities and challenges for the nutrition research in post-genome era. Novel omics technologies and bioinformatics tools offer enormous potential to investigate the complex relationship between nutrition and metabolism. An overview of omics technologies in nutrition research is presented in this paper, which focuses on recent applications of nutritranscriptomics, nutriproteomics and nutrimetabolomics in nutrition research. The potential limitations and future prospects are also discussed. The combination of various omics technologies, systems biology, will greatly facilitate the discovery of new biomarkers associated with specific nutrients or other dietary factors. It can be expected that the future omics-based human nutrition research can provide personalized dietary recommendations for disease prevention. PMID- 18164162 TI - Homespun remedy, homespun toxicity: baking soda ingestion for dyspepsia. AB - A 68-year-old man presented to the Emergency Department with a severe metabolic alkalosis after ingesting large quantities of baking soda to treat his dyspepsia. His underlying pulmonary disease and a progressively worsening mental status necessitated intubation for respiratory failure. Laboratory studies revealed a hyponatremic, hypochloremic, hypokalemic metabolic alkalosis. The patient was successfully treated after cessation of the oral bicarbonate, initiation of intravenous hydration, and correction of electrolyte abnormalities. PMID- 18164163 TI - A technique for temporary control of hemorrhage. AB - Traumatic injury to blood vessels is a common presentation in the Emergency Department. Control of hemorrhage is generally attempted using tight absorbent compression dressings. This may be a cause of inefficient control of hemorrhage and also contributes to tissue ischemia. We present a simple technique to control hemorrhage that does not cause tissue ischemia, which frequently accompanies the tight compression bandages applied for vascular injuries. PMID- 18164164 TI - Automatic external defibrillation around the world: the time has come. PMID- 18164165 TI - Altered antioxidant status and increased lipid peroxidation in children with acute gastroenteritis admitted to a pediatric emergency service. AB - Acute gastroenteritis is a common illness worldwide and has a great impact on children. Our aim was to examine possible alterations in the antioxidant defense in pediatric gastroenteritis. To comprehensively examine the reaction of the antioxidant system, all possible components of the system were measured. The whole blood malondialdehyde and reduced glutathione, serum beta-carotene, retinol, vitamin C, vitamin E, catalase, ceruloplasmin, albumin, total bilirubin, uric acid, erythrocyte superoxide dismutase, and glutathione peroxidase levels were studied. Superoxide dismutase and glutathione peroxidase antioxidant enzyme activities and malondialdehyde levels were found to be increased; however, beta carotene, retinol, vitamin C, vitamin E, reduced glutathione, and albumin levels were observed to be significantly decreased. Catalase activity remained unchanged, whereas some of the other non-enzymatic antioxidants such as ceruloplasmin, total bilirubin, and uric acid levels were increased compared to the control group. We have shown an association between antioxidant levels and gastroenteritis in children. Further study is needed to assess whether antioxidant supplementation will be beneficial as an adjunct to conventional relevant therapy of the disease. PMID- 18164166 TI - Emergency medicine residents and statistics: what is the confidence? AB - The objective of this study was to assess whether residents have the essential tools and a sense of competency when evaluating published studies, especially the statistics. Questionnaires were mailed to emergency medicine (EM) residency programs in the United States querying residents' demographics and training in statistics as well as their impressions and use of statistics in the current literature; a five-question statistical quiz was also included. Possible responses of-almost always, more than (1/2) time, (1/2) time, less than (1/2) time, almost never-were tallied individually as well as compared in groups of polarized answers: over 1/2 time (almost always + more than (1/2) time) vs. under (1/2) time (less than (1/2) time + almost never). There were 495 questionnaires returned from 42 centers. No significant difference was found when comparing quiz performance with participants' self-reported statistical knowledge. There were considerable differences in the polarized answers (Over vs. Under), whether statistics: were used appropriately (40% vs. 15%, respectively); were used to enhance weak data (54% vs. 13%, respectively); enhanced their understanding of information (38% vs. 24%, respectively); simplified complex data (26% vs. 41%, respectively); were understood by them (23% vs. 38%, respectively); confused them (37% vs. 24%, respectively); were skipped (52% vs. 23%, respectively). Participants felt there should be more statistical training (49% vs. 22%, Over vs. Under, respectively). There was no difference in respondents who did or did not read the statistics (39% vs. 34%, Over vs. Under, respectively). Many EM residents surveyed do not trust, read, or understand statistics presented in current journal articles. Residency programs may want to consider enhanced training in statistics. PMID- 18164167 TI - The role of fibrinolytics in the prehospital treatment of ST-elevation myocardial infarction (STEMI). AB - The efficacy of fibrinolytics in the treatment of ST-elevation myocardial infarction is directly related to the time of administration, with the first 2 h after symptom onset seen as a critical period for greatest improvement in cardiovascular parameters and mortality. The American College of Cardiology/American Heart Association recommends a medical contact to treatment time of 30 min for fibrinolysis in patients with ST-elevation myocardial infarction. In selected patients, reperfusion goals may be expedited with prehospital administration of fibrinolytics. In clinical trials, prehospital fibrinolysis markedly reduced the time from symptom onset to treatment, allowed earlier ST-segment resolution, and reduced short- and long-term mortality compared with in-hospital treatment. Prehospital fibrinolysis has become more feasible with the introduction of prehospital 12-lead electrocardiography, improved skills of emergency medical services personnel, improved communication with the Emergency Department, and the advent of bolus fibrinolysis. Rapid and accurate administration of a fibrinolytic is vital for the success of prehospital fibrinolysis. PMID- 18164168 TI - Framing effect debiasing in medical decision making. AB - OBJECTIVE: Numerous studies have demonstrated the robustness of the framing effect in a variety of contexts. The present study investigated the effects of a debiasing procedure designed to prevent the framing effect for young adults who made decisions based on hypothetical medical decision-making vignettes. METHODS: The debiasing technique involved participants listing advantages and disadvantages of each treatment prior to making a choice. One hundred and two undergraduate students read a set of three medical treatment vignettes that presented information in terms of different outcome probabilities under either debiasing or control conditions. RESULTS: The framing effect was demonstrated by the control group in two of the three vignettes. The debiasing group successfully avoided the framing effect for both of these vignettes. CONCLUSION: These results further support previous findings of the framing effect as well as an effective debiasing technique. This study improved upon previous framing debiasing studies by including a control group and personal medical scenarios, as well as demonstrating debiasing in a framing condition in which the framing effect was demonstrated without a debiasing procedure. PRACTICE IMPLICATIONS: The findings suggest a relatively simple manipulation may circumvent the use of decision making heuristics in patients. PMID- 18164169 TI - Talking matters: abused women's views on disclosure of partner abuse to the family doctor and its role in handling the abuse situation. AB - OBJECTIVE: We aimed to explore what women valued most in disclosing partner abuse to their doctor and whether disclosure played a role in handling their abuse situation. METHODS: A qualitative method was used to understand abused women's views and experiences with disclosure to their family doctor. Thirty-six women were interviewed within 4 weeks after disclosure to their family doctor. RESULTS: Most women went to see the doctor for some medical complaint, and only three women planned to disclose the abuse. Twenty-five women valued most their doctor's communicative approach with empathy or empowering and nine women valued most the instrumental approach. Eight women of the latter group wanted this combined with a communicative approach. After disclosure to the family doctor, a group of women (n=20) perceived a real change in their possibilities to handle their situation. They appeared to be in a position we named: 'in transition', a state in which they started or continued a process of change. Another group of women (n=13) appeared to be in a 'locked-up' position, a state without any prospect on change, feeling out of control and fearing the abuser. Three women reacted reserved towards change. CONCLUSION: A communicative approach, providing empathy and empowerment, is important to women in disclosing partner abuse. More than half of the women perceived possibilities for a change. PRACTICE IMPLICATIONS: Talking about abuse is an important step in a woman's process of change. Doctors should acknowledge the advantage of their position as a professional confidant and ask women about abuse. PMID- 18164170 TI - Improvement of the cold storage of blood vessels with a vascular preservation solution. Study in porcine aortic segments. AB - BACKGROUND: Cold-induced injury to various cell types has been shown to be mediated predominantly by chelatable iron. For endothelial cells, this type of injury has so far only been shown in cultured cells. Hypothesizing that this iron dependent cold-induced injury might also occur in the endothelium of intact vessels, we here set out to optimize the hypothermic storage of blood vessels. METHODS: Segments of porcine aorta were stored for 2 to 21 days in histidine tryptophan-ketoglutarate (HTK) solution or in modified solutions with or without the iron chelators deferoxamine or LK 614 at 4 degrees C. Parts of the segments were assayed immediately after cold storage, the other parts after subsequent rewarming. The percentage of dead (propidium iodide-positive) endothelial cells was assessed by "intravital" fluorescence microscopy, mitochondrial membrane potential was assessed by laser scanning microscopy after staining with tetramethylrhodamine methyl ester (TMRM) and thrombocyte adhesion was studied using 5-(and -6)-carboxy SNARF-1-stained thrombocytes. RESULTS: The endothelium of porcine aortic segments sustained moderate injury during the cold incubation itself, but major injury during rewarming. The addition of the iron chelator deferoxamine (1 mmol/L) significantly inhibited cold-induced endothelial cell injury irrespective of the solution used for cold storage (eg, 14 days of cold storage + 3 hours rewarming: HTK 66 +/- 7%, HTK + 1 mmol/L deferoxamine 40 +/- 10% propidium iodide-positive endothelial cells). An amino acid (glycine, alanine, aspartate)-containing base solution with N-acetylhistidine as buffer was optimized. The optimized base solution with pH 7.0 and potassium and chloride as main ions yielded a further decrease of endothelial cell injury. Combination of deferoxamine (in lower concentration, ie, 0.1 mmol/L) with the new, more membrane permeable iron chelator LK 614 (20 mumol/L) further improved preservation so that even after 3 weeks of cold storage plus 3 hours rewarming only 10 +/- 1% of endothelial cells were propidium iodide positive. In this optimized solution, both endothelial cell survival and mitochondrial membrane potential were significantly better preserved than in the clinically used solutions HTK, University of Wisconsin (UW) and Perfadex, or in physiological saline. Thrombocyte adhesion was also significantly reduced after cold storage in the optimized solution compared with HTK solution. CONCLUSION: Cold-induced injury to the endothelium of porcine aortic segments is, as the injury to cultured endothelial cells, mediated by chelatable iron. Thus, iron chelators, but also optimized base solutions, are options to improve the storage of vascular endothelium. The optimized solution should now be tested in in vivo animal experiments. PMID- 18164171 TI - Detection of intraplaque hemorrhage by magnetic resonance imaging in symptomatic patients with mild to moderate carotid stenosis predicts recurrent neurological events. AB - BACKGROUND: Carotid endarterectomy is beneficial in severe (>70%) symptomatic carotid stenosis. The risk of stroke in moderate carotid stenosis (50%-69%) is modest, and so the role of carotid endarterectomy in this group is unclear. Intraplaque hemorrhage is associated with advanced atherosclerosis and can be detected in the carotid arteries by magnetic resonance imaging. This study evaluates whether magnetic resonance imaging detected intraplaque hemorrhage (MR IPH) can identify patients with symptomatic mild to moderate carotid stenosis who are at higher risk of ipsilateral transient ischemic attack (TIA) and stroke. METHODS: Prospective longitudinal cohort study of symptomatic patients with mild to moderate (30%-69%) carotid stenosis followed up for 2 years after imaging for IPH using magnetic resonance imaging. RESULTS: Sixty four participants were followed up for a median of 28 months (interquartile range 26-30) after MRI of the carotid arteries. Thirty-nine (61%) ipsilateral arteries showed intraplaque hemorrhage. During follow-up, five ipsilateral strokes and a total of 14 ipsilateral ischemic events were observed. Thirteen of these ischemic events, of which five were strokes, occurred in those with ipsilateral carotid intraplaque hemorrhage (hazard ratio = 9.8, 95% confidence interval 1.3-75.1, P = .03). CONCLUSIONS: MR IPH is a good predictor of ipsilateral stroke and TIA in patients with symptomatic mild to moderate (30%-69%) carotid stenosis. This technique could help in the selection of patients for carotid endarterectomy. PMID- 18164172 TI - Prognostic relevance of proliferative activity evaluated by Mib-1 immunostaining in node negative breast cancer. AB - AIM: The purpose of this prospective observational study was to analyze the role of Mib-1 immunostaining as a proliferation index in breast cancer. Correlations between Mib-1 expression and clinico-pathological characteristics as well as its prognostic value have been studied in a series of 432 node negative breast cancers. METHODS: Mib-1 expression was evaluated by immunohistochemistry. Tumor sections from highly cellular invasive areas of cancer were stained by monoclonal antibody Mib-1 (Dako) and cells whose nuclei stained positive were counted in 10 randomly chosen HPFs and expressed as percentages of all epithelial cells. A minimum of 400 cells were counted. Correlation between Mib-1 staining and clinico pathological factors was investigated by means of univariate and multivariate analyses. The prognostic impact on actuarial disease free (DFS) and overall survival (OS) was evaluated by univariate analysis using the log-rank test and by multivariate analysis using Cox regression model. RESULTS: Tumors were considered as positive for Mib-1 expression when more than 15% of cells counted were stained. Mib-1 positivity was found in 190/432 cases and resulted in being significantly related to tumor grade, tumor size and absence of estrogen receptors at multivariate analysis. With a median follow-up of 66 months, Mib-1 positivity resulted in being the only independent predictor of OS (RR 2.92), and an independent predictor of DFS (RR 2.01) together with absence of estrogen receptors (RR 2.15). CONCLUSIONS: Mib-1 index of proliferative activity correlates well to other established prognostic factors of breast cancer. Mib-1 index may improve the tailoring of adjuvant therapy in early breast cancer, and our experience adds evidence to its effectiveness as prognostic factor. Efforts to reach uniformity in the methodology and in the scoring system should be done to warrant a standardized procedure and make Mib-1 determination definitively reliable in the current clinical practice. PMID- 18164173 TI - [Report of the 14th meeting of the Club of Anesthesia and Resuscitation in ORL (CARORL)]. PMID- 18164174 TI - [Evaluation of synthetic glue for the fixation of continuous peripheral nerve catheters]. AB - Accidental dislodgement of continuous peripheral nerve catheters remains a frequent problem that causes failure of postoperative analgesia. We have assessed the use of new synthetic glue (Mastisol) to secure and maintain catheters in the correct position among 60 patients. This method allowed securing an effective fixation in 94% of cases,resulting in efficient ambulatory orthopedic surgery postoperative analgesia. No nervous or infectious complications were observed. This technique offers a simple, complementary method to secure peripheral nerve catheters. PMID- 18164175 TI - [Ulnar neuropathy following venipuncture]. PMID- 18164176 TI - All medical eponyms should be abandoned. PMID- 18164177 TI - And yet they correlate: psychophysiological activation predicts self-report outcomes of exposure therapy in claustrophobia. AB - The study examines whether self-reported fear and physiological activation are concordant when claustrophobic patients are exposed to small spaces, whether the measures change in synchrony for individual patients and whether initial activation of measures can predict the outcome of an exposure treatment. Ten patients with claustrophobia participated in six in-vivo exposure sessions with continuous monitoring of self-reported fear and their EKG. Partial pressure of carbon dioxide (pCO(2)), a measure of hyperventilation, was available in a subsample of patients. While evidence for concordance of self-reported fear and heart rate was limited, the measures changed synchronously within subjects. Most importantly, higher heart rate at the beginning of the first exposure session predicted better treatment outcome. Because self-reported fear turned out not to be a reliable predictor of the outcome, this is interpreted as evidence for the incremental validity of physiological measures of fear. PMID- 18164178 TI - EnsemPro: an ensemble approach to predicting transcription start sites in human genomic DNA sequences. AB - Although several computational methods have been developed to identify transcription start sites (TSSs)/promoters, the computational prediction still needs improvement. Due to low performance, the promoter prediction programs can provide misleading results in functional genomic studies. To improve the prediction accuracy, we propose the use of an ensemble approach, EnsemPro (Ensemble Promoter), which combines the prediction results of the existing promoter predictors. We schematically compared the prediction performance of the currently available promoter prediction programs in an identical evaluating environment, and the results served as a guide for choosing the combined predictors. We applied three representative ensemble schemes-the majority voting, the weighted voting, and the Bayesian approach-for the TSS prediction of hundreds of human genomic sequences. EnsemPro identified the TSSs more precisely than other combining methods as well as the currently available individual predictor programs. The source code of EnsemPro is available on request from the authors. PMID- 18164179 TI - Locked nucleic acids in PCR primers increase sensitivity and performance. AB - The incorporation of locked nucleic acids (LNAs) into oligonucleotide primers has been shown to increase template binding strength and specificity for DNA amplification. Real-time PCR and DNA sequencing have been shown to be significantly enhanced by the use of LNAs. Theoretically, increasing primers' binding strength may also increase the sensitivity of conventional PCR, reducing minimum template requirements. We compared LNA-modified PCR primers with their standard DNA counterparts for amplification sensitivity with template amounts as low as 5 pg. Although the results are highly dependent on the design of the LNA primers, large increases in peak height can be achieved from as little as 75 pg, as well as clearer and more complete profiles. Increased amplification success with lower template amounts may also be seen. Additionally, the use of LNAs can enhance multiplexing. Thus, incorporating LNAs into PCR primers can increase amplification success, sensitivity, and performance under a wide range of conditions. PMID- 18164180 TI - The development of an instrument to evaluate treatment intrusiveness for individuals with severe and challenging behavior. AB - The current study examined the use of an instrument designed to evaluate the intrusiveness of treatments for addressing problem behaviors. Participants read case vignettes with proposed treatments and then completed a checklist of factors that could influence the degree of treatment intrusiveness. Results indicated that the participants were capable of using the instrument to differentiate among treatments that were considered to have varying levels of treatment intrusiveness. Implications of using a systematic method for evaluating the intrusiveness of treatments are discussed. PMID- 18164182 TI - Increased oxidative stress in epileptic children treated with valproic acid. AB - PURPOSE: To determine influence of Valproic Acid (VPA) treatment on oxidative status in non-obese and overweight epileptic children. METHODS: A prospective study was conducted at the Departments of Pediatrics, University of Chieti and Bologna. Thirty-one epileptic children were studied before and after 1 year of therapy with VPA. Also 31 sex-, age- and BMI-matched healthy controls were evaluated. Insulin and glucose serum levels and plasma Vitamin E, Lag phase and Malondialdehyde (MDA) levels were determined. RESULTS: Before the beginning of VPA therapy, insulin and glucose serum values and plasma Vitamin E, Lag phase and MDA levels were normal in all subjects. At the end of follow-up, 11 (35.5%) epileptic patients developed obesity. In obese VPA treated patients, we found lower serum levels of antioxidant (Vitamin E, p<0.001) and higher levels of oxidant markers (MDA, p<0.001; Lag phase, p<0.001) compared to VPA-treated non obese patients and controls. CONCLUSION: After 1 year of VPA therapy oxidative stress occurs only in overweight children. This increase in the levels of oxidant markers, probably caused by obesity, might contribute to the development of endothelial dysfunction and atherosclerosis later in life. PMID- 18164181 TI - Wnt signaling and skeletal development. AB - Wnt proteins are a family of secreted proteins that regulate many aspects of cellular functions. The discovery that mutations in low-density lipoprotein receptor-related protein 5, a putative Wnt coreceptor, could positively and negatively affect bone mass in humans generated an enormous amount of interest in the possible role of the Wnt signaling pathway in skeletal biology. Over the last decade, considerable progress has been made in determining the role of the canonical Wnt signaling pathway in various aspects of skeletal development. Furthermore, recent evidence indicates the important role of non-canonical Wnt signaling in skeletal development. In this review we discuss the current understanding of the role of Wnt signaling in chondrogenesis, osteoblastogenesis, and osteoclastogenesis. PMID- 18164183 TI - FEMS - Federation of European Microbiological Societies: past, present and future. PMID- 18164184 TI - Comparison of gyrA gene mutations between laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis strains and clinical isolates. AB - To understand the relationship between mutations in the quinolone resistance determining region (QRDR) of the gyrA gene and drug resistance to ofloxacin, 85 laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis mutant strains and 110 M. tuberculosis clinical isolates, screened by denaturing high performance liquid chromatography to contain mutations, were analysed for their mutation patterns by sequencing as well as their ofloxacin minimal inhibitory concentrations (MICs). All mutations detected occurred at the codons Ala74, Ala90, Ser91 and Asp94 in all strains. One of the five different forms of missense mutation in Asp94 occurred in 60% of the laboratory-selected strains and 78% of the clinical isolates. However, 53 clinical isolates (48%) and only 2 laboratory-selected strains (2.4%) harboured double point mutations. The mutation Ala74Ser occurred only in the clinical isolates and only in combination with the Asp94Gly mutation. The ofloxacin MIC for the clinical isolates ranged from 0.5microg/mL to 20microg/mL, whilst the MICs for the laboratory-selected strains were > or =10microg/mL. The differences in gyrA gene mutation patterns and MICs between the laboratory-selected resistant strains and clinically isolated resistant strains identified here might help to understand the mechanisms involved in fluoroquinolone resistance. PMID- 18164185 TI - Sodium butyrate, an epigenetic interventional strategy, attenuates a stress induced alteration of MK-801's pharmacologic action. AB - Twenty-four hours after mice are exposed to a single session of forced swimming in cold water, the ability of MK-801 (dizocilpine), a noncompetitive NMDA receptor antagonist, to antagonize electrically precipitated seizures is reduced. Conceivably, this reduction in MK-801's antiseizure efficacy reflects a stress induced alteration in NMDA receptor-mediated neurotransmission due to changes in gene expression 24 h after a single stress. Recently, epigenetic interventional strategies impacting expression of genes whose regulation is controlled by the acetylation status of histone proteins in the nucleosome, an octomeric complex of histone proteins and promoter regions of double-stranded DNA, have been tested in preclinical models of various neuropsychiatric disorders, including Huntington disease and major depression. These strategies have been studied extensively in cancer biology. In the current investigation, the severity of the stress-induced reduction of MK-801's ability to raise the threshold voltage for the elicitation of tonic hindlimb extension was reduced when sodium butyrate (1.5 g/kg, ip) was administered around the time of stress. Prior research showed that this dose of sodium butyrate reliably increased the acetylation status of H3 and H4 histone proteins in the hippocampus and cerebral cortex of mice. Thus, the attenuation of the stress-induced reduction of MK-801's antiseizure efficacy may be due to the increased acetylation of histone proteins in the nucleosomal core and promotion of gene expression. These data encourage development of epigenetic strategies to prevent some of the deleterious consequences of stress. PMID- 18164186 TI - Osteogenic properties of human myogenic progenitor cells. AB - Here, we identified human myogenic progenitor cells coexpressing Pax7, a marker of muscle satellite cells and bone-specific alkaline phosphatase, a marker of osteoblasts, in regenerating muscle. To determine whether human myogenic progenitor cells are able to act as osteoprogenitor cells, we cultured both primary and immortalized progenitor cells derived from the healthy muscle of a nondystrophic woman. The undifferentiated myogenic progenitors spontaneously expressed two osteoblast-specific proteins, bone-specific alkaline phosphatase and Runx2, and were able to undergo terminal osteogenic differentiation without exposure to an exogenous inductive agent such as bone morphogenetic proteins. They also expressed the muscle lineage-specific proteins Pax7 and MyoD, and lost their osteogenic characteristics in association with terminal muscle differentiation. Both myoblastic and osteoblastic properties are thus simultaneously expressed in the human myogenic cell lineage prior to commitment to muscle differentiation. In addition, C3 transferase, a specific inhibitor of Rho GTPase, blocked myogenic but not osteogenic differentiation of human myogenic progenitor cells. These data suggest that human myogenic progenitor cells retain the capacity to act as osteoprogenitor cells that form ectopic bone spontaneously, and that Rho signaling is involved in a critical switch between myogenesis and osteogenesis in the human myogenic cell lineage. PMID- 18164187 TI - Microbial surfactant-enhanced mineral oil recovery under laboratory conditions. AB - Microbial enhanced oil recovery (MEOR) is potentially useful to recover incremental oil from a reservoir being beyond primary and secondary recovery operations. Effort has been made to isolate and characterize natural biosurfactant produced by bacterial isolates collected from various oil fields of ONGC in Assam. Production of biosurfactant has been considered to be an effective major index for the purpose of enhanced oil recovery. On the basis of the index, four promising bacterial isolates: Pseudomonas aeruginosa (MTCC7815), P. aeruginosa (MTCC7814), P. aeruginosa (MTCC7812) and P. aeruginosa (MTCC8165) were selected for subsequent testing. Biosurfactant produced by the promising bacterial isolates have been found to be effective in the recovery of crude oil from saturated column under laboratory conditions. Two bacterial strains: P. aeruginosa (MTCC7815) and P. aeruginosa (MTCC7812) have been found to be the highest producer of biosurfactant. Tensiometer studies revealed that biosurfactants produced by these bacterial strains could reduce the surface tension (sigma) of the growth medium from 68 to 30 mN m(-1) after 96 h of growth. The bacterial biosurfactants were found to be functionally stable at varying pH (2.5-11) conditions and temperature of 100 degrees C. The treatment of biosurfactant containing, cell free culture broth in crude oil saturated sand pack column could release about 15% more crude oil at 90 degrees C than at room temperature and 10% more than at 70 degrees C under laboratory condition. PMID- 18164188 TI - [Juvenile dermatomyositis: diagnosis and treatment]. AB - Juvenile dermatomyositis is a rare inflammatory myopathy with characteristic skin manifestations and muscular weakness. Muscle biopsy is important for establishing the diagnosis. Four patients with juvenile dermatomyositis were studied retrospectively. Steroids remain the first line treatment. Corticosteroids resistance is the primary indication for the use of intravenous immunoglobulins or immunosuppressive drugs. Further studies are necessarily aimed at finding biological markers to select and guide new therapeutical approaches for those patients. PMID- 18164189 TI - Berberine prevents UV-induced MMP-1 and reduction of type I procollagen expression in human dermal fibroblasts. AB - Matrix metalloproteinases (MMPs) induction and type I procollagen reduction in photoaging of the skin due to exposure to ultraviolet (UV) irradiation. Therefore, regulation of these genes has been suggested to be a useful tool to abolish skin aging. In this study, antioxidative plant ingredients used in traditional Chinese medicine, berberine (BBR) was investigated for their capacity to regulate MMP-1 and type I procollagen expression in human dermal fibroblasts. Our results showed that both basal and UV-induced MMP-1 expression was decreased by BBR. On the other hand, type I procollagen expression was dose-dependently increased by it. In addition, UV-induced reduction of type I procollagen expression is recovered by it. Therefore, we suggest that BBR may be a possible candidate for anti-skin aging agent. PMID- 18164190 TI - Bacterial extracts for the prevention of acute exacerbations in chronic obstructive pulmonary disease: a point of view. AB - Given the high prevalence of chronic obstructive pulmonary disease (COPD), the impact of exacerbations on quality of life, and the costs incurred, effective ways for the prevention of exacerbations, and for reductions in the severity and duration of COPD symptoms are needed. Bacterial immunostimulation has been advocated as a management strategy in COPD for the purposes of preventing acute exacerbations. In particular, it suggests that the use of oral multicomponent vaccines may reduce the severity and duration of acute episodes. The way in which bacterial extracts may exert their effects is not fully understood although a number of possible specific mechanisms have been suggested. Given the high prevalence of COPD worldwide and the high cost of acute exacerbations, some cost effectiveness analyses suggest that bacterial immunostimulants may become a key element in the improved control of this condition. Nonetheless, larger and longer clinical trials are needed to investigate efficacy before oral vaccination could be recommended as part of the routine clinical management of COPD, mainly in advanced COPD. It remains also to be investigated whether this protective effect may be additive to the other treatments. In any case, it is well known that for Streptococcus pneumoniae, non-typable Haemophilus influenzae and Moraxella catarrhalis, recurrent infections occur because of strain heterogeneity. Therefore, a single or even multiple strain vaccine with a killed whole cell formulation is possibly not the ideal vaccine. Moreover, the method of inactivation can affect the immunogenicity of essential antigens through denaturation. For this reason, the efficacy of bacterial immunostimulants should not only be assessed but also compared. PMID- 18164191 TI - An innovative maintenance follow-up program after a first inpatient pulmonary rehabilitation. AB - Although the benefits of pulmonary rehabilitation (PR) have been demonstrated in patients with COPD, most studies suggest that short-term programs are insufficient to maintain the benefits beyond a post-discharge period of 6 months to 1 year. We were interested to evaluate the effects of an innovative maintenance intervention compared with a usual after-care. Forty moderate to severe COPD patients, who had just completed their first inpatient PR, were consecutively included in either a maintenance group (MG) or a standard after care group. The maintenance program was coordinated within a health-care network including self-help associations, and offered weekly activities. We measured the 6-min walk distance (6MWD), the quality of life using the St George Respiratory Questionnaire (SGRQ), the dyspnea, the maximal workload and the health-care utilization. Data were collected at respiratory clinic admission and discharge, and at 6- and 12-month visits after the PR. After 12 months, we found statistically and clinically significant differences in favor of the MG in 6MWD (74 m; p < or = 0.01) and in the three domains of SGRQ: symptom (19%; p < or = 0.01), activity (27%; p < or = 0.01) and impact (32%; p < or = 0.01). The results showed no difference between groups in dyspnea and maximal workload. We also found that the number of days spent in hospital for respiratory disorders was significantly lower in the MG after 12 months (p < or = 0.03). The multidisciplinary management of COPD patients in the post-rehabilitation period within a health-care network including self-help associations seems to be an effective strategy for maintaining, and even improving, the benefits of a first initial structured program. PMID- 18164192 TI - Different expression ratio of S100A8/A9 and S100A12 in acute and chronic lung diseases. AB - Calgranulins are a family of powerful chemoattractants, which have been implicated as biomarkers in inflammatory diseases. To determine how different respiratory diseases affect the expression of calgranulins, we measured the expression of S100A8/A9 and S100A12 in bronchoalveolar lavage fluid (BALF) of acute respiratory distress syndrome (ARDS) patients and healthy volunteers by ELISA. Analysis of calgranulin expression revealed a high level of S100A12 in the lavages of patients suffering from ARDS compared to controls (p<0.001). Based on the hypothesis that the increased expression of S100A12 relative to the S100A8/A9 heterodimer was a characteristic of respiratory diseases with neutrophilic inflammation, we measured calgranulin expression in BALF of cystic fibrosis (CF) patients. Despite similarly elevated levels of S100A8/A9, S100A12 was significantly higher in ARDS compared to CF BALF (p<0.001). The differential expression of calgranulins was unique for inflammatory markers, as an array of cytokines did not differ between CF and ARDS patients. Since ARDS is an acute event and CF a chronic inflammation with acute exacerbations, we compared calgranulin expression in sputum obtained from CF and patients with chronic obstructive lung disease (COPD). Levels of S100A12 and S100A8/9 were elevated in CF sputum compared to COPD sputum, but the ratio of S100A12 to S100A8/A9 was similar in COPD and CF and reflected more closely than seen in healthy controls. The results indicate that the regulation of human calgranulin expression and the ratio of S100A8/A9 to S100A12 may provide important insights in the mechanism of respiratory inflammation. PMID- 18164193 TI - Opioid dependent patients' experiences of and attitudes towards having their injecting sites examined. AB - BACKGROUND: This study explored the attitude towards, and experiences of, injection site examination among injecting drug users in opioid treatment and the potential impact of this routine examination on information disclosure and future injection practices. METHODS: A self-complete, anonymous, cross-sectional questionnaire was used with 153 patients recruited from three public clinics in Sydney, Australia. RESULTS: The vast majority (97%) had ever injected in their upper limb, 19% in their leg, 16% in their neck, and 7% in their groin. The majority were 'happy to have their sites inspected' (78%), and felt it was an 'appropriate part of routine examination' (72%). Seventy-seven percent said they would be more honest about recent injecting, and 25% would inject in other sites if upper limb inspection occurred at every clinical review. CONCLUSIONS: The examination of injecting sites can provide useful corroboration of self-reported injecting drug use and an opportunity to offer harm reduction advice. The inspection of injecting sites was acceptable to most patients and should form part of routine clinical reviews. PMID- 18164194 TI - Role of surface adsorption and porosity features in the molecular recognition ability of imprinted sol-gels. AB - Organically modified molecularly imprinted silicas (MIS) for nafcillin recognition were prepared using a simple sol-gel procedure. Molecular recognition of the template was observed by tuning the chemical and structural properties of the MIS. The relative amounts of organically modified alkoxysilane precursors were found to be key in the textural and morphological characteristics of the MIS as well as for developing an imprinting effect in the materials. The recognition properties of the imprinted materials were found to be strongly influenced by the hydrolytic stability of the alkoxysilanes and their inductive effects during sol gel hydrolysis/condensation stages. The concept was to combine properties of organic groups with those of glass-like materials in order to develop synergetic properties through variations in the composition. Results from batch rebinding experiments as well as from the thorough study of the N(2) adsorption properties and the textural and structural characteristics of the MIS revealed that an imprint effect could be attributed to the presence of the template during the synthesis of MIS. PMID- 18164195 TI - Organ weight and serum triglyceride responses of older (80 week) commercial laying hens fed an alfalfa meal molt diet. AB - Since one of the costs in the commercial egg industry is that of replacement pullets, commercial egg layer managers have opted to induce molt older hens in order to extend their productive life for additional egg laying cycles. Conventional molt induction involves the complete removal of feed for several days. However, this management practice can lead to deleterious physiological responses by the hen and subsequent susceptibility to infection by pathogens. Consequently less stressful molting regimens involving the feeding of low energy diets such as alfalfa have been developed. In this study, 80 week old laying hens that were deprived of feed or fed alfalfa meal during a nine day induced molt. Full fed hens were used as the control. On day 8 serum triglycerides were quantified and on day 9 hens were euthanized and the liver, spleen, heart, intestine, pancreas, ovary, and kidney were collected and weighed. Intestinal weight were highest in the non-molted hens, lower in the hens fed alfalfa, and lower still in the hens deprived of feed. Molted hens exhibited reduced weights of liver, heart, ovary, and pancreas compared to the non-molted hens. Serum triglycerides were highest in the non-molted hens, less in feed deprived hens, and the lowest in alfalfa fed hens. These results suggest that a comparable molt could be achieved with feeding alfalfa meal to 80 week hens compared to feed deprivation. PMID- 18164196 TI - Development of applications of industrial enzymes from Malaysian indigenous microbial sources. AB - Malaysian enzyme industry is considered almost non-existence, although the import volume is large. Realizing the importance of enzymes, encompassing a wide range of applications in bioindustry, the development of home grown technologies for enzyme production and applications becomes one of the national priorities in industrial biotechnology. Enzyme production from indigenous microbial isolates was performed either by submerged or solid state fermentation processes. Based on its wide and unique spectrum of properties, enzymes have been developed for wide applications in various industrial processes. The development of the enzyme catalysed applications is based on the modification of the reaction systems to enhance their catalytic activities. Some of the applications of the industrial enzymes include the fine chemicals production, oleochemicals modification, detergent formulation, enzymatic drinking of waste papers, animal feed formulation and effluent treatment processes. Enzymes have also shown to be successfully used as analytical tool in the determination of compounds in body fluids. Although, most of these enzyme catalysed reactions were performed in aqueous phase, the use of enzymes in organic solvents was found to be significant for the production of new chemicals. PMID- 18164197 TI - Antimitotic quinoid triterpenes from Maytenus chuchuhuasca. AB - Four cytotoxic quinoid triterpenes, tingenone (1), 22 beta-hydroxytingenone (2), pristimerin (3), and celastrol (4), isolated from Maytenus chuchuhuasca, potently inhibit the polymerization of tubulin. PMID- 18164198 TI - Synthesis and anti-HIV activity of GS-9148 (2'-Fd4AP), a novel nucleoside phosphonate HIV reverse transcriptase inhibitor. AB - GS-9148 (2'-Fd4AP, 4) has been identified as a nucleoside phosphonate reverse transcriptase (RT) inhibitor with activity against wild-type HIV (EC(50)=12 microM). Unlike many clinical RT inhibitors, relevant reverse transcriptase mutants (M184V, K65R, 6-TAMs) maintain a susceptibility to 2'-Fd4AP that is similar to wild-type virus. The 2'-fluorine group was rationally designed into the molecule to improve the selectivity profile and in preliminary studies using HepG2 cells, compound 4 showed no measurable effect on mitochondrial DNA content indicating a low potential for mitochondrial toxicity. PMID- 18164199 TI - Potent heteroarylpiperidine and carboxyphenylpiperidine 1-alkyl-cyclopentane carboxamide CCR2 antagonists. AB - This report describes replacement of the 4-(4-fluorophenyl)piperidine moiety in our CCR2 antagonists with 4-heteroaryl piperidine and 4-(carboxyphenyl) piperidine subunits. Some of the resulting analogs retained potency in our CCR2 binding assay and had improved selectivity versus the I(Kr) channel; poor selectivity against I(Kr) had been a liability of earlier analogs in this series. PMID- 18164200 TI - Synthesis and protective effects of coumarin derivatives against oxidative stress induced by doxorubicin. AB - The use of doxorubicin (DOX) in the treatment of solid tumors is limited by cardiotoxicity essentially due to oxidative stress generation. The aim of this study was to identify coumarin derivatives displaying a protective antioxidant activity without affecting DOX antitumoral efficiency. A set of eighteen coumarinic derivatives was synthesized. Their antioxidant power was evaluated in vitro with the FRAP (ferric reducing ability of plasma) method and in human breast adenocarcinoma MCF7 cells using H(2)DCFDA (2',7' dichlorodihydrofluorescein diacetate) in a cytometric analysis. 4-Methyl-7,8 dihydroxycoumarin was found to exhibit an important antioxidant strength, a low cytotoxicity, and could decrease ROS (reactive oxygen species) production generated by DOX treatment without affecting DOX cytotoxicity in MCF7 cells. PMID- 18164201 TI - Exponential decline of deep-sea ecosystem functioning linked to benthic biodiversity loss. AB - BACKGROUND: Recent investigations suggest that biodiversity loss might impair the functioning and sustainability of ecosystems. Although deep-sea ecosystems are the most extensive on Earth, represent the largest reservoir of biomass, and host a large proportion of undiscovered biodiversity, the data needed to evaluate the consequences of biodiversity loss on the ocean floor are completely lacking. RESULTS: Here, we present a global-scale study based on 116 deep-sea sites that relates benthic biodiversity to several independent indicators of ecosystem functioning and efficiency. We show that deep-sea ecosystem functioning is exponentially related to deep-sea biodiversity and that ecosystem efficiency is also exponentially linked to functional biodiversity. These results suggest that a higher biodiversity supports higher rates of ecosystem processes and an increased efficiency with which these processes are performed. The exponential relationships presented here, being consistent across a wide range of deep-sea ecosystems, suggest that mutually positive functional interactions (ecological facilitation) can be common in the largest biome of our biosphere. CONCLUSIONS: Our results suggest that a biodiversity loss in deep-sea ecosystems might be associated with exponential reductions of their functions. Because the deep sea plays a key role in ecological and biogeochemical processes at a global scale, this study provides scientific evidence that the conservation of deep-sea biodiversity is a priority for a sustainable functioning of the worlds' oceans. PMID- 18164202 TI - TORC1 is essential for NF1-associated malignancies. AB - Inactivating mutations in NF1 underlie the prevalent familial cancer syndrome neurofibromatosis type 1 [1]. The NF1-encoded protein is a Ras GTPase-activating protein (RasGAP) [2]. Accordingly, Ras is aberrantly activated in NF1-deficient tumors; however, it is unknown which effector pathways critically function in tumor development. Here we provide in vivo evidence that TORC1/mTOR activity is essential for tumorigenesis. Specifically, we show that the mTOR inhibitor rapamycin potently suppresses the growth of aggressive NF1-associated malignancies in a genetically engineered murine model. However, in these tumors rapamycin does not function via mechanisms generally assumed to mediate tumor suppression, including inhibition of HIF-1alpha and indirect suppression of AKT, but does suppress the mTOR target Cyclin D1 [3]. These results demonstrate that mTOR inhibitors may be an effective targeted therapy for this commonly untreatable malignancy. Moreover, they indicate that mTOR inhibitors do not suppress all tumor types via the same mechanism, suggesting that current biomarkers that rely on HIF-1alpha suppression may not be informative for all cancers. Finally, our results reveal important differences between the effects of mTOR inhibition on the microvasculature in genetically engineered versus xenograft models and indicate that the former may be required for effective preclinical screening with this class of inhibitors. PMID- 18164203 TI - Core-modified porphyrins. Part 6: Effects of lipophilicity and core structures on physicochemical and biological properties in vitro. AB - Thiaporphyrins 2-8 were prepared as analogues of 5,20-diphenyl-10,15-bis[4 (carboxymethyleneoxy)-phenyl]-21,23- dithiaporphyrin (1) to examine the effect of structural modifications: substituent changes in meso aryl groups of dithiaporphyrins with one water-solubilizing group (2-5), dihydroxylation of a pyrrole double bond and reduction to dihydroxychlorins (6 and 7), and the removal of two meso aryl groups to give unsubstituted meso positions (8). The impact of these structural modifications was measured in both physicochemical (UV spectra, generation of singlet oxygen, lipophilicity, and aggregate formation) and biological properties (dark toxicity and phototoxicity, cellular uptake, and subcellular localization). Mono-functionalized porphyrins had much higher lipophilicity than di-functionalized porphyrin 1 and, consequently, formed more aggregates in aqueous media. The formation of aggregates might lower the efficiency of lipophilic porphyrins as photosensitizers. Interestingly, dihydroxylation of a core pyrrole group in the dithiaporphyrin core did not affect either the absorption spectrum or the efficiency for generating singlet oxygen. The phototoxicity of dihydroxydithiachlorins mainly depended on their intracellular uptake. The potent phototoxicity of 6, IC(50)=0.18muM, was attributed to the extraordinarily high uptake. The intracellular uptake of 6 was about 7.6 times higher than 1. In contrast, thiaporphyrin 8 with only two meso aryl groups was less effective as a photosensitizer, perhaps due to poorer uptake and a lower quantum yield for the generation of singlet oxygen. PMID- 18164204 TI - Discovery of piperazinylimidazo[1,2-a]pyridines as novel S4 binding elements for orally active factor Xa inhibitors. AB - We have recently reported the discovery of orally active sulfonylalkylamide Factor Xa (FXa) inhibitors, as typified by compound 1 (FXa IC(50)=0.061 microM). Since the pyridylpiperidine moiety was not investigated in our previous study, we conducted detailed structure-activity relationship studies on this S4 binding element. This investigation led to the discovery of piperazinylimidazo[1,2 a]pyridine 2b as a novel and potent FXa inhibitor (FXa IC(50)=0.021 microM). Further modification resulted in the discovery of 2-hydroxymethylimidazo[1,2 a]pyridine 2e (FXa IC(50)=0.0090 microM), which was found to be a selective and orally bioavailable FXa inhibitor with reduced CYP3A4 inhibition. PMID- 18164205 TI - Pyrimidinone-peptoid hybrid molecules with distinct effects on molecular chaperone function and cell proliferation. AB - The Hsp70 molecular chaperones are ATPases that play critical roles in the pathogenesis of many human diseases, including breast cancer. Hsp70 ATP hydrolysis is relatively weak but is stimulated by J domain-containing proteins. We identified pyrimidinone-peptoid hybrid molecules that inhibit cell proliferation with greater potency than previously described Hsp70 modulators. In many cases, anti-proliferative activity correlated with inhibition of J domain stimulation of Hsp70. PMID- 18164206 TI - Synthesis and cytotoxic activity of diosgenyl saponin analogues. AB - Diosgenyl saponins are steroidal glycosides that are often found as major components in many traditional oriental medicines. Recently, a number of naturally occurring diosgenyl saponins have been shown to exert cytotoxic activity against several strains of human cancer cells. Use of these saponin compounds for cancer treatment is hampered due to the lack of understanding of their action mechanism as well as limited access to such structurally complicated molecules. In the present paper, we have prepared a group of diosgenyl saponin analogues which contain a beta-D-2-amino-2-deoxy-glucopyranose residue having different substituents at the amino group. Moderate cytotoxic activity is found for most analogues against neuroblastoma (SK-N-SH) cells, breast cancer (MCF-7) cells, and cervical cancer (HeLa) cells. The analogue 13 that contains an alpha lipoic acid residue exhibits the highest potency against all three cancer cell lines with IC(50) ranging from 4.8 microM in SK-N-SH cells to 7.3 microM in HeLa cells. Preliminary mechanistic investigation with one saponin analogue (10) shows that the compound induces cell cycle arrest at G(1) phase in SK-N-SH cells, but the same compound induces cell cycle arrest at G(2) phase in MCF-7 cells. This result suggests that the cytotoxic activity of these saponin analogues may involve different action mechanisms in cell lines derived from different cancer sites. PMID- 18164207 TI - Use of a bioindicator system in the study of the mutagenetical effects in the neighborhoods of deposits of radioactive waste. AB - The purpose of the research described in this paper was to analyze the biological mutational effects caused by low doses of ionizing radiation on biological samples placed nearby and around deposits of radioactive waste, as a way of monitoring the environment close to them. In order to do this, the plant Tradescantia pallida was chosen, and through micronuclei tests the sensitivity of the dose/response to bio-monitoring could be observed. The plants were exposed for a period of 24h in previously chosen sites around Brazil, within the proximity of nuclear waste deposits. In each location, three points were chosen for bio-monitoring. The results obtained at these locations showed a small increment in the frequency of micronuclei per cell of the biosensor. From these data, a scale of mutagenesis effects due to low-dose radiation was built up. T. pallida is a good alternative for environmental bio-monitoring in tropical climates, as it is an excellent alternative tool in the studies of the effects of ionizing radiation on the environment. PMID- 18164208 TI - Exclusion of the non-functioning right ventricle in children with pulmonary atresia and intact ventricular septum. AB - OBJECTIVE: A group of patients with pulmonary atresia and intact ventricular septum (PAIVS) have to undergo single ventricle repair. In these patients, the presence of the non-functioning right ventricle (RV) may lead to the aggravation of RV to coronary connections and left ventricular (LV) dysfunction. To prevent these deleterious effects, the RV was excluded surgically. METHODS: Between December 2000 and February 2006, 10 patients with PAIVS underwent RV exclusion in conjunction with cavo-pulmonary anastomosis (n=6) or systemic-to-pulmonary artery shunt (n=4). Median age at surgery was 5 months (range, 0.2-13.8). Median z-value of the tricuspid valve was -4.0 (range, -6.5 to -1.3). None had RV dependent coronary circulation. The tricuspid valve was closed directly or using a patch. Thrombotic materials were inserted into the RV cavity in eight patients. RESULTS: There was no mortality. Follow-up was completed in all patients for up to 79.1 months (median, 30.3). Seven patients underwent a Fontan procedure and the other three are waiting. Eight patients showed completely obliterated RV after exclusion. One of the two patients who had residual RV cavity underwent re exclusion after a Fontan operation. Postoperative echocardiography revealed that LV end-diastolic dimension increased with borderline significance (p=0.050), whereas LV end-systolic dimension showed no significant changes. During follow up, LV showed no evidence of regional ischemia or global dysfunction, and fractional shortening significantly increased compared with the pre-exclusion value (p=0.017). CONCLUSIONS: This study demonstrates that RV exclusion may be performed safely in a selected group of patients with PAIVS. This procedure may have beneficial effects on LV systolic function, by preventing possible ischemia, and on LV diastolic function, by obliterating the non-functioning cavity and improving the geometry of the ventricular septum. PMID- 18164209 TI - An attempt to correlate a "Multidimensional Geriatric Assessment" (MGA), treatment assignment and clinical outcome in elderly cancer patients: results of a phase II open study. AB - OBJECTIVES: To establish a correlation between a specific MGA category, an appropriate preventively established treatment and clinical outcome in a population of elderly cancer patients. The ultimate goal was to verify whether the appropriate treatment given to elderly cancer patients according to their MGA category could translate into a better clinical outcome assessed as clinical response and toxicity, i.e whether this process might achieve a clinically meaningful impact. PATIENTS AND METHODS: We carried out a phase II open, prospective non-randomized study in 75 elderly cancer patients (lung, head and neck, colorectal, gynecologic and breast) hospitalized at the Department of Medical Oncology, University of Cagliari, Italy. All patients underwent MGA evaluation and were assigned to three different categories: fit, intermediate and frail. Thereafter, an appropriate preventively established treatment was administered and the clinical outcome was assessed. The clinical outcome after 3 month treatment was defined on the basis of objective clinical response and toxicity. The difference of clinical outcome in the MGA categories was assessed by ANOVA test. Moreover, the correlation between MGA category and the clinical outcome (clinical response and toxicity) was assessed by Spearman's correlation test. RESULTS: A better clinical response was observed in fit patients as compared both to intermediate and frail patients. Treatment toxicity was comparable in the different MGA categories. The correlation analysis between MGA category, clinical response to treatment and toxicity showed that there was a significant direct correlation with clinical response and no correlation with toxicity. Overall, the regression analysis showed that MGA was predictive of clinical outcome, in the sense that it is truly predictive for clinical response and no predictive for toxicity. CONCLUSION: Our study demonstrates that the MGA, although time-consuming, is a useful and cost-benefit effective tool to appropriately select elderly cancer patients to be treated effectively in terms of a survival advantage and those who would benefit mainly in terms of improvement of quality of life. Moreover, the treatment preventively established for each MGA category was shown to be adequate and accomplished the most appropriate performances in terms of effectiveness and toxicity. PMID- 18164210 TI - Phosphodiesterase-Ialpha/autotaxin's MORFO domain regulates oligodendroglial process network formation and focal adhesion organization. AB - Development of a complex process network by maturing oligodendrocytes is a critical but currently poorly characterized step toward myelination. Here, we demonstrate that the matricellular oligodendrocyte-derived protein phosphodiesterase-Ialpha/autotaxin (PD-Ialpha/ATX) and especially its MORFO domain are able to promote this developmental step. In particular, the single EF hand-like motif located within PD-Ialpha/ATX's MORFO domain was found to stimulate the outgrowth of higher order branches but not process elongation. This motif was also observed to be critical for the stimulatory effect of PD Ialpha/ATX's MORFO domain on the reorganization of focal adhesions located at the leading edge of oligodendroglial protrusions. Collectively, our data suggest that PD-Ialpha/ATX promotes oligodendroglial process network formation and expansion via the cooperative action of multiple functional sites located within the MORFO domain and more specifically, a novel signaling pathway mediated by the single EF hand-like motif and regulating the correlated events of process outgrowth and focal adhesion organization. PMID- 18164211 TI - Expression and purification of cysteine introduced recombinant saporin. AB - Saporin, a ribosome inactivating protein is widely used for immunotoxin construction. Here we describe a mutation of saporin (sap)-3 DNA by introducing a cysteine residue, followed by protein expression and purification by ion exchange chromatography. The purified Cys255sap-3, sap-3 isomer and commercially purchased saporin, were tested for toxicity using assays measuring inhibition for protein synthesis. The IC(50) values showed that the toxicity of the Cys255sap-3 is equivalent to the sap-3 isomer and commercial saporin. Reactivity of Cys255sap-3 was confirmed by labeling with a thio-specific fluorescent probe as well as conjugation with a nonspecific mouse IgG. We have found that a single cysteine within saporin provides a method for antibody conjugation that ensures a uniform and reproducible modification of a saporin variant retaining high activity. PMID- 18164212 TI - Evidence for positive selection in the TLR9 gene of teleosts. AB - Toll-like receptors (TLRs) have been identified as key sensors of invading microbes by identifying pathogen-associated molecular patterns and activating innate immune responses. Whereas purifying selection has been suggested in mammalian TLR9, evolutionary features of TLR9 in teleosts have not been investigated in detail. We therefore analysed TLR9 DNA sequences of eight teleost species, including zebrafish (Danio rerio), Japanese flounder (Paralichthys olivaceus), pufferfish (Takifugu rubripes), and five seabreams. Eleven sites subjected to positive selection were identified using the codon-substitution models of PAML 3.15. Ten of these 11 sites were found to be associated with leucine-rich repeats (LRRs). Seven of these 10 positively selected sites were associated with the convex surface of the LRR solenoids, leading to variations of the structures of the LRRs possibly by the introduction of flexibility into the LRR solenoids. The positive selection of LRRs in TLR9 may indicate the adaptation of teleosts to different oligodeoxynucleotides present in different bacterial species. PMID- 18164213 TI - Neural correlates of envisioning emotional events in the near and far future. AB - Being able to envision emotional events that might happen in the future has a clear adaptive value. This study addressed the functional neuroanatomy of this process and investigated whether it is modulated by temporal distance. Participants imagined positive and negative events pertaining to the near future or far future while their brain activity was measured with fMRI. The results demonstrate that the anterior part of the ventromedial prefrontal cortex (vmPFC) was more active in envisioning emotional events in the far future than in the near future, whereas the caudate nucleus was engaged in envisioning emotional (especially positive) situations in the near future. We argue that the anterior part of the vmPFC might assign emotional values to mental representations of future events that pertain to long-term goals. On the other hand, the caudate might support more concrete simulations of action plans to achieve rewarding situations in the near future. PMID- 18164214 TI - Localization of correlated network activity at the cortical level with MEG. AB - In both hemodynamic and neurophysiological imaging methods, analysis of functionally interconnected networks has typically focused on brain areas that show strong activation in specific tasks. Alternatively, connectivity measures may be used directly to localize network nodes, independent of their level of activation. This approach requires initial cortical reference areas which may be identified based on their high level of activation, their coherence with an external reference signal, or their strong connectivity with other brain areas. Irrespective of how the nodes have been localized the mathematical complexity of the analysis methods precludes verification of the accuracy and completeness of the network structure by direct comparison with the measured data. Therefore, it is critical to understand how the choices of parameters and procedures used in the analysis affect the network identification. Here, using simulated and measured magnetoencephalography (MEG) data, and Dynamic Imaging of Coherent Sources (DICS) for connectivity analysis, we quantify the veracity of network detection at the individual and group level as a function of relevant parameter choices. Using simulations, we demonstrate that coupling measures enable accurate identification of the network structure even without external reference signals, and illustrate the applicability of this approach to real data. We show that a valid estimate of interindividual variability is critical for reliable group level analysis. Although this study focuses on application of DICS to MEG data, many issues considered here, especially those regarding individual vs. group level analysis, are likely to be relevant for other neuroimaging methods and analysis approaches as well. PMID- 18164215 TI - Effects of topiramate on seizure susceptibility in kainate-kindled rats: involvement of peripheral-type benzodiazepine receptors. AB - This study was aimed to quantitatively evaluate the effects of topiramate (TPM) on seizure susceptibility and hippocampal peripheral-type benzodiazepine receptors (PBRs) in the kainic acid (KA) model of temporal lobe epilepsy. Male rats were randomized into saline control group, KA group, KA/TPM low dose group and KA/TPM high dose group. Three weeks after single injection of KA (10 mg kg( 1), sc), the effects of TPM were tested at two doses (10 and 30 mg kg(-1), sc) once a day for 1 week in KA/TPM low dose group or KA/TPM high dose group, respectively. Rats in KA group received comparable injections of saline. Four weeks after initial KA injection, a subconvulsant dose KA (5 mg kg(-1), sc) was administered in rats in these three groups. Rats in saline control group received equal volume of saline. All animals were decapitated and hippocampus synaptosomes were purified 180 min after behavioral observation. PBRs specific binding sites were assessed by an in vitro binding technique utilizing the highly selective ligand [(3)H]PK11195. Seizure threshold was elevated and specific PBRs binding in hippocampus was decreased by TPM in dose-dependent manner. Specific PBRs binding in hippocampus was significantly related to seizure latency and seizure intensity. These results suggest that TPM can reduce the susceptibility to seizures in KA-kindled rats and its anticonvulsant effect seems resulting from, at least in part, the reduced PBRs binding after treatment. These results also support the hypothesis that PBRs represent a novel target for antiepileptic drug development. PMID- 18164216 TI - Disorders of reproduction in patients with epilepsy: antiepileptic drug related mechanisms. AB - Epilepsy, antiepileptic drugs (AEDs), and the reproductive system have complex interactions. Fertility is lower in both men and women with epilepsy than in the general population. Moreover, reproductive endocrine disorders are more common among patients with epilepsy than among the population in general. These disorders have been attributed both to epilepsy itself and to AEDs. The use of the liver enzyme inducing AEDs phenobarbital, phenytoin and carbamazepine increases serum sex hormone binding globulin (SHBG) concentrations in both men and women with epilepsy. Over time the increase in serum SHBG levels leads to diminished bioactivity of testosterone and estradiol, which may result in diminished potency in men and menstrual disorders in some women, and, thus, to reduced fertility. Valproate (VPA) medication may have effects on serum androgen concentrations and it reduces serum follicle stimulating hormone levels in men with epilepsy. However, the clinical significance of the VPA related reproductive endocrine changes in men is unknown. On the other hand, in women the use of VPA is associated with a frequent occurrence of reproductive endocrine disorders characterized by polycystic changes in the ovaries, high serum testosterone concentrations (hyperandrogenism) and menstrual disorders. Young women with epilepsy seem to be especially vulnerable to the effects of VPA on serum androgen levels. The endocrine effects of the new AEDs have not been widely studied. However, it seems they may offer an alternative if reproductive endocrine problems emerge during treatment with the older antiepileptic drugs. On the other hand, it seems that in many cases the reproductive endocrine effects of the AEDs are reversible, if the medication is discontinued. PMID- 18164217 TI - Does menopause affect the epilepsy? AB - Hormonal fluctuations in the menstrual cycle have been shown to have an impact on the epilepsy in women with catamenial epilepsy. The epilepsy frequency is higher in the elderly, and this part of the population is steadily increasing. It is therefore of interest whether the hormonal changes in the menopausal transition affects the epilepsy. The perimenopause is associated with hormonal fluctuations and may worsen the epilepsy in women with catamenial epilepsy. It has been suggested that seizures may improve after menopause, particularly in the women with catamenial epilepsy. There is still however, limited data on the effects of menopause on the epilepsy. Prospective studies imply a long time span, and research in this field is scarce. More research on the relationship between hormonal fluctuations and seizures may enable us to predict the effect on the epilepsy. Special awareness should be given to a woman with epilepsy in the menopausal transition. PMID- 18164218 TI - Decision-making in temporal lobe epilepsy surgery: the contribution of basic non invasive tests. AB - PURPOSE: We studied the extent to which the widely used diagnostic tests contribute to the decision whether or not to perform temporal lobe epilepsy (TLE) surgery in The Netherlands. METHODS: This nation-wide, retrospective study included 201 consecutive patients referred for TLE surgery screening. The individual and combined contribution of nine index tests to the consensus decision to perform surgery was investigated. The contribution of each test was quantified using multivariable logistic regression and ROC curves. RESULTS: Surgery was performed in 119 patients (59%). Patient history and routine EEG findings were hardly contributory to decision-making, whereas a convergence of MRI with long-term interictal and ictal EEG findings correctly identified the candidates considered eligible for surgery (25% of total). Videotaped seizure semiology contributed less to the results. The area under the ROC curve of the combination of basic tests was 0.75. Ineligibility was never accurately predicted with any test combination. CONCLUSIONS: In the Dutch presurgical work-up, when MRI and long-term EEG findings were concordant, a decision for TLE surgery could be reached without further ancillary tests. Videotaped seizure semiology contributed less than expected to the final clinical decision. In our study, basic test findings alone were insufficient to exclude patients from surgery. PMID- 18164219 TI - Aluminum ions inhibit phospholipase D in a microtubule-dependent manner. AB - Aluminum is a highly cytotoxic metal to plants, but the molecular base and the primary target of Al toxicity are still unknown. The most important physiological consequence of Al toxicity is a cessation of root growth and changes in root morphology suggesting a role of the root cytoskeleton as a target structure. The important role of phospholipid degrading enzyme phospholipase D in regulation of cytoskeleton remodelling in both animal and plant organisms is now evident. Both the phospholipid pathway and the cytoskeleton are influenced by Al(3+), but their relationship with Al stress remains to be explored. Therefore, we tested the possibility that Al stress could be sensed by plants through microtubules in close interaction with phospholipases. We have shown that Al(3+) reduced the formation of phosphatidic acid in vivo, inhibited activity of phosphatidylinositol-4,5-bisphosphate-dependent phospholipase D in vitro and that the phosphatidic acid production is modified by microtubule dynamics. PMID- 18164220 TI - Constitutive endocytosis in characean internodal cells is independent of an intact actin cytoskeleton. AB - We have investigated constitutive endocytosis in internodal cells of the characean green algae. The endocytic tracer FM1-43 accumulated in distinct plasma membrane domains that are probably enriched in sterol-like substances. Internalization of the dye was active but independent of an intact actin or microtubule cytoskeleton. PMID- 18164221 TI - S1P1 receptor signaling overrides retention mediated by G alpha i-coupled receptors to promote T cell egress. AB - The mechanism by which sphingosine-1-phosphate receptor-1 (S1P1) acts to promote lymphocyte egress from lymphoid organs is not defined. Here, we showed that CCR7 deficient T cells left lymph nodes more rapidly than wild-type cells did, whereas CCR7-overexpressing cells were retained for longer. After treatment with FTY720, an agonist that causes downmodulation of lymphocyte S1P1, CCR7-deficient T cells were less effectively retained than wild-type T cells. Moreover, treatment with pertussis toxin to inactivate signaling via G alpha i-protein-coupled receptors restored egress competence to S1P1-deficient lymphocytes. We also found that T cell accumulation in lymph node cortical sinusoids required intrinsic S1P1 expression and was antagonized by CCR7. These findings suggest a model where S1P1 acts in the lymphocyte to promote lymph node egress by overcoming retention signals mediated by CCR7 and additional G alpha i-coupled receptors. Furthermore, by simultaneously upregulating S1P1 and downregulating CCR7, T cells that have divided multiple times switch to a state favoring egress over retention. PMID- 18164222 TI - T helper 17 lineage differentiation is programmed by orphan nuclear receptors ROR alpha and ROR gamma. AB - T cell functional differentiation is mediated by lineage-specific transcription factors. T helper 17 (Th17) has been recently identified as a distinct Th lineage mediating tissue inflammation. Retinoic acid receptor-related orphan receptor gamma (ROR gamma) was shown to regulate Th17 differentiation; ROR gamma deficiency, however, did not completely abolish Th17 cytokine expression. Here, we report Th17 cells highly expressed another related nuclear receptor, ROR alpha, induced by transforming growth factor-beta and interleukin-6 (IL-6), which is dependent on signal transducer and activator of transcription 3. Overexpression of ROR alpha promoted Th17 differentiation, possibly through the conserved noncoding sequence 2 in Il17-Il17f locus. ROR alpha deficiency resulted in reduced IL-17 expression in vitro and in vivo. Furthermore, ROR alpha and ROR gamma coexpression synergistically led to greater Th17 differentiation. Double deficiencies in ROR alpha and ROR gamma globally impaired Th17 generation and completely protected mice against experimental autoimmune encephalomyelitis. Therefore, Th17 differentiation is directed by two lineage-specific nuclear receptors, ROR alpha and ROR gamma. PMID- 18164223 TI - Measurement of urinary hepcidin levels by SELDI-TOF-MS in HFE-hemochromatosis. AB - INTRODUCTION: Insufficient production of hepcidin, the master regulator of iron metabolism, is recognized as the key pathogenetic feature of HFE-related hereditary hemochromatosis (HH). There is a growing interest in measuring the hepcidin levels, which may improve the diagnosis, prognostic evaluation and clinical management of HH. Nevertheless, few investigative tools are available: an immunodot method for urinary hepcidin developed by a single centre (UCLA), not yet ready for large-scale diffusion, and mass spectrometry (MS) based assays, such as surface-enhanced laser desorption/ionization time-of-flight (SELDI-TOF MS). The latter is well suited to small peptides like hepcidin, and can rapidly analyze crude samples with high throughput. This study measured urinary hepcidin levels by SELDI-TOF-MS in a large group of HH patients at diagnosis and during treatment, including both C282Y homozygous and C282Y/H63D compound heterozygotes. METHODS: We used a protocol based on PBSIIc mass spectrometer and Normal Phase chips. Urinary samples from 30 control subjects were compared to those obtained from 80 HH patients (57 C282Y homozygotes, 23 C282Y/H63D compound heterozygotes). Eighteen C282Y homozygotes and 11 C282Y/H63D compound heterozygotes were analyzed at diagnosis, the remainder during maintenance phlebotomy. RESULTS: C282Y homozygotes either at diagnosis, or after phlebotomy had significantly lower urinary hepcidin levels than controls (P<0.05). C282Y/H63D compound heterozygotes had hepcidin levels at diagnosis higher than controls, while the hepcidin/ferritin ratio was significantly decreased (P<0.001) suggesting inadequate hepcidin production. After phlebotomy, mean hepcidin levels in the compound heterozygotes were significantly lower than in controls (P<0.001). Samples from 12 randomly selected control subjects were sent to UCLA for duplicate measurement by the immunodot method, yielding a significant correlation (rho=0.64; P=0.024). CONCLUSIONS: This study supports the use of SELDI-TOF-MS for measuring hepcidin levels in research and clinical applications. Our results in phlebotomized patients suggest that the depletion of iron stores may further exacerbate the HFE-related hepcidin defect. PMID- 18164224 TI - The exp1 gene essential for pileus expansion and autolysis of the inky cap mushroom Coprinopsis cinerea (Coprinus cinereus) encodes an HMG protein. AB - The homobasidiomycete Coprinopsis cinerea is a member of the fungi known as inky cap mushrooms, and its fruiting-body pileus autolyzes soon after completion of the development. During the last 3h of the development, the pileus exhibits umbrella-like expansion: the pileal tissue is cracked at the base of each gill and then each gill tissue is split to form a V-shape, as seen in a cross section. We identified two C. cinerea mutants defective in both pileus expansion and autolysis. The defects in both mutants are due to recessive mutations in a single gene, designated exp1. The exp1 gene is predicted to encode an HMG1/2-like protein with two HMG domains. The transcription of exp1 is strongly induced in the pileus 3h before pileus expansion. This result, together with the fact that the exp1 mutations cause a specific developmental phenotype, suggest that Exp1 is a novel, transcriptional regulator controlling the final phase of fruiting-body morphogenesis. PMID- 18164225 TI - Comparison of psychological and physical function in neuropathic pain and nociceptive pain: implications for cognitive behavioral pain management programs. AB - Research has increased our understanding of the psychological and physical functioning associated with persistent pain and has facilitated the development of cognitive behavioral pain management programs to help improve people's physical function and decrease their distress in the presence of persistent pain. The majority of this research has focused on nociceptive pain or pain of mixed etiology. There has been less focus on these aspects of neuropathic pain. It is possible that differences exist in the function and difficulties associated with nociceptive and neuropathic pain. These differences may be associated with our clinical observation that some people with neuropathic pain have difficulty applying some aspects of the theory and practice of cognitive behavioral pain management. The purpose of this study was to compare a single neuropathic pain condition (post-herpetic neuralgia) with a persistent pain of nociceptive origin (low back pain) and determine whether differences exist in: (1) physical and psychological function; (2) factors that increase difficulties; (3) responses to pain; (4) beliefs about pain and (5) problems experienced. The results suggest that the differences between the two groups were not on the major variables of pain, mood, cognition and physical function. The main differences were in factors that increase pain, people's responses to pain, their beliefs about diagnosis and the cause of pain and the problems they reported as a result of experiencing pain. The implications of our findings for the development of cognitive behavioral pain management programs for people with neuropathic pain are discussed. PMID- 18164226 TI - Confinement exposure induces glucose regulated protein 75 (GRP75/mortalin/mtHsp70/PBP74/HSPA9B) in the hepatic tissue of gilthead sea bream (Sparus aurata L.). AB - Glucose regulated protein 75 (GRP75/mortalin/mtHsp70/PBP74/HSPA9B) is a molecular chaperone that was partially cloned and sequenced in gilthead sea bream (Sparus aurata L.) using a RT-PCR and 3'RACE approach. The deduced amino acid sequence supported the early vertebrate divergence of the heat shock protein 70 family into cytoplasmic Hsp70/Hsc70 group, endoplasmic reticulum-resident group and the mitochondrial-type group of GRP75. The tissue-specific regulation of GRP75 was analyzed by real-time PCR and Western blot after acute (24 h, 120 kg/m(3)) and prolonged confinement exposure (3 weeks-trial, 45-50 kg/m(3)). In both experiments, GRP75 gene expression was not significantly altered in brain, head kidney and gills. By contrast, hepatic transcripts of GRP75 were up-regulated and the magnitude of the response was dependent on the intensity of stressor. Furthermore, similar increments in hepatic transcripts and protein levels of GRP75 were found after prolonged confinement exposure. In addition, these stressed animals exhibited a 10% reduction in feed efficiency, significantly increased glycaemia and plasma peroxidases, and their plasma transaminases and respiratory burst of circulating leucocytes were significantly decreased. This stress-mediated response may act in concert with the increased production of hepatic GRP75 to protect metabolically active tissues against oxidative damage. PMID- 18164227 TI - Reduction of acetate accumulation in Escherichia coli cultures for increased recombinant protein production. AB - The culture of Escherichia coli for the commercial production of recombinant proteins has increased significantly in recent years. The production of acetate as a byproduct retards cell growth, inhibits protein formation, and diverts carbon from biomass to protein product. Our approach to reducing acetate accumulation was to disable the phosphoenolpyruvate:sugar phosphotransferase system (PEP-PTS) by deleting the ptsHI operon in the wild-type E. coli strain GJT001. The mutation caused a severe reduction in growth rate and glucose uptake rate in glucose-supplemented M9 minimal medium, which confirmed the mutation, and eliminated acetate accumulation. The mutant strain (TC110) apparently metabolized glucose by a non-PTS mechanism that we are currently investigating, followed by phosphorylation by glucokinase. In complex medium such as 2xLB broth with 2% glucose, TC110 was able to grow quickly and still retained the phenotype of significantly reduced acetate accumulation (9.1+/-6.6 vs. 90.4+/-1.6mM in GJT001, P<0.05). The reduced acetate accumulation resulted in a significant improvement in final OD (23.5+/-0.7 in TC110 vs. 8.0+/-0.1 in GJT001, P<0.05). We tested the strains for the production of model recombinant proteins such as green fluorescent protein (GFP) and beta-galactosidase. TC110 had a 385-fold improvement in final volumetric productivity of GFP over GJT001 in shake flasks with 2xLB broth with 2% glucose. The distribution of GFP fluorescence in the cell population, as determined by flow cytometry, was much broader in GJT001 (coefficient of variation=466+/-35%) than in TC110 (coefficient of variation=55+/ 1%). In corn steep liquor medium with 2% glucose, we observed a 28.5-fold improvement in final volumetric production of GFP in TC110 over GJT001. TC110 had a 7.5-fold improvement in final volumetric productivity of beta-galactosidase over GJT001 in 2xLB broth with 2% glucose medium. When tested in a batch bioreactor cultures with 2xLB broth with 2% glucose medium, the volumetric production of GFP by TC110 was 25-fold higher than that of GJT001. In summary, the ptsHI mutant of GJT001 resulted in reduced acetate accumulation, which led to significant improvements in recombinant protein production in batch bioreactors. PMID- 18164228 TI - Spectrum of MMACHC mutations in Italian and Portuguese patients with combined methylmalonic aciduria and homocystinuria, cblC type. AB - Methylmalonic aciduria (MMA) and homocystinuria, cblC type (MIM 277400) is the most frequent inborn error of vitamin B(12). The recent identification of the disease gene, MMACHC, has permitted preliminary genotype-phenotype correlations. We studied 24 Italian and 17 Portuguese patients with cblC defect to illustrate the spectrum of mutations in a southern European population and discuss the impact that mutation identification has on routine diagnostic procedures. Since the metabolic defect raises the serum levels of homocysteine, we also tested if variants in MTHFR-playing a key role in homocysteine remethylation pathway-could act as genetic modifier in cblC defect. We found that the c.271dupA (accounting for 55% of the MMACH alleles in our cohort) followed by c.394C>T (16%) and c.331C>T (9%) were the most frequent mutations. In our study we also identified a novel mutation (c.544T>C). On the other hand, the MTHFR genotype did not appear to influence age at onset, the clinical phenotype and outcome of patients with cblC defect. This study shows that mutation screening for the most common MMACH mutations occurring in early-onset forms (c.271dupA and c.331C>T) seems to have a high diagnostic yield in a southern European population with cblC defect. Although the identification of the gene defect per se does not predict completely time and severity of disease appearance, our data corroborate the importance of a molecular testing to offer accurate prenatal diagnosis to couples at high risk of having affected children. PMID- 18164229 TI - Correct terminology for electrodiagnostic studies. PMID- 18164230 TI - [Perilunate dislocations and fracture-dislocations of the wrist, a review of 14 cases]. AB - OBJECTIVES: We report a retrospective series of 14 dislocations or perilunate fracture-dislocations. The results of our series are compared with the data of the literature and we discuss epidemiology, types of lesions, surgical treatment, complications and prognosis of this pathology. METHODS: The series included seven pure dislocations and seven fracture-dislocations including three trans-scapho lunate forms (including one Fenton's syndrome). The displacement of all these lesions was posterior. The mean age was 35 years. Sixty-four percent were manual workers. All 14 patients had undergone surgical treatment through a dorsal approach in the first seven days following the injury. They were reviewed clinically and radiologically with a mean follow-up of 25 months. RESULTS: The average Cooney functional score was 72/100 with two excellent, six good, four fair and two poor results. Average flexion-extension motion arc was 74%, the grip strength was 77% compared to the other wrist. Persistent wrist pain was almost constant. One carpal instability was observed and one patient required a four corner arthrodesis for SLAC wrist. Eighty-five percent of all patients were employed at least. CONCLUSIONS: Early diagnosis and anatomical reduction can provide satisfactory functional results. Emergency surgical treatment is required. We prefer a dorsal approach and we do not perform primary closed reductions. PMID- 18164231 TI - Effect of ultrasound sonication on electroplating of iridium. AB - Effect of ultrasound sonication was examined on the electroplating of iridium in aqueous hexabromoiridate(III) solution. The electrodeposits were evaluated by observing the defects of the iridium deposits by means of voltammetry, in which the current-potential curves of the iridium deposits on copper were measured. Applying ultrasound sonication to the electroplating of iridium decreased the defects including the cracks in the deposit whenever the glycerol as the additives was contained or not in the electrolyte. PMID- 18164232 TI - Nrf2 signaling: an adaptive response pathway for protection against environmental toxic insults. AB - Human exposures to environmental toxicants have been associated with development of a number of diseases. Animal experiments have identified a number of cytoprotective enzymes under the transcriptional control of NF-E2-related factor 2 (Nrf2) including electrophile conjugation and antioxidative enzymes and enzymes responsible for the production of antioxidants, reducing equivalents and cofactors. The up-regulation of these enzymes represents an adaptive response which occurs in the face of exposure to electrophilic or oxidative compounds thereby leading to enhanced metabolism of these molecules or their reactive metabolites. This adaptive response is regulated by an interaction between Keap1 and Nrf2 in which the exposure to reactive molecules is sensed either directly by Keap1 or indirectly by cellular signaling cascades resulting in activation of Nrf2 transcriptional regulation. The Nrf2-mediated adaptive response has been shown to attenuate toxicity and carcinogenesis during electrophile or oxidative stress as well as inflammation in rodent models. The cytoprotective attributes of the Nrf2 signaling pathway have been targeted for chemoprevention as administration of Nrf2-inducing agents has been shown to result in decreased carcinogenesis in animal models and altered carcinogen metabolism in humans. On the other hand, polymorphisms in the Nrf2 signaling pathway can lead to differential susceptibility to disease while mutations in the Nrf2 signaling pathway have been shown to an effective mechanism for cancer cells to evade chemotherapy. Overall, the Nrf2 cytoprotective adaptive response has evolved to be a powerful protective strategy for organisms against exposure to environmental toxicants and may provide insight into differential disease susceptibilities across populations and responses to therapies designed to alleviate these conditions. PMID- 18164233 TI - Human respiratory syncytial virus and other viral infections in infants receiving palivizumab. AB - BACKGROUND: Palivizumab is a humanized monoclonal antibody that prevents severe human respiratory syncytial virus (HRSV) infections. OBJECTIVES: We determined the etiology of respiratory viral infections in palivizumab recipients, and monitored the clinical outcome and HRSV genotype in HRSV-infected infants. STUDY DESIGN: Nasopharyngeal aspirates (NPAs) were collected from children receiving palivizumab who consulted or were hospitalized for acute respiratory tract infection (ARTI) during the 2004-2005 season. Viral cultures and multiplex RT-PCR for influenza A/B, HRSV and human metapneumovirus were performed. The fusion (F) gene of HRSV amplicons was also sequenced. RESULTS: Among 116 enrolled patients, 51 (44%) had > or = 1 episode of ARTI for a total of 93 visits. At least one virus was identified in 33 (36%) of the 93 NPA samples; HRSV accounted for 11 (33%) of confirmed viral etiologies. Compared to subjects who had other viral ARTI, HRSV-positive subjects had less fever (p=0.01) and tended to have more bronchiolitis (p=0.07). Ten subjects (11 visits) developed HRSV infection, although only one was hospitalized. HRSV was detected after a median of 5.5 palivizumab doses and a median of 14 days after the last dose. One of the 11 HRSV strains tested had a F mutation located in the palivizumab-binding site. CONCLUSION: HRSV is still a major cause of ARTI in children receiving palivizumab, although the outcome of infected children appears mild. PMID- 18164234 TI - Rapid routine detection of enterovirus RNA in cerebrospinal fluid by a one-step real-time RT-PCR assay. AB - BACKGROUND AND OBJECTIVES: This study provides a one-step transcription/real-time (TaqMan probe) PCR assay (TM-PCR) with new consensus primer and probe sequences for generic detection of human pathogenic enteroviruses including difficult to detect ones like for instance Echovirus 30. The amplicon included parts of domain IV and V of the highly conserved internal ribosomal entry site. Generic detection was confirmed by testing a panel of 41 prototypes representing all five human enterovirus/poliovirus species. STUDY DESIGN AND RESULTS: The 95% detection limit was found to be 100 copies per run using in vitro transcribed coxsackievirus B3 RNA. TM-PCR was compared to an in house nested-PCR assay implemented in detecting enterovirus RNA from CSF samples of patients suffering from meningitis and encephalitis. Concordant results were obtained in all samples (11 positive, 101 negative). Specificity was confirmed with laboratory strains of other neurotropic viruses, and by testing 76 CSF samples of patients with encephalomyelitis disseminata, which all gave negative results. CONCLUSIONS: The new TM-PCR is a convincing alternative to conventional PCR protocols for the diagnosis of enterovirus meningitis. The one-step strategy limits hands on time and cross contamination risk combined with accelerated assay procedure of only 100 min. PMID- 18164235 TI - HCMV microinfections in inflammatory diseases and cancer. AB - Human cytomegalovirus (HCMV) is a wide-spread human virus that was mainly known to cause disease in immunocompromised patients. A new entity of infection can be diagnosed with high sensitive techniques; HCMV microinfections that often exhibit an altered pattern of IE protein expression. We have recently discovered that HCMV microinfections are very common in patients with inflammatory diseases and certain cancers. The discovery of active HCMV infections in tissue specimens from patients with inflammatory diseases raises the question of whether the infection is an epiphenomenon or whether the virus plays a causative role in disease development. After a primary infection, which is generally asymptomatic in immunocompetent individuals, HCMV establishes latency and persists in its host. In infected cells, the virus can produce over 250 proteins, but only about 50-60 are believed to be essential for viral replication. Thus, the vast majority of these viral proteins enable the virus to co-exist with its host. Such proteins act through highly sophisticated mechanisms to control different cellular and immunological functions in order to facilitate viral production and to avoid detection and elimination of the virus by the immune system. These proteins may also contribute to the development of common inflammation-related diseases. PMID- 18164236 TI - Diagnostic criteria for amyotrophic lateral sclerosis (ALS). PMID- 18164237 TI - Abnormal corticomotor excitability assessed in biceps brachii preceding pronator contraction post-stroke. AB - OBJECTIVE: To assess corticomotor (CM) excitability of the antagonist biceps brachii (BB) post-stroke in preparation for pronator contraction. In healthy subjects, we previously demonstrated that prior to pronator contraction CM excitability of the antagonist BB was suppressed. METHODS: Transcranial magnetic stimulation (TMS) was used to assess pre-contraction changes in motor evoked potential (MEP) amplitude of the BB, when BB was acting either as an antagonist or an agonist. TMS was applied 100-200ms prior to rhythmic isometric BB or pronator contractions in chronic stroke survivors and age/gender matched healthy control subjects. RESULTS: Prior to pronator contraction, MEPs in BB were elicited in the stroke group but were absent in healthy controls indicating that CM excitability of the antagonist BB was increased post-stroke. The extent of the abnormal increase in excitability positively correlated with the extent of upper limb motor impairment. CONCLUSIONS: Our results suggest that an alteration of cortical control mechanisms regulating motor excitability of the antagonist BB may contribute to the impairment of upper limb motor coordination post-stroke. SIGNIFICANCE: This study offers a unique approach to study the potential for a cortical origin of post-stroke motor discoordination. PMID- 18164238 TI - The topography of the scalp-recorded visual N700. AB - OBJECTIVE: To describe the topography of the N700 component of the scalp-recorded visual event-related potential (ERP) and to provide fundamental knowledge of the conditions under which it occurs. METHODS: We examined the time-course of visual ERP in response to the short (100ms) and prolonged (7s) presentation of simple salient visual stimuli separated by long interstimulus intervals employing high resolution 64-channel DC-EEG. Current source density (CSD) mapping and spatio temporal dipole source analysis were performed. RESULTS: CSD analysis revealed highly significant bilateral current sinks over occipito-temporal areas from about 450ms up to 1s after stimulus offset (visual N700). CSD topography and dipole source analysis suggested late prolonged activation of extrastriate visual areas which did not depend merely upon a stimulus offset response, afterimages or blinking, as confirmed by control conditions. CONCLUSIONS: Our findings provide basic knowledge about the time-course of sensory activation. We found that passive watching of rare salient short stimuli automatically evoked sustained activity in the extrastriate visual cortex up to 1s after stimulus offset. SIGNIFICANCE: Visual N700 provides a promising tool for important insights into the cortical mechanisms of stimulus post-processing. Its role in associative learning of temporally non-overlapping stimuli (automatic ultra-short-term sensory memory) should be explored. PMID- 18164239 TI - A novel approach for enhancing the functional specificity of TMS: revealing the properties of distinct neural populations within the stimulated region. PMID- 18164240 TI - Magnetoencephalographic pattern of epileptiform activity in children with early onset autism spectrum disorders. AB - OBJECTIVE: To provide further data around magnetoencephalographic (MEG) findings in early-onset autism spectrum disorders (ASD). METHODS: Thirty-six children (mean age 7 years) diagnosed of PDD (DSM-IV, ICD-10) were studied. There were 22 children with autistic disorder, 9 with Asperger's syndrome, and 5 with pervasive developmental disorder not otherwise specified (PDD-NOS). According to the Childhood Autism Rating Scale (CARS), the autistic disorder was mild to moderate in 11, and severe in 11. Neuroimaging studies using three-dimensional MRI as well as simultaneous MEG-EEG and fusion techniques through magnetic source imaging (MSI) were performed, with the aid of anesthesia in non-cooperative patients. RESULTS: Most patients had no EEG abnormalities. All ASD children showed common specific abnormalities in the shape of low amplitude monophasic and biphasic spikes (isolated or short bursts) as well as acute waves, predominantly distributed in the perisylvian areas. In Asperger's syndrome, epileptiform spikes were mostly found in the right hemisphere. No lateralized epileptiform activity was observed in non-Asperger's autistic patients. CONCLUSIONS: MEG epileptiform activity is frequently documented in children with early-onset ASD. SIGNIFICANCE: Subclinical epileptiform activity is present especially in the perisylvian regions for many patients with ASD. PMID- 18164241 TI - Giant somatosensory evoked potentials as indicator of nonconvulsive status epilepticus. PMID- 18164242 TI - Electrodiagnostic criteria for diagnosis of ALS. AB - A consensus meeting was held to determine the best use and interpretation of electrophysiological data in the diagnosis of ALS. The utility of needle EMG and nerve conduction studies was affirmed. It is recommended that electrophysiological evidence for chronic neurogenic change should be taken as equivalent to clinical information in the recognition of involvement of individual muscles in a limb. In addition, in the context of a suspected clinical diagnosis of ALS, fasciculation potentials should be taken as equivalent to fibrillation potentials and positive sharp waves in recognising denervation. The importance of searching for instability in fasciculation potentials and in motor unit potentials in ALS is stressed. These changes in the interpretation of electrophysiological data render obsolete the category Probable Laboratory Supported ALS in the modified El Escorial diagnostic criteria for ALS. Methods for detection of upper motor neuron abnormality appear sensitive but require further study, particularly regarding their value when clinical signs of upper motor neuron lesion are uncertain. PMID- 18164243 TI - Focal transcranial magnetic stimulation of motor cortex evokes bilateral and symmetrical silent periods in human masseter muscles. AB - OBJECTIVE: To determine whether a single hemisphere exerts distinct inhibitory influences over masseter muscles on each side, and to compare features of the masseter cortical silent period (CSP) evoked by transcranial magnetic stimulation (TMS) with previous reports from limb and other cranial muscles. METHODS: Focal TMS was applied over the motor cortex jaw area in 14 normal subjects. In one experiment, TMS intensity was constant (1.1 or 1.3x active motor threshold, T) and masseter muscle activation varied from 10% to 100% of maximal. In another experiment, muscle activation was constant (20% maximal) and TMS intensity varied from 0.7 to 1.3T. RESULTS: In all subjects, TMS evoked a silent period of similar duration in masseter muscles on both sides. Masseter CSP duration increased at higher TMS intensities, but was not affected by muscle activation level or the size of the excitatory response evoked by TMS. Weak TMS produced a bilateral CSP without short-latency excitation. The masseter CSP was short ( approximately 100ms at 1.3T), yet this was not due to maintenance of excitatory drive from the unstimulated hemisphere, as the masseter CSP was not prolonged with dual hemisphere TMS. CONCLUSIONS: Intracortical inhibitory circuits activated by TMS have a relatively weak effect on corticotrigeminal neurons supplying masseter, and effects are equivalent for corticobulbar efferents directed to contralateral and ipsilateral masseter motoneuron pools. SIGNIFICANCE: Trigeminally innervated masseter muscles exhibit weak, bilaterally symmetric inhibition following focal TMS. This method can be used to investigate abnormalities of intracortical inhibition in movement disorders or focal lesions affecting the masticatory muscles in humans. PMID- 18164244 TI - On the neural basis of fMRI signals. PMID- 18164245 TI - A randomised trial of a pre-synaptic stimulator of DA2-dopaminergic and alpha2 adrenergic receptors on morbidity and mortality in patients with heart failure. AB - BACKGROUND: By pre-synaptic stimulation of DA(2)-dopaminergic and alpha(2) adrenergic receptors, nolomirole inhibits norepinephrine secretion from sympathetic nerve endings. We performed a clinical study with nolomirole in patients with heart failure (HF). METHODS: The study was designed as a multicentre, double blind, parallel group trial of 5 mg b.i.d. of nolomirole (n=501) versus placebo (n=499) in patients with severe left ventricular systolic dysfunction, recently in New York Heart Association (NYHA) class III/IV. The primary endpoint was time to all cause death or hospitalisation for HF, whichever came first. The study was event driven and required 420 primary events. The study was completed as scheduled. RESULTS: Mean age of patients was 70 years, and 73% were male. Heart rate and blood pressure were not different in the two treatment groups. There were no changes in blood pressure. There were 233 primary events in the nolomirole group versus 208 in the placebo group (p=0.1). There were 142/145 deaths and 369/374 all cause hospitalisations in the nolomirole/placebo groups. There were no differences in walking distance, quality of life or NYHA class. CONCLUSION: A dose of 5 mg b.i.d. of nolomirole was not beneficial (or harmful) in patients with heart failure. PMID- 18164246 TI - The volume-sensitive chloride channel inhibitors prevent both contractile dysfunction and apoptosis induced by doxorubicin through PI3kinase, Akt and Erk 1/2. AB - Contractile dysfunction and cardiomyopathies secondary to apoptotic cell death are limiting factors for treating cancer with doxorubicin. Inhibition of volume sensitive chloride currents (I(Cl,vol)) has been reported to blunt doxorubicin induced apoptosis in cardiomyocytes. To investigate cellular contractility during acute induction of apoptosis by doxorubicin and to determine whether I(Cl,vol) inhibitors are able to prevent the subsequent contractile dysfunction, electrically paced ventricular myocytes freshly isolated from adult rabbits were acutely exposed to doxorubicin in the presence and absence of I(Cl,vol) inhibitors IAA-94 or DIDS. Doxorubicin induced increases in both annexin V labelling and caspase-3 activity and decreases in cell volume. Alteration in cardiac contractility was observed after doxorubicin exposure. Both IAA-94 and DIDS abolished the doxorubicin-induced decreases in peak shortening and cell volume as well as the increases in caspase-3 activity and annexin V labelling. These protective effects of I(Cl,vol) inhibitors were abolished by previous inhibition of PI(3)kinase, Akt and Erk 1/2. Thus, I(Cl,vol) inhibitors prevent doxorubicin-induced apoptosis and subsequent contractile dysfunction through PI(3)kinase/Akt and Erk 1/2. Inhibition of I(Cl,vol) may represent a new pharmacological strategy for developing cytoprotective drugs against apoptotic cell death and contractile dysfunction. PMID- 18164247 TI - High-nitrate levels in drinking water may be a risk factor for thyroid dysfunction in children and pregnant women living in rural Bulgarian areas. AB - Nitrate inhibits the accumulation of iodide in thyroid gland. The aim of present study was to evaluate the influence of this ion on the iodine status of two risk population groups. Subjects of study were pregnant women and children aged between 3 and 6 years from two villages in Bulgaria with high- and low-nitrate levels in drinking water. The relative risk of thyroid disorders for the pregnant women living in the village with high-nitrate levels in drinking water expressed as an odds ratio was 5.294 (95% confidence intervals 1.003-27.939; P=0.0454) and was considered as significant. Statistically significant differences were found between the goiter rate in exposed and non-exposed pregnant women. The relative risk of thyroid dysfunction for the children exposed to a high-nitrate level, expressed as an odds ratio was 2.333 (95% confidence intervals 0.8491-6.412; P=0.1396) and was considered as not significant; the goiter prevalence in the exposed children was also not statistically different. The results of the study confirmed the role of high-nitrate level in drinking water as a risk factor for thyroid dysfunction in vulnerable population groups. PMID- 18164248 TI - Can transgenic mosquitoes afford the fitness cost? AB - In a recent study, SM1-transgenic Anopheles stephensi, which are resistant partially to Plasmodium berghei, had higher fitness than non-transgenic mosquitoes when they were maintained on Plasmodium-infected blood. This result should be interpreted cautiously with respect to malaria control using transgenic mosquitoes because, despite the evolutionary advantage conferred by the transgene, a concomitant cost prevents it from invading the entire population. Indeed, for the spread of a resistance transgene in a natural situation, the transgene's fitness cost and the efficacy of the gene drive will be more crucial than any evolutionary advantage. PMID- 18164249 TI - Spouses' conceptions of the pre-hospital phase when their partners suffered an acute myocardial infarction--a qualitative analysis. AB - BACKGROUND: Delay from onset of acute myocardial infarction symptoms to the delivery of medical care is a major determinant of prognosis. Although studies have explored patient reasons for delay, there are only limited data concerning experiences of the spouse. AIM: Was to describe spouses' conceptions of the pre hospital phase when their partners suffered an acute myocardial infarction. METHOD: A phenomenographic approach was applied. Fifteen spouses were interviewed <48 h after the partner's hospital admittance. FINDINGS: Two categories with underlying sub-categories conceptualised the spouses' experiences. The category being resourceful contained: sharing the experience, having knowledge, understanding the severity, being rational, and consulting others. The category respecting independence contained: accepting the need for control, marital roles and experiences, restraining emotions, and seeking agreement. CONCLUSION: Our findings suggest that spouses have a strong influence on the course of events. When accepting the partner's need for control through following earlier marital roles and experiences, restraining own emotions and seeking agreement, this seemed to contribute to delay. However, when the spouse was resourceful by sharing the experience, having knowledge, understanding the severity, being rational and consulting others when needed, this seemed to have a positive influence on the pre-hospital time. PMID- 18164250 TI - Psychogenic nonepileptic seizures: answers and questions. AB - Psychogenic nonepileptic seizures (PNES) superficially resemble epileptic seizures, but are not associated with ictal electrical discharges in the brain. PNES constitute one of the most important differential diagnoses of epilepsy. However, despite the fact they have been recognized as a distinctive clinical phenomenon for centuries and that access to video/EEG monitoring has allowed clinicians to make near-certain diagnoses for several decades, our understanding of the etiology, underlying mental processes, and, subsequently, subdifferentiation, nosology, and treatment remains seriously deficient. Emphasizing the clinical picture throughout, the first part of this article is intended to "look and look again" at what we know about the epidemiology, semiology, clinical context, treatment, and prognosis of PNES. The second part is dedicated to the questions that remain to be answered. It argues that the most important reason our understanding of PNES remains limited is the focus on the visible manifestations of PNES or the seizures themselves. In contrast, subjective seizure manifestations and the biographic or clinical context in which they occur have been relatively neglected. PMID- 18164251 TI - Perceived impact of epilepsy in teenagers and young adults: an international survey. AB - OBJECTIVE: The purpose of this study was to survey the impact of epilepsy and its treatment on the quality of life, development, and opportunities for children/teenagers with the condition. METHODS: We asked three groups children/teenagers, parents and caregivers, and health care professionals-to complete specifically designed questionnaires containing clinical and demographic information and perceptions of the impact of epilepsy and its treatment on aspects of daily living, including education and leisure. For health care professionals, the survey included questions on area of speciality and volume of children with epilepsy seen in their clinic. Questionnaires were translated into the local written language in each participating country. Prepaid envelopes were provided for return of completed questionnaires. Responses were analyzed by an independent data analysis company. RESULTS: Two hundred twelve young people with epilepsy and 507 parents or carers of children/teenagers from 16 different countries completed the questionnaire. The majority of children were over the age of 12 (74%) and attended mainstream schools (64%). Approximately two-thirds (65%) of the children/teenagers who responded reported that their seizures caused them to lose, on average, 7 school days per year. More than one-third of the children/teenagers who responded to the survey (36%) have kept their epilepsy a secret from other people at some time for fear of being treated differently and a belief that people should not know (47%). The majority (87%) of respondents were taking epilepsy medications. More than a third of the children/teenagers who responded had experienced side effects in some form or another, with weight change (49%), headaches (46%), dizziness (41%), and shaking (33%) most commonly reported. More than one-third of the children/teenagers expected the condition to hinder their lives in the future, with impact on employment opportunities (73%), traveling and exploring (37%), and education (36%) most commonly reported. CONCLUSIONS: This survey documents some of the real-life consequences of childhood epilepsy and highlights important challenges and issues faced by children/teenagers with epilepsy and their families. Although there are a number of limitations to this study relating to sample selection, ensuring that children are as free from seizures as possible and minimizing the side effects of treatment must be the primary goal of epilepsy management, but considering the wider implications of epilepsy and treatment is also important. PMID- 18164252 TI - Recent developments of enantioseparation techniques for adrenergic drugs using liquid chromatography and capillary electrophoresis: a review. AB - This review provides the achievements of enantioseparation of adrenergic drugs and application of these methods in clinical and pharmaceutical analysis. The adrenergic agonist and antagonist drugs are analyzed in the direct and indirect modes by liquid chromatography (LC) and capillary electrophoresis (CE). Other chromatographic enantioseparation methods including super- and sub-critical fluid chromatography (SFC), and capillary electrochromatography (CEC) are presented likewise to analyse the cited compounds. The different separation processes for these drugs are briefly discussed and some applications are presented. PMID- 18164253 TI - Chest wall reconstruction in a patient with Cantrell syndrome. AB - Cantrell syndrome is a very rare congenital anomaly with up to five features: a midline, upper abdominal wall abnormality, lower sternal defect, anterior diaphragmatic defect, diaphragmatic pericardial defect, and congenital abnormalities of the heart. This report describes our experience of performing a reconstruction of a chest wall defect in a Cantrell syndrome case with herniation of the heart. The patient was a 1-month-old female infant who received surgical patch repair of a ventricular septal defect (VSD) and atrial septal defect (ASD) at the Department of Cardiac Surgery. Subsequently, the patient underwent reconstruction at the second-stage surgery. A rhomboid skin flap with an inferior pedicle was used to close the defect. In this process the flap, including portions of the rectus abdominis muscles, was elevated and transferred into the defect. The sectioned ends of the divided pectoralis major muscles were sutured together to simultaneously reconstruct the muscles. It has been 2 years since the surgery, and the defect is covered with normal skin, and the protrusion of the heart from the chest wall and the externally visible pulsation have been resolved. The progress has been very good functionally and cosmetically. PMID- 18164254 TI - Contaminating insert degradation by preincubation colony PCR: a method for avoiding false positives in transformant screening. AB - Colony PCR is a convenient alternative to conventional plasmid isolation and restriction digestion for high-throughput screening of recombinant colonies. However, an insert carryover from the ligation mix, adhered to colony or agar plate, generates a substantial number of false positives. To avoid this, a simple single-tube technique involving pre-PCR nuclease incubation has been developed by optimizing a buffer system that provides nuclease action and PCR amplification sequentially. Results presented in this work provide a technique that is amenable for high-throughput screening of recombinant colonies. PMID- 18164255 TI - A method of isolating the collagen (I) alpha2 chain carboxytelopeptide for species identification in bone fragments. AB - We present a novel method for the isolation and analysis of the bone collagen (I) alpha2 chain carboxytelopeptide as a species biomarker. Conventional methods for the analysis and sequencing of mixtures of proteins and peptides commonly involve using the protease trypsin to cleave proteins present in the sample. However, in the study of collagen, these methods result in very complex mixtures of peptides that are difficult to analyze and the acquired results are not reproducible. Here we use bacterial collagenase (from Clostridium histolyticum) for its ability to cleave the highly unusual Gly-Xaa-Yaa repeating sequence of collagen, where Xaa usually is Pro and Yaa often is Hyp. Followed by a simple isolation step using a reverse phase solid phase extraction cartridge, the alpha2 (I) chain carboxytelopeptide can be readily analyzed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and the results can be used to distinguish between different species of origin. PMID- 18164256 TI - Mutation in keratin 18 induces mitochondrial fragmentation in liver-derived epithelial cells. AB - Microtubules (MTs) and microfilaments (MFs) are known to modulate mitochondrial morphology, distribution and function. However, little is known evidence about the role of intermediate filaments (IFs) in modulating mitochondria except desmin. To investigate whether or not the IFs regulate mitochondrial morphology, distribution, and function, we manipulated the IFs of cultured epithelial cells to express a mutant keratin 18 (K18). In contrast to the filamentous expression of wild K18, mutant K18 induced aggregation of K8/18, showing no fine IF network in the cells. In mutant K18-transfected cells, the mitochondria were fragmented into small spheroids, although they were observed as mitochondrial fibers in un transfected or wild K18-transfected cells. Fluorescence recovery after photobleaching of fluorescence-labeled mitochondria was markedly less in the mutant K18-transfected cells, although a significant recovery was confirmed in wild K18-transfected cells. These findings suggest that the IFs are important for the maintenance of normal mitochondrial structures. PMID- 18164257 TI - Tauroursodeoxycholic acid reduces bile acid-induced apoptosis by modulation of AP 1. AB - Ursodeoxycholic acid (UDCA) is used in the therapy of cholestatic liver diseases. Apoptosis induced by toxic bile acids plays an important role in the pathogenesis of liver injury during cholestasis and appears to be mediated by the human transcription factor AP-1. We aimed to study if TUDCA can decrease taurolitholic acid (TLCA)-induced apoptosis by modulating AP-1. TLCA (20 microM) upregulated AP 1 proteins cFos (26-fold) and JunB (11-fold) as determined by quantitative real time PCR in HepG2-Ntcp hepatoma cells. AP-1 transcriptional activity increased by 300% after exposure to TLCA. cFos and JunB expression as well as AP-1 transcriptional activity were unaffected by TUDCA (75 microM). However, TUDCA significantly decreased TLCA-induced upregulation of cFos and JunB. Furthermore, TUDCA inhibited TLCA-induced AP-1 transcriptional activity and reduced TLCA induced apoptosis. These data suggest that reversal of bile acid-induced AP-1 activation may be relevant for the antiapoptotic effect of TUDCA in liver cells. PMID- 18164258 TI - Up-regulation of the Cbl family of ubiquitin ligases is involved in ATRA and bufalin-induced cell adhesion but not cell differentiation. AB - The Casitas B-lineage Lymphoma (Cbl) family of ubiquitin ligases is multifunctional proteins that play important roles in different cell signaling pathways. It has been reported that c-Cbl and Cbl-b mRNAs are up-regulated during TPA-induced U937 and HL-60 cell differentiation. But the mechanism of the up regulation and the roles of the Cbl family of ubiquitin ligases still remain unclear. In the present study, we demonstrated that bufalin enhanced all-trans retinoic acid (ATRA) induced differentiation of HL-60 cells, accompanied by up regulation of the Cbl family of ubiquitin ligases. CsA, an inhibitor of calcium mobilization, reversed this up-regulation. Pretreatment with CsA and PS-341 did not affect the expression of CD11b, but suppressed the percentage of adherent cells. Lipid raft localization of Cbl-b enhanced cell adhesion, while C-terminal deletion partially suppressed the effect. Moreover, the expression of the adhesion-related kinases Pyk2 and Paxillin was up-regulated in parallel with the increase of Cbl proteins. These results suggested that up-regulation of c-Cbl and Cbl-b was involved in the regulation of ATRA and bufalin-induced HL-60 cell adhesion rather than cell differentiation, which might be mediated by lipid raft localization, ubiquitin ligase activity and C-terminal structure of Cbl proteins. Meanwhile, up-regulation of proline-rich tyrosine kinase (Pyk2) and Paxillin might also be implicated in this regulation. PMID- 18164259 TI - Quantitative differences in chromatin accessibility across regulatory regions can be directly compared in distinct cell-types. AB - Transcriptional activation in eukaryotes is often accompanied by alterations to chromatin structure at specific regulatory sites while other genomic regions may remain unchanged. In this study, we have examined the correlation between expression and chromatin accessibility of the human CR2 gene in a panel of cell lines (U937, REH, Ramos, and Raji) using the CHART-PCR assay with the accessibility agent micrococcal nuclease (MNase). To validate the use of this assay for comparing multiple cell-types, we first tested a series of genomic regions to determine if we could observe consistent, site-specific levels of MNase chromatin accessibility. Promoter regions of the ubiquitously expressed genes GAPDH and beta-actin were similar and showed high accessibility to MNase digestion in each of the cell lines, while on the other hand, promoter regions of developmentally restricted genes PAX-7 and SP-A2 showed consistently reduced chromatin accessibility. Since CHART-PCR detected site-specific differences in chromatin accessibility in a manner that could be compared between cell-types, we next examined chromatin accessibility over the CR2 core promoter in the panel of cell lines representing either CR2 expressing or CR2 non-expressing cell-types. Our data revealed significantly enhanced accessibility over the -289 to -101 and the -115 to -12 regions of the CR2 promoter in expressing B-cells (Ramos, Raji) compared to non-expressing cells (U937, REH). Thus, CHART-PCR assays detected a correlation between chromatin accessibility and expression of the human CR2 gene, while the accessibility of other genomic regions was site-specific, but not altered between cell-types. PMID- 18164260 TI - Molecular cloning and developmental expression of plakophilin 2 in zebrafish. AB - Armadillo proteins are involved in providing strength and support to cells and tissues, nuclear transport, and transcriptional activation. In this report, we describe the identification and characterisation of the cDNA of the desmosomal armadillo protein plakophilin 2 in zebrafish. The 2448bp coding sequence encodes a predicted 815 amino acid protein, with nine armadillo repeats characteristic of the p120-catenin subfamily. It shares conserved N-glycosylation, myristoylation, and glycogen synthase kinase 3, casein kinase 2, and protein kinase C phosphorylation sites with mammalian armadillo proteins including plakoglobin and beta-catenin. Semi-quantitative reverse transcription polymerase chain reaction and whole mount in situ hybridisation show that it is expressed both maternally and zygotically. It is ubiquitously expressed during blastula stages but becomes restricted to epidermal and cardiac tissue during gastrulation. These results provide evidence that zebrafish plakophilin 2 is developmentally regulated with potential roles in cell adhesion, signalling, and cardiac and skin development. PMID- 18164261 TI - Competition potency of siRNA is specified by the 5'-half sequence of the guide strand. AB - Small-interfering RNAs (siRNAs) execute specific cellular gene silencing by exploiting the endogenous RNA interference (RNAi) pathway. Therefore, excess amounts of siRNAs can saturate cellular RNAi machineries. Indeed, some siRNAs saturate the RNA-induced silencing complex (RISC) and competitively inhibit silencing by other siRNAs. However, the molecular feature of siRNAs that specifies competition potency has been undetermined. While previous reports suggested a correlation between the competition potency and silencing efficiency of siRNAs, we found that the silencing efficiency was insufficient to explain the competition potency. Instead, we show that the nucleotide sequence of the 5'-half of the guide strand determines the competition potency of an siRNA. Our finding provides important information for understanding the mechanistic basis of competition in combinatorial RNAi treatment. PMID- 18164262 TI - Phosphorylation of eIF-4E positively regulates formation of the eIF-4F translation initiation complex following DNA damage. AB - The eukaryotic translation initiation factor 4E (eIF-4E) is essential for cap dependent protein translation. However, the role of eIF-4E phosphorylation in protein translation is still unclear. In this study, the function of eIF-4E phosphorylation in the formation of the translational initiation complex eIF-4F following DNA damage was investigated. Our results show that etoposide treatment caused a rapid increase in eIF-4E phosphorylation. The addition of CGP57380, a specific inhibitor of the eIF-4E kinase Mnk, not only inhibited eIF-4E phosphorylation but also resulted in reduced interaction between eIF-4E and eIF 4G. Furthermore, neither the p38 MAPK inhibitor nor the ERK inhibitor caused significant inhibition in eIF-4E phosphorylation induced by etoposide. However, a JNK-specific inhibitor, SP600125, strongly suppressed etoposide-induced eIF-4E phosphorylation. Our results provide the first evidence indicating that phosphorylation of eIF-4E by Mnk, possibly mediated by JNK or JNK-like kinases, is critical for formation of the translational initiation complex eIF-4F following DNA damage. PMID- 18164263 TI - Performance characteristics of the LiBASys des-gamma-carboxy prothrombin assay. PMID- 18164264 TI - Effects of ABCA1 SNPs, including the C-105T novel variant, on serum lipids of Brazilian individuals. AB - BACKGROUND: ABCA1 plays an important role in HDL metabolism. Single nucleotide polymorphisms (SNPs) in ABCA1 gene were associated with variation in plasma HDL c. METHODS: The effect of the ABCA1 SNPs C-14T, R219K and of a novel variant C 105T on serum lipids was investigated in 367 unrelated Brazilian individuals (224 hypercholesterolemic and 143 normolipidemic). The relation between ABCA1 SNPs and the lipid-lowering response to atorvastatin (10 mg/day/4 weeks) was also evaluated in 141 hypercholesterolemic (HC) individuals. The polymorphisms were detected by PCR-RFLP and confirmed by DNA sequencing. RESULTS: Linkage disequilibrium was found between the SNPs C-105T and C-14T in the HC group. HC individuals carrying -105CT/TT genotypes had higher serum HDL-c and lower triglyceride and VLDL-c concentrations as well as lower TG/HDL-c ratio compared to the -105CC carriers (p<0.05). The R219K SNP was associated with reduced serum triglyceride, VLDL-c and TG/HDL-c ratio in the HC group (p<0.05), and with an increased serum apoAI in NL individuals. The effects of ABCA1 SNPs on basal serum lipids of HC individuals were not modified by atorvastatin treatment. CONCLUSIONS: The ABCA1 SNPs R219K and C-105T were associated with a less atherogenic lipid profile but not with the lowering-cholesterol response to atorvastatin in a Brazilian population. PMID- 18164265 TI - Prominent increase in plasma ganglioside GM3 is associated with clinical manifestations of type I Gaucher disease. AB - BACKGROUND: Patients with Gaucher disease show signs of insulin resistance. The ganglioside GM3 has recently shown to be a negative regulator of insulin sensitivity. In fibroblasts of Gaucher patients, deficient in degradation of glucosylceramide, an increased anabolism of this lipid to gangliosides occurs. The goal of the current study was to establish whether GM3 is elevated in plasma of type I Gaucher disease patients, and is related to disease manifestations. METHODS: Plasma GM3, glucosylceramide, and ceramide were determined and compared to overall severity of disease, hepatomegaly, and plasma chitotriosidase activity. RESULTS: The ceramide concentration in plasma of untreated Gaucher patients was slightly but not significantly lower than in controls (median: 9.8 micromol/L, range: 5.7-14.7 micromol/L, (n=40) vs. median: 11.0 micromol/L, range: 5.1-18.0 micromol/L, (n=30)). Glucosylceramide was significantly (p<0.0001) elevated. GM3 was also significantly (p<0.0001) increased (median: 10.2 micromol/L, range: 4.3-19.1 micromol/L, (n=40) vs. median: 3.6 micromol/L, range: 2.7-5.4 micromol/L, (n=30)). Plasma GM3 concentrations correlated with those of plasma chitotriosidase activity (rho=0.45, p=0.0036), overall severity of disease (rho=0.39, p=0.012), and hepatomegaly (rho=0.49, p=0.0015). CONCLUSIONS: GM3 is strikingly elevated in plasma of most Gaucher patients. The increase is comparable to that of glucosylceramide, the primary storage lipid. The marked elevations in GM3 may play a role in the insulin resistance of Gaucher patients. PMID- 18164266 TI - Differential roles of p80- and p130-angiomotin in the switch between migration and stabilization of endothelial cells. AB - We have previously shown that angiomotin (Amot) plays an important role in growth factor-induced migration of endothelial cells in vitro. Genetic knock-down of Amot in zebrafish also results in inhibition of migration of intersegmental vessels in vivo. Amot is expressed as two different isoforms, p80-Amot and p130 Amot. Here we have analyzed the expression of the two Amot isoforms during retinal angiogenesis in vivo and demonstrate that p80-Amot is expressed during the migratory phase. In contrast, p130-Amot is expressed during the period of blood vessel stabilization and maturation. We also show that the N-terminal domain of p130-Amot serves as a targeting domain responsible for localization of p130-Amot to actin and tight junctions. We further show that the relative expression levels of p80-Amot and p130-Amot regulate a switch between a migratory and a non-migratory cell phenotype where the migratory function of p80-Amot is dominant over the stabilization and maturation function of p130-Amot. Our data indicates that homo-oligomerization of p80-Amot and hetero-oligomerization of both isoforms are critical for this regulation. PMID- 18164267 TI - Cartilage-hair hypoplasia-associated mutations in the RNase MRP P3 domain affect RNA folding and ribonucleoprotein assembly. AB - Cartilage-hair hypoplasia (CHH) is caused by mutations in the gene encoding the RNA component of RNase MRP. Currently it is unknown how these mutations affect the function of this endoribonuclease. In this study we investigated the effect of mutations in the P3 domain on protein binding and RNA folding. Our data demonstrate that a number of P3 nucleotide substitutions reduced the efficiency of its interaction with Rpp25 and Rpp20, two protein subunits binding as a heterodimer to this domain. The CHH-associated 40G>A substitution, as well as the replacement of residue 47, almost completely abrogated Rpp25 and Rpp20 binding in different assays. Also other CHH-associated P3 mutations reduced the efficiency by which the RNase MRP RNA is bound by Rpp25-Rpp20. These data demonstrate that the most important residues for binding of the Rpp25-Rpp20 dimer reside in the apical stem-loop of the P3 domain. Structural analyses by NMR not only showed that this loop may adopt a pseudo-triloop structure, but also demonstrated that the 40G>A substitution alters the folding of this part of the P3 domain. Our data are the first to provide insight into the molecular mechanism by which CHH associated mutations affect the function of RNase MRP. PMID- 18164268 TI - The functional implications of Akt activity and TGF-beta signaling in tamoxifen resistant breast cancer. AB - Development of acquired resistance to tamoxifen is a major clinical problem during endocrine treatment in estrogen receptor positive breast cancer. Transforming growth factor-beta1 (TGF-beta) has been implicated in tamoxifen induced cellular signaling in breast cancer, and increased Akt activation is associated with tamoxifen-resistant cell types. We hypothesized that the relationship between TGF-beta and Akt signaling may be involved in the development and progression of tamoxifen resistance. Tamoxifen-resistant (Tam-R) cells were established from parental MCF-7 cells by continuously exposing them to 4-hydroxytamoxifen (4-OHT). Tam-R cells were associated with a decrease in TGF beta1 secretion, TGF-beta-mediated transcriptional response, and growth inhibitory effects of 4-OHT. Tam-R cells expressed significantly higher levels of phosphorylated Akt and lower levels of phosphorylated Smad 3 in both the absence and presence of 4-OHT when compared to MCF-7 cells treated with 4-OHT. Ectopic expression of constitutively active Akt (Myc-Akt(Myr)) rendered MCF-7 cells resistant to activation by TGF-beta and the growth inhibitory effects of 4-OHT, while over-expression of kinase-dead Akt (Myc-Akt(K179M)) or LY294002 treatment of Tam-R cells enhanced TGF-beta activation and blocked cell growth. These results suggest that suppression of TGF-beta signaling by activated Akt is correlated with the development of tamoxifen resistance in breast cancer. PMID- 18164269 TI - Thioredoxin 2 haploinsufficiency in mice results in impaired mitochondrial function and increased oxidative stress. AB - The mitochondrial form of thioredoxin, thioredoxin 2 (Txn2), plays an important role in redox control and protection against ROS-induced mitochondrial damage. To evaluate the effect of reduced levels of Txn2 in vivo, we measured oxidative damage and mitochondrial function using mice heterozygous for the Txn2 gene (Txn2(+/-)). The Txn2(+/-) mice showed approximately 50% decrease in Trx-2 protein expression in all tissues without upregulating the other major components of the antioxidant defense system. Reduced levels of Txn2 resulted in decreased mitochondrial function as shown by reduced ATP production by isolated mitochondria and reduced activity of electron transport chain complexes (ETCs). Mitochondria isolated from Txn2(+/-) mice also showed increased ROS production compared to wild type mice. The Txn2(+/-) mice showed increased oxidative damage to nuclear DNA, lipids, and proteins in liver. In addition, we observed an increase in apoptosis in liver from Txn2(+/-) mice compared with wild type mice after diquat treatment. Our results suggest that Txn2 plays an important role in protecting the mitochondria against oxidative stress and in sensitizing the cells to ROS-induced apoptosis. PMID- 18164270 TI - Oxidation of mitochondrial peroxiredoxin 3 during the initiation of receptor mediated apoptosis. AB - It is hypothesized that activation of death receptors disrupts the redox homeostasis of cells and that this contributes to the induction of apoptosis. The redox status of the peroxiredoxins, which are extremely sensitive to increases in H2O2 and disruption of the thioredoxin system, were monitored in Jurkat T lymphoma cells undergoing Fas-mediated apoptosis. The only detectable change during the early stages of apoptosis was oxidation of mitochondrial peroxiredoxin 3. Increased H2O2 triggers peroxiredoxin overoxidation to a sulphinic acid; however during apoptosis peroxiredoxin 3 was captured as a disulfide, suggesting impairment of the thioredoxin system responsible for maintaining peroxiredoxin 3 in its reduced form. Peroxiredoxin 3 oxidation was an early event, occurring within the same timeframe as increased mitochondrial oxidant production, caspase activation and cytochrome c release. It preceded other major apoptotic events including mitochondrial permeability transition and phosphatidylserine exposure, and glutathione depletion, global thiol protein oxidation and protein carbonylation. Peroxiredoxin 3 oxidation was also observed in U937 cells stimulated with TNF-alpha. We hypothesize that the selective oxidation of peroxiredoxin 3 leads to an increase in mitochondrial H2O2 and that this may influence the progression of apoptosis. PMID- 18164272 TI - Depression-related differences in learning and forgetting responses to unrelated cues. AB - Using the think/no-think paradigm, we examined the effect of a meaningful connection between emotionally neutral cues and targets on initial learning and later recall by students in dysphoric or nondysphoric mood states. Compared to meaningfully connected cue-target pairs, unrelated pairs were less easily learned and more easily forgotten, even when initial learning was controlled. Depressive deficits were obtained in initial learning (only marginally) and final recall. When examined separately within each cuing condition, the recall deficit associated with depressed mood was restricted to the unrelated condition, but when initial learning differences were controlled this deficit was only marginally significant. Results are discussed in relation to other recent findings concerning depression-related performance in this paradigm and to findings of depression and memory more generally. PMID- 18164273 TI - Forgetting unwanted memories: directed forgetting and thought suppression methods. AB - Experimental psychopathologists have tested hypotheses regarding mechanisms that ought to be operative if victims possess skills for forgetting material related to trauma. In this article, we review research on directed forgetting and thought suppression paradigms, concentrating on laboratory studies involving attempts by individuals reporting trauma histories to forget emotionally negative material. Most studies have shown that trauma survivors, especially those with post traumatic stress disorder, are characterized by a breakdown in the ability to forget disturbing material. Studies on individuals reporting repressed or recovered memories of trauma have not confirmed predictions regarding heightened forgetting skills for trauma-related words. However, recent research on suppressing disturbing autobiographical memories suggests that people who report spontaneously recalling childhood abuse outside of psychotherapy may, indeed, possess skills for not thinking about disturbing material. PMID- 18164271 TI - NADPH oxidases in lung biology and pathology: host defense enzymes, and more. AB - The deliberate production of reactive oxygen species (ROS) by phagocyte NADPH oxidase is widely appreciated as a critical component of antimicrobial host defense. Recently, additional homologs of NADPH oxidase (NOX) have been discovered throughout the animal and plant kingdoms, which appear to possess diverse functions in addition to host defense, in cell proliferation, differentiation, and in regulation of gene expression. Several of these NOX homologs are also expressed within the respiratory tract, where they participate in innate host defense as well as in epithelial and inflammatory cell signaling and gene expression, and fibroblast and smooth muscle cell proliferation, in response to bacterial or viral infection and environmental stress. Inappropriate expression or activation of NOX/DUOX during various lung pathologies suggests their specific involvement in respiratory disease. This review summarizes the current state of knowledge regarding the general functional properties of mammalian NOX enzymes, and their specific importance in respiratory tract physiology and pathology. PMID- 18164274 TI - [Humoral hypercalcemia revealing a malignant non hodgkin lymphoma]. AB - INTRODUCTION: Hypercalcemia is a rare complication of non-Hodgkin lymphoma. Usually, hypercalcemia occurs late in the disease course, except for high-grade lymphoma. Most often hypercalcemia is related to excessive level of circulating PTH-rP or sometimes, 1,25(OH)2D3. Concomitant high plasmatic concentration of PTH rP and 1,25(OH)2D3 is uncommon. EXEGESIS: We report the case of a 82-year-old man who presented with abdominal pain and weight loss, leading to the diagnosis of diffuse large-B-cell lymphoma (high-grade lymphoma) associated with symptomatic hypercalcemia (3.21mmol/l). PTH-rP and 1,25(OH)2D3 plasmatic levels were high. Calcium concentration was normalized with glucocorticoids and sequential chemotherapy. CONCLUSION: This case report confirms that hypercalcemia, as consequence of excessive plasmatic level of PTH-rP secreted by tumoral cells, can occur early in the course of high-grade lymphoma. Glucocorticoids and chemotherapy are the best treatment options. PMID- 18164275 TI - Effects of variable lighting intensities and colour temperatures on sulphatoxymelatonin and subjective mood in an experimental office workplace. AB - Workplace illumination is of paramount importance in determining the employee's productivity and well-being. Moreover, light exerts non-visual effects with respect to biological rhythms. In this study, we investigated the impact of different lighting conditions (500-1800 lx, 6500 K; 500 lx, 4000 K) on sulphatoxymelatonin (aMT6-s) and subjective mood in an experimental office accommodation. Urinary aMT6-s concentrations were significantly decreased at all days of the experiment in both lights. On day 3, differences between aMT6-s concentrations in specimen collected at 05:00 p.m. and at 09:00 a.m. were significantly higher under variable lighting conditions. Analyses of a mood rating inventory revealed a benefit of variable light with respect to the dimensions of "Activity", while "Deactivation" and "Fatigue" were increased in regular light on day 1. "Activity", "Concentration", and "Deactivation" changed in opposite directions when comparing variable with regular illumination on two consecutive days. In conclusion, variable light exerts a potential advantage in indoor office accommodations with respect to subjective mood, although no unequivocal differences in the profile of aMT6-s were found as compared to regular light. PMID- 18164276 TI - The feature-positive effect and hypochondriacal concerns. AB - There is evidence to suggest that people have more problems with processing information that is absent (negative information) than with present (positive) information. This bias of overweighing positive and underweighing negative information has been termed the "feature-positive effect" (FPE). Typically, hypochondriasis is characterized by excessive focusing on bodily complaints (cf. positive information), and at the same time discarding negative medical test results (cf. negative information). It was explored whether the FPE is involved in this pattern. Fifty-three undergraduates completed a measure of hypochondriacal concerns and a general, domain-free test of the FPE. As expected, a positive correlation between the FPE and hypochondriacal concerns was observed. Implications for cognitive-behavioural theory are discussed. PMID- 18164277 TI - Cell shape, cell-cell contact, cell-extracellular matrix contact and cell polarity are all required for the maximum induction of CYP2B1 and CYP2B2 gene expression by phenobarbital in adult rat cultured hepatocytes. AB - The effect of cell shape, cell density, contact with extracellular matrix and cell polarity on the phenobarbital (PB)-induced gene expression of CYP2B1 and CYP2B2 (CYP2B1/2B2) in adult rat hepatocytes was investigated. High cell density enhanced the induction of CYP2B1/2B2 gene expression by PB. Hepatocytes cultured on EHS gel showed a spherical cell shape and highly enhanced the induction of CYP2B1/2B2 gene expression by PB. Although monolayer hepatocytes cultured on type I collagen (TIC) and type IV collagen exhibited poor induction of CYP2B1/2B2 gene expression by PB, monolayer cells on laminin showed substantial induction. The addition of soluble laminin to media did not show any effect on induction in monolayer hepatocytes cultured on TIC. Dishes coated with different concentrations of immovable laminin demonstrated complicated effects. Coating with higher concentrations of laminin resulted in greater induction of CYP2B1/2B2 gene expression by PB. On the other hand, when hepatocytes were cultured on dishes coated with lower concentrations of laminin, they became round and greater induction of CYP2B1/2B2 gene expression by PB was observed. Spherical hepatocytes cultured on low concentrations of TIC also showed highly enhanced induction of CYP2B1/2B2 gene expression by PB. EHS gel overlay to hepatocytes cultured on TIC and collagen sandwich configurations that are known to induce cell polarity enhanced the induction by PB. The induction of CYP2B1/2B2 gene expression needed cytoskeleton organization, such as actin filament, microtubule filament and intermediate filament. These results demonstrate that cell shape, cell density, contact with extracellular matrix and cell polarity all play critical roles in the induction of CYP2B1/2B2 gene expression by PB. PMID- 18164278 TI - Gene expression profiling elucidates a specific role for RARgamma in the retinoic acid-induced differentiation of F9 teratocarcinoma stem cells. AB - The biological effects of all-trans-retinoic acid (RA), a major active metabolite of retinol, are mainly mediated through its interactions with retinoic acid receptor (RARs alpha, beta, gamma) and retinoid X receptor (RXRs alpha, beta, gamma) heterodimers. RAR/RXR heterodimers activate transcription by binding to RA response elements (RAREs or RXREs) in the promoters of primary target genes. Murine F9 teratocarcinoma stem cells have been widely used as a model for cellular differentiation and RA signaling during embryonic development. We identified and characterized genes that are differentially expressed in F9 wild type (Wt) and F9 RARgamma-/- cells, with and without RA treatment, through the use of oligonucleotide-based microarrays. Our data indicate that RARgamma, in the absence of exogenous RA, modulates gene expression. Genes such as Sfrp2, Tie1, Fbp2, Emp1, and Emp3 exhibited higher transcript levels in RA-treated Wt, RARalpha-/- and RARbeta2-/- lines than in RA-treated RARgamma-/- cells, and represent specific RARgamma targets. Other genes, such as Runx1, were expressed at lower levels in both F9 RARbeta2-/- and RARgamma-/- cell lines than in F9 Wt and RARalpha-/-. Genes specifically induced by RA at 6h with the protein synthesis inhibitor cycloheximide in F9 Wt, but not in RARgamma-/- cells, included Hoxa3, Hoxa5, Gas1, Cyp26a1, Sfrp2, Fbp2, and Emp1. These genes represent specific primary RARgamma targets in F9 cells. Several genes in the Wnt signaling pathway were regulated by RARgamma. Delineation of the receptor specific actions of RA with respect to cell proliferation and differentiation should result in more effective therapies with this drug. PMID- 18164279 TI - A rhesus monkey model of self-injury: effects of relocation stress on behavior and neuroendocrine function. AB - BACKGROUND: Self-injurious behavior (SIB), a disorder that afflicts many individuals within both clinical and nonclinical populations, has been linked to states of heightened stress and arousal. However, there are no published longitudinal data on the relationship between increases in stress and changes in the incidence of SIB. This study investigated the short- and long-term behavioral and neuroendocrine responses of SIB and control monkeys to the stress of relocation. METHODS: Twenty adult male rhesus macaques were exposed to the stress of relocation to a new housing arrangement in a newly constructed facility. Daytime behavior, sleep, and multiple measures of hypothalamic-pituitary adrenocortical (HPA) axis function were investigated before and after the move. RESULTS: Relocation induced a complex pattern of short- and long-term effects in the animals. The SIB animals showed a long-lasting increase in self-biting behavior, as well as evidence of sleep disturbance. Both groups exhibited elevated cortisol levels in saliva, serum, and hair, and also an unexpected delayed increase in circulating concentrations of corticosteroid binding globulin (CBG). CONCLUSIONS: Our results indicate that relocation is a significant stressor for rhesus macaques and that this stressor triggers an increase in self biting behavior as well as sleep disturbance in monkeys previously identified as suffering from SIB. These findings suggest that life stresses may similarly exacerbate SIB in humans with this disorder. The HPA axis results underscore the potential role of CBG in regulating long-term neuroendocrine responses to major stressors. PMID- 18164280 TI - Interindividual sleep spindle differences and their relation to learning-related enhancements. AB - We reported earlier that overnight change in explicit memory is positively related to the change in sleep spindle activity (between a control and a learning night). However, it remained unclear whether this effect was restricted to good memory performers and whether a general association of sleep spindles and a "sleep-related learning trait" may not account for this effect. Here we now present a secondary and more detailed analysis of our randomized multicenter study. Subjects were studied over a 4-week study period (including actigraphy and daily sleep diaries), including three overnight stays in the sleep laboratory. In the course of the study, subjects completed test-batteries of memory (Wechsler Memory-Scale-revised; WMS) and other cognitive abilities (Raven's Advanced Progressive-Matrices; APM) and were asked to study 160 word pairs in the evening before being tested by cued-recall. Afterwards, subjects went to bed in the laboratory with full polysomnographic montages. Additionally, subjects participated on another occasion in a non-learning control (perceptual priming) task that was counterbalanced either before or after the learning condition. Slow as well as fast spindle activities were analyzed at frontopolar and central topographies. Although it was found that spindle activity is generally (in learning as well as control nights) elevated in highly gifted subjects, spindle analyses revealed that spindle increase (control to learning night) is specifically related to explicit memory improvement overnight, independent of individual learning traits. Together these findings suggest that the spindle increase after learning is related to elaborate encoding before sleep, whereas an individual's general learning ability is well reflected in interindividual (and trait-like) differences of absolute sleep spindle activity. PMID- 18164281 TI - Chronic corticosterone injections induce a decrease of ATP levels and sustained activation of AMP-activated protein kinase in hippocampal tissues of male mice. AB - Chronic corticosterone injections induce hippocampus tissue damage and depression like behavior in rodent animals, the cause of which is not known. Nevertheless, increasing evidence shows that adenylate kinase (AK) and AMP-activated protein kinase (AMPK) play a very important role in intracellular energy metabolism and are especially critical for neurons which are known to have very small energy reserves and narrow margin of safety between the energy that can be generated and the energy required for maximum activity. Abnormalities of AK or AMPK system have detrimental effects on neurons or brain function especially at times of increased activity. In this study, we investigated the effects of chronic corticosterone exposure on energy metabolism, as well as AK and AMPK in hippocampal tissues in male C57BL/6N mice. Our results show that chronic corticosterone injection induced depression-like behavior in male mice, significantly decreased the energy levels and caused a sustained increase of AMP:ATP ratio in hippocampal tissues. Interestingly, chronic corticosterone injections did not produce obvious effects on AK1 protein and mRNA levels, but caused a sustained activation of AMPK. The results indicate that sustained AMPK activation might be a mechanism by which chronic corticosterone treatment causes depression-like behavior in male mice. PMID- 18164282 TI - Evidence for a non-linguistic distinction between singular and plural sets in rhesus monkeys. AB - Set representations are explicitly expressed in natural language. For example, many languages distinguish between sets and subsets (all vs. some), as well as between singular and plural sets (a cat vs. some cats). Three experiments explored the hypothesis that these representations are language specific, and thus absent from the conceptual resources of non-linguistic animals. We found that rhesus monkeys spontaneously discriminate sets based on a conceptual singular-plural distinction. Under conditions that do not elicit comparisons based on approximate magnitudes or one-to-one correspondence, rhesus monkeys distinguished between singular and plural sets (1 vs. 2 and 1 vs. 5), but not between two plural sets (2 vs. 3, 2 vs. 4, and 2 vs. 5). These results suggest that set-relational distinctions are not a privileged part of natural language, and may have evolved in non-linguistic species to support domain general quantitative computations. PMID- 18164283 TI - Visualization of monoaminergic neurons and neurotoxicity of MPTP in live transgenic zebrafish. AB - We describe an enhancer trap transgenic zebrafish line, ETvmat2:GFP, in which most monoaminergic neurons are labeled by green fluorescent protein (GFP) during embryonic development. The reporter gene of ETvmat2:GFP was inserted into the second intron of vesicular monoamine transporter 2 (vmat2) gene, and the GFP expression pattern recapitulates that of the vmat2 gene. The GFP positive neurons include the large and pear-shaped tyrosine hydroxylase positive neurons (TH populations 2 and 4) in the posterior tuberculum of ventral diencephalon (PT neurons), which are thought to be equivalent to the midbrain dopamine neurons in mammals. We found that these PT neurons and two other GFP labeled non-TH type neuronal groups, one in the paraventricular organ of the posterior tuberculum and the other in the hypothalamus, were significantly reduced after exposure to MPTP, while the rest of GFP-positive neuronal clusters, including those in telencephalon, pretectum, raphe nuclei and locus coeruleus, remain largely unchanged. Furthermore, we showed that the effects of hedgehog signaling pathway inhibition on the development of monoaminergic neurons can be easily visualized in individual living ETvmat2:GFP embryos. This enhancer trap line should be useful for genetic and pharmacological analyses of monoaminergic neuron development and processes underlying Parkinson's disease. PMID- 18164284 TI - Comparative analysis of embryonic cell lineage between Caenorhabditis briggsae and Caenorhabditis elegans. AB - Comparative genomic analysis of important signaling pathways in Caenorhabditis briggsae and Caenorhabditis elegans reveals both conserved features and also differences. To build a framework to address the significance of these features we determined the C. briggsae embryonic cell lineage, using the tools StarryNite and AceTree. We traced both cell divisions and cell positions for all cells through all but the last round of cell division and for selected cells through the final round. We found the lineage to be remarkably similar to that of C. elegans. Not only did the founder cells give rise to similar numbers of progeny, the relative cell division timing and positions were largely maintained. These lineage similarities appear to give rise to similar cell fates as judged both by the positions of lineally equivalent cells and by the patterns of cell deaths in both species. However, some reproducible differences were seen, e.g., the P4 cell cycle length is more than 40% longer in C. briggsae than that in C. elegans (p<0.01). The extensive conservation of embryonic development between such divergent species suggests that substantial evolutionary distance between these two species has not altered these early developmental cellular events, although the developmental defects of transpecies hybrids suggest that the details of the underlying molecular pathways have diverged sufficiently so as to not be interchangeable. PMID- 18164285 TI - Mirror-imaged doublets of Tetmemena pustulata: implications for the development of left-right asymmetry. AB - Ciliated protozoa possess cellular axes reflected in the arrangement of their ciliature. Upon transverse fission, daughter cells develop an identical ciliary pattern, ensuring perpetuation of the cellular phenotype. Experimentally manipulated cells can be induced to form atypical phenotypes, capable of intraclonal propagation and regeneration after encystment. One such phenotype in the ciliate Tetmemena pustulata (formerly Stylonychia pustulata) is the mirror imaged doublet. These cells possess two distinct sets of ciliature, juxtaposed on the surfaces in mirror image symmetry, with a common anterior-posterior axis. We have examined whether individual ciliary components of Tetmemena mirror-image doublets are mirror imaged. Ultrastructural analysis indicates that despite global mirror imaging of the ciliature, detailed organization of the membranelles is reversed in the mirror-image oral apparatus (OA), such that the ciliary effective stroke propels food away from the OA. Assembly of compound ciliary structures of both OAs starts out identically, but as the structures associated with the mirror-image OA continue to form, the new set of membranelles undergoes a 180 degrees planar rotation on the ventral surface relative to the same structures in the typical OA. The overall symmetry of the OA thus appears to be separable from the more localized assembly of individual basal bodies. True mirror imagery of the membranelles would require new enantiomorphic forms of the individual ciliary components, particularly the basal bodies, which is never observed. These observations suggest a mechanistic hypothesis with implications for the development of left-right asymmetry not only in ciliates, but perhaps also in development of left-right asymmetry in general. PMID- 18164286 TI - Oral administration of MA-2029, a novel selective and competitive motilin receptor antagonist, inhibits motilin-induced intestinal contractions and visceral pain in rabbits. AB - The pharmacological properties of MA-2029, a novel motilin receptor antagonist, were investigated. In vitro, MA-2029 (1 to 30 nM) competitively inhibited motilin induced contractions in isolated rabbit duodenal longitudinal muscle strips, with a pA2 value of 9.17+/-0.01 (n=5). However, contractile responses to acetylcholine and substance P were unaffected even at 1 microM of MA-2029. MA-2029 concentration-dependently inhibited the binding of [125 I]motilin to motilin receptors in a homogenate of rabbit colon smooth muscle tissue and membranes of HEK 293 cells expressing human motilin receptors. The pKi of MA-2029 was 8.58+/ 0.04 in the rabbit colon homogenate (n=4) and 8.39 in the HEK 293 cells (mean of duplicate experiments). In vivo, orally-administered MA-2029 (3 to 30 mg/kg) dose dependently inhibited colonic contractions induced by motilin (3 microg/kg, i.v.) in conscious rabbits. Inhibition was caused by all doses at 30 min after administration and by 10 mg/kg or more at 4 h after administration. The plasma concentration of MA-2029 correlated with its inhibitory effect. Furthermore, the oral administration of MA-2029 (0.3 to 3 mg/kg) also inhibited abdominal muscle contractions (an index of the visceral pain) induced by intravenous infusion of motilin (3 microg/kg/h) during colorectal distension in conscious rabbits. These results indicate that MA-2029 is an orally active, selective and competitive motilin receptor antagonist. It is suggested that this compound may be useful for gastrointestinal disorders associated with disturbed gastrointestinal motility such as irritable bowel syndrome. PMID- 18164287 TI - Seizure susceptibility alteration following reversible cholestasis in mice: Modulation by opioids and nitric oxide. AB - There is an increasing body of evidence that the central nervous system is affected by cholestatic liver disorders. Cholestasis has been shown to result in a decreased seizure propensity which is believed to be mediated by an increased opioidergic tone and nitric oxide (NO) signaling pathway. In this study, we used a reversible chemically-induced cholestasis model in mice to investigate the changes in seizure susceptibility. The cholestasis was induced by intragastric administration of alpha-naphthylisothiocyanate (ANIT) (100 mg/kg) or vehicle (corn oil). The threshold to generalized clonic seizures induced by timed intravenous infusion of pentylenetetrazole (PTZ) was used as an index of seizure propensity. The role of opioid receptors and NO pathway in the changes of seizure threshold, and the responsiveness to the anticonvulsant effect of opioid agonist, morphine, during and after the resolution of cholestasis was studied in this reversible paradigm of cholestatic disease. A significant increase in cholestasis related biochemical markers as well as in clonic seizure threshold was observed; it was maximal at day 3 after cholestasis induction and slowly decreased to normal thereafter. Seizure threshold rise was inhibited by chronic administration of the opioid antagonist naltrexone or acute administration of N-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO production. Co-administration of subeffective doses of L-NAME and naltrexone showed an additive effect. Injection of an anticonvulsant dose of morphine on day 7 after cholestasis induction did not increase seizure threshold, suggestive of a downregulation of receptors even after cholestasis resolution. These data shows that ANIT-induced cholestasis leads to a reversible increased resistance to PTZ-induced seizures through an opioid/NO-mediated pathway, and is probably accompanied by downregulation of opioid receptors. PMID- 18164288 TI - Acute administration of l-arginine restores nitric oxide-mediated relaxation in isolated pulmonary arteries from pulmonary hypertensive exercise trained rats. AB - Hypoxia-induced pulmonary hypertension is associated with an impairment of nitric oxide-mediated vasorelaxation in the pulmonary circulation that is not prevented by exercise training. The present study was designed to test the hypothesis that a decrease in l-arginine bioavailability could be involved in this blunted response to exercise training. Male Wistar rats were randomly assigned to 4 groups: normotensive sedentary, normotensive trained, pulmonary hypertensive sedentary, pulmonary hypertensive trained. Pulmonary hypertension was induced by chronic exposure to hypobaric hypoxia (PIO(2) approximately 90 mmHg). Endothelium dependent vasorelaxation to acetylcholine (10(-8)-10(-4) M) with or without l arginine (10(-3) M) and/or nitro-l-arginine methyl ester (5.10(-6) M) was assessed on isolated pulmonary arterial rings. Maximal relaxation to acetylcholine was impaired in both pulmonary hypertensive groups. Acute l arginine supplementation improved acetylcholine-induced vasorelaxation in the pulmonary hypertensive trained rats (P<0.01), to the level obtained in the normotensive sedentary ones, but not in the pulmonary hypertensive sedentary rats. This improvement was abolished when nitro-l-arginine methyl ester was added to the organ bath and was accounted for by an increase in eNOS protein content. These results confirm that the potential beneficial effect of exercise on nitric oxide-mediated pulmonary artery vasorelaxation is partly blunted by deleterious effects of hypoxia on l-arginine bioavailability. Further studies are needed to evaluate the benefit of the combination of exercise training and l-arginine supplementation for the treatment of pulmonary hypertension. PMID- 18164289 TI - Dual localization of the RNA binding protein CUGBP-1 to stress granule and perinucleolar compartment. AB - The mRNA-binding protein CUGBP-1 is a multi-faceted factor, involved in a wide range of biological processes including splicing, translation initiation and mRNA degradation. Here we show that CUGBP-1 is a novel constituent of stress granule (SG), the translational silencing machinery assembled in response to environmental stress. CUGBP-1 was rapidly routed to SGs upon exposure to a variety of environmental stress, and actively shuttles between the nucleus and SGs. The linker domain located between the second and third RNA recognition motifs (RRMs) was found to be essential for the recruitment of CUGBP-1 to SGs. Importantly, we discovered that the linker domain is also required to direct CUGBP-1 to another subcellular structure, perinucleolar compartment (PNC). These results demonstrate the dynamic behavior of CUGBP-1 during stress response and that the linker region, in concert with RRMs, plays a significant role in defining its subcellular localization and dynamics. PMID- 18164290 TI - Screening of genes involved in cell migration in Dictyostelium. AB - A single cell of wild-type Dictyostelium discoideum forms a visible colony on a plastic dish in several days, but due to enhanced cell migration, amiB-null mutant cells scatter over a large area and do not form noticeable colonies. Here, with an aim to identify genes involved in cell migration, we isolated suppresser mutants of amiB-null mutants that restore the ability to form colonies. From REMI (restriction enzyme-mediated integration)-mutagenized pool of double-mutants, we identified 18 responsible genes from them. These genes can be categorized into several biological processes. One cell line, Sab16 (Suppressor of amiB) was chosen for further analysis, which had a disrupted phospholipase D pldB gene. To confirm the role of pldB gene in cell migration, we knocked out the pldB gene and over-expressed gfp-pldB in wild-type cells. GFP-PLDB localized to plasma membrane and on vesicles, and in migrating cells, at the protruding regions of pseudopodia. Migration speed of vegetative pldB-null cells was reduced to 73% of that of the wild-type. These results suggest that PLDB plays an important role in migration in Dictyostelium cells, and that our screening system is useful for the identification of genes involved in cell migration. PMID- 18164291 TI - Heterogeneity and timing of translocation and membrane-mediated assembly of different annexins. AB - Many cell types, including neurons and epithelial cells, express a variety of annexins. Although the overall function has only been partially unravelled, a dominant feature is the formation of two-dimensional assemblies under the plasma membrane in a calcium-dependent manner. Here we show that fluorescently tagged annexins A1, A2, A4, A5, and A6 translocate and assemble at the plasma membrane and the nuclear envelope, except annexin A2, which only attaches to the plasma membrane. All annexins have different response times to elevated calcium levels as was shown by the translocation of co-expressed proteins. Fluorescence recovery after photobleaching revealed the static nature of all annexin assemblies. Analysis of the assemblies by Foerster resonance energy transfer (FRET) using acceptor bleaching demonstrated mostly annexin-specific self-assembly. Heterogeneous assembly formation was shown between annexins A5 and A1, and A5 and A2. The formation of homo- and heterogeneous annexin assemblies may play an important role when high increases in calcium occur, such as after disruption of the plasma membrane. PMID- 18164292 TI - Accommodation-induced changes in iris curvature. AB - Eyes were imaged using anterior segment ultrasound biomicroscopy to compare alterations in iris contour following the onset of accommodation in eyes with narrow angles, pigment dispersion syndrome, and controls. A radial perpendicular image in the horizontal temporal meridian was obtained for one eye while the subject focused on a distant target (~6m, unaccommodated state) with the fellow eye. The subject then focused steadily on a near target (~0.33 m, accommodated state) for 3 min. Images were acquired at 0, 1, 2, and 3 min. Iris curvature was determined by measuring the maximum distance between the posterior iris surface and a line from the iris root to the first point of contact between the iris and lens. In control subjects (n=22), iris curvature decreased immediately after the onset of accommodation, but not significantly (p=0.49), from 246+/-37 microm (mean+/-SEM) to 205+/-82 microm; curvature increased after 3 min of accommodation to 298+/-57 microm (p=0.10 vs. onset of accommodation). Eyes with pigment dispersion syndrome (n=15) exhibited curvatures of 60+/-79 microm when unaccommodated, -3+/-83 microm immediately after accommodation (p=0.12), and 146+/-94 microm (p=0.01) 3 min later. Eyes with narrow angles (n=16) exhibited curvatures of 449+/-45 microm when unaccommodated, 414+/-46 microm immediately after accommodation (p=0.37), and 523+/-40 microm (p<0.01) 3 min later. The results confirm the time-dependent nature of iris contour response, with significant differences observed between the initial observation after accommodation and the observation 3 min later. The largest drop in curvature immediately after accommodation and the most rapid increase in curvature during subsequent observation were seen in the PDS subjects. We suspect that the more rapid increase in curvature in the PDS subjects is due to the elevated anterior chamber pressure caused by the "reverse pupillary block" effect. PMID- 18164293 TI - Electrical stimulation of intact peripheral sensory axons in rats promotes outgrowth of their central projections. AB - A lesion of a peripheral nerve before a second injury (conditioning lesion, CL), enhances peripheral and central regeneration of dorsal root ganglion (DRG) axons. This effect is mediated by elevated neuronal cAMP. Here we wanted to investigate whether electrical stimulation (ES) of an intact nerve, which has been shown to accelerate peripheral axon outgrowth, is also effective in promoting axon regeneration of injured DRG axons in vitro and of the central DRG axons in vivo and, whether this effect is mediated by elevation of cAMP. For the in vitro assay, the intact sciatic nerve of adult rats was stimulated at 20 Hz for 1 h, 7 days before harvest and primary culture of DRG neurons on a growth permissive substrate. In the in vivo study, the central axons of the lumbosacral DRGs were cut in the Th8 dorsal column, and the sciatic nerve was either cut or left intact, and subjected to 1 h ES at 20 Hz or 200 Hz. In vitro, ES increased neurite outgrowth 4-fold as compared to non-stimulated DRG neurons. In vivo, ES at 20 Hz significantly increased axon outgrowth into the central lesion site as compared to the Sham control. The 20 Hz ES was as effective as the CL in increasing axon outgrowth into the lesion site but not in promoting axonal elongation even though 20 Hz ES increased intracellular cAMP levels in DRG neurons as effectively as the CL. Thus elevation of cAMP may account for the central axonal outgrowth after ES and a CL. PMID- 18164294 TI - Voluntary running attenuates age-related deficits following SCI. AB - Over the past few decades, the average age at time of spinal cord injury (SCI) has increased. Here we examined locomotor recovery and myelin pathology in both young and aged adult rats following contusion SCI. Our assessment indicates that the rate of locomotor recovery following SCI is significantly delayed in aged rats as compared to young rats, and is associated with a greater degree of pathology and demyelination. Additionally, we examined the effect of voluntary exercise, pre- and post-injury, on locomotor recovery and myelin pathology following contusion SCI. Our data indicate that exercise improves the locomotor recovery of injured aged rats such that it is comparable to the recovery rate of injured young rats, and is associated with a decreased area of pathology and amount of demyelination. Interestingly, the rate of locomotor recovery and myelin pathology in the aged exercised rats was similar to that of the young sedentary rats after injury, indicating that exercise attenuates the delayed recovery of function and associated histopathology in aged rats. These data indicate that there is an age-related delay in locomotor recovery following SCI, and an age related increase in histopathology following SCI. Importantly, our data indicate that exercise attenuates these age-related deficits following SCI. PMID- 18164295 TI - Relationship of phosphorylated alpha-synuclein and tau accumulation to Abeta deposition in the cerebral cortex of dementia with Lewy bodies. AB - Alpha-synuclein accumulated in the brain of dementia with Lewy bodies (DLB) is phosphorylated at serine129 (palpha-synuclein). We investigated the accumulation of palpha-synuclein in the brains of patients with DLB and Alzheimer's disease (AD). We employed 18 DLB patients with neocortical Lewy body type pathology (nLBTP) with or without AD. We also employed the same number of AD cases without significant nLBTP. We refer to the former group as the nLBTP group and to the latter as the AD type pathology (ADTP) group. In the nLBTP group, palpha synuclein positive neurite pathology such as threads and dots occurs in all layers of the temporal neocortex. It was comparable in degree with tau pathology in AD. Fifteen cases in the nLBTP group were associated with Abeta deposition that meets the CERAD plaque score "C" and one case with a score "B". In these plaque-associated cases, the severity of palpha-synuclein pathology was related to the degree of Abeta deposition. In the cases with relatively moderate Abeta deposition, tau pathology was disproportionately mild in the nLBTP group, while the total of tau and palpha-synuclein pathology was proportionate to Abeta deposition in both the nLBTP and ADTP groups. Our results support the ideas that there is an overlap in the pathology between AD and DLB and that Abeta promotes accumulation of both alpha-synuclein and tau. The procession from Abeta to neurite pathology in the cerebral cortex of AD and DLB may be unifiable. PMID- 18164296 TI - Insulin resistance in chronic hepatitis C: association with genotypes 1 and 4, serum HCV RNA level, and liver fibrosis. AB - BACKGROUND & AIMS: Our study was designed to test the association between insulin resistance (IR) and hepatitis C virus (HCV) genotypes, serum HCV RNA level and liver fibrosis stage in a large prospective cohort of chronic hepatitis C (CHC) patients. METHODS: Six hundred consecutive patients (CHC, n = 500; chronic hepatitis B (CHB), n = 100) were evaluated on the day of liver biopsy. IR (Homeostasis Model for Assessment of Insulin Resistance) and all components of the metabolic syndrome were assessed. By logistic regression, independent factors associated with IR and those associated with significant fibrosis were assessed in nondiabetic and noncirrhotic CHC, respectively. Parameters of IR were compared between hepatitis B and 240 CHC matched by epidemiologic, metabolic, and histologic features. RESULTS: IR was present in 32.4% of the 462 nondiabetic CHC and associated with the metabolic syndrome, genotypes 1 and 4, significant fibrosis, and severe steatosis. IR was diagnosed in 15% of 145 CHC without metabolic syndrome or significant fibrosis, and associated with genotypes 1 and 4, high serum HCV RNA level, and moderate-severe necroinflammation. Significant fibrosis was present in 51.1% of the 454 noncirrhotic CHC patients and associated with male sex, age >40 years, IR, moderate-severe necroinflammation, and severe steatosis. IR was less frequent in CHB than in matched CHC (5% vs 35%, respectively, P < .001). CONCLUSIONS: IR is a specific feature of CHC, associated with genotypes 1 and 4 and high serum HCV RNA level. Significant fibrosis is associated with IR independent from steatosis. PMID- 18164297 TI - Spatial and temporal dynamics of corticosterone and corticosterone binding globulin are driven by environmental heterogeneity. AB - The question of whether changes in glucocorticoid concentrations reflect consistent changes in physiology associated with transitions between different stages of reproduction, or whether they reflect responses to environmental conditions, is one the central issues in field endocrinology studies. We examined the temporal and spatial dynamics of corticosterone (CORT, baseline, and acute stress-induced) and corticosterone binding globulin (CBG) concentrations in blood of Black-legged Kittiwakes (Rissa tridactyla) breeding at four major colonies in the Bering Sea, Alaska, during 1999-2005. We found that total CORT, free CORT, and CBG capacity varied inconsistently among reproductive stages, colonies, and years. Total CORT levels were positively correlated with CBG capacity. Variation in free CORT was largely driven by variation in total CORT. Results suggest that the adrenocortical function and CBG in breeding kittiwakes do not vary as a consequence of stage-specific modulation associated with a particular reproductive stage as in some short-lived passerine birds. Rather, in accord with predictions for a long-lived species, the lack of consistent colony, year, and reproductive stage patterns in baseline and maximum CORT, and CBG indicates that environmental factors, probably local dynamics of food availability, drive variation in these factors. PMID- 18164298 TI - Results of the T2 humeral nailing system with special focus on compression interlocking. AB - BACKGROUND: The outcome of interlocking nailing of humeral shaft fractures is controversial. Variations in implants, operative technique and follow-up parameters hinder comparative studies. The aim of this investigation was to evaluate the T2 humeral nailing system according to clinical results, and to recognise advantages and disadvantages of compression interlocking. METHODS: A total of 36 consecutive humeral shaft fractures were treated with an interlocking humeral nail. In 23 cases the antegrade and in 14 cases the retrograde approach was used. Compression locking was performed in 21 and static locking in 15 cases. RESULTS: At 22-month follow-up, bone consolidation was present in 35 cases; in 5 cases complications developed. There was no significant difference between compression and static locking nor between the antegrade and retrograde approach regarding complications, Constant shoulder score, mean Morrey elbow score, SF-12 physical score or SF-12 mental score. CONCLUSION: Antegrade and retrograde interlocking nailing of humeral shaft fractures with the T2 nailing system can result in good functional outcome and unimpaired quality of life. Compression interlocking can minimise the fracture gap and increase the biomechanical stiffness. Potential disadvantages of compression interlocking include possible bending or loosening of the locking screw in the dynamic oblong hole. An additional static locking screw should be used on the humerus after compression interlocking. PMID- 18164299 TI - The burden of terrorism: high rate of recurrent hospital referrals. AB - BACKGROUND: Recurrent emergency room referrals and re-hospitalisation of terror victims (external cause of injury E990-E998 and selected cases from E970-E978) [International Classification of Diseases, 9th revision. Clinical modification, 5th ed. (ICD-9-CM). Los Angeles, CA: Practice Management Information Corporation; 1998] have not as yet been examined in the literature. Our objective was to evaluate the extent of hospital services' usage following a terror event and to characterise the casualties who return for hospitalisation and rehabilitation following their discharge. METHODS: A retrospective longitudinal study including all terror victims who were hospitalised at our level I trauma centre between October 2000 and March 2004. Data on the first hospitalisation of these victims (n=497 cases) were retrieved from the hospital's trauma registry. Data on recurrent emergency room referrals and re-hospitalisation of the 464 cases who survived were taken from the hospital's administrative computerised database. RESULTS: Four hundred and ninety-seven terror victims were hospitalised, of which 464 survived their first hospitalisation. Two hundred and nineteen (47%) were subsequently re-referred to the hospital. The total number of recurrent hospital referral days amounted to 77% of the total first hospitalisation days for all casualties. A strong association was found with regard to severity of injury, length of stay in the intensive care unit (ICU) and total length of stay. Logistic regression analysis found total length of stay of initial hospitalisation as the only significant variable. CONCLUSIONS: The recurrent hospitalisation of terror victims places a heavy burden on the health system. Further studies should be conducted to determine the reasons for these recurrent referrals and to explore whether the number of recurrent referrals can be reduced or at least planned for. PMID- 18164300 TI - Pleural decompression and drainage during trauma reception and resuscitation. AB - This review examines pleural decompression and drainage during initial hospital adult trauma reception and resuscitation, when it is indicated for haemodynamically unstable patients with signs of pneumothorax or haemothorax. The relevant historical background, techniques, complications and current controversies are highlighted. Key findings of this review are that: 1. Needle thoracocentesis is an unreliable means of decompressing the chest of an unstable patient and should only be used as a technique of last resort. 2. Blunt dissection and digital decompression through the pleura is the essential first step for pleural decompression, as decompression of the pleural space is a primary goal during reception of the haemodynamically unstable patient with a haemothorax or pneumothorax. Drainage and insertion of a chest tube is a secondary priority. 3. Techniques to prevent tube thoracostomy (TT) complications include aseptic technique, avoidance of trocars, digital exploration of the insertion site and guidance of the tube posteriorly and superiorly during insertion. 4. Whenever possible, blunt thoracic trauma patients should undergo definitive CT imaging after TT to check for appropriate tube position. PMID- 18164301 TI - Gender differences in outcome in patients with hypotension and severe traumatic brain injury. AB - BACKGROUND: Animal studies have identified hormonal influences on responses to injury and recovery, creating a potential gender effect on outcome. Progesterone and oestrogen are thought to afford protection in the immediate post-injury period, suggesting females have an advantage, although there has been limited evidence of this in human outcome studies. METHODS: This study examined the influence of gender on outcome in 229 adults (151 males), aged >17 years, with severe blunt head trauma, initial GCS <9 and hypotension, recruited into a randomised controlled trial of pre-hospital hypertonic saline resuscitation versus conventional fluid management. Outcome was measured by survival and Glasgow Outcome Scale-Extended version (GOS-E) scores at 6 months post-injury. RESULTS: Females recruited into the study had a higher mean age. Females were more likely to be injured as passengers and pedestrians and males as drivers or motorcyclists. There were no gender differences in GCS or injury severity scores, ICP, cerebral perfusion pressure, gas exchange (PaO2/FiO2 ratio), or duration of mechanical ventilation. After controlling for GCS, age and cause of injury, females had a lower rate of survival. They also showed a lower rate of good outcome (GOS-E score >4) at 6 months, but this appeared to reflect the lower rate of initial survival. Those females surviving had similar outcomes to males. CONCLUSIONS: The study provides no evidence that females fare better than males following severe TBI, suggesting rather that females may fare worse. PMID- 18164302 TI - Acute care surgery and trauma: a marriage of convenience. PMID- 18164303 TI - Goserelin versus leuprolide before hysterectomy for uterine fibroids. AB - OBJECTIVE: To compare goserelin and leuprolide given before hysterectomy for symptomatic large fibroid uteri. METHODS: A randomized study of 66 premenopausal women with fibroid uteri at least 14 weeks of gestation in a gravid uterus. Women were randomized to receive either subcutaneous depot 3.6 mg goserelin or 3.75 mg leuprolide every 4 weeks for a total of 3 doses. Hysterectomy was performed within 1 month of the last dose. RESULTS: A total of 34 women randomized to the goserelin group and 31 women to the leuprolide group were available for analysis. Preoperative hemoglobin level (P=0.89), operative blood loss (P=0.72), and operating time (P=0.39) were not different between the 2 groups. Postoperative hemoglobin was higher in the leuprolide group (P=0.003), but blood transfusion requirement was not different between the groups (P=1.0). Other outcomes and side effects of the drugs were similar. CONCLUSIONS: Goserelin and leuprolide administered before hysterectomy for uterine fibroids have similar perioperative outcomes. PMID- 18164304 TI - A randomized comparative study of prophylactic oxytocin versus ergometrine in the third stage of labor. AB - OBJECTIVE: To compare the effect of prophylactic use of oxytocin and ergometrine in management of the third stage of labor. METHODS: A prospective randomized study of 600 women assigned to receive either oxytocin or ergometrine in the third stage of labor. Outcome measures were the predelivery and 48-hour postdelivery hematocrit, duration of the third stage, specific side effects, and incidence of postpartum hemorrhage. Statistical analyses were done using the t test for continuous variables and chi2 test for categorical variables. The level of significance was set at P<0.05. RESULTS: There were no significant differences between the 2 groups in maternal age, gestational age, duration of third stage, birth weights, risk for retained placenta, manual removal of placenta, or need for additional oxytocics. Patients in the ergometrine group were at significant risk for nausea, vomiting, headaches, and elevated blood pressure (P=0.0001). CONCLUSION: Oxytocin is as effective as ergometrine at reducing the incidence of postpartum hemorrhage, but without the undesirable side effects of nausea, vomiting, and elevated blood pressure associated with ergometrine. PMID- 18164305 TI - Zygote score and status 1 or 2 days after cleavage and assisted reproduction outcome. AB - OBJECTIVE: To determine whether pronuclear morphology is a useful additional criterion when selecting embryos for transfer. METHOD: A modified Tesarik and Greco scoring system was used in this retrospective study to assess the pronuclear morphology of 883 zygotes from 214 in vitro fertilization or intracytoplasmic sperm injection cycles. Pronucleus size and presence of cytoplasmic halo were also noted. Embryo selection was performed on Day 2 or 3 following oocyte retrieval. The chi2, analysis of variance, Kruskal-Wallis, and Fisher exact tests were used where appropriate. RESULTS: A stepwise logistic regression revealed that a better embryo quality was associated with both pattern 0 of the scoring system and pronuclei of equal size on Days 2 or 3. Higher pregnancy and multiple gestation rates were obtained when at least 1 embryo classified as pattern 0 was included in the set of embryos transferred. CONCLUSION: Combined with embryo morphology evaluation on Days 2 and 3, a scoring system based on pronuclear morphology seems to provide a good criterion when selecting embryos for transfer. PMID- 18164306 TI - Serum total homocysteine concentrations and risk of mortality from stroke and coronary heart disease in Japanese: The JACC study. AB - Evidence for association between serum total homocysteine (tHcy) level and cardiovascular disease is limited in Asian populations. We conducted a nested case-control study under JACC Study. A total of 39,242 subjects aged 40-79 years provided serum samples at baseline surveys between 1988 and 1990. Control subjects were selected by matching for sex, age, community and year of serum storage. Serum tHcy levels were measured by high-performance liquid chromatography. During the 10-year follow-up, there were 444 deaths due to total cardiovascular disease, including 310 total stroke (131 hemorrhage and 101 ischemic strokes) and 134 coronary heart diseases. The risks of mortality from ischemic stroke, coronary heart disease, and total cardiovascular disease were significantly higher in individuals with the highest serum tHcy quartile (>or=15.3micromol/L) than in those with the lowest quartile (<10.5micromol/L); the respective multivariable odds ratios (95% CI) were 4.35 (1.12-16.9), 3.40 (1.17-9.86), and 1.68 (1.02-2.77). The multivariable odds ratios associated with a 5-micromol/L increase in tHcy were 1.49 (1.01-2.18), 2.01 (1.21-3.35), and 1.15 (1.00-1.32), respectively. High serum tHcy levels were associated with increased mortality from ischemic stroke, coronary heart disease and total cardiovascular disease among Japanese. PMID- 18164307 TI - The effect of coenzyme Q10 on microcirculatory endothelial function of subjects with type 2 diabetes mellitus. PMID- 18164308 TI - Calorimetric study of the alpha-tocopherol solubility in reversed AOT micelles. AB - The experimental data of heat of mixing (Q) for heterogeneous system alpha tocopherol/AOT/n-heptane with and without water at 25 degrees C are presented. The Q dependence on AOT (sodium bis (2-ethylhexyl) sulfosuccinate) concentration, and R parameter defined as R=[H2O]/[AOT] with flow calorimetric method were investigated. Using the D'Aprano model (which is formally identical to that used earlier by Magid et al.) the binding constant (K), the distribution constant of alpha-tocopherol (K distr) between hydrocarbon and the micellar phase, and the standard enthalpy of transfer (DeltaH tr 0) of alpha-tocopherol from the hydrocarbon to AOT reversed micelles were calculated. The solubility of alpha tocopherol in AOT reversed micelles explored with the calorimetric technique was compared to the literature data obtained respectively with UV spectrophotometry for reversed micelles and by other techniques for the phospholipid bilayer. PMID- 18164309 TI - Effect of shear rate on aggregate size and morphology investigated under turbulent conditions in stirred tank. AB - Aggregation and breakage of aggregates produced from fully destabilized polystyrene latex particles in turbulent flow was studied experimentally in both batch and continuous stirred tank. Detailed investigation of the initial kinetics showed that the collision efficiency, alpha, depends on the shear rate according to alpha proportional to G(-b), with a power law exponent, b, equal to 0.18. After steady state was reached the dynamic response of the system on a change in stirring speed and solid volume fraction was investigated. It was found that the steady-state values of two measured moments of the cluster mass distribution (CMD) are fully reversible upon a change in stirring speed. This indicates that although the moments of CMD at steady-state depend on the applied shear rate, the aggregate structure is independent of the shear rate in the given range of stirring speeds. This was proved by independent measurement of the fractal dimension, d(f), using image analysis which provided a d(f) equal to 2.62 +/- 0.18 independent of applied stirring speed. The critical aggregate size, below which breakage is negligible, determined by dilution experiments was consequently used to evaluate the aggregate cohesive force holding the aggregate together, which was found to be independent of the aggregate size and equal to 6.2 +/- 1.0 nN. PMID- 18164310 TI - Adult honeybees (Apis mellifera L.) abandon hemocytic, but not phenoloxidase based immunity. AB - Hemocytes and the (prophenol-) phenoloxidase system constitute the immediate innate immune system in insects. These components of insect immunity are present at any post-embryonic life stage without previous infection. Differences between individuals and species in these immune parameters can reflect differences in infection risk, life expectancy, and biological function. In honeybees which show an age-related division of labor within the worker caste, previous studies demonstrated that foragers show a strongly reduced number of hemoctyes compared to the younger nurse bees. This loss of immune competence has been regarded advantageous with respect to an already high mortality rate due to foraging and to redistribution of energy costs at the colony level. Based on the idea that abandoning hemocytes in all adults would be a reasonably direct regulatory mechanism, we posed the hypothesis that abandoning hemocytic immunity is not restricted to worker honeybees. We tested our hypotheses by performing a comprehensive analysis of hemocyte number and phenoloxidase (PO)-activity levels in immunologically naive workers, queens, and drones. We found that in all three adult phenotypes hemocyte number is dramatically reduced in early adult life. In contrast, we found that the dynamics of PO-activity levels have sex and caste specific characteristics. In workers, PO activity reached a plateau within the first week of adult life, and in queens enzyme levels continuously increased with age and reached levels twice as high as those found in workers. PO-activity levels slightly declined with age in drones. These data support our hypothesis, from which we infer that the previously reported reduction of hemocyte in foragers is not worker specific but represents a general phenomenon occurring in all honeybee adult phenotypes. PMID- 18164311 TI - Crystal structure of long-chain alkane monooxygenase (LadA) in complex with coenzyme FMN: unveiling the long-chain alkane hydroxylase. AB - LadA, a long-chain alkane monooxygenase, utilizes a terminal oxidation pathway for the conversion of long-chain alkanes (up to at least C(36)) to corresponding primary alcohols in thermophilic bacillus Geobacillus thermodenitrificans NG80-2. Here, we report the first structure of the long-chain alkane hydroxylase, LadA, and its complex with the flavin mononucleotide (FMN) coenzyme. LadA is characterized as a new member of the SsuD subfamily of the bacterial luciferase family via a surprising structural relationship. The LadA:FMN binary complex structure and a LadA:FMN:alkane model reveal a hydrophobic cavity that has dual roles: to provide a hydrogen-bond donor (His138) for catalysis and to create a solvent-free environment in which to stabilize the C4a-hydroperoxyflavin intermediate. Consequently, LadA should catalyze the conversion of long-chain alkanes via the acknowledged flavoprotein monooxygenase mechanism. This finding suggests that the ability of LadA to catalyze the degradation of long-chain alkanes is determined by the binding mode of the long-chain alkane substrates. The LadA structure opens a rational perspective to explore and alter the substrate binding site of LadA, with potential biotechnological applications in areas such as petroleum exploration and treatment of environmental oil pollution. PMID- 18164312 TI - Structural analysis of the PP2C phosphatase tPphA from Thermosynechococcus elongatus: a flexible flap subdomain controls access to the catalytic site. AB - The homologue of the phosphoprotein PII phosphatase PphA from Thermosynechococcus elongatus, termed tPphA, was identified and its structure was resolved in two different space groups, C222(1) and P4(1)2(1)2, at a resolution of 1.28 and 3.05 A, respectively. tPphA belongs to a large and widely distributed subfamily of Mg(2+)/Mn(2+)-dependent phosphatases of the PPM superfamily characterized by the lack of catalytic and regulatory domains. The core structure of tPphA shows a high degree of similarity to the two PPM structures identified so far. In contrast to human PP2C, but similar to Mycobacterium tuberculosis phosphatase PstP, the catalytic centre exhibits a third metal ion in addition to the dinuclear metal centre universally conserved in all PPM members. The fact that the third metal is only liganded by amino acids, which are universally conserved in all PPM members, implies that the third metal could be general for all members of this family. As a specific feature of tPphA, a flexible subdomain, previously recognized as a flap domain, could be revealed. Comparison of different structural isomers of tPphA as well as site-specific mutagenesis implied that the flap domain is involved in substrate binding and catalytic activity. The structural arrangement of the flap domain was accompanied by a large side-chain movement of an Arg residue (Arg169) at the basis of the flap. Mutation of this residue strongly impaired protein stability as well as catalytic activity, emphasizing the importance of this amino acid for the regional polysterism of the flap subdomain and confirming the assumption that flap domain flexibility is involved in catalysis. PMID- 18164313 TI - Membrane fusogenic activity of the Alzheimer's peptide A beta(1-42) demonstrated by small-angle neutron scattering. AB - Amyloid-beta peptide (A beta) is considered a triggering agent of Alzheimer's disease. In relation to a therapeutic treatment of the disease, the interaction of A beta with the cell membrane has to be elucidated at the molecular level to understand its mechanism of action. In previous works, we had ascertained by neutron diffraction on stacked lipid multilayers that a toxic fragment of A beta is able to penetrate and perturb the lipid bilayer. Here, the influence of A beta(1-42), the most abundant A beta form in senile plaques, on unilamellar lipid vesicles of phospholipids is investigated by small-angle neutron scattering. We have used the recently proposed separated form factor method to fit the data and to obtain information about the vesicle diameter and structure of the lipid bilayer and its change upon peptide administration. The lipid membrane parameters were obtained with different models of the bilayer profile. As a result, we obtained an increase in the vesicle radii, indicating vesicle fusion. This effect was particularly enhanced at pH 7.0 and at a high peptide/lipid ratio. At the same time, a thinning of the lipid bilayer occurred. A fusogenic activity of the peptide may have very important consequences and may contribute to cytotoxicity by destabilizing the cell membrane. The perturbation of the bilayer structure suggests a strong interaction and/or insertion of the peptide into the membrane, although its localization remains beyond the limit of the experimental resolution. PMID- 18164314 TI - Mechanism of dTTP inhibition of the bifunctional dCTP deaminase:dUTPase encoded by Mycobacterium tuberculosis. AB - Recombinant deoxycytidine triphosphate (dCTP) deaminase from Mycobacterium tuberculosis was produced in Escherichia coli and purified. The enzyme proved to be a bifunctional dCTP deaminase:deoxyuridine triphosphatase. As such, the M. tuberculosis enzyme is the second bifunctional enzyme to be characterised and provides evidence for bifunctionality of dCTP deaminase occurring outside the Archaea kingdom. A steady-state kinetic analysis revealed that the affinity for dCTP and deoxyuridine triphosphate as substrates for the synthesis of deoxyuridine monophosphate were very similar, a result that contrasts that obtained previously for the archaean Methanocaldococcus jannaschii enzyme, which showed approximately 10-fold lower affinity for deoxyuridine triphosphate than for dCTP. The crystal structures of the enzyme in complex with the inhibitor, thymidine triphosphate, and the apo form have been solved. Comparison of the two shows that upon binding of thymidine triphosphate, the disordered C-terminal arranges as a lid covering the active site, and the enzyme adapts an inactive conformation as a result of structural changes in the active site. In the inactive conformation dephosphorylation cannot take place due to the absence of a water molecule otherwise hydrogen-bonded to O2 of the alpha-phosphate. PMID- 18164315 TI - The buried diversity of bovine seminal ribonuclease: shape and cytotoxicity of the swapped non-covalent form of the enzyme. AB - Bovine seminal ribonuclease exists in the native state as an equilibrium mixture of a swapped and an unswapped dimer. The molecular envelope and the exposed surface of the two isomers are practically indistinguishable and their diversity is almost completely buried in the interior of the protein. Surprisingly, the cytotoxic and antitumor activity of the enzyme is a peculiar property of the swapped dimer. This buried diversity comes into light in the reducing environment of the cytosol, where the unswapped dimer dissociates into monomers, whereas the swapped one generates a metastable dimeric form (NCD-BS) with a quaternary assembly that allows the molecule to escape the protein inhibitor of ribonucleases. The stability of this quaternary shape was mainly attributed to the combined presence of Pro19 and Leu28. We have prepared and fully characterized by X-ray diffraction the double mutant P19A/L28Q (PALQ) of the seminal enzyme. While the swapped and unswapped forms of the mutant have structures very similar to that of the corresponding wild-type forms, the non covalent form (NCD-PALQ) adopts an opened quaternary structure, different from that of NCD-BS. Moreover, model building clearly indicates that NCD-PALQ can be easily sequestered by the protein inhibitor. In agreement with these results, cytotoxic assays have revealed that PALQ has limited activity, whereas the single mutants P19A and L28Q display cytotoxic activity against malignant cells almost as large as the wild-type enzyme. The significant increase in the antitumor activity, brought about by the substitution of just two residues in going from the double mutant to the wild-type enzyme, suggests a new strategy to improve this important biological property by strengthening the interface that stabilizes the quaternary structure of NCD-BS. PMID- 18164316 TI - Molecular basis for the unique deubiquitinating activity of the NF-kappaB inhibitor A20. AB - Nuclear factor kappaB (NF-kappaB) activation in tumor necrosis factor, interleukin-1, and Toll-like receptor pathways requires Lys63-linked nondegradative polyubiquitination. A20 is a specific feedback inhibitor of NF kappaB activation in these pathways that possesses dual ubiquitin-editing functions. While the N-terminal domain of A20 is a deubiquitinating enzyme (DUB) for Lys63-linked polyubiquitinated signaling mediators such as TRAF6 and RIP, its C-terminal domain is a ubiquitin ligase (E3) for Lys48-linked degradative polyubiquitination of the same substrates. To elucidate the molecular basis for the DUB activity of A20, we determined its crystal structure and performed a series of biochemical and cell biological studies. The structure reveals the potential catalytic mechanism of A20, which may be significantly different from papain-like cysteine proteases. Ubiquitin can be docked onto a conserved A20 surface; this interaction exhibits charge complementarity and no steric clash. Surprisingly, A20 does not have specificity for Lys63-linked polyubiquitin chains. Instead, it effectively removes Lys63-linked polyubiquitin chains from TRAF6 without dissembling the chains themselves. Our studies suggest that A20 does not act as a general DUB but has the specificity for particular polyubiquitinated substrates to assure its fidelity in regulating NF-kappaB activation in the tumor necrosis factor, interleukin-1, and Toll-like receptor pathways. PMID- 18164317 TI - Acceleration of diabetic wound healing with chitosan-crosslinked collagen sponge containing recombinant human acidic fibroblast growth factor in healing-impaired STZ diabetic rats. AB - In order to develop a better wound-dressing to enhance diabetic wound healing, we have examined the biochemical and biophysical features of chitosan-crosslinked collagen sponge (CCCS) and pre-clinically evaluated the CCCS containing recombinant human acidic fibroblast growth factor (CCCS/FGF) in accelerating diabetic wound healing as compared to collagen sponge alone and FGF alone. Collagen crosslinked with chitosan showed several advantages required for wound dressing, including the uniform and porous ultrastructure, less water-imbibition, small interval porosity, high resistance to collagenase digestion and slow release of FGF from CCCS/FGF. Therapeutic effect of the new wound-dressing containing FGF (i.e.: CCCS/FGF) on diabetic wound healing was examined in type 1 diabetic rat model in which hyperglycemia was induced by single dose of streptozotocin (STZ) and persisted for two months. The CCCS/FGF provided the most efficiently therapeutic effect among various treatments, showing the shortest healing time (14 days in the CCCS/FGF-treated group as compared to 18~21 days in other treatment groups), the quickest tissue collagen generation, the earliest and highest TGF-beta1 expression and dermal cell proliferation (PCNA expression). All these results suggest that CCCS/FGF is an ideal wound-dressing to improve the recovery of healing-impaired wound such as diabetic skin wound, which provides a great potential use in clinics for diabetic patients in the future. PMID- 18164318 TI - Reactivation of methylation-silenced tumor suppressor gene p16INK4a by nordihydroguaiaretic acid and its implication in G1 cell cycle arrest. AB - Phytoestrogens, including the two major groups isoflavones and lignans, are chemicals with weak estrogenic activity which occur naturally in many foods and herbs. Recently, several intriguing studies reported that some isoflavones can affect DNA methylation status. However, little is known about the effect of plant lignans on epigenetic modification. Using cultured T47D and RKO human cancer cells as a model, we studied the modulating effects of nordihydroguaiaretic acid (NDGA), a member of the lignan family, on the methylation status of the gene promoter region. Our results indicated that NDGA reverses p16INK4a CpG island hypermethylation, and restores its transcription and expression in both cell lines. Cytometric analysis showed that NDGA significantly affects cell cycle progression by arresting cells at the G1 phase. Consistent with the reacquisition of p16INK4a expression, we also found that NDGA induces cellular senescence in cancer cells. This is the first study demonstrating that a member of the lignan family can induce demethylation in human cancer cell lines, suggesting a novel epigenetic mechanism in the prevention or treatment of cancer. PMID- 18164319 TI - Non-steroidal anti-inflammatory drugs have potent anti-fibrillogenic and fibril destabilizing effects for alpha-synuclein fibrils in vitro. AB - The aggregation of alpha-synuclein (alphaS) in the brain has been implicated as a critical step in the development of Lewy body diseases (LBD) [Parkinson's disease (PD)/dementia with Lewy bodies (DLB)] and multiple system atrophy (MSA). The involvement of neuroinflammation and microglial activation has been emphasized in the pathogenesis of PD. Recent epidemiological studies have revealed that therapeutic use of non-steroidal anti-inflammatory drugs (NSAIDs) reduces the risk of developing PD. Here, we examined the effects of NSAIDs, such as ibuprofen, aspirin, acetaminophen, meclofenamic acid sodium salt, sulindac sulfide, ketoprofen, flurbiprofen, diclofenac sodium salt, naproxen, and indomethacin, on the formation and destabilization of alphaS fibrils (falphaS) at pH 7.5 and 37 degrees C in vitro, using fluorescence spectroscopy with thioflavin S and electron microscopy. All examined NSAIDs, except for naproxen and indomethacin, inhibited the formation of falphaS in a dose-dependent manner. Moreover, these molecules dose-dependently destabilized preformed falphaS. The overall activity was in the order: ibuprofen approximately aspirin approximately acetaminophen approximately meclofenamic acid sodium salt approximately sulindac sulfide>ketoprofen approximately flurbiprofen approximately diclofenac sodium salt>naproxen approximately indomethacin. These findings indicate that NSAIDs could be key molecules for the development of therapeutic or preventive agents for LBD and MSA. PMID- 18164320 TI - Free recall in autism spectrum disorder: the role of relational and item-specific encoding. AB - Autism spectrum disorders (ASDs) are characterised by a relatively specific pattern of typical and atypical memory functioning. Convergent behavioural and neuroscientific evidence indicates that this pattern of functioning may be the result of specific impairments in hippocampally mediated relational memory processes, whilst brain-mechanisms mediating item-specific memory processes remain intact. In the current paper we draw on a behavioural paradigm developed by Hunt and Seta [Hunt, R. R., & Seta, C. E. (1984). Category size effects in recall--The roles of relational and individual item information. Journal of Experimental Psychology: Learning, Memory and Cognition, 10, 454-464], which not only allowed us to determine whether individuals with ASD did indeed experience selective difficulties in relational processes, but in addition enabled us to gain insights into the severity of this impairment. Our results suggest that whilst individuals with ASD employ relational memory processes atypically, this impairment seems restricted to situations in which such processes need to be deployed spontaneously to facilitate memory. Under situations that provide environmental support for the processing of relational information, individuals with ASD did demonstrate the ability to employ such processes relatively effectively. These findings provide further support for the 'Task Support Hypothesis' and suggest that relational memory processes may in principle be functionally intact despite not being triggered by the same environmental situations as in typical development. PMID- 18164321 TI - Stimulus (polyphenol, IFN-gamma, LPS)-dependent nitric oxide production and antileishmanial effects in RAW 264.7 macrophages. AB - The effects of interferon (IFN-gamma), lipopolysaccharide (LPS), and some polyphenols as individual stimuli, as well as in various combinations on NO production in non-infected and infected macrophage-like RAW 264.7 cells were investigated, with emphasis on the NO/parasite kill relationship. In non-infected and in Leishmania parasitized cells, gallic acid significantly inhibited the IFN gamma and LPS-induced NO detected in the supernatant. This effect was less prominent in IFN-gamma- than in LPS-stimulated cells. Interestingly, and in contrast to non-infected cells, gallic acid inhibited NO production only when added within 3h after IFN-gamma+LPS. Addition of gallic acid following prolonged incubation with IFN-gamma+LPS periods (24 h) no longer inhibited, sometimes even enhanced NO release. Notably, an excellent NO/parasite kill relationship was evident from all the experiments. This study was extended to a series of polyphenols (3-O-shikimic acid, its 3,5-digalloylated analogue, catechin, EGCG, and a procyanidin hexamer) with proven immunostimulatory activities. Although these compounds themselves were found to be weak NO-inducers, the viability of intracellular Leishmania parasites was considerably reduced. Furthermore, their dose-dependent effects on macrophage NO release was determined in the presence of IFN-gamma and/or LPS. Again, non-infected and infected cells differed significantly in the NO response, while inhibition of IFN-gamma and/or LPS induced NO production by the tested polyphenols strongly depended on the given time of exposure and the sequence of immunological stimuli. A strong inverse correlation between NO levels and intracellular survival rates of Leishmania parasites supported the assumption that the observed inhibition of NO was not simply due to interference with the Griess assay used for detection. PMID- 18164322 TI - Physiatrist 2007: who are we and where are we going? AB - The field of physical medicine and rehabilitation has evolved greatly over the last half century. Although practice patterns continue to change, the unifying concepts of physiatry remain the same. Awareness of the unique aspects of physiatry is still not optimal. It is incumbent on the physiatric community to educate our colleagues, our patients, and the public about our unique field. PMID- 18164323 TI - Moral and ethical decisions: to be or not to be. The 39th Walter J. Zeiter Lecture. AB - Throughout history, some societies have short-changed the weak, the elderly, and children with disabilities. In this lecture, I will explore some of the sad statistics related to dealing with people with disabilities, as well as attitudes toward, and treatments of, children and young adults with disabilities during the past 70 years. I will follow this with some suggestions for change in our society's attitudes toward this vulnerable population. PMID- 18164324 TI - Team training and stroke rehabilitation outcomes: a cluster randomized trial. AB - OBJECTIVE: To test whether a team training intervention in stroke rehabilitation is associated with improved patient outcomes. DESIGN: A cluster randomized trial of 31 rehabilitation units comparing stroke outcomes between intervention and control groups. SETTING: Thirty-one Veterans Affairs medical centers. PARTICIPANTS: A total of 237 clinical staff on 16 control teams and 227 staff on 15 intervention teams. Stroke patients (N=487) treated by these teams before and after the intervention. INTERVENTION: The intervention consisted of a multiphase, staff training program delivered over 6 months, including: an off-site workshop emphasizing team dynamics, problem solving, and the use of performance feedback data; and action plans for process improvement; and telephone and videoconference consultations. Control and intervention teams received site-specific team performance profiles with recommendations to use this information to modify team process. MAIN OUTCOME MEASURES: Three patient outcomes: functional improvement as measured by the change in motor items of the FIM instrument, community discharge, and length of stay (LOS). RESULTS: For both the primary (stroke only) and secondary analyses (all patients), there was a significant difference in improvement of functional outcome between the 2 groups, with the percentage of stroke patients gaining more than a median FIM gain of 23 points increasing significantly more in the intervention group (difference in increase, 13.6%; P=.032). There was no significant difference in LOS or rates of community discharge. CONCLUSIONS: Stroke patients treated by staff who participated in a team training program were more likely to make functional gains than those treated by staff receiving information only. Team based clinicians are encouraged to examine their own team. (ClinicalTrials.gov identifier NCT00237757). PMID- 18164325 TI - Biochemicals associated with pain and inflammation are elevated in sites near to and remote from active myofascial trigger points. AB - OBJECTIVES: To investigate the biochemical milieu of the upper trapezius muscle in subjects with active, latent, or absent myofascial trigger points (MTPs) and to contrast this with that of the noninvolved gastrocnemius muscle. DESIGN: We used a microanalytic technique, including needle insertions at standardized locations in subjects identified as active (having neck pain and MTP), latent (no neck pain but with MTP), or normal (no neck pain, no MTP). We followed a predetermined sampling schedule; first in the trapezius muscle and then in normal gastrocnemius muscle, to measure pH, bradykinin, substance P, calcitonin gene related peptide, tumor necrosis factor alpha, interleukin 1beta (IL-1beta), IL-6, IL-8, serotonin, and norepinephrine, using immunocapillary electrophoresis and capillary electrochromatography. Pressure algometry was obtained. We compared analyte concentrations among groups with 2-way repeated-measures analysis of variance. SETTING: A biomedical research facility. PARTICIPANTS: Nine healthy volunteer subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Preselected analyte concentrations. RESULTS: Within the trapezius muscle, concentrations for all analytes were higher in active subjects than in latent or normal subjects (P<.002); pH was lower (P<.03). At needle insertion, analyte concentrations in the trapezius for the active group were always higher (pH not different) than concentrations in the gastrocnemius muscle. At all times within the gastrocnemius, the active group had higher concentrations of all analytes than did subjects in the latent and normal groups (P<.05); pH was lower (P<.01). CONCLUSIONS: We have shown the feasibility of continuous, in vivo recovery of small molecules from soft tissue without harmful effects. Subjects with active MTPs in the trapezius muscle have a biochemical milieu of selected inflammatory mediators, neuropeptides, cytokines, and catecholamines different from subjects with latent or absent MTPs in their trapezius. These concentrations also differ quantitatively from a remote, uninvolved site in the gastrocnemius muscle. The milieu of the gastrocnemius in subjects with active MTPs in the trapezius differs from subjects without active MTPs. PMID- 18164326 TI - Cost-effectiveness of C-leg compared with non-microprocessor-controlled knees: a modeling approach. AB - OBJECTIVE: To estimate the costs and health outcomes of C-Leg and non microprocessor-controlled (NMC) knees using a decision-analytic model. DESIGN: Data on costs, rates and duration of problems, knee survival, and health-related quality of life were obtained from interviews with patients and prosthetists with experience of both C-Leg and NMC knees. Interview data were assessed in a decision-analytic Markov model to estimate cost-effectiveness from a health care perspective. SETTING: Outpatient. PARTICIPANTS: A population sample of 20 patients currently using the C-Leg and prior experience of nonmicroprocessor knees, and 5 prosthetists. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Incremental cost per quality-adjusted life year (QALY). RESULTS: The mean incremental cost (in 2006 Euros) and QALYs for the C-Leg was 7657 euros and 2.38, respectively, yielding a cost per QALY gained of 3218 euros. CONCLUSIONS: It is important to provide decision-makers with relevant information on costs and health outcomes of different treatment strategies on actual decision problems despite limited evidence. The results of the study, taking into account both costs and a broadly defined health outcome in terms of QALY, show that given existing albeit limited evidence the C-Leg appears to yield positive health outcomes at an acceptable cost. PMID- 18164327 TI - Sexual functioning in adolescents and young adults with spina bifida. AB - OBJECTIVE: To assess sexual education and sexual functioning in adolescents and young adults with spina bifida. DESIGN: Survey, inception cohort. SETTING: The community. PARTICIPANTS: A cohort of adolescents and young adults (N=121; range, 15-35 y; 58% women) enrolled in a longitudinal pediatric database. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Questions on sexual function, reproductive function, bladder and bowel continence, the Perceived Quality of Life Scale, and the Satisfaction With Life Scale. RESULTS: Almost all adolescents and young adults with spina bifida in our study received sexual education at school, less at home, or by physicians. Twenty-five percent of men and 68% of women were informed about reproductive function by their physicians. Participants who reported that they smoked were 10 times more likely to report being sexually active and women were 2.3 times more likely to be sexually active than men. Hydrocephalus was a significant predictor of sexual activity among women but not men. Participants with urinary incontinence were less likely to be sexually active. Women without hydrocephalus were significantly more satisfied with life than women with hydrocephalus. CONCLUSIONS: Adolescents and young adults with spina bifida in this sample were only slightly satisfied with life and sexual activity was only associated with life satisfaction among women. Dissatisfaction with life often leads to engagement in health-risk behaviors, which may, in part, account for the association between sexual activity and smoking behavior observed in these data. Further studies of health risk behaviors among youth with spina bifida are warranted and interventions aimed at reducing health risk behaviors among adolescents and young adults should specifically include spina bifida as a target group. PMID- 18164328 TI - Elevated C-reactive protein associated with decreased high-density lipoprotein cholesterol in men with spinal cord injury. AB - OBJECTIVES: To determine if people with spinal cord injury (SCI) have elevated C reactive protein (CRP), to examine the association of CRP with high-density lipoprotein cholesterol (HDL-C), and to assess the influence of completeness and level of injury on these parameters. DESIGN: Cross-sectional. SETTING: Urban university. PARTICIPANTS: Men with SCI (n=129) who were free of infection and/or recent anti-inflammatory medication use as well as their 1:1 age- and race matched able-bodied counterparts from the 1999-2002 National Health and Nutrition Examination Surveys. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: High CRP was defined as 3 mg/L or higher and low HDL-C as less than 1.04 mmol/L. RESULTS: Men with SCI were more likely to have high CRP (odds ratio [OR]=2.29; 95% confidence interval [CI], 1.33-3.95) and low HDL-C (OR=1.81; 95% CI, 1.01 3.27). The OR for low HDL-C in SCI was no longer significant when high CRP was controlled. CRP was higher in complete versus incomplete injury (median, 3.7 mg/L vs 1.2mg/L; P=.005), and this elevation was independent of age, smoking, physical activity, waist circumference, and weight. No conclusion can be made on the association of injury level and CRP because of a lack of power. CONCLUSIONS: The elevated CRP, possibly the major risk factor, together with decreased HDL-C may contribute to greater incidence for cardiovascular disease in the SCI population. PMID- 18164329 TI - Effect of severity of post-traumatic confusion and its constituent symptoms on outcome after traumatic brain injury. AB - OBJECTIVE: To investigate the prognostic significance of severity of post traumatic confusion (PTC) and its constituent symptoms for early and late outcome after traumatic brain injury (TBI). DESIGN: Prospective cohort study. SETTING: Inpatient brain injury rehabilitation program. PARTICIPANTS: A total of 168 patients meeting study criteria from 195 consecutive Traumatic Brain Injury Model Systems neurorehabilitation admissions. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Employability at neurorehabilitation discharge and productivity status at 1 year postinjury. RESULTS: More severely confused patients had poorer outcomes for both employability and productivity. Multivariable logistic regression revealed that after adjustment for all other predictors, time to follow commands, and confusion severity predicted employability at discharge and age and confusion severity predicted productivity status at 1 year. Each symptom showed an unadjusted effect on discharge employability. All symptoms except nighttime sleep disturbance or daytime decreased arousal had effects on productivity at 1 year. Presence of psychotic-type symptoms was associated with especially poor productivity outcomes. CONCLUSIONS: PTC constituent symptoms and severity predict outcome after TBI. Presence or absence of psychotic-type symptoms on a single evaluation at approximately 21 days postinjury may have particular prognostic significance for productivity outcome. PMID- 18164330 TI - Relationships among premorbid alcohol use, acute intoxication, and early functional status after traumatic brain injury. AB - OBJECTIVE: To investigate the relationships among intoxication at time of injury, preinjury history of problem drinking, and early functional status in patients with traumatic brain injury (TBI). DESIGN: Prospective cohort study. SETTING: Acute inpatient TBI rehabilitation. PARTICIPANTS: Participants were 1748 persons with TBI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Blood alcohol levels (BALs) were obtained at admission to the emergency department, and a history of problem drinking was obtained through interview. Study outcomes, Disability Rating Scale (DRS), and FIM instrument scores were gathered at admission to inpatient rehabilitation. RESULTS: Multivariate regression analysis revealed that BAL and a history of binge drinking were predictive of DRS, but not FIM, scores. A higher BAL was associated with poorer functional status on the DRS. Paradoxically, a history of binge drinking was associated with more intact functional status on the DRS. CONCLUSIONS: The relationships among intoxication at time of injury, history of problem drinking, and early outcome after TBI were modest. Injury severity had a more significant association with TBI functional status. PMID- 18164331 TI - Effect of botulinum toxin injection in the rectus femoris on stiff-knee gait in people with stroke: a prospective observational study. AB - OBJECTIVE: To study the effect of botulinum toxin type A (BTX-A) injection in the rectus femoris on the decreased knee flexion during the swing phase of gait (stiff-knee gait) in people with stroke. DESIGN: Intervention study (before-after trial) with an observational design. SETTING: Outpatient rehabilitation clinic and gait laboratory. PARTICIPANTS: Nineteen chronic hemiparetic adults presenting with stiff-knee gait. INTERVENTION: Injection of 200 U of BTX-A (Botox) into the rectus femoris. MAIN OUTCOME MEASURES: Before and 2 months after BTX-A rectus femoris injection: Stroke Impairment Assessment Set (SIAS), Duncan-Ely test, and an instrumented gait analysis. RESULTS: Median SIAS score improved from 53 (range, 36-65) to 57 (range, 42-70) (signed-rank test, P=.005) and the Duncan-Ely score from 3 (range, 1-3) to 1 (range, 0-3) (P<.001). In gait analysis, mean (+/- standard deviation) maximum knee flexion improved from 26 degrees +/-13 degrees to 31 degrees +/-14 degrees during the swing phase (paired t test, P<.001), knee flexion speed at toe-off improved from 82 degrees +/-63 degrees to 112 degrees +/ 75 degrees/s (P=.009), and knee negative joint power (eccentric muscular contraction) improved from -.27+/-.23 to -.37+/-.26 W/kg (P<.001). The 4 patients who almost did not flex the knee (<10 degrees) before the BTX-A rectus femoris injection did not improve after the injection. The other 14 patients who flexed the knee more than 10 degrees before the BTX-A rectus femoris injection decreased the walking energy cost from 5.4+/-1.6 to 4.6+/-1.3 J x kg(-1) x m(-1) (P=.006). CONCLUSIONS: BTX-A rectus femoris injection may be beneficial in patients with a stiff-knee gait after stroke, particularly in patients with some knee flexion (>10 degrees). PMID- 18164332 TI - Functional significance of ipsilesional motor deficits after unilateral stroke. AB - OBJECTIVE: To determine whether ipsilesional motor skills, which have been related to independent functioning, are present chronically after unilateral stroke and are more common in people with apraxia than in those without apraxia. DESIGN: Observational cohort comparing the performance of an able-bodied control group, stroke patients with left- or right-hemisphere damage matched for lesion volume, and left-hemisphere stroke patients with and without ideomotor limb apraxia. SETTING: Primary care Veterans Affairs and private medical center. PARTICIPANTS: Volunteer right-handed sample; stroke patients with left- or right hemisphere damage about 4 years poststroke; a control group of demographically matched, able-bodied adults. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Total time to perform the (1) Williams doors test and the (2) timed manual performance test (TMPT), which includes parts of the Jebsen-Taylor Hand Function Test. RESULTS: Ipsilesional motor deficits were present after left- or right-hemisphere stroke when using both measures, but deficits were consistently more common in patients with limb apraxia only for the TMPT. CONCLUSIONS: These findings add to a growing literature that suggests that ipsilesional motor deficits may have a functional impact in unilateral stroke patients, especially in patients with ideomotor limb apraxia. PMID- 18164333 TI - Psychometric properties of the Neck Disability Index and Numeric Pain Rating Scale in patients with mechanical neck pain. AB - OBJECTIVE: To examine the psychometric properties including test-retest reliability, construct validity, and minimum levels of detectable and clinically important change for the Neck Disability Index (NDI) and the numeric rating scale (NRS) for pain in a cohort of patients with neck pain. DESIGN: Single-group repeated-measures design. SETTING: Outpatient physical therapy (PT) clinics. PARTICIPANTS: Patients (N=137) presenting to PT with a primary report of neck pain. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: All patients completed the NDI and the NRS at the baseline examination and at a follow-up. At the time of the follow-up, all patients also completed the global rating of change, which was used to dichotomize patients as improved or stable. Baseline and follow-up scores were used to determine the test-retest reliability, construct validity, and minimal levels of detectable and clinically important change for both the NDI and NRS. RESULTS: Test-retest reliability was calculated using an intraclass correlation coefficient (ICC) (NDI ICC=.50; 95% confidence interval [CI], .25-.67; NRS ICC=.76; 95% CI, .51-.87). The area under the curve was .83 (95% CI, .75-.90) for the NDI score and .85 (95% CI, .78-.93) for the NRS score for determining between stable and improved patients. Thresholds for the minimum clinically important difference (MCID) for the NDI were 19-percentage points and 1.3 for the NRS. CONCLUSIONS: Both the NDI and NRS exhibit fair to moderate test-retest reliability in patients with mechanical neck pain. Both instruments also showed adequate responsiveness in this patient population. However, the MCID required to be certain that the change in scores has surpassed a level that could be contributed to measurement error for the NDI was twice that which has previously been reported. Therefore the ongoing analyses of the properties of the NDI in a patient population with neck pain are warranted. PMID- 18164334 TI - Therapeutic use of botulinum toxin type A in treating neck and upper-back pain of myofascial origin: a pilot study. AB - OBJECTIVE: To determine the efficacy of botulinum toxin type A (BTX-A) in treating neck and upper-back pain of myofascial origin. DESIGN: A randomized, double-blind, placebo-controlled pilot study. SETTING: Outpatient physical medicine and rehabilitation clinic of a university-affiliated tertiary hospital. PARTICIPANTS: A total of 29 subjects enrolled from among 45 screened patients. No subject withdrawal due to serious adverse events occurred. INTERVENTION: Subjects were evaluated at baseline, received a 1-time injection of either BTX-A (treatment group) or saline (control group), and were followed up at 2 weeks and at months 1, 2, 3, 4, and 6. MAIN OUTCOME MEASURES: Visual analog scale (VAS) for pain, the Neck Disability Index (NDI), and the Medical Outcome Study 36-Item Short-Form Health Survey (SF-36). RESULTS: Improvements in the VAS and NDI scores were seen in the treatment group but were not significant when compared with the controls. Statistically significant improvements for the treatment group were seen in the SF-36 bodily pain (at months 2 and 4) and mental health (at month 1) scales but not in the other scales, nor in the summary measures. No serious adverse events were reported. CONCLUSIONS: Trends toward improvements in VAS and NDI scores of the BTX-A group are encouraging, but they were possibly due to a placebo effect and were not statistically significant. The BTX-A subjects, at certain time points, showed statistically significant improvements in the bodily pain and mental health scales of the SF-36 compared with controls. Our study had limited power and population base, but the results could be used to properly power follow-up studies to further investigate this topic. PMID- 18164335 TI - Influence of acuity on physical therapy outcomes for patients with cervical disorders. AB - OBJECTIVES: To assess the influence of symptom acuity on functional outcomes, pain, and patient perception of recovery after a physical therapy (PT) program for cervical disorders and to determine what variables are associated with patient function at discharge. DESIGN: Retrospective case series. SETTING: Outpatient settings at a tertiary care facility. PARTICIPANTS: Patients (N=220) who were seen for PT between June 2003 and November 2005. INTERVENTIONS: A customized rehabilitation program was developed for each patient based on examination findings and included a combination of the following interventions: mobilization or manipulation, flexibility exercises, strengthening exercises, endurance exercises, massage techniques, and heat and cold modalities. MAIN OUTCOME MEASURES: Functional outcome, functional improvement, perceived pain, and perceived improvement scores in the CareConnections Outcomes System (formerly TAOS) database. RESULTS: Persons whose symptom duration was greater than 6 months (chronic group) had significantly less functional improvement than persons whose symptom duration was less than 1 month (acute group). The median percentage improvement score for patient perceived recovery was also significantly lower for the chronic group than for the acute group. There was no significant difference in the percentage decrease in pain among the acute, subacute (symptom duration, 1 6 mo), and chronic groups. In regression analyses, a model with age (P=.001), symptom duration (P=.05), and inclusion of mobilization and manipulation interventions (P=.02) fit the data well and explained 35.6% of the variance in functional outcome score for all 3 groups combined. CONCLUSIONS: Patients showed improvements in function after a rehabilitation program for cervical disorders. Patient functional score at discharge is influenced by age, symptom duration, and inclusion of mobilization or manipulation treatments. PMID- 18164336 TI - Three-dimensional motions of trunk and pelvis during transfemoral amputee gait. AB - OBJECTIVES: To identify characteristics of upper-body kinematics and torque transmission to the ground during locomotion in a group of patients with transfemoral amputation as compared with a group of asymptomatic subjects; and to investigate the influence of walking velocity and residual limb length on several characteristics of upper-body motion. DESIGN: Three-dimensional gait analysis with an optoelectronic device. SETTING: Gait laboratory. PARTICIPANTS: Twenty seven patients with transfemoral amputation and a control group of 33 nondisabled subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Three-dimensional kinematics of the pelvis and the thorax and ground reaction force for amputees and control subjects. RESULTS: For subjects with transfemoral amputation, it was observed that upper-body angular ranges of motion (ROMs) increased globally as walking velocity decreased. For these subjects, specific patterns of pelvic rotation and torque transmission by the lower limbs around the vertical axis were found. The counter-rotation between the pelvic and scapular girdles was reduced. This reduction proved to be linked with the decrease of walking velocity. Walking velocity also affected all the parameters describing the motion of upper body. Pelvic ROM increased with the length of the limb decreasing. CONCLUSIONS: The huge differences found between subjects with and without amputation suggest that the motion of the upper body must be considered to enhance gait. PMID- 18164337 TI - The feasibility of measuring joint angular velocity with a gyro-sensor. AB - OBJECTIVES: To determine the reliability of an assessment of joint angular velocity using a gyro-sensor and to examine the relationship between ankle angular velocity and physical functions. DESIGN: Cross-sectional. SETTING: Kinesiology laboratory. PARTICIPANTS: Twenty healthy young adults (mean age, 22.5 y) and 113 community-dwelling older adults (mean age, 75.1 y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Maximal ankle joint velocity was measured using a gyro-sensor during heel-rising and jumping with knee extended. The intraclass correlation coefficient (ICC) was used to determine the intertester and intratester reliability. The Pearson correlation coefficient was used to examine the relationships between maximal ankle joint velocity and isometric muscle strength and isokinetic muscle power in young adults and also to examine the relationships between maximal ankle joint velocity and functional performance measurements such as walking time in older adults. RESULTS: High reliability was found for intertester (ICC=.96) and intratester reliability (ICC=.96). The data from the gyro-sensor highly correlated with muscle strength (r range, .62-.68; P<.01) and muscle power (r range, .45-.79; P range, .01-.05). In older subjects, mobility functions significantly correlated with the angular velocity of ankle plantarflexion. CONCLUSIONS: Measurement of ankle angular velocity using a gyro sensor is both reliable and feasible, with the results representing a significant correlation to muscle power and performance measurements. PMID- 18164338 TI - Self-reported difficulty in climbing up or down stairs in nondisabled elderly. AB - OBJECTIVE: To examine clinical and functional correlates of self-reported difficulty in climbing up or climbing down stairs in older adults. DESIGN: Cross sectional survey. SETTING: Community sample. PARTICIPANTS: Older adults (N=310; mean age, 79.7 y; 62% women), without disability or dementia. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Clinical and functional status as well as activity limitations (able to perform activities of daily living [ADLs] with some difficulty). RESULTS: Of the 310 subjects, 140 reported difficulties in climbing up and 83 in climbing down stairs (59 both). Self-reported difficulty in climbing up stairs was associated with hypertension, arthritis, and depressive symptoms. Difficulty in climbing up stairs was also associated with poor balance and grip strength as well as neurologic gait abnormalities. Subjects with difficulty climbing down stairs had more falls. Both activities were associated with leg claudication, fear of falling, non-neurologic gait abnormalities, and slow gait. Examined individually, self-reported difficulty climbing down stairs captured a wider spectrum of ADL limitations than climbing up stairs. However, combined difficulty in both phases of stair climbing had a stronger association with activity limitations (vs no difficulty; odds ratio, 6.58; 95% confidence interval, 3.35-12.91) than difficulty in any one phase alone. CONCLUSIONS: Self reported difficulty in climbing up and down stairs revealed commonalities as well as differences in related clinical correlates. Difficulty in both climbing up and down stairs should be separately assessed to better capture clinical and functional status in older adults. PMID- 18164339 TI - Rehabilitation professionals and human immunodeficiency virus care: results of a national Canadian survey. AB - OBJECTIVE: To describe rehabilitation professionals' practices, knowledge and training, professional views, and service delivery issues for people living with human immunodeficiency virus (HIV) or acquired immune deficiency syndrome (PHAs) in Canada. DESIGN: Nationwide cross-sectional postal survey. SETTING: Canada. PARTICIPANTS: Random sample (N=2105) of occupational therapists, physical therapists, speech-language pathologists, and physiatrists who had practiced in the past year. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Survey items on current practices, HIV knowledge and training, professional views on rehabilitation and HIV, and HIV rehabilitation service delivery issues. RESULTS: Seventy-four percent (1492/2006) of the traceable sample responded, 53% (n=1058) of whom yielded completed surveys. Sixty-one percent of survey respondents never knowingly had served an HIV-positive patient. Of this group, 27% indicated these were patients they would like to work with, 27% indicated they were unwilling, and 46% were unsure. The 39% who knowingly had served PHAs had served an average of 4 PHAs in the last year, and less than 25% of their HIV patients' rehabilitation issues were HIV-related. CONCLUSIONS: Despite the role rehabilitation professionals have to play in the care of PHAs, only a minority serves PHAs. Results of this survey show a potential gap between the documented rehabilitative needs of PHAs and services provided by the rehabilitation professional community. PMID- 18164340 TI - Determinants of utilization of physical rehabilitation services for persons with chronic and disabling conditions: an exploratory study. AB - OBJECTIVE: To determine which variables influence the receipt of physical rehabilitation services (ie, physical, occupational, speech therapy) for a population of people with chronic and disabling conditions. DESIGN: A convenience sample of 502 adults with cerebral palsy (CP), multiple sclerosis (MS), and spinal cord injury (SCI), drawn from diverse parts of the United States. SETTING: Respondents were surveyed in the general community. PARTICIPANTS: Persons (91% under the age of 65 y) with CP, MS, or SCI who responded to the 1999 component of a national longitudinal survey. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Self-reported need for physical rehabilitation services. Bivariate and multivariate analyses were used to examine variables that influenced utilization of services. RESULTS: Some 53% of respondents did not receive self-reported needed physical rehabilitation services. Respondents who had Medicaid were more likely than those with Medicare or private insurance to receive physical rehabilitation services. Respondents having a lower household income and poorer health were less likely to receive services. CONCLUSIONS: Our findings indicate that health care funding sources provide widely disparate coverage for physical rehabilitation services to persons with 3 specific chronic and disabling conditions. Policy-makers and health plan administrators should re-evaluate their coverage of physical rehabilitation services designed to enhance quality of life and reduce the burden of lost independence. PMID- 18164341 TI - Self-reported health status and quality of life in youth with cerebral palsy and typically developing youth. AB - OBJECTIVE: To describe self-reported health status and quality of life (QOL) of ambulatory youths with cerebral palsy (CP) compared with sex- and age-matched typically developing youth (TDY). DESIGN: Prospective cross-sectional cohort comparison. SETTING: Community-based. PARTICIPANTS: A convenience sample of 81 youth with CP (age range, 10-13 y) with Gross Motor Function Classification System (GMFCS) levels I through III and 30 TDY participated. They were recruited from 2 regional children's hospitals and 1 regional military medical center. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Participants completed the Child Health Questionnaire-Child Form (CHQ-CF87) for health status and the Youth Quality of Life for QOL. RESULTS: Youth with CP reported significantly lower health status than age- and sex-matched TDY in the following CHQ-CF87 subscales: role/social behavioral physical, bodily pain, physical function, and general health (CP mean rank, 46.8-55.2; TDY mean rank, 62.2-80.9). There were significant differences across GMFCS levels. There were no significant differences in self-reported QOL. CONCLUSIONS: Self-reported health status, but not QOL, appears sensitive to the functional health issues experienced by ambulatory youth with CP. Pain management and psychosocial support may be indicated for them. PMID- 18164342 TI - The impact of limitations in physical, executive, and emotional function on health-related quality of life among adult survivors of childhood cancer: a report from the Childhood Cancer Survivor Study. AB - OBJECTIVE: To examine associations between limitations in physical performance, executive function, and emotional health (activity domains) and either social role attainment or health-related quality of life (HRQOL) in adult survivors of childhood cancer. DESIGN: Cross-sectional analysis. SETTING: Cancer survivors living in the community; previously treated for childhood cancer at one of 26 institutions. PARTICIPANTS: Subjects included 7147 (76.8%) of 9307 eligible adult members of the Childhood Cancer Survivor Study who completed a follow-up questionnaire between 2002 and 2004. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Demographic information was used to classify social roles and the Medical Outcomes Survey 36-Item Short-Form Health Survey to ascertain HRQOL. Questions from the National Health Interview Survey were used to represent physical performance; from the Brief Symptom Inventory to classify emotional health; and from the Behavioral Rating of Executive Function to describe executive function. Multivariate logistic regression was used to examine the association between limitations in activity domains, role attainment, and HRQOL. RESULTS: In this cohort, 18.1% reported deficits in physical performance, 10.5% in emotional health, and 14.0% in executive function. In adjusted models, when compared with survivors who reported no limitations, those with physical performance, executive function, or emotional health deficits were less likely to be employed, married, or have incomes greater than $20,000 a year. Limitations in executive function or emotional health were associated with no health insurance. Limitations in any activity domain were associated with poor HRQOL. Emotional health limitations had the most impact, with odds ratios from 3.18 (physical performance summary) to 25.81 (mental health). CONCLUSIONS: The results of these analyses show the need for development and testing of interventions to remediate limitations in activity domains, because they negatively impact role attainment and HRQOL. PMID- 18164343 TI - Wavelet-based spectrum analysis of sacral skin blood flow response to alternating pressure. AB - OBJECTIVES: To provide insight into the physiologic mechanisms associated with alternating pressure, using wavelet analysis of skin blood flow (SBF) oscillations, and to determine whether the application of alternating pressure induces myogenic responses, thereby enhancing SBF as compared with constant loading. DESIGN: Repeated-measures design. SETTING: University research laboratory. PARTICIPANTS: Healthy, young adults (N=10; 5 men, 5 women; mean age +/- standard deviation, 30.0+/-3.1 y). INTERVENTION: Alternating pressure for 20 minutes (four 5-min cycles with either 60 mmHg or 3 mmHg) and constant loading for 20 minutes at 30 mmHg on the skin over the sacrum. MAIN OUTCOME MEASURES: A laser Doppler flowmeter was used to measure sacral SBF response to both alternating pressure and constant loading. Wavelet-based spectrum analysis of SBF oscillations was used to assess underlying physiologic mechanisms including endothelium-related metabolic (.008-.02 Hz), neurogenic (.02-.05 Hz), and myogenic (.05-.15 Hz) controls. RESULTS: Alternating pressure stimulated an increase in sacral SBF of compressed soft tissues as compared with constant loading (P<.01). SBF during the high-pressure phase of 4 alternating pressure cycles showed an increasing trend. An increase in power in metabolic frequency range and a decrease in power in the myogenic frequency range during alternating pressure were observed compared with SBF prior to loading. Power increased in the myogenic frequency range during the low-pressure phase of alternating pressure and decreased during the high-pressure phase. CONCLUSIONS: SBF control mechanisms, as assessed by the characteristic frequencies embedded in SBF oscillations, show different responses to 2 loading pressures with the same average pressure but different patterns. Our study suggests that optimization of operating parameters and configurations of alternating pressure support surfaces to compensate for impaired SBF control mechanisms in pathologic populations may be possible using wavelet analysis of blood flow oscillations. PMID- 18164344 TI - An unusual cause of S1 radicular pain presenting as early phantom pain in a transfemoral amputee: a case report. AB - Recent epidemiologic studies have shown back pain to be a significant cause of pain in lower-limb amputees, but only a handful of cases have reported sciatica in amputees. The symptoms are usually described as a phantom pain or neuropathic pain in the residual limb that is often refractory to conventional treatments. These symptoms typically occur with back pain and are distinct from the patient's usual symptoms. Interestingly, back pain is not a universal finding. We present a patient with presumed phantom limb pain subsequently discovered to be caused by an S1 radiculopathy. This patient's supposed phantom pain persisted despite multiple medication trials. Initial work-up revealed a sciatic neuroma at the stump. Treatments targeting this neuroma were unsuccessful. Further evaluation found that a sacroiliac joint screw placed to stabilize a pelvic fracture had intruded into the S1 neuroforamen. A diagnostic S1 nerve block temporarily relieved the patient's pain, and the screw was removed. Pain persisted and a spinal cord stimulator was placed resulting in improvement of his pain. Because conventional diagnostic tests are limited, including physical exam and electromyography, a fluoroscopically guided selective spinal nerve block proved to be a useful diagnostic tool in this patient. PMID- 18164345 TI - Transitional cell carcinoma of the urethra [correction of ureter] in a patient with buttock pain: a case report. AB - This case reports on a patient with an unusual presentation of a rare tumor: urethral transitional cell carcinoma (TCC). Urethral TCC occurs in approximately 0.7% to 4.0% of patients who have had primary bladder cancer. The initial symptoms usually involve hematuria, with approximately a third of patients reporting flank area pain. Buttock pain and the absence of hematuria are uncommon with this disorder. The patient was initially suspected to have piriformis syndrome, but when he did not respond as expected to treatment, and because of his history of primary bladder cancer, further evaluation was undertaken and the diagnosis was made. The patient responded well to radiation and chemotherapy. Musculoskeletal physicians should be particularly suspicious of the presence of urethral TCC in a patient with a history of primary bladder cancer who reports low back or buttock pain, particularly if the patient does not respond quickly to treatment. PMID- 18164346 TI - Ultrasound-guided intra-articular injection of the trapeziometacarpal joint: description of technique. AB - OBJECTIVE: To describe a new technique to perform an ultrasound-guided intra articular injection of the trapeziometacarpal (TMC) joint. DESIGN: Ultrasound guided injection of the TMC joint was completed on fresh frozen cadaver hand specimens using diatriazoate meglumine contrast. A fluoroscopic posteroanterior image of the TMC joint was then obtained to verify intra-articular placement of the contrast. SETTING: Anatomy lab in a medical college. SPECIMENS: Seventeen fresh frozen cadaver hand specimens. INTERVENTIONS: Not applicable. MAIN OUTCOMES MEASURE: Verification of this technique was confirmed using fluoroscopy and contrast. RESULTS: Sixteen (94%) of 17 joints injected showed contrast material within the TMC joint with a single cutaneous puncture. One intra-articular injection was initially misplaced into the scaphotrapeziotrapezoid joint. CONCLUSIONS: Ultrasound may be used to accurately perform intra-articular TMC injections. Ultrasound provides a viable alternative to fluoroscopy when accurate injection into the TMC joint is required for diagnostic or therapeutic purposes. PMID- 18164347 TI - New views of myofascial trigger points: etiology and diagnosis. AB - Two studies appearing in Archives, one by Shah and colleagues and another one by Chen and colleagues, present groundbreaking findings that can reduce some of the controversy surrounding myofascial trigger points (MTPs). Both author groups recognize the ubiquity of this disease and the importance to patients of health care professionals becoming better acquainted with the cause and identification of MTPs. The integrated hypothesis is the most credible and most complete proposed etiology of MTPs. However, the feedback loop suggested in this hypothesis has a few weak links, and studies by Shah and colleagues in particular supply a solid link for one of them. The feedback loop connects the hypothesized energy crisis with the milieu changes responsible for noxious stimulation of local nociceptors that causes the local and referred pain of MTPs. Shah's reports quantify the presence of not just 1 noxious stimulant but 11 of them with outstanding concentrations of immune system histochemicals. The results also strongly place a solid histochemical base under the important clinical distinction between active and latent MTPs. The study by Chen on the use of magnetic resonance elastography (MRE) imaging of the taut band of an MTP in an upper trapezius muscle may open a whole new chapter in the centuries-old search for a convincing demonstration of the cause of MTP symptoms. MRE is a modification of existing magnetic resonance imaging equipment, and it images stress produced by adjacent tissues with different degrees of tension. This report seems to present an MRE image of the taut band that shows the chevron signature of the increased tension of the taut band compared with surrounding tissues. PMID- 18164348 TI - "To care for him who shall have borne the battle and for his widow and his orphan" (Abraham Lincoln): the Department of Veterans Affairs polytrauma system of care. AB - The initiation of combat in Iraq and Afghanistan has resulted in a new cohort of active-duty service members and veterans seeking rehabilitation care through the U.S. Department of Veterans Affairs (VA). Service members injured in combat most often sustain multiple injuries (polytrauma) and require a unique service delivery model to meet their needs. The VA recognized this need and responded with the development of the Polytrauma System of Care (PSC). This national system of care balances access and expertise to provide specialized life-long care to the combat injured. The PSC is comprised of: 4 specialized regional rehabilitation centers that are accredited in brain injury by the Commission on Accreditation of Rehabilitation Facilities; 21 specialized outpatient and subacute rehabilitation programs; designated polytrauma teams at smaller, more remote VA facilities; and a point of contact at all other VA facilities. In addition, the PSC has developed a proactive case-management model, a specialized telehealth network, guidelines for long-term follow-up, and services for those individuals who are unable to return home. The following commentary and articles provide additional detail on this new and unique system of care. PMID- 18164349 TI - Characteristics and rehabilitation outcomes among patients with blast and other injuries sustained during the Global War on Terror. AB - OBJECTIVE: To describe characteristics and rehabilitation outcomes among patients who received inpatient rehabilitation for blast and other injuries sustained in Iraq and Afghanistan during the Global War on Terror. DESIGN: Observational study based on chart review and Department of Veterans Affairs (VA) administrative data. SETTING: The 4 VA polytrauma rehabilitation centers (PRCs). PARTICIPANTS: Service members (N=188) admitted to a PRC during the first 4 years of the Global War on Terror for injuries sustained during Operation Iraqi Freedom or Operation Enduring Freedom. INTERVENTION: Multidisciplinary comprehensive rehabilitation program. MAIN OUTCOMES MEASURES: Cognitive and motor FIM instrument gain scores and length of stay (LOS). RESULTS: Most war-injured patients had traumatic brain injury, injuries to several other body systems and organs, and associated pain. Fifty-six percent had blast-related injuries, and the pattern of injuries was unique among those with injuries secondary to blasts. Soft tissue, eye, oral and maxillofacial, otologic, penetrating brain injuries, symptoms of post-traumatic stress disorder, and auditory impairments were more common in blast-injured patients than in those with war injuries of other etiologies. The mechanism of the injury did not predict functional outcomes. LOS was variable, particularly for those with blast injuries. Patients with low levels of independence at admissions made the most progress but remained more dependent at discharge compared with other PRC patients. The rate of gain was slower in this low functioning group. CONCLUSIONS: Blasts produce a unique constellation of injuries but do not make a unique contribution to functional gain scores. Findings underscore the need for assessment and treatment of pain and mental health problems among patients with polytrauma and blast-related injuries. Patients with polytrauma have lifelong needs, and future research should examine needs over time after community re-entry. PMID- 18164350 TI - Provider perspectives on rehabilitation of patients with polytrauma. AB - OBJECTIVES: To describe, from the perspective of U.S. Department of Veterans Affairs (VA) polytrauma rehabilitation providers, (1) patients with combat related polytrauma and their rehabilitation, (2) polytrauma patient family member involvement in rehabilitation, and (3) the impact on providers of providing polytrauma rehabilitation. DESIGN: Qualitative study based on rapid assessment process methodology, which included semistructured interviews, observation, and use of a field liaison. SETTING: The 4 VA polytrauma rehabilitation centers (PRCs). PARTICIPANTS: Fifty-six purposefully selected PRC providers and providers from consulting services. INTERVENTIONS: Not applicable. MAIN OUTCOMES MEASURES: Provider self-report of polytrauma patient characteristics, polytrauma patient family member involvement in rehabilitation, and the impact of polytrauma rehabilitation on providers themselves. RESULTS: According to PRC providers, polytrauma patients are younger than VA rehabilitation patients. Strong military identities affect rehabilitation needs and reactions to severe injury. The public and the media have particular interest in war-injured patients. Patients with blast-related polytrauma have unique constellations of visible (including amputations, craniectomies, and burns) and invisible (including traumatic brain injury, pain, and posttraumatic stress disorder) injuries. Providers have adjusted treatment strategies and involved services outside of rehabilitation because of this clinical complexity. Family members are intensely involved in rehabilitation and have service needs that may surpass those of families of rehabilitation patients without polytrauma. Sources of provider stress include new responsibilities, media attention, increased oversight, and emotional costs associated with treating severely injured young patients and their families. Providers also described the work as deeply rewarding. CONCLUSIONS: The VA should prioritize the identification or development and implementation of strategies to address family member needs and to monitor and ensure that PRC providers have access to appropriate resources. Future research should determine whether findings generalize to patients injured in other wars and to people who sustain polytraumatic injuries outside of a war zone, including victims of terrorist attacks. PMID- 18164351 TI - The interdisciplinary team and polytrauma rehabilitation: prescription for partnership. AB - Optimal outcomes for polytrauma survivors depend on the integration of complex medical, psychosocial, financial, educational, and vocational resources across diverse specialties and multiple medical centers, programs, and organizations and all in a setting of high public visibility and family involvement. Well functioning teams are critical to service integration, and teams are more effective in supportive hospital environments. Here, we offer a model of team functioning relevant to polytrauma and outline a team training program to improve services. Furthermore, we propose a partnership among the team, hospital administrators, and national leaders and with patients and their families. Integrated care requires partnerships among the various stakeholders, and those working in polytrauma have a unique opportunity to create an updated paradigm of the team approach responsive to the complexities of contemporary health care. PMID- 18164352 TI - Enhancing access of combat-wounded veterans to specialist rehabilitation services: the VA Polytrauma Telehealth Network. AB - Operations Iraqi Freedom and Enduring Freedom have resulted in U.S. military personnel sustaining combat wounds of unprecedented severity and complexity that necessitate long-term rehabilitation. To meet what are often conflicting requirements in providing severely wounded veterans with timely and convenient access to specialist rehabilitation care, and to enable them to return to their local communities, the Veterans Health Administration has developed a state-of the-art Polytrauma Telehealth Network that enhances access to such services by linking Veterans Administration rehabilitation facilities. This article describes the clinical, technical, and business process issues involved in the development of this network. PMID- 18164353 TI - Beyond war and military medicine: social factors in the development of prosthetics. AB - Polytrauma is an immediate outcome of current warfare, and the need to investigate this condition is equally immediate. The value of historical analysis in this endeavor should not be underestimated. It is among the best tools we have to help ensure that current research and practice involve engagement with the social contexts of polytrauma as well as with the medical science of its treatment. This special communication provides historical perspective on certain aspects of the polytraumatic condition--namely, limb loss, prosthetic rehabilitation, and community reintegration after receiving a prosthesis. It discusses the influential role of societal factors in these areas to encourage greater understanding that the care of persons with polytrauma must involve critical thinking about their relationships to and participation in society as well as their treatment by medical science. This special communication also provides historical perspective to enrich appreciation of the value of history for the field of physical medicine and rehabilitation (PM&R), the PM&R clinician, and the PM&R clinical researcher. Readers will learn that historical knowledge puts PM&R research and practice into perspective, reminding us that rehabilitation should involve critical thinking not only about medicine, but also about social roles and the participation of people in society despite physical and psychologic challenges. PMID- 18164354 TI - Acupuncture study hypotheses should rely on scientific, not imaginary, models. PMID- 18164356 TI - Proteomic analysis of dog tears for potential cancer markers. AB - The first reference map of the proteome of pooled normal dog tears was created using 2-dimensional polyacrylamide gel electrophoresis and the identity of a number of the major species determined using matrix-assisted laser desorption time of flight mass spectrometry (MALDI-TOF) and peptide mass fingerprint matching on protein sequence databases. In order to understand the changes in protein expression in the tear film of dogs with cancer, tears from such animals were similarly examined. A number of differences were found between the tears of healthy dogs and the dogs with cancer. Differences were found in levels of actin and albumin and in an unidentified protein which may be analogous to human lacryglobulin. These findings suggest that it may be possible to develop tear film analysis to provide a simple non-invasive test for the diagnosis and/or management of canine cancers. PMID- 18164357 TI - Effects of high dietary vitamin A supplementation on tibial dyschondroplasia, skin pigmentation and growth performance in avian broilers. AB - An experiment was conducted to study high dietary vitamin A on tibial dyschondroplasia, growth performance and skin pigmentation in broilers. One hundred and twenty Avian commercial broilers were randomly allotted to three treatments: group C (control group), in which broilers were fed basic diet containing vitamin A 5512IU/kg diet; group A, in which broilers were fed basic diet with addition vitamin A 35512IU/kg; group B, broilers were fed basic diet with supplement vitamin A 65512IU/kg. The experiment lasted 35d and at the end of the trial, broilers were killed and the right proximal tibiotarsi were dissected in longitudinal section for the assessment of TD incidence and TD index, skin from the same area of breast and tibia in broilers were collected to determine pigmentation. The results showed that a high level vitamin A significantly increased the rate of TD incidence and TD index, but middle level vitamin A did not have a significant effect on that. Both low and high retinoic acid decreased growth performance and skin pigmentation in broilers. It suggests that a high dietary vitamin A cause tibial dyschondroplasia in broilers, decreased growth performance and skin pigmentation. It is likely that the effect of vitamin A on TD is mediated through a depression of vitamin D status. PMID- 18164358 TI - Dual roles of brain serine hydrolase KIAA1363 in ether lipid metabolism and organophosphate detoxification. AB - Serine hydrolase KIAA1363 is an acetyl monoalkylglycerol ether (AcMAGE) hydrolase involved in tumor cell invasiveness. It is also an organophosphate (OP) insecticide-detoxifying enzyme. The key to understanding these dual properties was the use of KIAA1363 +/+ (wildtype) and -/- (gene deficient) mice to define the role of this enzyme in brain and other tissues and its effectiveness in vivo in reducing OP toxicity. KIAA1363 was the primary AcMAGE hydrolase in brain, lung, heart and kidney and was highly sensitive to inactivation by chlorpyrifos oxon (CPO) (IC50 2 nM) [the bioactivated metabolite of the major insecticide chlorpyrifos (CPF)]. Although there was no difference in hydrolysis product monoalkylglycerol ether (MAGE) levels in +/+ and -/- mouse brains in vivo, isopropyl dodecylfluorophosphonate (30 mg/kg) and CPF (100 mg/kg) resulted in 23 51% decrease in brain MAGE levels consistent with inhibition of AcMAGE hydrolase activity. On incubating +/+ and -/- brain membranes with AcMAGE and cytidine-5' diphosphocholine, the absence of KIAA1363 activity dramatically increased de novo formation of platelet-activating factor (PAF) and lyso-PAF, signifying that metabolically-stabilized AcMAGE can be converted to this bioactive lipid in brain. On considering detoxification, KIAA1363 -/- mice were significantly more sensitive than +/+ mice to ip-administered CPF (100 mg/kg) and parathion (10 mg/kg) with increased tremoring and mortality that correlated for CPF with greater brain acetylcholinesterase inhibition. Docking AcMAGE and CPO in a KIAA1363 active site model showed similar positioning of their acetyl and trichloropyridinyl moieties, respectively. This study establishes the relevance of KIAA1363 in ether lipid metabolism and OP detoxification. PMID- 18164359 TI - Toxicological effects of cinnabar in rats by NMR-based metabolic profiling of urine and serum. AB - Cinnabar, an important traditional Chinese mineral medicine, has been widely used as a Chinese patent medicine ingredient for sedative therapy. However, the pharmaceutical and toxicological effects of cinnabar, especially in the whole organism, were subjected to few investigations. In this study, an NMR-based metabolomics approach has been applied to investigate the toxicological effects of cinnabar after intragastrical administration (dosed at 0.5, 2 and 5 g/kg body weight) on male Wistar rats. Liver and kidney histopathology examinations and serum clinical chemistry analyses were also performed. The 1H NMR spectra were analyzed using multivariate pattern recognition techniques to show the time- and dose-dependent biochemical variations induced by cinnabar. The metabolic signature of urinalysis from cinnabar-treated animals exhibited an increase in the levels of creatinine, acetate, acetoacetate, taurine, hippurate and phenylacetylglycine, together with a decrease in the levels of trimethyl-N-oxide, dimethylglycine and Kreb's cycle intermediates (citrate, 2-oxoglutarate and succinate). The metabolomics analyses of serum showed elevated concentrations of ketone bodies (3-d-hydroxybutyrate and acetoacetate), branched-chain amino acids (valine, leucine and isoleucine), choline and creatine as well as decreased glucose, lipids and lipoproteins from cinnabar-treated animals. These findings indicated cinnabar induced disturbance in energy metabolism, amino acid metabolism and gut microflora environment as well as slight injury in liver and kidney, which might indirectly result from cinnabar induced oxidative stress. This work illustrated the high reliability of NMR-based metabolomic approach on the study of the biochemical effects induced by traditional Chinese medicine. PMID- 18164360 TI - Single whole-body exposure to sarin vapor in rats: long-term neuronal and behavioral deficits. AB - Freely moving rats were exposed to sarin vapor (34.2+/-0.8 microg/l) for 10 min. Mortality at 24 h was 35% and toxic sings in the surviving rats ranged from sever (prolonged convulsions) through moderate to almost no overt signs. Some of the surviving rats developed delayed, intermittent convulsions. All rats were evaluated for long-term functional deficits in comparison to air-exposed control rats. Histological analysis revealed typical cell loss at 1 week post inhalation exposure. Neuronal inflammation was demonstrated by a 20-fold increase in prostaglandin (PGE(2)) levels 24 h following exposure that markedly decreased 6 days later. An additional, delayed increase in PGE(2) was detected at 1 month and continued to increase for up to 6 months post exposure. Glial activation following neural damage was demonstrated by an elevated level of peripheral benzodiazepine receptors (PBR) seen in the brain 4 and 6 months after exposure. At the same time muscarinic receptors were unaffected. Six weeks, four and six months post exposure behavioral evaluations were performed. In the open field, sarin-exposed rats showed a significant increase in overall activity with no habituation over days. In a working memory paradigm in the water maze, these same rats showed impaired working and reference memory processes with no recovery. Our data suggest long lasting impairment of brain functions in surviving rats following a single sarin exposure. Animals that seem to fully recover from the exposure, and even animals that initially show no toxicity signs, developed some adverse neural changes with time. PMID- 18164361 TI - Transcriptional responses in thyroid tissues from rats treated with a tumorigenic and a non-tumorigenic triazole conazole fungicide. AB - Conazoles are azole-containing fungicides that are used in agriculture and medicine. Conazoles can induce follicular cell adenomas of the thyroid in rats after chronic bioassay. The goal of this study was to identify pathways and networks of genes that were associated with thyroid tumorigenesis through transcriptional analyses. To this end, we compared transcriptional profiles from tissues of rats treated with a tumorigenic and a non-tumorigenic conazole. Triadimefon, a rat thyroid tumorigen, and myclobutanil, which was not tumorigenic in rats after a 2-year bioassay, were administered in the feed to male Wistar/Han rats for 30 or 90 days similar to the treatment conditions previously used in their chronic bioassays. Thyroid gene expression was determined using high density Affymetrix GeneChips (Rat 230_2). Gene expression was analyzed by the Gene Set Expression Analyses method which clearly separated the tumorigenic treatments (tumorigenic response group (TRG)) from the non-tumorigenic treatments (non-tumorigenic response group (NRG)). Core genes from these gene sets were mapped to canonical, metabolic, and GeneGo processes and these processes compared across group and treatment time. Extensive analyses were performed on the 30-day gene sets as they represented the major perturbations. Gene sets in the 30-day TRG group had over representation of fatty acid metabolism, oxidation, and degradation processes (including PPARgamma and CYP involvement), and of cell proliferation responses. Core genes from these gene sets were combined into networks and found to possess signaling interactions. In addition, the core genes in each gene set were compared with genes known to be associated with human thyroid cancer. Among the genes that appeared in both rat and human data sets were: Acaca, Asns, Cebpg, Crem, Ddit3, Gja1, Grn, Jun, Junb, and Vegf. These genes were major contributors in the previously developed network from triadimefon-treated rat thyroids. It is postulated that triadimefon induces oxidative response genes and activates the nuclear receptor, Ppargamma, initiating transcription of gene products and signaling to a series of genes involved in cell proliferation. PMID- 18164362 TI - Anticancer activity of botanical alkyl hydroquinones attributed to topoisomerase II poisoning. AB - Cytotoxic alkyl hydroquinone compounds have been isolated from many plants. We previously isolated 3 structurally similar cytotoxic alkyl hydroquinone compounds from the sap of the lacquer tree Rhus succedanea L. belonging to the sumac family, which have a long history of medicinal use in Asia. Each has an unsaturated alkyl chain attached to the 2-position of a hydroquinone ring. One of these isolates, 10'(Z),13'(E),15'(E)-heptadecatrienylhydroquinone [HQ17(3)], being the most cytotoxic, was chosen for studying the anticancer mechanism of these compounds. We found that HQ17(3) was a topoisomerase (Topo) II poison. It irreversibly inhibited Topo IIalpha activity through the accumulation of Topo II DNA cleavable complexes. A cell-based assay showed that HQ17(3) inhibited the growth of leukemia HL-60 cells with an EC50 of 0.9 microM, inhibited the topoisomerase-II-deficient cells HL-60/MX2 with an EC50 of 9.6 microM, and exerted no effect on peripheral blood mononuclear cells at concentrations up to 50 microM. These results suggest that Topo II is the cellular drug target. In HL 60 cells, HQ17(3) promptly inhibited DNA synthesis, induced chromosomal breakage, and led to cell death with an EC50 about one-tenth that of hydroquinone. Pretreatment of the cells with N-acetylcysteine could not attenuate the cytotoxicity and DNA damage induced by HQ17(3). However, N-acetylcysteine did significantly reduce the cytotoxicity of hydroquinone. In F344 rats, intraperitoneal injection of HQ17(3) for 28 days induced no clinical signs of toxicity. These results indicated that HQ17(3) is a potential anticancer agent, and its structural features could be a model for anticancer drug design. PMID- 18164363 TI - A machine learning predictor of facial attractiveness revealing human-like psychophysical biases. AB - Recent psychological studies have strongly suggested that humans share common visual preferences for facial attractiveness. Here, we present a learning model that automatically extracts measurements of facial features from raw images and obtains human-level performance in predicting facial attractiveness ratings. The machine's ratings are highly correlated with mean human ratings, markedly improving on recent machine learning studies of this task. Simulated psychophysical experiments with virtually manipulated images reveal preferences in the machine's judgments that are remarkably similar to those of humans. Thus, a model trained explicitly to capture a specific operational performance criteria, implicitly captures basic human psychophysical characteristics. PMID- 18164364 TI - Levels and distribution of polybrominated diphenyl ethers in plant, shellfish and sediment samples from Laizhou Bay in China. AB - Polybrominated diphenyl ethers (PBDEs) are potentially harmful and persistent environmental pollutants. PBDEs concentrations are reported in plant, shellfish and sediment samples collected from Chinese Laizhou Bay. The summation operator(11)PBDE concentrations in plant and shellfish samples from Laizhou Bay were in the range of 70-5900ngg(-1) and 230-720ngg(-1) lipid, respectively. The summation operator(11)PBDEs concentrations in river sediment intervals samples ranged from 1.3 to 1800ng g(-1) dry weight. Pearson correlation analyses were performed on concentrations of PBDEs congeners of different sediment sample intervals. There were significant positive correlations for BDE 28 and BDE 100 (r=0.945, p<0.01), BDE 47 and BDE 99 (r=0.879, p<0.01), BDE 153 and BDE 154 (r=0.934, p<0.01), nona-BDEs and BDE 209 (r>0.934, p<0.01). BDE 209 was the predominant congener in all analyzed samples, consistent with the fact that deca BDE technical mixtures are the dominant PBDEs product in Laizhou Bay. Data showed that PBDEs should be considered as an increasing pollution problem in the Laizhou Bay region. PMID- 18164365 TI - Influence of metal resistant-plant growth-promoting bacteria on the growth of Ricinus communis in soil contaminated with heavy metals. AB - The metal resistant-plant growth-promoting bacterial (PGPB) strains PsM6 and PjM15 isolated from a serpentine soil were characterized as Pseudomonas sp. and Pseudomonas jessenii, respectively, on the basis of their morphological, physiological, biochemical characteristics and 16S rDNA sequences. Assessment of plant growth-promoting parameters revealed the intrinsic ability of the strains for the utilization of 1-aminocyclopropane-1-carboxylic acid as the sole N source, solubilization of insoluble phosphate and production of indole-3-acetic acid (IAA). Further, a pot experiment was conducted to elucidate the effects of inoculating metal resistant PGPB on the plant growth and the uptake of Ni, Cu and Zn by Ricinus communis. Inoculation of Pseudomonas sp. PsM6 or P. jessenii PjM15 increased the shoot and root biomass of R. communis grown in non-contaminated and contaminated soil. However, the maximum biomass was observed in the plants inoculated with strain PjM15. This effect can be attributed to the solubilization of phosphate and production of IAA. Inoculation of Pseudomonas sp. PsM6 and PjM15 did not greatly alter the organ metal concentrations except Zn which concentration was higher in root, stem and leaf of inoculated plants. The results of metal extraction with PGPB strains showed that PsM6 was more efficient at solubilizing Zn than PjM15, and that PjM15 was better at solubilising Ni and Cu than PsM6. Owing to its wide action spectrum, the metal resistant PGPB could serve as an effective metal sequestering and growth-promoting bioinoculant for plants in metal-stressed soil. The present study has provided a new insight into the phytoremediation of metal-contaminated soil. PMID- 18164366 TI - Distribution characteristics of nonylphenolic chemicals in Masan Bay environments, Korea. AB - To understand the distribution characteristics of nonylphenolics and sterols, samples such as in creek water, sea surface water, waste water treatment plant (WWTP) effluent water, sediment and mussel were collected and analyzed. The principal analytes are nonylphenol (NP), nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), coprostanol (5beta) and cholestanol (5alpha). All these target pollutants showed 100% detection frequency in all of the samples analyzed. Total concentration of nonylphenolic compounds ranged from 334 to 3628ngl(-1) (average: 1331ngl(-1)) in creek water, from 15 to 36400ngl(-1) (average: 1013ngl(-1)) in sea surface water, from 131 to 2811ngg(-1) dry weight (average: 581ngg(-1) dry weight) in sediment and from 50.5 to 289ngg(-1) dry weight (average: 139ngg(-1) dry weight) in mussel. For water samples, levels of nonylphenolics determined in summer season were higher than those in spring season. Among them, nonylphenol and NP1EO was dominant in creek water and seawater, respectively. The highest concentration was recorded in sediment near a WWTP effluent outlet. And high levels of nonylphenolics and sterols were found in about 3km area surrounding WWTP effluent outlet. Coefficient of linear regression (R(2)) for NP in mussel and in sediment was 0.90. Similarly good correlation (R(2)=0.98) was obtained between concentration in water and in mussel indicating that a steady state has been reached in this bay. The calculated bio concentration factor (BCF=2990) for NP in Masan Bay agrees well with reported values in the literature. PMID- 18164367 TI - Genotoxicity of the organochlorine pesticides 1,1-dichloro-2,2- bis(p chlorophenyl)ethylene (DDE) and hexachlorobenzene (HCB) in cultured human lymphocytes. AB - The possible genotoxic potential of 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), which is a metabolite of dichlorobiphenyltrichloroetane (DDT), and hexachlorobenzene (HCB), which are organochlorine pesticides have been evaluated in vitro by using human lymphocytes as test system. Genetic damage was determined by scoring the frequency of micronuclei (MN) in primary lymphocyte cultures obtained from different donors. The results indicated that, under the experimental conditions used, the DDT metabolite DDE was able to induce significant increases in the frequency of micronucleated cells, which indicate a certain clastogenic and/or aneugenic potential. DDE was tested in the range of 10 80 mM, but the only concentration producing a significant genotoxic effect was 80 mM. On the other hand, HCB was unable to induce a significant increase in the MN frequency in the range of concentrations assayed, from 0.005 to 0.1mM. The selected concentrations of DDE and HCB were chosen according to their toxicity in cell blood cultures; higher concentrations reduced significantly cell proliferation and produced a low frequency of binucleated cells. In conclusion, the results indicate that a genotoxic risk is associated with the exposure to DDE at concentrations 80 mM and above. PMID- 18164369 TI - Atypical Spitz tumors: facts and opinions on intranodal melanocytes. PMID- 18164370 TI - Human cranial diversity and evidence for an ancient lineage of modern humans. AB - This study examines the genetic affinities of various modern human groupings using a multivariate analysis of morphometric data. Phylogenetic relationships among these groupings are also explored using neighbor-joining analysis of the metric data. Results indicate that the terminal Pleistocene/early Holocene fossils from Australasia exhibit a close genetic affinity with early modern humans from the Levant. Furthermore, recent human populations and Upper Paleolithic Europeans share a most recent common ancestor not shared with either the early Australasians or the early Levantine humans. This pattern of genetic and phylogenetic relationships suggests that the early modern humans from the Levant either contributed directly to the ancestry of an early lineage of Australasians, or that they share a recent common ancestor with them. The principal findings of the study, therefore, lend support to the notion of an early dispersal from Africa by a more ancient lineage of modern human prior to 50 ka, perhaps as early as OIS 5 times (76-100 ka). PMID- 18164371 TI - Arsenic uptake and speciation in rice plants grown under greenhouse conditions with arsenic contaminated irrigation water. AB - The accumulation of arsenic (As) by rice (Oryza sativa L.) is of great interest considering the dietary intake of rice is potentially a major As exposure pathway in countries where rice is irrigated with As contaminated groundwater. A small scale rice paddy experiment was conducted to evaluate the uptake of As by rice. Arsenic concentrations in rice tissue increased in the order grain<1000 mL, omental extension to spleen >1 cm, parenchymal liver disease >1 cm, porta hepatis involvement >1 cm, diaphragmatic disease >1 cm, carcinomatosis >1 cm, and suprarenal adenopathy >1 cm) were selected for further study. RESULTS: A total of 180 consecutive patients had disease meeting conventional criteria for unresectability at =1 site(s). Optimal cytoreduction (residual disease=1 cm) was achieved in 166 patients (92.2%). Optimal resection rates according to the most common individual unresectable disease criteria were as follows: ascites >1000 mL=91.3% (116/127), carcinomatosis >1 cm=91.0% (81/89), and splenic involvement >1 cm=84.9% (45/53). For patients with ascites >1000 mL alone, optimal cytoreduction was achieved in 95.8% (46/48) of cases. Optimal resection rates according to the total number of unresectable disease sites were as follows: 1 site=95.0% (19/20), 2 sites=93.8% (61/65), 3 sites=81.5% (22/27), 4 sites=93.3% (14/15), and 5 sites=80.0% (4/5). CONCLUSIONS: These data suggest that commonly accepted criteria of surgically unresectable disease for women with advanced ovarian cancer lack the necessary precision to guide clinical management. Pre-operative assessment of resectability should be made by an experienced surgical team prior to deferring the initial attempt at surgical cytoreduction. PMID- 18164381 TI - Risks with dental materials. AB - OBJECTIVES: Dental materials are among the most used substitutes for biological tissue in the human body. The possible health effects of amalgam have been studied extensively and have sometimes been replaced with other less well examined materials. The aim was to study the hazards of other dental materials than amalgam. METHODS: Toxicological information was collected from the Swedish Dental Materials Register 2003 (DentMr). The material safety data sheets (MSDS) included in the DentMR was examined regarding the given composition of the products, the occurrence of CAS-numbers and risk and safety phrases of the substances. Furthermore, complementary information about risk and safety phrases for substances with missing information in the MSDS was collected by using two easily available databases. RESULTS: There were 482 products that included 377 substances of which 219 could be identified. Only 26% of 219 substances had risk and safety phrases. However, via the Swedish Chemicals Inspectorate Classification Register and the Sigma-Aldrich product register, risk and safety phrases could be found for 37 substances. A literature search indicated that some of the listed substances had possible hazards, e.g. substances with embryotoxic and neurotoxic potential. SIGNIFICANCE: The information about hazards with dental materials seems insufficiently described in MSDS and there might be materials with side effects unknown to both patients and dental personnel. There is a need for stronger regulation of dental materials, at least substances that the patients will be exposed to for decades. PMID- 18164382 TI - Effect of blue light on the proliferation of human gingival fibroblasts. AB - OBJECTIVES: Previous studies have reported that blue light, under conditions similar to those used for orthodontic bonding, influences several aspects of cellular physiology. The purpose of this study was to investigate the effect of the exposure to blue light curing sources, i.e. halogen, light emitting diode (LED) and plasma arc irradiation, on the proliferation of human gingival fibroblasts. METHODS: Primary cultures of human gingival fibroblasts were exposed to halogen, LED and plasma arc irradiation for 240, 180 and 120 s, respectively. The effect of blue light on DNA synthesis and cell proliferation was estimated by tritiated thymidine incorporation and direct cell counting, respectively. The possible involvement of an oxidative stress on the effect of blue light irradiation was studied by using N-acetyl-cysteine. Finally the formation of DNA double-strand breaks after irradiation was studied by immunofluorescence with an antibody against histone H2A.x phosphorylated in Ser139. RESULTS: Blue light showed no immediate effect on the regulation of DNA synthesis. However, exposure of cells to these light sources inhibits cell proliferation measured one week after irradiation. This phenomenon is not attributed to the formation of DNA double strand breaks and cannot be annulled by N-acetyl-cysteine. SIGNIFICANCE: The results presented here indicate a mild inhibition of gingival fibroblasts' proliferation after exposure to blue light and necessitate further study to clarify the exact mechanism underlying this effect. PMID- 18164383 TI - Testing phenanthrene distribution properties of virgin plastic pellets and plastic eroded pellets found on Lesvos island beaches (Greece). AB - Plastic pellets have been characterized as toxic pollutant carriers throughout the world oceans and coastal zones. However, their sorptive properties are not yet well understood. In the present study, virgin pellets and plastic eroded pellets (PEP) are used to elucidate their distribution characteristics through distribution kinetic studies. Distribution occurs through diffusion into the pellet for all materials (polyethylene, polyoxymethylene, and PEP) except polypropylene (PP). Although diffusion into the polymer happens with similar rates for both freshwater and saltwater external solutions, apparent diffusion is dependent on the solution salinity because it results in higher equilibrium distribution coefficients. Distribution coefficient into the PEP is higher and diffusion is slower than into the virgin materials. This is attributed to increased crystallinity of the PEP due to weathering. PP demonstrates diffusion rates that are increased by salinity and is apparently faster than into the other polymers suggesting a surface diffusion process. PMID- 18164384 TI - Quantification of granulocyte-macrophage colony-stimulating factor hypersensitivity in juvenile myelomonocytic leukemia by 3H-thymidine assay. AB - Recent molecular studies have revealed that the GM-CSF/RAS signaling pathway plays a central role in the pathogenesis of juvenile myelomonocytic leukemia (JMML). CFU-GM colony assay is an important test for GM-CSF hypersensitivity in patients with JMML, but requires specific skills. We established a simple and easy quantification method to test GM-CSF hypersensitivity, using a (3)H thymidine assay. With this quantification method, JMML patients with RAS mutations showed significantly higher GM-CSF sensitivity than JMML patients with PTPN11 mutations. This method will be useful not only in the diagnosis of JMML, but also to evaluate the difference of GM-CSF sensitivity among patients. PMID- 18164385 TI - Functional mental capacity is not independent of the severity of psychosis. AB - BACKGROUND: Function-specific mental capacities are the legal criteria for competence. These are regarded as superior to clinical assessments of mental state and general function. AIMS: To determine whether tests of fitness to plead and capacity to consent are independent of each other and independent of mental state and global function in psychosis. METHOD: The MacCAT-T and MacCAT-FP, PANSS and GAF were administered to 102 compulsorily detained forensic patients with psychosis. Criteria for incompetence were inability to express a preference concerning treatment, and independent rating as unfit to plead. RESULTS: MacCAT T, MacCAT-FP totals and sub-scales correlated with each other and with PANSS and GAF. Those independently rated unfit to plead or who were incapable of making a treatment choice scored significantly worse on all rating scales. No test had satisfactory sensitivity or specificity. CONCLUSIONS: Legal definitions of mind and of functional capacity offer a basis for structured clinical judgement regarding decision-making capacity. However, function-specific measures of understanding, reasoning and appreciation generate much the same results as measures of mental state and global functioning. PMID- 18164386 TI - Detection of gastrointestinal bleeding by using multislice computed tomography- acute and chronic hemorrhages. AB - Nowadays, computed tomography (CT) is established for diagnosing gastrointestinal bleeding. In this retrospective study, the use of CT in diagnosing gastrointestinal bleeding was evaluated. Fifty-three patients received a contrast medium-enhanced helical multislice CT (MSCT) to locate the bleeding site. Seventy nine percent of the hemorrhage were acute gastrointestinal bleedings. Fifty-five percent of the acute hemorrhages were located via helical MSCT, 45% of the chronic bleeding sites were detected. Notably, bleeding of diverticula, tumors, and angiodysplasias were well demonstrated. In conclusion, contrast-medium enhanced MSCT may be used effectively as a noninvasive diagnostic tool for detecting gastrointestinal bleedings. PMID- 18164387 TI - Rate of liver volume increase: can it predict recipient prognosis after left lobe transplantation from living related donors? AB - PURPOSE: The purpose of this study was to determine whether liver volume increase can predict recipient outcome. MATERIALS AND METHODS: Size-matched left lobe transplantation was performed for 16 patients. Computed tomography was performed in Week 1 after transplantation. Rate of liver volume increase was compared between survivors and deceased subjects. RESULTS: Mean rate of liver volume increase was significantly higher for survivors than for fatalities. CONCLUSION: Rate of liver volume increase might be useful for predicting outcome of living donor liver transplantation. PMID- 18164388 TI - The benefits of comparing conventional sonography, real-time spatial compound sonography, tissue harmonic sonography, and tissue harmonic compound sonography of hepatic lesions. AB - This study aimed to compare conventional sonography, tissue harmonic imaging (THI), spatial compound sonography (SONOCT), and SONOCT+THI for overall image quality, lesion conspicuity, and elimination of artifacts of hepatic lesions. Forty-five patients were randomly selected, and 51 different hepatic lesions were scanned using each of the four techniques. The combined images of SONOCT+THI exhibited the best image quality for solid and cystic lesions, while conventional images were the worst for most hepatic lesions (P<.001). SONOCT was the best for fatty liver. PMID- 18164389 TI - Pathologies of the lower abdomen and pelvis: PET/CT reduces interpretation errors due to urinary contamination. AB - AIM: The aim of this study was to assess the frequency of indeterminate (18)F fluorodeoxyglucose (FDG) accumulations on positron emission tomography (PET) and PET/computed tomography (CT) images in patients with pelvic pathologies. METHODS: We retrospectively evaluated 536 focal FDG accumulations of 166 PET/CT examinations. A consensus reading of PET/CT images, clinical data, and other imaging tests was the standard of reference to assess sensitivities and specificities of PET and PET/CT. Frequencies of indeterminate findings and intraobserver agreement were evaluated. CONCLUSION: PET/CT improves the anatomic delineation and the correct assignment of physiologic and pathologic uptake. PMID- 18164390 TI - Characteristics of typical and atypical meningiomas on ADC maps with respect to schwannomas. AB - The differences in apparent diffusion coefficient (ADC) between typical and atypical meningiomas and schwannomas were investigated, with 41 patients included in the study. There were no significant differences in ADC values or ADC ratios between typical and atypical meningiomas. The discrimination between schwannomas and the typical and atypical meningiomas on ADC maps was reliable, with significant differences in ADC values and ratios and with the narrow range of ADC values in meningiomas. PMID- 18164391 TI - Sonographically occult screen detected breast masses: a retrospective analysis of cases undergoing biopsy. AB - OBJECTIVE: The purpose of this study was to evaluate nonpalpable breast masses identified on mammograms that are sonographically occult. METHODS: The pathology data base at the Woman's Place for Breast Care of the Woman's Hospital of Texas was searched to identify patients who had undergone biopsy for a nonpalpable breast mass identified on mammograms in which a mass was not visible at sonography. RESULTS: There were 32 of 231 such nonpalpable masses that were sonographically occult. Twenty-eight of the masses were histologically benign; four were malignant. CONCLUSION: A small percentage of mammographically visible nonpalpable breast masses are sonographically occult; a majority of these masses are benign. However, biopsy of such masses should be considered and the decision to biopsy based on mammographic features and interval change. PMID- 18164392 TI - Comparison of whole-body 18F-FDG SPECT and posttherapeutic 131I scintigraphy in the detection of metastatic thyroid cancer. AB - PURPOSE: The usefulness of fluorine 18 fluorodeoxyglucose ((18)F-FDG) imaging in differentiated thyroid cancer (DTC) has been demonstrated by investigators. The aim of this study is to compare the ability of fluorodeoxyglucose (FDG)-single photon emission computed tomography (SPECT) to detect metastatic DTC with posttherapeutic iodine131 ((131)I) scintigraphy. METHODS: 239 patients (78 men, 161 women; age range, 23-76 years, mean 45 years); All patients underwent FDG SPECT and (131)I whole-body scan. RESULTS: The sensitivity of (18)F-FDG and (131)I imaging in the patients with high hTg levels is 48.7% (114/234) and 50.4% (118/234) respectively. However, the combined sensitivity of both protocols is 89.7% (210/234). In the patients with (131)I Imaging negative, the sensitivity of (18)F-FDG is 79.3%,but in the patients with (131)I imaging positive, the sensitivity of (18)F-FDG is only 18.6%. CONCLUSION: This study with (131)I and (18)F-FDG in FDG-SPECT further demonstrates the single study of (131)I whole body scan or (18)F-FDG imaging can not provide a high enough sensitivity for the detection of metastatic thyroid cancer. But, the combined protocols can get better results for the staging of thyroid carcinoma with the alternating uptake of (131)I and (18)F-FDG. PMID- 18164393 TI - F-18 FDG PET in detecting uterine leiomyoma. AB - PURPOSE: Uterine leiomyoma, benign tumors of the human uterus, are clinically apparent in about 25% of women and the most common solid pelvic tumors. The purpose of this study was to investigate the F-18 2-fluoro-2-deoxy-D-glucose (FDG) uptake in the uterine leiomyoma and assess the correlation between the intensity of FDG uptake in the uterine leiomyomas and menstrual cycle. METHODS: A total of 589 charts of healthy females examined by whole body FDG positron emission tomography (PET) for health screening examination were reviewed retrospectively. Twenty-two of them were suspected gynacecological tumors and referred to the department of gynacecology to ascertain the nature of the causes. Final diagnosis as uterine leiomyomas were made based on uterine sonography, pelvic computed tomography, or pelvic magnetic resonance imaging scans. We defined FDG uptake as Grade I when FDG uptake was less than liver uptake, Grade II when FDG uptake was equal to liver uptake, and Grade III when FDG uptake was greater than liver uptake. The menstrual cycle was recorded on the day of performing FDG PET in premenopausal women. RESULTS: The FDG uptake in the uterine region is Grade I in three of these 22 females (13.65%), Grade II in 16 (72.7%), and Grade III in 3 (13.65%). CONCLUSION: There is no significant correlation between the intensity of FDG uptake in the uterine leiomyomas and menstrual cycle (P=.914). PMID- 18164394 TI - CT and MR characteristics of hibernoma: six cases. AB - PURPOSE: The purpose of this study was to describe the computed tomography (CT) and magnetic resonance (MR) imaging findings of hibernoma. MATERIALS AND METHODS: We retrospectively evaluated imaging findings of CT and MR examinations of six patients (three men and three woman, aged 27-48 years) with histopathological diagnosis of hibernoma. RESULTS: On CT examination, the lesions were slightly hyperdense, and on T1- and T2-weighted MR images, they were isointense or slightly hypointense compared to the subcutaneous fat. All of these lesions showed contrast enhancement and one out of the six lesions had internal linear septations. CONCLUSION: Hibernoma has a wide spectrum of CT and MR imaging findings, which should be considered in differential diagnosis, especially with other lipomatous lesions. PMID- 18164395 TI - Fetal MRI of lung hypoplasia: imaging findings. AB - Fetal magnetic resonance imaging has become a useful noninvasive modality to examine the human fetus in case of suspicious sonography. Normally, central nervous system structures of the fetus in utero indicate the need for further evaluation. Fast T(2)-weighted sequences are used, which minimize artifacts and allow high contrast between different structures. Cases in which a diaphragmatic hernia was diagnosed via ultrasound will be shown in the magnetic resonance images of the fetuses in this study. PMID- 18164396 TI - Idiopathic isolated laryngotracheobronchial cartilage calcification in a child. AB - Laryngeal, tracheal, and bronchial cartilage calcification is quite rare in children. It was reported to occur in congenital cardiovascular diseases, Keutel syndrome, chondroplasia punctata, warfarin embryopathy, and warfarin sodium therapy. It can occur idiopathically as well. Laryngotracheobronchial cartilage calcification is demonstrated in this report by chest radiography and multiplanar three-dimensional CT examination in a 2.5-year-old boy with nonspecific cough. PMID- 18164397 TI - Nodular thymic lymphoid follicular hyperplasia mimicking thymoma. AB - In this report, we describe a case of thymic lymphoid follicular hyperplasia that was incidentally found as a small thymic nodule in the health screening program including a low-dose chest CT. The CT and MRI findings of the nodule were similar to those of thymoma, and it was difficult to differentiate the lesion from thymoma. PMID- 18164398 TI - Primary malignant melanoma of the anterior mediastinum: CT and MR findings. AB - Malignant melanoma presenting in the mediastinum without an extrathoracic primary is extremely rare. To our knowledge, its imaging findings have not been described in the radiologic literature. We report a case of primary malignant melanoma of the anterior mediastinum and discuss its imaging findings on computed tomography and magnetic resonance imaging. PMID- 18164399 TI - Pulmonary varix mimicking arteriovenous malformation. AB - We report here a case of a 44-year-old female with a pulmonary varix. The patient was asymptomatic; a routine chest X-ray incidentally revealed a serpiginous mass in the left hilar region. Computed tomography revealed a serpiginous, dilated vascular structure in the left upper lobe that continued into the lower lobe and was suggestive of an arteriovenous malformation. Pulmonary angiography revealed no arterial abnormality; however, the filling of an abnormally dilated pulmonary vein was observed. Angiography is considered to be the most useful method for the confirmatory diagnosis of pulmonary varices. PMID- 18164400 TI - Malrotation with volvulus of the transverse colon and duodenal obstruction secondary to Ladd's bands. AB - Colonic volvulus is a relatively uncommon cause of large bowel obstruction, accounting for 10% of colonic obstructions. Volvulus of the transverse colon is quite rare, accounting for only 4-11% of all reported cases of colonic volvulus (Kerry RL, Ransom HK. Volvulus of the colon: etiology, diagnosis, and treatment. Arch Surg 1969;99:215-221). We report an unusual case of documented volvulus of the transverse colon in a young man with intestinal malrotation and concomitant duodenal obstruction by Ladd's bands. PMID- 18164401 TI - Conventional intramedullary chondrosarcoma with subarticular involvement. AB - A 78-year-old female patient with a lesion in the distal tibia is shown to have conventional intramedullary chondrosarcoma. Typical considerations for this end of-bone neoplastic lesion are giant cell tumor, clear cell chondrosarcoma, and chondroblastoma. Salient clinical and radiographic features and behavior of this lesion broaden the typical view of conventional intramedullary chondrosarcoma. PMID- 18164402 TI - Intraosseous ganglion of the phalanx. AB - Intraosseous ganglions occurring in the hand are rare. Most reported cases involve the lunate or the scaphoid. This report presents a very extremely rare case of intraosseous ganglion arising in the middle phalanx of an index finger. Multi-modality imaging features using plane radiograph, CT, and MRI of this ganglion are presented for the first time. PMID- 18164403 TI - Speciation and surface structure of inorganic arsenic in solid phases: a review. AB - Accurate determination of individual arsenic species is critical because the toxicology, mobility, and adsorptivity of arsenic vary substantially with its chemical forms and oxidation states. Separation techniques together with techniques for chemical identification make it possible to determine the combinational forms and oxidation states of arsenic in solid phases. Selective sequential extraction is often employed to determine operationally defined fractions, but it has a poor precision and selectivity. Direct methods, based on X-ray techniques and vibrational spectroscopy, have been developed to analyze the valence, local coordination, protonation, and other properties of arsenic in solid phases. Extensive research studies in the literature have been performed to elucidate the interfacial reactions between inorganic arsenic and solid surfaces of sulfides, and Fe, Al, and Mn (hydro)oxides. Outer-sphere and inner-sphere complex (monodentate mononuclear, bidentate mononuclear, and bidentate binuclear complex) models have been proposed to interpret the sorption mechanisms. The nature of the surface complexes has been inspected by spectroscopic methods but remains controversial. This paper focuses on the recent advancement in arsenic speciation in solid phases and covers relevant methodological, analytical and modeling aspects. The identification of arsenic species in natural materials, however, is complicated by the presence of multiple species, and the applications of instrumental methods are usually limited due to their comparatively high detection limits. Development of advanced in-situ methods with high sensitivity, therefore, is required. PMID- 18164404 TI - Outcomes of same-sizing versus oversizing donor trephines in keratoconic patients undergoing first penetrating keratoplasty. AB - PURPOSE: To investigate whether use of same-size donor and recipient trephines reduced myopia after penetrating keratoplasty for keratoconus. DESIGN: Retrospective cohort study. PARTICIPANTS: Eight hundred seventy-eight first grafts for keratoconus were reported to UK Transplant between April 1999 and December 2003. There were 234 and 644 grafts in the same-size and oversize donor trephine groups, respectively. At 1 year, mean spherical equivalent (SE) data were available for 116 eyes (50%) and 295 eyes (46%) in the same-size and oversize groups. At 2 years, mean SE data were available for 64 eyes (27%) and 148 eyes (23%) in the same-size and oversize groups. METHODS: Logistic regression and univariate analysis of follow-up data submitted to UK Transplant. MAIN OUTCOME MEASURES: At 1 and 2 years postoperatively, mean SE, magnitude of the cylindrical component of the refraction, postoperative uncorrected visual acuity (VA), postoperative best-corrected VA, and postoperative complications were recorded. RESULTS: The mean SEs for the same-size and oversize donor trephine groups, respectively, were -1.45 diopters (D) and -1.41 D at 1 year (P = 0.6) and -1.74 D and -2.19 D at 2 years postoperatively (P = 0.3). Although there were no differences in graft survival between the groups at 1 and 2 years, there was a higher incidence of postoperative wound leaks in the same-size group (P = 0.03). CONCLUSION: Use of same-size donor and recipient trephines did not reduce myopia and was associated with an increased risk of postoperative complications. PMID- 18164405 TI - Bilirubin injury to neurons: contribution of oxidative stress and rescue by glycoursodeoxycholic acid. AB - It is well established that high levels of unconjugated bilirubin (UCB) can be toxic to the central nervous system, and oxidative stress is emerging as a relevant event in the mechanisms of UCB encephalopathy. In contrast, the hydrophilic bile acid, ursodeoxycholic acid (UDCA), has been reported as a cytoprotective and antioxidant molecule. In this study, we investigated if exposure of rat neurons in primary culture to clinically relevant concentrations of UCB leads to oxidative injury. The contribution of oxidative stress in UCB neurotoxicity was further investigated by examining whether the reduction of NO production by NAME, an inhibitor of nitric oxide synthase, prevents the disruption of the redox status and neuronal damage. Moreover, we evaluated the ability of glycoursodeoxycholic acid (GUDCA), the most relevant conjugated derivative in the serum of patients treated with UDCA, to abrogate the UCB induced oxidative damage. Cultured rat neurons were incubated with 50 or 100microM UCB in the presence of 100microM human serum albumin, alone or in combination with 100microM NAME or with 50microM GUDCA, for 4h at 37 degrees C. Protein carbonyls, 4-hydroxy-2-nonenal-protein adducts, intracellular glutathione content and cell death were determined. The results obtained showed that UCB induces protein oxidation and lipid peroxidation, while diminishes the thiol antioxidant defences, events that were correlated with the extent of cell death. Moreover, these events were counteracted by NAME and abrogated in the presence of GUDCA. Collectively, this study shows that oxidative stress is one of the pathways associated with neuronal viability impairment by UCB, and that GUDCA significantly prevents such effects from occurring. These findings corroborate the antioxidant properties of the bile acid and point to a new therapeutic approach for UCB-induced neurotoxicity due to oxidative stress. PMID- 18164406 TI - Tuning the Au(I)-mediated inhibition of cathepsin B through ligand substitutions. AB - It has been over 80 years since the antiarthritic properties of gold(I) complexes were first recognized. However, a detailed understanding of their mechanism of action has been slow to develop. One likely biological target of gold(I) is the cathepsin family of lysosomal cysteine proteases, enzymes involved in the inflammation and joint destruction that are hallmarks of rheumatoid arthritis (RA). We have previously shown that analogs of auranofin, a clinically available antiarthritic drug, inhibit cathepsin B. In this study, the extent to which the steric and electronic properties of the phosphine ligand can be modified to obtain enhanced potency against cathepsin B is investigated. PMID- 18164407 TI - Mixed serous carcinoma of the endometrium with trophoblastic differentiation: analysis of the p53 tumor suppressor gene suggests stem cell origin. AB - The pathogenesis of mixed endometrial adenocarcinoma with trophoblastic differentiation is quite unclear at times. The present study examines a serous carcinoma with choriocarcinomatous differentiation. p53 staining was seen in the serous component and the cytotrophoblastic cells of the choriocarcinomatous component, but not in the syncytiotrophoblastic cells. p53 mutational analysis showed a heterozygotic mutation at exon 8 for the choriocarcinomatous component and a homozygote deletion at exon 7 for the serous component. These alterations suggest that the multidirectional tumor differentiation might occur from a common stem cell in these malignancies. PMID- 18164408 TI - Basaloid carcinoma of the breast: a review of 9 cases, with delineation of a possible clinicopathologic entity. AB - Basaloid carcinoma of the breast (BCB) is an unusual neoplasm composed of basal type neoplastic cells similar to those found in adenoid cystic carcinoma (ACC), although lacking distinctive features such as a cribriform pattern, a dual neoplastic population (epithelial-myoepithelial/basaloid), and stromal deposits of basement membrane-like material. In this article, we present 9 cases of breast cancer showing overall/predominant basaloid morphology. Patients' ages ranged from 47 to 75 years (mean, 61.4 years). Surgical treatment included mastectomy or quadrant excision with or without axillary dissection. Most tumors had a circumscribed outline and ranged in size from 1.3 to 5.5 cm (mean, 2.5 cm). Microscopically, they featured sheets, nests, and cords of proliferating basaloid tumor cells with ovoid, hyperchromatic nuclei with inconspicuous nucleoli and scant cytoplasm. No foci with characteristics of ACC were found in any of the tumors. Transition into pleomorphic basaloid carcinoma with foci of high-grade ductal carcinoma in situ plus infiltrating ductal carcinoma (IDC) and admixture with grade 3 ductal and sarcomatoid carcinoma was seen in 2 cases. Tumor cells were positive for wide-spectrum keratins and epithelial membrane antigen (9/9) and high-molecular-weight keratins (7/9). They were negative for smooth muscle actin, p63, calponin, and CD10 in all tested cases. Estrogen receptor, progesterone receptor, and HER-2 were negative. Axillary lymph node metastases were seen in 3 cases. At follow-up (range, 10-169 months), 5 patients were alive, 1 with evidence of contralateral breast cancer. Three patients died: one of disseminated BCB metastases, another of liver cirrhosis, and one of disseminated estrogen receptor/progesterone receptor-positive contralateral IDC. One patient was lost to follow-up. We concluded that BCB has some phenotypic and immunohistochemical features enabling its distinction from ACC or IDC. It appears to be a morphological and possibly a clinical entity. Compared with ACC, BCB appears to be more aggressive and may entail a more guarded prognosis. PMID- 18164409 TI - Ovarian thecoma: clinicopathological analysis of 50 cases. AB - The clinicopathological findings of 50 thecoma cases were studied to establish the most useful diagnostic criteria helpful in characterizing this ovarian stromal tumor. Patient age ranged from 21 to 77 years (median, 57.5 years). In this study, thecoma appears to be associated with endometrial diseases (15 patients) as an incidental finding in a gynecologic routine examination (14 patients) and in a cesarean delivery (1 patient). Arteries, veins, lymphatics, and mast cells are normally present in the ovarian medulla and are absent in the cortical area. The 50 thecomas studied showed proliferation of stromal cells and presence of arteries, lymphatics, and mast cells. Thecoma tumoral growth appeared to cause atrophy or compression of the cortical area. These findings are significant for diagnosis; thus, thecoma is proposed as a tumor originating in the ovarian medulla. Fibroma and thecoma seem to be different neoplasms and should be considered distinct, separate entities because they have different origin, morphology, and potential functionality. PMID- 18164410 TI - Significance of disease extent in high-grade cervical intraepithelial neoplasia excised with negative margins by loop electrosurgical excision procedure. AB - Approximately 15% of patients with high-grade cervical intraepithelial neoplasia (CIN 2-3), treated by the loop electrosurgical excision procedure (LEEP) will experience a persistence or recurrence of their disease. The full spectrum of pathologic factors that may be predictive of an increased probability of this event remains to be elucidated. In this work, we evaluate the significance of disease extent in a group of patients with CIN 2 to 3 that was completely excised by LEEP (ie, endocervical and ectocervical margins were negative). From 102 patients whose LEEP specimens had negative margins during a 5-year period, at least 1 cytologic follow-up was available in 87 patients (median follow-up, 23 months). For patients with multiple follow-up cytologic samples, the most severe cytologic abnormality was used for the purpose of this analysis (1 sample per patient). Extent of disease was determined semiquantitatively by calculating for every case the proportion of processed slides on which any CIN 2 to 3 could be identified. The 87 cases were classified into 3 groups (<33%, 33%-66%, >66%) for ease of statistical comparison (Fisher exact test). There were 10 recurrences of high-grade squamous intraepithelial lesion (HGSIL) among the 87 patients with follow-up. There were no statistically significant differences between the aforementioned 3 groups regarding the follow-up diagnostic frequencies of HGSIL (10% in the <33% group, 40% in the 33%-66% group, and 50% in the >66% group) and "negative for intraepithelial lesion or malignancy" (32.25% in the <33% group, 25.8% in the 33%-66% group, and 35.48% in the >66% group). If confirmed, our findings suggest that in an LEEP specimen with CIN 2 to 3 that was excised with negative margins, the extent of disease lacks prognostic significance and need not be reported. PMID- 18164411 TI - Fibroosseous [corrected] pseudotumor of the digit: a clinicopathologic study of 43 new cases. AB - Myositis ossificans (MO) is a reactive zonal fibroosseous lesion in skeletal muscle of the proximal extremities and trunk of young patients. It generally matures over several weeks to form a peripheral rim of bone. Fibroosseous pseudotumor of the digits (FOPD) is a similar reactive lesion of the digits, which is reportedly less well organized. Cases up to year 1980 were previously reported from our institution. We examined new cases of FOPD since 1980 and addressed the relationship of this lesion to both myositis ossificans and other distal extremity fibroosseous lesions. Fifty-two cases coded as FOPD or MO limited to the hands and feet, from 1980 to the present, were retrieved from our files. Nine cases were excluded due to incomplete material or rediagnosis as other lesion. Materials reviewed included radiologic images and pathologic material. Forty-three cases of FOPD were included. All cases were in the fingers, except for 1 toe case, with the proximal phalanx of the index finger the most commonly affected site. Duration of lesions was radiologically estimated to range from 2 to 6 weeks. Age of patients ranged from 10 to 64 years (mean and median of 40 years). Sixty percent of cases occurred in women. Morphologically, half of these lesions had a zonal organization, with mature woven bone peripherally and immature woven bone centrally; all bone demonstrated osteoblastic rimming. Clinically, 6 patients had known antecedent trauma, yet edema and pain of the digit were noted in all. Novel findings were that patients often had occupations requiring repetitive manual labor. Cases of FOPD were diagnosed and treated by simple excision. Three patients had residual disease, subsequently resected, without any recurrences. Fibroosseous pseudotumor of the digits is a distinctive, benign, reactive myofibroblastic and osseous zonal lesion that occurs in the finger of mainly adult patients, many with occupations that require repetitive manual use. Imaging can date these lesions, determine their exact location, and help with diagnosis. These lesions are the superficial and distal counterparts of MO. It is important to understand the morphology of FOPD to distinguish this type of lesion from other benign and malignant fibroosseous lesions. PMID- 18164412 TI - Ectopic Leydig cells of testis An immunohistochemical study on tissue microarray. AB - The incidence of ectopic Leydig cells (LCs) has been reported to be 40% to 90%. Several theories have been proposed to account for ectopic LCs, including in situ differentiation, migration from testicular interstitium, and trapping of peritubular LCs in the tunica propria of the seminiferous tubule during its thickening. To document the nature of ectopic LCs and to compare them with interstitial LCs, Sertoli cells, epididymal cells, mesothelial cells, and epithelial cells in rete testis, immunostainings with calretinin, CD10, inhibin, CK7, and CK20 were performed in 40 cases of orchiectomy specimens using tissue microarray sections. In addition, the frequency of ectopic LCs was evaluated. Of the 40 orchiectomy specimens, 14 cases demonstrated ectopic LCs with an incidence of 35%. Inhibin was positive in more than half of ectopic LCs (9/14, 64.3%) and in almost all interstitial LCs. Calretinin was positive in most of ectopic LCs (12/14, 85.7%) as well as in most of normally located interstitial LCs and mesothelial cells and some of rete testis epithelial cells. The ectopic and interstitial LCs as well as Sertoli cells were negative for CK7, CK20, and CD10. CK7 was positive in all epididymal cells and in most mesothelial cells and epithelial cells in rete testis. CD10 was positive in some of epididymal cells and epithelial cells in rete testis. CK20 was negative in all cells in the testis and epididymis. Ectopic LCs showed similar staining patterns to interstitial LCs with positive immunoreactivity for calretinin and inhibin. In this study, the frequency of ectopic LCs was 35%. The lower incidence in this study was most likely because of the limited sampling. Immunohistochemically, ectopic LCs showed identical immunohistochemical patterns with those of interstitial LCs. Calretinin appeared to be more sensitive but less specific than inhibin for LCs. Because calretinin is frequently positive in cells other than ectopic or interstitial LCs, a precaution is required to differentiate LCs from mesothelial cells and rete testis epithelial cells. Based on our study, we could not add anything else of what is known in regard to the histogenesis of ectopic LCs. The current theories including in situ differentiation, migration from testicular interstitium, and trapping of peritubular LCs in the tunica propria of the seminiferous tubules during its thickening seem to be valid histogenetic theories. PMID- 18164413 TI - AlphaB-crystallin: a novel marker of invasive basal-like and metaplastic breast carcinomas. AB - Basal-like tumors are a newly recognized estrogen receptor (ER) negative and HER2 negative breast cancer subtype that express basal epithelial genes and are associated with poor survival. Metaplastic carcinomas are thought to belong within the basal-like group. We have recently demonstrated that the small heat shock protein alphaB-crystallin is commonly expressed in basal-like tumors and contributes to their aggressive phenotype. The current study examined the rates and patterns of alphaB-crystallin expression in whole tissue sections of human breast, including normal tissue, proliferative lesions, in situ and invasive carcinomas (ER positive, HER2 positive, basal-like, and metaplastic cancers). In normal breast tissue, proliferative lesions and in situ carcinomas, alphaB crystallin expression was restricted to the myoepithelial cell compartment of ductal and lobular units. Most basal-like and metaplastic carcinomas demonstrated cytoplasmic expression of alphaB-crystallin (81% and 86%, respectively). Conversely, no staining for alphaB-crystallin was observed in nonbasal-like (ie, ER positive or HER2 positive) breast carcinomas. Taken together, our results indicate that alphaB-crystallin is a sensitive (81%) and specific (100%) marker for basal-like breast carcinomas. Moreover, the high rates of expression of alphaB-crystallin in metaplastic breast carcinomas (86%) suggest that these tumors may represent a histologically distinctive subset of basal-like breast tumors with a similar underlying molecular etiology. PMID- 18164414 TI - Extramedullary plasmacytoma of the tonsil diagnosed by fine-needle aspiration cytology. AB - A case of tonsillar extramedullary plasmacytoma in a 53-year-old man with a complaint of lump sensation in the throat is presented. Examination of the oral cavity showed enlargement of the left tonsil. Magnetic resonance imaging demonstrated a solid mass, measuring 3.2 x 2.0 x 3.8 cm, in the left tonsil. Cytologic smear obtained by fine-needle aspiration biopsy appeared highly cellular and was composed of clusters of plasma cells with varying maturity. Atypical plasma cells had prominent eccentric nuclei with nucleoli and finely granular cytoplasm. Binucleated cells and mitotic figures were also identified. The cytoplasm of mature-looking small plasma cells was also finely granular without a perinuclear halo. A cytologic diagnosis of plasmacytoma was made. Excisional biopsy showed sheets of plasmacytoid cells with abundant eosinophilic granular cytoplasm. Occasional binucleated and pleomorphic cells with giant nuclei and prominent nucleoli were observed. These plasmacytoid cells were diffusely immunoreactive for lambda light chain and IgG, partially positive for epithelial membrane antigen. Metastatic examination finding was negative for multiple myeloma, and the patient was diagnosed as having extramedullary plasmacytoma. Although the diagnosis of plasmacytoma on cytologic smear may be difficult, in the current case, fine-needle aspiration cytology provided a rapid and accurate diagnosis. PMID- 18164415 TI - Inflammatory pseudotumor of the peroneal nerve: case report and literature review. AB - Inflammatory pseudotumors (IPTs) are a group of lesions of obscure etiology. Although they are presumably reactive, their exact pathogenesis is unknown. Inflammatory pseudotumors are identified in different organs; however, IPTs of the peripheral nerves have been rarely reported in the English literature. We report a 44-year-old woman who presented with left foot drop. Examination revealed a thickening in the posterolateral aspect of the popliteal fossa and features consistent with peroneal nerve palsy. Magnetic resonance imaging revealed a mass that may represent a Baker cyst or perineural sheath mass that was excised. After pathological examination, the case was diagnosed as IPT of peroneal nerve, and the patient received no further therapy. Inflammatory pseudotumors of the peripheral nerves, although rare, should always be of consideration in the differential diagnosis of peripheral nerve nodular mass lesions. They can mimic neoplastic nerve sheath lesions, clinically and radiologically. However, they are amenable to surgical cure with nerve-sparing surgery. Thus, despite their rarity, on dealing with neural S100 negative lesion, their inclusion is mandatory to avoid an unnecessary and a rather aggressive surgery. PMID- 18164416 TI - Application of tissue microarrays in placental research. AB - A complementary tool to gene-based arrays are tissue microarrays, which allow the validation of DNA/RNA-derived profiling results on the protein level and offer the possibility of exact location within the tissue structures. Here we describe the technical details of the application of tissue microarray in placental research. Although the placenta represents a very inhomogeneous organ, this technique is also usable in placental research. PMID- 18164417 TI - An unusual clinical presentation of a rare tumor of the thyroid gland: report on one case of leiomyosarcoma and review of literature. AB - Primary muscle tumors of the thyroid gland are exceedingly rare. We report on the case of a patient with primary leiomyosarcoma of the thyroid gland and review the literature. An 83-year-old woman complaining of neuropathic pain in her left arm and enlargement of her anterior neck underwent multiple surgical biopsies of the thyroid gland. The tumor was composed of interlacing fascicles of spindle-shaped cells that expressed smooth muscle actin and vimentin but were negative for cytokeratins and thyroglobulin. Ultrastructurally, bundles of myofilaments were present. Magnetic resonance imaging showed a thyroid tumor that directly extended to the adjacent vertebra with an associated pachymeningitis. The patient died 2 months after surgery. The diagnosis of primary leiomyosarcoma of the thyroid gland is difficult and requires numerous clinical, radiologic, and pathologic data. To our knowledge, this case is the first one with such a locoregional extension. PMID- 18164418 TI - Metachronus malignant rhabdoid tumor of the ileum and adenocarcinoma of lung: a unique case report. AB - Malignant extrarenal rhabdoid tumors had been described in a variety of anatomic locations including gastrointestinal tract. Carcinomas of small intestine are quite uncommon neoplasms, and those with rhabdoid differentiation are exceedingly rare. These poorly differentiated tumors pose a great deal of difficulty in diagnosis as well as in deciding whether they are primary or metastatic in origin because malignant rhabdoid tumors or carcinomas with rhabdoid differentiation of other sites can metastasize to the small intestine. They behave more aggressively than the typical adenocarcinomas of the same location. Herein, we report a 52 year-old patient with primary small bowel malignant rhabdoid tumor, who was completely disease-free after the initial presentation and management. Five years later, he was found to have a lung mass proved to be adenocarcinoma, exhibiting focal giant cell differentiation without rhabdoid features. PMID- 18164419 TI - Cloacal dysgenesis sequence. AB - Cloacal dysgenesis sequence is a rare congenital malformation seen in 1:50000 to 250000 neonates. It shows a phallus-like structure, smooth perineum, and absence of urethral, vaginal, and anal openings as primary malformations. We report a case of a 4-day-old preterm female neonate. The prenatal ultrasound showed singleton fetus in breech presentation with echogenic bowel, dilated colon, perineal mass, dilated bladder, and fetal hydrops. The complete autopsy revealed smooth perineum without patent urethral, vaginal, and anal openings, and enlarged wrinkled clitoral-like structure. Other anomalies included renal hydronephrosis, hydroureter, megacolon, and urethral agenesis. The chromosomal analysis showed normal female karyotype. These findings are consistent with cloacal dysgenesis sequence, and it results from the defect in cloaca formation during the first 50 days of gestation. It is usually fatal and should be differentiated from other conditions having better prognosis such as fetal obstructive uropathies and persistent cloaca. PMID- 18164420 TI - Breast metastasis by medullary thyroid carcinoma detected by FDG positron emission tomography. AB - Medullary thyroid carcinoma (MTC) is an uncommon thyroid cancer comprising 5% to 8% of thyroid neoplasms. In contrast to common thyroid tumors, this tumor originates from the calcitonin-producing C cells. Regional metastases to cervical lymph nodes occur early in the disease, whereas distant metastasis occurs late. Common metastatic sites include the liver, bone, brain, and adrenal medulla. We present a case of MTC metastatic to the breast. We report on this case for the following reasons: (1) metastasis to the breast is an extremely rare occurrence and could be easily confused clinically and pathologically with a primary breast neoplasm and (2) this is the first reported case of detection of breast metastasis by an MTC using FDG ((18)F-fluoro-2-deoxy-D-glucose) positron emission tomography with an accompanying histologic description. PMID- 18164421 TI - Immunohistochemical approaches to the diagnosis of undifferentiated malignant tumors. AB - Undifferentiated malignant neoplasms are a daunting diagnostic problem for anatomical pathologists, calling for a tour de force in morphological skill, clinicopathologic correlation, and application of adjunctive laboratory studies. The most useful approach to these lesions begins with generic classification into 1 of 4 histologic categories: small round cell; spindle cell; large polygonal cell (epithelioid); and pleomorphic neoplasms. Once that step has been accomplished, one can systemically apply corresponding groups of antibody reagents in immunohistologic studies and interpret the results in an algorithmic fashion. This review presents the tumor markers that are the most useful in this contextual approach, as well as the specific algorithmic structures that can be applied to the 4 specified tumor groups. Other selected problems in the diagnosis of morphologically ambiguous tumors are considered as well. PMID- 18164422 TI - Understanding neuronal molecular networks builds on neuronal cellular network architecture. AB - The central nervous system (CNS) is a nested network at all levels of its organization. In particular, neuronal cellular networks (the neuronal circuits), interconnected to form neuronal systems, are formed by neurons, which operate thanks to their molecular networks. Proteins are the main components of the molecular networks and via protein-protein interactions can be assembled in multimeric complexes, which can work as micro-devices. On this basis, we have introduced the term "fractal logic" to describe networks of networks where at the various levels of the nested organization the same rules (logic) to perform operations are used. If this assumption is true, the description of the information handling at one of the nested levels sheds light on the way in which similar operations are carried out at other levels. This conceptual frame has been used to deduce from some features of neuronal networks the features of the molecular networks as far as modes for inter-node communication and their architecture. It should be noted that these features are such to allow a highly regulated cross-talk between signalling pathways, hence preserving selectivity and privacy. To investigate these aspects, the protein-protein interactions in beta2 Adrenergic Receptor (beta2AR) and Epidermal growth factor receptor (EGFR) signalling pathways have been analysed. The presence of disordered sequences in interacting domains can favour via the "fly-casting mechanism" protein-protein interactions, in addition it favours an induced-fitting rather than a lock-key type of interactions. Thus, by means of a computer assisted analysis the presence of disorder sequences in the main streams of the molecular networks that have beta2AR and EGFR as input proteins leading to MAP kinase activation has been evaluated. PMID- 18164423 TI - CNS inflammation and neuronal degeneration is aggravated by impaired CD200-CD200R mediated macrophage silencing. AB - Multiple sclerosis is a chronic disabling CNS disorder, characterized by autoimmune inflammatory demyelination and neurodegeneration. CD200, broadly expressed on neurons and endothelial cells, mediates inhibitory signals through its receptor, CD200R, on cells of myeloid origin. Antibody-mediated blockade of CD200R leads to an aggravated clinical course of rodent experimental autoimmune encephalomyelitis in vivo, accompanied by profoundly augmented cellular infiltrates consisting of T cells and activated iNOS(+) macrophages in inflammatory spinal cord lesions. In vitro blockade of CD200R on macrophages leads to enhanced IFN-gamma-induced release of IL6 and neuronal cell death in co cultures with hippocampal neurons expressing CD200. CD200 and its receptor could also be detected on neurons and macrophages in human MS plaques. Therefore the CD200-CD200R pathway seems of critical relevance for macrophage-mediated damage in autoimmune inflammation of the CNS. PMID- 18164424 TI - Production and functions of IL-17 in microglia. AB - Interleukin (IL)-17-producing helper T cells may play a pivotal role in the pathogenesis of multiple sclerosis. Here, we examined the effects of IL-17 on microglia, which are known to be critically involved in multiple sclerosis. Treatment with IL-17 upregulated the microglial production of IL-6, macrophage inflammatory protein-2, nitric oxide, adhesion molecules, and neurotrophic factors. We also found that IL-17 was produced by microglia in response to IL-23 or IL-1beta. Because microglia produce IL-1beta and IL-23, these cytokines may act in an autocrine manner to induce IL-17 expression in microglia, and thereby contribute to autoimmune diseases, such as MS, in the central nervous system. PMID- 18164425 TI - Dynamic behavior of hepatitis C virus quasispecies in a long-term culture of the three-dimensional radial-flow bioreactor system. AB - Hepatitis C virus (HCV) exists in infected individuals as quasispecies, usually consisting of a dominant viral isolate and a variable mixture of related, yet genetically distinct, variants. A prior HCV infection system was developed using human hepatocellular carcinoma cells cultured in the three-dimensional radial flow bioreactor (RFB), in which the cells retain morphological appearance and their differentiated hepatocyte functions for an extended period of time. This report studies the selection and alteration of the viral quasispecies in the RFB system inoculated with pooled serum derived from HCV carriers. Monitoring the viral RNA and core protein in the culture supernatants, together with nucleotide sequencing of hypervariable region 1 of the HCV genome, demonstrated that (1) the virus production intermittently fluctuated in the cultures, (2) the viral genetic diversity was markedly reduced 3 days post-infection (p.i.), and (3) dominant species changed on days 19-33p.i., suggesting that the virus populations can be selected according to susceptibility to the viral infection and replication. A therapeutic effect of interferon-alpha also demonstrated the inhibition of HCV expression. Thus, this HCV infection model in the RFB system should be useful for investigating the dynamic behavior of HCV quasispecies in cultured cells and evaluating anti-HCV compounds. PMID- 18164426 TI - Postpartum hemorrhage prevention: a case study in northern rural Honduras. AB - Postpartum hemorrhage (PPH) is the leading cause of maternal mortality globally. Safe Motherhood policies have been directed towards the reduction of PPH by recommending active management of third-stage labor as the standard of care. One component of active management involves routine use of a uterotonic agent within 1 minute of the delivery of the baby. A case study at Clinica Materno-Infantil, a free-standing public birth center in Honduras, is presented, focusing on methods to reduce PPH. The nursing staff was trained to estimate blood loss and in methods to manage PPH, including elements of active management of the third stage of labor. Medical records were reviewed and an analysis of PPH management compared to estimated blood loss (EBL) was conducted. There was no significant correlation between PPH management techniques and EBL (r = .060; P = .368). There was a statistically significant (P < .001) correlation between oxytocin administration and lower EBL (r = -.232), indicating that there was less blood loss when oxytocin was administered. At Clinica Materno-Infantil, routine use of a uterotonic agent appears beneficial and further implementation of active management of the third stage of labor appears warranted. PMID- 18164427 TI - The evolution of a journal. PMID- 18164428 TI - Injury: a major cause of pregnancy-associated morbidity in Massachusetts. AB - Hospital visits (inpatient, observation, and emergency department) for injury occurring during pregnancy and 1 year postpartum (the pregnancy-associated period) were examined to determine groups at risk for injuries. The dataset included maternally linked vital records and hospital visit data for a population based cohort of women residing in Massachusetts who delivered between 2002 and 2003 (n = 100,051). Injury morbidity (injury visits with International Classification of Diseases, Ninth Revision, Clinical Modification codes 800 999.99 or selected E-codes) was evaluated by individual woman- and visit-based analyses. Overall, one in seven women sought hospital care for pregnancy associated injuries, and rates were as high as one in four for some subgroups. Most pregnancy-associated injury visits (91%) occurred in emergency departments. More than 4% of women had a motor vehicle collision, which was the leading cause of injury. The risk for injury was significantly higher among women who were adolescents (relative risk [RR] = 1.88; 95% confidence interval [CI], 1.78-1.98), black non-Hispanic (RR = 1.88; 95% CI, 1.80-1.97), those who had public insurance (RR = 2.50; 95% CI, 2.41-2.56), or those who had less than a high school education (RR = 2.48; 95% CI, 2.39-2.58) when compared with referent groups. Clinical guidelines for preconception and pregnancy-associated periods should include recommendations for injury history assessment and preventative counseling for women. PMID- 18164429 TI - Cervical cancer screening practices of certified nurse-midwives in the United States. AB - The purpose of this study was to determine how closely certified nurse-midwives in the United States follow contemporary cervical cancer screening guidelines. A survey was sent to 264 randomly selected certified nurse-midwives. Survey questions included demographics and clinical scenarios addressing initiation, frequency, and cessation of screening. Responses were received from 60% of the sampled certified nurse-midwives who had valid mailing addresses; 127 were eligible for the analytic sample. Many nurse-midwives initiate cervical cancer screening earlier than guidelines recommend; 72% would initiate screening in an 18-year-old within 1 month of coitarche, while 36% would begin screening virginal girls at age 18, and many continue cervical cancer screening after guidelines recommend cessation. More than 60% of the respondents would continue screening a woman who had undergone total hysterectomy for symptomatic fibroids who had no history of dysplasia, and half would continue to screen a 70-year-old woman with a 30-year history of previous normal Pap tests. In addition, despite guidelines which recommend less frequent screening, more than one-quarter (28%) would continue annual screening in a 35-year-old woman with three or more normal tests. Certified nurse-midwives are performing cervical cancer screening more frequently than current guidelines recommend. Comparisons to the practice of other providers are offered. Education to limit unnecessary testing is needed. PMID- 18164430 TI - Malpractice liability burden in midwifery: a survey of Michigan certified nurse midwives. AB - A statewide survey was conducted among 282 nurse-midwives in Michigan to examine the extent of their current medical liability burden. Two hundred ten responses were received for an adjusted response rate of 76.9%. Data from 145 certified nurse-midwives (CNMs) who were currently engaged in clinical practice in Michigan were used for this analysis. Sixty-nine percent of CNMs reported that liability concerns had a negative impact on their clinical decision making. Most CNMs (88.1%) acquired malpractice insurance coverage through an employer, whereas 4.9% were practicing "bare" due to difficulty in obtaining coverage. Thirty-five percent of the respondents had been named in a malpractice claim at least once in their career, and 15.5% had at least one malpractice payment of $30,000 or more made on their behalf. CNMs who purchased malpractice insurance coverage themselves or were going bare were significantly less likely to include obstetrics in their practice than their counterparts covered through an employer (70.6% versus 87.2%; P = .04). These findings among Michigan CNMs call for further investigation into the consequences of the current malpractice situation surrounding nurse-midwifery practice and its influence on obstetric care, particularly among women from disadvantaged populations. PMID- 18164431 TI - It's not all about birth: biomechanics applied to pelvic organ prolapse prevention. AB - Pelvic organ prolapse is a common and costly women's health problem. Prevention of prolapse continues to play a role in the debate about the risks and benefits of elective cesarean section, making this an important topic for midwives to understand. While some women appear to be at higher risk for prolapse because of intrinsic anatomic and physiologic factors, others have modifiable risk factors that can be addressed by health care providers. This article discusses the current knowledge related to the etiology of prolapse and related components of pelvic anatomy. Biomechanical principles are then applied to enhance the understanding of prolapse development and prevention. Clinical recommendations are based on current evidence regarding topics such as physical activity during pregnancy, treatment of vaginal atrophy, and optimal pelvic floor muscle exercises. PMID- 18164432 TI - Nutritional counseling for vegetarians during pregnancy and lactation. AB - A woman's nutritional status directly affects pregnancy outcome and the quality of breast milk after birth. Clinicians who provide prenatal care have an important role in assessing the nutritional status of women and directing them to appropriate resources while respecting their choices. Vegetarian and vegan diets may present with unique nutrient deficiencies that can be addressed during prenatal nutritional counseling. PMID- 18164433 TI - Differences in contraceptive choice among female adolescents at a state-funded family planning clinic. AB - Our objective was to examine differences in contraceptive choice among female adolescents from low socioeconomic backgrounds both before and after an appointment for reproductive health care at a community-based, state-funded, family planning clinic. This study utilized data collected from 4237 charts from young women attending a family planning clinic. Logistic regression was utilized to examine variables associated with the following main questions: (1) the selection of use or nonuse of contraception; (2) the selection of barrier versus hormonal contraception; and (3) within those who selected hormonal contraception, the preference for injectable versus oral hormonal methods. We found that race, age, school status, and type of health insurance were all associated with contraceptive decision-making among female adolescents. Some but not all of these associations remained after the clinic visit, which included no-cost contraception. PMID- 18164434 TI - A qualitative analysis of women's short accounts of labour and birth in a Western Australian public tertiary hospital. AB - This paper reports the findings of the postnatal qualitative arm of a larger study, which investigated women's prenatal and postnatal levels of childbirth fear. Women's expectations and experiences of labour and birth in a Western Australian public tertiary hospital were identified following thematic analysis of short written accounts from 141 participants who had given birth in the previous 6 to 14 weeks. Four major categories emerged to describe features and mediating factors in the trajectory of childbirth and the early puerperium. "Anticipating Labour and Birth," "Labour and Birth Depicted," "Mediating Factors and their Consequences," and "Evaluating, Resolving, and Looking Ahead" portray women's comparative reflections on expectations and realities of birth, on mediating influences, and on moving on from their experience. These findings will provide maternity care professionals with insight into the personal and environmental features of the childbirth setting which colours women's recollections. Being aware of what women value during labour and birth will reinforce the need for professionals to provide care using a mindful approach that considers the potential psychological, emotional, and behavioural implications of events. PMID- 18164435 TI - Cultural aspects in the care of the orthodox Jewish woman. AB - This article offers an overview and explanation of some of the main customs and laws in the Jewish religion surrounding the reproductive health care of the Torah observant woman. By understanding the religious and spiritual needs and preferences of a patient, the midwife is better able to provide optimal, culturally-competent care. Some of the aspects discussed include procreation, menstruation, modesty, contraception, abortion, genetic testing, induction, the Sabbath, Kosher diet, circumcision, and naming of the child. PMID- 18164436 TI - Tobacco intervention training in American College of Nurse-Midwives accredited education programs. AB - The objective of this study was to assess the content, amount of time, and educational techniques used in tobacco intervention training in American College of Nurse-Midwives (ACNM) accredited programs. All 43 ACNM-accredited programs were surveyed; 34 (79%) responded. Almost one in three programs (29%) offered fewer than 3 hours of instruction in tobacco education. The programs were more likely to offer training in the clinical science areas and least likely to offer training in the sociopolitical areas. Few programs (38%) required students to be taught smoking cessation techniques with patients in a clinical setting. The main barriers to teaching more (>3 h) tobacco education were not enough time in the curriculum (28%) and not having staff who are adequately trained (15%). Midwife education programs need to increase their instructional efforts, especially in the clinical science and sociopolitical areas, if midwives are to meet their goals of keeping women healthy, and in the case of pregnancies, making it a healthy experience for the woman and her newborn. This may require the development of a model core tobacco curriculum for all ACNM-accredited programs. PMID- 18164437 TI - Tending communities, crossing cultures: midwives in 19th-century California. AB - This article addresses the social history of midwifery and nursing on the California frontier between 1835 and 1885. Drawing on the published interviews of Juana Machado de Ridington (1814-1901) and Apolinaria Lorenzana (born in 1795), this study addresses the practice of midwifery in this culturally diverse frontier setting. The identity of the nineteenth-century Californiana midwife was based on the multiple roles she played in the communities she served. In addition to midwife (as we know it today), these identities included the role of lay minister, godmother, foster mother, and interpreter/translator. Previous histories have tended to marginalize these women and have failed to recognize their importance on the frontier both as women and as caretakers. The primary source material available tended to give only secondary attention to these midwives and their experiences. It is important that we preserve a place in history for these midwives and understand as best we can the ways in which they served their communities. PMID- 18164438 TI - Management of the second stage of labor in women with epidural analgesia. PMID- 18164439 TI - Broken wing. PMID- 18164446 TI - The challenge of preterm birth. PMID- 18164447 TI - Inhibition of murine AIDS by pro-glutathione (GSH) molecules. AB - Antioxidant molecules can be used both to replenish the depletion of reduced glutathione (GSH) occurring during HIV infection, and to inhibit HIV replication. The purpose of this work was to assess the efficacy of two pro-GSH molecules able to cross the cell membrane more easily than GSH. We used an experimental animal model consisting of C57BL/6 mice infected with the LP-BM5 viral complex; the treatments were based on the intramuscular administration of I-152, a pro-drug of N-acetylcysteine and S-acetyl-beta-mercaptoethylamine, and S-acetylglutathione, an acetylated GSH derivative. The results show that I-152, at a concentration of 10.7 times lower than GSH, caused a reduction in lymph node and spleen weights of about 55% when compared to infected animals and an inhibition of about 66% in spleen and lymph node virus content. S-acetylglutathione, at half the concentration of GSH, caused a reduction in lymph node weight of about 17% and in spleen and lymph node virus content of about 70% and 30%, respectively. These results show that the administration of pro-GSH molecules may favorably substitute for the use of GSH as such. PMID- 18164448 TI - A supermarket approach to the evidence-informed management of chronic low back pain. PMID- 18164449 TI - A systematic review of low back pain cost of illness studies in the United States and internationally. AB - BACKGROUND CONTEXT: The economic burden of low back pain (LBP) is very large and appears to be growing. It is not possible to impact this burden without understanding the strengths and weaknesses of the research on which these costs are calculated. PURPOSE: To conduct a systematic review of LBP cost of illness studies in the United States and internationally. STUDY DESIGN/SETTING: Systematic review of the literature. METHODS: Medline was searched to uncover studies about the direct or indirect costs of LBP published in English from 1997 to 2007. Data extracted for each eligible study included study design, population, definition of LBP, methods of estimating costs, year of data, and estimates of direct, indirect, or total costs. Results were synthesized descriptively. RESULTS: The search yielded 147 studies, of which 21 were deemed relevant; 4 other studies and 2 additional abstracts were found by searching reference lists, bringing the total to 27 relevant studies. The studies reported on data from Australia, Belgium, Japan, Korea, the Netherlands, Sweden, the UK, and the United States. Nine studies estimated direct costs only, nine indirect costs only, and nine both direct and indirect costs, from a societal (n=18) or private insurer (n=9) perspective. Methodology used to derive both direct and indirect cost estimates differed markedly among the studies. Among studies providing a breakdown on direct costs, the largest proportion of direct medical costs for LBP was spent on physical therapy (17%) and inpatient services (17%), followed by pharmacy (13%) and primary care (13%). Among studies providing estimates of total costs, indirect costs resulting from lost work productivity represented a majority of overall costs associated with LBP. Three studies reported that estimates with the friction period approach were 56% lower than with the human capital approach. CONCLUSIONS: Several studies have attempted to estimate the direct, indirect, or total costs associated with LBP in various countries using heterogeneous methodology. Estimates of the economic costs in different countries vary greatly depending on study methodology but by any standards must be considered a substantial burden on society. This review did not identify any studies estimating the total costs of LBP in the United States from a societal perspective. Such studies may be helpful in determining appropriate allocation of health-care resources devoted to this condition. PMID- 18164450 TI - Evidence-informed management of chronic low back pain with adjunctive analgesics. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164451 TI - Evidence-informed management of chronic low back pain with back schools, brief education, and fear-avoidance training. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164452 TI - Evidence-informed management of chronic low back pain with cognitive behavioral therapy. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164453 TI - Evidence-informed management of chronic low back pain with epidural steroid injections. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to chronic low back pain, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164454 TI - Evidence-informed management of chronic low back pain with facet injections and radiofrequency neurotomy. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164455 TI - Evidence-informed management of chronic low back pain with functional restoration. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164456 TI - Evidence-informed management of chronic low back pain with herbal, vitamin, mineral, and homeopathic supplements. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164457 TI - Evidence-informed management of chronic low back pain with intradiscal electrothermal therapy. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164458 TI - Evidence-informed management of chronic low back pain with lumbar extensor strengthening exercises. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of to The Spine Journal, titled Evidence Informed Management of Chronic Low Back Pain Without Surgery. Articles in this supplement were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164459 TI - Evidence-informed management of chronic low back pain with lumbar stabilization exercises. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164460 TI - Evidence-informed management of chronic low back pain with massage. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-informed management of chronic low back pain without surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164461 TI - Evidence-informed management of chronic low back pain with the McKenzie method. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164462 TI - Evidence-informed management of chronic low back pain with medicine-assisted manipulation. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence Informed Management of Chronic Low Back Pain Without Surgery. Articles in this supplement were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164463 TI - Evidence-informed management of chronic low back pain with minimally invasive nuclear decompression. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164464 TI - Evidence-informed management of chronic low back pain with needle acupuncture. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue to The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164465 TI - Evidence-informed management of chronic low back pain with nonsteroidal anti inflammatory drugs, muscle relaxants, and simple analgesics. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164466 TI - Evidence-informed management of chronic low back pain with opioid analgesics. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164467 TI - Evidence-informed management of chronic low back pain with physical activity, smoking cessation, and weight loss. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164468 TI - Evidence-informed management of chronic low back pain with prolotherapy. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164469 TI - Evidence-informed management of chronic low back pain with spinal manipulation and mobilization. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164470 TI - Evidence-informed management of chronic low back pain with transcutaneous electrical nerve stimulation, interferential current, electrical muscle stimulation, ultrasound, and thermotherapy. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164471 TI - Evidence-informed management of chronic low back pain with traction therapy. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164472 TI - Evidence-informed management of chronic low back pain with trigger point injections. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing amongst available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164473 TI - Evidence-informed management of chronic low back pain with watchful waiting. AB - The management of chronic low back pain (CLBP) has proven very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-Informed Management of Chronic Low Back Pain Without Surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Each of the articles contains five sections (description, theory, evidence of efficacy, harms, and summary) with common subheadings to facilitate comparison across the 24 different interventions profiled in this special focus issue, blending narrative and systematic review methodology as deemed appropriate by the authors. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164474 TI - A brief overview of evidence-informed management of chronic low back pain with surgery. AB - The management of chronic low back pain (CLBP) has proven to be very challenging in North America, as evidenced by its mounting socioeconomic burden. Choosing among available nonsurgical therapies can be overwhelming for many stakeholders, including patients, health providers, policy makers, and third-party payers. Although all parties share a common goal and wish to use limited health-care resources to support interventions most likely to result in clinically meaningful improvements, there is often uncertainty about the most appropriate intervention for a particular patient. To help understand and evaluate the various commonly used nonsurgical approaches to CLBP, the North American Spine Society has sponsored this special focus issue of The Spine Journal, titled Evidence-informed management of chronic low back pain without surgery. Articles in this special focus issue were contributed by leading spine practitioners and researchers, who were invited to summarize the best available evidence for a particular intervention and encouraged to make this information accessible to nonexperts. Although this special focus issue was focused on nonoperative care, it was deemed important to provide an overview of the surgical management of CLBP. This is intended to inform stakeholders of surgical options that are available to them should nonsurgical interventions prove ineffective or contraindicated. It is hoped that articles in this special focus issue will be informative and aid in decision making for the many stakeholders evaluating nonsurgical interventions for CLBP. PMID- 18164475 TI - What have we learned about the evidence-informed management of chronic low back pain? PMID- 18164476 TI - Domain interplay concept in animal models of neuropsychiatric disorders: a new strategy for high-throughput neurophenotyping research. AB - Genetic and environmental factors play a key role in psychiatric disorders. While some disorders display exceptionally high heritability, others show gene x experience x personality interactions, contributing complexity to psychiatric phenotypes. As some brain disorders frequently overlap and co-occur (representing a continuum or spectrum of phenomena), modern psychiatry is shifting from "artificial" heterogeneity to the recognition of common elements in the pathogenesis of emotional, personality and behavioral disorders. Genetic animal models of these disorders represent an important direction of research, and are widely used to explore the role of different genes in brain mechanisms. Several concepts (such as endophenotypes, gene x environment interactions, and cross species trait genetics) have been suggested for animal experimentation in this field. Here we develop a new concept based on targeting the complex interplay between different behavioral domains, meant to foster high-throughput phenotyping and integrative modeling of psychiatric disorders. PMID- 18164477 TI - Pharmacodynamic consequences of P-glycoprotein-dependent pharmacokinetics of risperidone and haloperidol in mice. AB - Efflux transporters, like P-glycoprotein (P-gp), may limit the access of drugs to the brain via the blood-brain barrier. The antipsychotic drug risperidone and its active metabolite 9-hydroxyrisperidone (paliperidone) are substrates of P-gp. Motor behavior of P-gp deficient mice (mdr1a/1b (-/-, -/-)) and wild type animals on a rotarod after acute doses of risperidone or haloperidol, a nonsubstrate of P gp, were analysed aiming to show that P-gp substrate properties of an antipsychotic drug have functional consequences. Behavioral tests revealed dose dependent effects of 0.3-3 mg/kg risperidone in wild type animals 0.5-12 h after i.p. injection of the drug. In knockout mice the 0.3 mg/kg dose of risperidone was as effective as the 3 mg/kg dose in wild type mice. A dose of 0.3 mg/kg haloperidol, however, exhibited similar pharmacodynamic effects in both genotypes. Brain concentrations of risperidone plus 9-hydroxyrisperidone were 10 fold higher in knockout than in wild type animals whereas brain concentrations of haloperidol did not differ between the two genotypes. P-gp-dependent brain distribution kinetics and behavioral effects of risperidone give evidence that the expression of P-gp has an impact on psychotropic drug actions when treating patients with drugs that are substrates of P-gp. PMID- 18164479 TI - The history and future of deep brain stimulation. AB - The advancement of electrical stimulation of the central nervous system has been a story of fits and bursts with numerous setbacks. In many ways, this history has paralleled the history of medicine and physics. We have moved from anecdotal observation to double-blinded, prospective randomized trials. We have moved from faradic stimulation to systems that lie completely under the skin and can deliver complex electrical currents to discrete areas of the brain while controlled through a device that is not much bigger than a PDA. This review will discuss how deep brain stimulation has developed into its current form, where we see the field going and the potential pitfalls along the way. PMID- 18164480 TI - Mechanisms of deep brain stimulation in movement disorders as revealed by changes in stimulus frequency. AB - Deep brain stimulation (DBS) is an established treatment for symptoms in movement disorders and is under investigation for symptom management in persons with psychiatric disorders and epilepsy. Nevertheless, there remains disagreement regarding the physiological mechanisms responsible for the actions of DBS, and this lack of understanding impedes both the design of DBS systems for treating novel diseases and the effective tuning of current DBS systems. Currently available data indicate that effective DBS overrides pathological bursts, low frequency oscillations, synchronization, and disrupted firing patterns present in movement disorders, and replaces them with more regularized firing. Although it is likely that the specific mechanism(s) by which DBS exerts its effects varies between diseases and target nuclei, the overriding of pathological activity appears to be ubiquitous. This review provides an overview of changes in motor symptoms with changes in DBS frequency and highlights parallels between the changes in motor symptoms and the changes in cellular activity that appear to underlie the motor symptoms. PMID- 18164481 TI - The treatment of movement disorders by deep brain stimulation. AB - It has been understood, for some time, that modulation of deep brain nuclei within the basal ganglia and thalamus can have a therapeutic effect in patients with movement disorders. Because of its reversibility and adjustability, deep brain stimulation (DBS) has largely come to replace traditional ablation procedures. The clinical effects of DBS vary, depending both on the target being stimulated and on the parameters of stimulation. Both aspects are currently the subject of substantial research and discovery. The most common targets for DBS treatment include the subthalamic nucleus for the treatment of advanced Parkinson's disease, the ventral intermediate nucleus of the thalamus for the treatment of medically refractory essential tremor, and the globus pallidus interna for the treatment of both cervical and generalized dystonias and Parkinson's disease. We review the current indications, targets, outcomes, and general procedure of DBS for essential tremor, Parkinson's disease, and dystonia. PMID- 18164482 TI - Motor cortex stimulation for pain and movement disorders. AB - Since initial reports in the early 1990s, stimulation of the M1 region of the cortex (MCS) has been used to treat chronic refractory pain conditions and a variety of movement disorders. A Medline search of literature between 1991 and 2007 revealed 512 cases using MCS. Although most of these relate to the treatment of pain (422), 84 of them involve movement disorders. More recently, several studies have specifically looked at treating Parkinson's disease (PD) with MCS. We report here several of our own cases using MCS to treat poststroke and non poststroke pain syndromes and movement disorders (n = 8), PD (n = 4), ET (n = 2), and cortico-basal degeneration (n = 1). We also cover the essential history of this procedure and our current research using computational modeling to understand further the underlying mechanisms of MCS. PMID- 18164483 TI - Deep brain stimulation for psychiatric disorders. AB - Surgery for psychiatric disorders first began in the early part of the last century when the therapeutic options for these patients were limited. The introduction of deep brain stimulation (DBS) has caused a new interest in the surgical treatment of these disorders. DBS may have some advantage over lesioning procedures used in the past. A critical review of the major DBS targets under investigation for Tourette's syndrome, obsessive-compulsive disorder, and major depression is presented. Current and future challenges for the use of DBS in psychiatric disorders are discussed, as well as a rationale for referring to this subspecialty as limbic disorders surgery based on the parallels with movement disorders surgery. PMID- 18164484 TI - Deep brain stimulation for epilepsy. AB - Many patients who suffer from medically refractory epilepsy are not candidates for resective brain surgery. Success of deep brain stimulation (DBS) in relieving a significant number of symptoms of various movement disorders paved the way for investigations into this modality for epilepsy. Open-label and small blinded trials have provided promising evidence for the use of DBS in refractory seizures, and the first randomized control trial of DBS of the anterior thalamic nucleus is currently underway. There are multiple potential targets, because many neural regions have been implicated in seizure propagation. Thus, it is difficult as yet to make any definitive judgments about the efficacy of DBS for seizure control. Future study is necessary to identify a patient population for whom this technique would be indicated, the most efficacious target, and optimal stimulation parameters. PMID- 18164485 TI - Responsive cortical stimulation for the treatment of epilepsy. AB - Epilepsy is a common chronic neurological disorder affecting approximately 1-2% of the population. Despite the available treatment options (pharmacotherapy, surgery, and vagus nerve stimulation), a large percentage of patients continue to have seizures. With the success of deep brain stimulation for treatment of movement disorders, brain stimulation has received renewed attention as a potential treatment option for epilepsy. Responsive stimulation aims to suppress epileptiform activity by delivering stimulation directly in response to electrographic activity. Animal and human data support the concept that responsive stimulation can abort epileptiform activity, and this modality may be a safe and effective treatment option for epilepsy. Responsive stimulation has the advantage of specificity. In contrast to the typically systemic administration of pharmacotherapy, with the concomitant possibility of side effects, electrical stimulation can be targeted to the specific brain regions involved in the seizure. In addition, responsive stimulation provides temporal specificity. Treatment is provided as needed, potentially reducing the likelihood of functional disruption or habituation due to continuous treatment. Here we review current animal and human research in responsive brain stimulation for epilepsy and then discuss the NeuroPace RNS System, an investigational implantable responsive neurostimulator system that is being evaluated in a multicenter, randomized, double-blinded trial to assess the safety and efficacy of responsive stimulation for the treatment of medically refractory epilepsy. PMID- 18164486 TI - Vagus nerve stimulation for epilepsy and depression. AB - Many patients with epilepsy suffer from persistent seizures despite maximal antiepileptic drug (AED) therapy. Chronic, intermittent vagus nerve stimulation (VNS) has proven to be a safe, effective option for patients suffering from refractory seizures who are not candidates for surgical resection. Although only a small minority of patients will be entirely seizure-free, VNS as an adjunct to medical therapy does appear to provide a significant amount of improvement in quality of life. Reports of antidepressant effects independent of seizure control, along with the use of multiple AEDs in the treatment of depression, has led to the investigation of VNS as a potential adjunctive treatment for major depressive disorder. Both the number of severely depressed patients refractory to available pharmacologic options and the need for repeated treatments and significant side effects associated with electroconvulsive therapy have heightened the interest in VNS for this patient population. Pilot studies of VNS for depression have shown impressive response rates; however, the effect appears to be gradual in onset, as demonstrated by the lack of a favorable response in a short-term, randomized controlled study. Investigation is thus needed to establish the potential role of VNS as an adjunctive treatment for severe depression. PMID- 18164487 TI - Spinal cord stimulation: an update. AB - Spinal cord stimulation has been used in the treatment of many chronic pain disorders since 1967. In this update, the indications for spinal cord stimulation are reviewed with attention to recent publications. A focused review of the literature on abdominal and visceral pain syndromes is also provided. Furthermore, the technology has evolved from the use of monopolar electrodes to complex electrode arrays. Similarly, the power source has changed from a radio frequency-driven system to a rechargeable impulse generator. These topics are covered, along with a short discussion of implant technique. Finally, we include a review of complications of such therapy. SCS as a technology and therapy continues to evolve. PMID- 18164488 TI - Peripheral nerve stimulation for neuropathic pain. AB - Peripheral nerve stimulation (PNS) has been used for treatment of neuropathic pain for more than 40 years. Recent resurgence of interest to this elegant surgical modality came from the introduction of less invasive implantation techniques and the wider acceptance of neuromodulation as a treatment of medically refractory cases. This article reviews the literature on the use of PNS for neuropathic pain and describes current indications and hardware choices in frequent use. Published experience indicates that neuropathic pain responds to PNS in many patients. PNS works well in both established indications, such as post-traumatic and postsurgical neuropathy, occipital neuralgia, and complex regional pain syndromes, and in relatively new indications for neuromodulation, such as migraines and daily headaches, cluster headaches, and fibromyalgia. Future research and growing clinical experience will help in identifying the best candidates for PNS, choosing the best procedure and best hardware for each individual patient, and defining adequate expectations for patients and pain specialists. PMID- 18164489 TI - Sacral nerve stimulation: neuromodulation for voiding dysfunction and pain. AB - Voiding dysfunction, which includes incontinence, retention, and chronic pelvic pain, is a relatively frequent problem that can be difficult to manage. Neuromodulation via stimulation of the sacral nerves has been shown to improve these symptoms, although the exact mechanisms remain elusive. Techniques for nerve stimulation may vary, depending on the disease, location of pain, and the patient's anatomy. In addition to placement of electrodes on the sacral nerve roots, modulation has also been reported by peripheral branches of the sacral nerves including the pudendal and posterior tibial nerves. Newer surgical techniques have significantly decreased the morbidity of the procedures and increased the probability of a successful outcome. PMID- 18164490 TI - Intrathecal pumps. AB - Direct neuroaxis drug delivery has expanded the physician's armamentarium to provide treatment options to those who have failed more conservative interventions. Starting from Bier's 'cocainization of the spinal cord' in 1898, direct nervous system pharmacy delivery has long been recognized as an effective means to treat pain. Intrathecal pump systems are now commonplace in the management of numerous pain states, as well as of neuromuscular sequelae of central nervous system injury. There has been much advancement in the physiologic and pharmacologic understanding of direct neuromodulation, providing a growing number of treatment options depending on the specific disease state. As well, surgical techniques and catheter systems have undergone refinements providing improved long-term safety and efficacy. We present a review of the historical evolution to current intrathecal therapies, as well as a dialog regarding patient selection, drug options, and side effects. Also, included is a discussion of surgical techniques, current delivery options and complications concerning pump placement. PMID- 18164491 TI - Image-guided convection-enhanced delivery platform in the treatment of neurological diseases. AB - Convection-enhanced delivery (CED) of substances within the human brain is becoming a more frequent experimental treatment option in the management of brain tumors, and more recently in phase 1 trials for gene therapy in Parkinson's disease (PD). Benefits of this intracranial drug-transfer technology include a more efficient delivery of large volumes of therapeutic agent to the target region when compared with more standard delivery approaches (i.e., biopolymers, local infusion). In this article, we describe specific technical modifications we have made to the CED process to make it more effective. For example, we developed a reflux-resistant infusion cannula that allows increased infusion rates to be used. We also describe our efforts to visualize the CED process in vivo, using liposomal nanotechnology and real-time intraoperative MRI. In addition to carrying the MRI contrast agent, nanoliposomes also provide a standardized delivery vehicle for the convection of drugs to a specific brain-tissue volume. This technology provides an added level of assurance via visual confirmation of CED, allowing intraoperative alterations to the infusion if there is reflux or aberrant delivery. We propose that these specific modifications to the CED technology will improve efficacy by documenting and standardizing the treatment volume delivery. Furthermore, we believe that this image-guided CED platform can be used in other translational neuroscience efforts, with eventual clinical application beyond neuro-oncology and PD. PMID- 18164492 TI - Auditory brainstem implants. AB - The development of cochlear implantation has allowed the majority of patients deafened after the development of language to regain significant auditory benefit. In a subset of patients, however, loss of hearing results from destruction of the cochlear nerves, rendering cochlear implantation ineffective. The most common cause of bilateral destruction of the cochlear nerves is neurofibromatosis type 2 (NF2). The hallmark of this genetic disorder is the development of bilateral acoustic neuromas, the growth or removal of which causes deafness in most patients. Patients with NF2 may benefit from direct stimulation of the cochlear nucleus. We describe the development, use, and results of the auditory brainstem implant (ABI), which is typically implanted via craniotomy at the time of tumor removal. Most patients with the implant have good appreciation of environmental sounds, but obtain more modest benefit with regard to speech perception. The majority of patients make use of the implant to facilitate lip reading; some can, to varying degrees, comprehend speech directly. We discuss future directions in central implants for hearing, including the penetrating ABI, the use of ABI in nontumor patients, and the auditory midbrain implant. PMID- 18164493 TI - The development of brain-machine interface neuroprosthetic devices. AB - The development of brain-machine interface technology is a logical next step in the overall direction of neuroprosthetics. Many of the required technological advances that will be required for clinical translation of brain-machine interfaces are already under development, including a new generation of recording electrodes, the decoding and interpretation of signals underlying intention and planning, actuators for implementation of mental plans in virtual or real contexts, direct somatosensory feedback to the nervous system to refine actions, and training to encourage plasticity in neural circuits. Although pre-clinical studies in nonhuman primates demonstrate high efficacy in a realistic motor task with motor cortical recordings, there are many challenges in the clinical translation of even simple tasks and devices. Foremost among these challenges is the development of biocompatible electrodes capable of long-term, stable recording of brain activity and implantable amplifiers and signal processors that are sufficiently resistant to noise and artifact to faithfully transmit recorded signals to the external environment. Whether there is a suitable market for such new technology depends on its efficacy in restoring and enhancing neural function, its risks of implantation, and its long-term efficacy and usefulness. Now is a critical time in brain-machine interface development because most ongoing studies are science-based and noncommercial, allowing new approaches to be included in commercial schemes under development. PMID- 18164494 TI - Spinal cord injury: present and future therapeutic devices and prostheses. AB - A range of passive and active devices are under development or are already in clinical use to partially restore function after spinal cord injury (SCI). Prosthetic devices to promote host tissue regeneration and plasticity and reconnection are under development, comprising bioengineered bridging materials free of cells. Alternatively, artificial electrical stimulation and robotic bridges may be used, which is our focus here. A range of neuroprostheses interfacing either with CNS or peripheral nervous system both above and below the lesion are under investigation and are at different stages of development or translation to the clinic. In addition, there are orthotic and robotic devices which are being developed and tested in the laboratory and clinic that can provide mechanical assistance, training or substitution after SCI. The range of different approaches used draw on many different aspects of our current but limited understanding of neural regeneration and plasticity, and spinal cord function and interactions with the cortex. The best therapeutic practice will ultimately likely depend on combinations of these approaches and technologies and on balancing the combined effects of these on the biological mechanisms and their interactions after injury. An increased understanding of plasticity of brain and spinal cord, and of the behavior of innate modular mechanisms in intact and injured systems, will likely assist in future developments. We review the range of device designs under development and in use, the basic understanding of spinal cord organization and plasticity, the problems and design issues in device interactions with the nervous system, and the possible benefits of active motor devices. PMID- 18164495 TI - A protein of the leucine-rich repeats (LRRs) superfamily is implicated in antimony resistance in Leishmania infantum amastigotes. AB - Pentavalent antimonial containing drugs (SbV) are the mainstay for the control of the protozoan parasite Leishmania but resistance to this class of drug is now prevalent in several endemic areas. We describe here the use of functional cloning where an expression cosmid bank derived from Leishmania infantum was transfected in L. infantum axenic amastigotes and selected for potassium antimonyl tartrate (SbIII) resistance. This strategy allowed the isolation of a cosmid encoding for a novel resistance protein, LinJ34.0570, which belongs to the superfamily of leucine-rich repeat (LRR) proteins. Parasites overexpressing this LRR protein, which is part of the LRR_CC subfamily, were resistant to SbIII as axenic amastigotes and to SbV as intracellular parasites. This work pinpoints a novel protein that can contribute to antimonial resistance in Leishmania. PMID- 18164496 TI - Porcine glucagon-like peptide-2: structure, signaling, metabolism and effects. AB - Mass spectrometry of HPLC-purified porcine glucagon-like peptide-2 (pGLP-2)(1) revealed a 35 amino acid sequence with C-terminal Ser and Leu, in contrast to the 33 amino acids of human, cow, rat and mouse GLP-2. Synthetic pGLP-2 stimulated cAMP-production in COS-7 cells expressing human GLP-2 (hGLP-2) receptor with the same potency and efficacy as hGLP-2. In anesthetized pigs (n=9) given intravenous pGLP-2 infusions, the half life (t1/2) of intact pGLP-2 (8.4+/-0.9 min) was shorter (p<0.01) than that of the primary metabolite pGLP-2 (3-35) (34.0+/-5.2 min), generated by dipeptidyl peptidase-4 (DPP-4) cleavage. Adding the DPP-4 inhibitor valine-pyrrolidide prolonged t1/2 of intact pGLP-2 (p<0.05). The metabolic clearance rate (MCR) of intact pGLP-2 (23.9+/-3.82 mL/(kg x min)) was greater (p<0.0001) than that of pGLP-2 (3-35) (6.36+/-1.45 mL/(kg x min)) and larger than the previously reported MCR of hGLP-2 in pig. The MCR of intact pGLP 2 was reduced by valine-pyrrolidide (p<0.05), but was still greater than that of intact hGLP-2 previously reported. In the isolated perfused porcine pancreas, pGLP-2 stimulated glucagon release (p<0.05), but had no effect on insulin or somatostatin release. Exocrine secretion was unaffected and there was no apparent vasoactive effect. In mice (n=8), both subcutaneous hGLP-2 and pGLP-2 given twice daily for 10 days, significantly and equally increased small intestinal weight, length and cross-sectional area of proximal ileum. In conclusion, pGLP-2 and hGLP 2 have similar effects in vivo and in vitro in spite of the structural differences. However, pGLP-2 is cleared more rapidly in pigs than hGLP-2. PMID- 18164497 TI - Ventricular late potentials among thalassemia patients. AB - BACKGROUND: Iron induced cardiac abnormalities remain the number one cause of death among thalassemia major (TM) patients. Signal averaged ECG (SAECG) was suggested to predict ventricular tachycardia as the underlying substrate for up to 5% incidence of sudden cardiac death among TM patients. The prevalence of ventricular late potentials (VLP) among different TM populations varied (3-31%); therefore to further clarify this we here describe the incidence of VLP among TM patients over a 7 year follow up period (1997 to 2004). METHODS: 26 TM patients were randomly selected from a group of 240 TM patients. SAECG, regular ECG, echocardiography-Doppler were analyzed during the study period. Ferritin levels and cardiac complaints were registered from an interview and chart review. RESULTS: Mean QRS duration increased from 89.23 (+/-10.60) ms in 1997 to 94.27 (+/-10.91) in 2004 (p<0.01), mean late amplitude signal (LAS) duration increased from 23.04 (+/-7.68) ms in 1997 to 27.69 (+/-6.82) ms in 2004 (p=0.01), whereas mean root mean square voltage RMS decreased from 80.85 (+/-51.19) mV in 1997 to 45.12 (+/-21.42) mV in 2004 (p<0.01). Changes in QRS duration and RMS voltage were found to be linearly correlated with average of ferritin over years (r=0.38, p=0.03 and r=-0.47, p=0.01 respectively); and only 1 patient developed VLP over 7 years. CONCLUSION: The incidence of VLP is 3.8% in the TM population over 7 years, despite the presence of significant changes in all SAECG criteria. RMS voltage and QRS duration changes over time seem to be related to iron overload measured by ferritin level. PMID- 18164498 TI - Coronary cyclic flow variations following primary angioplasty is associated with poor short-term prognosis. AB - AIMS: Recent studies have shown that coronary cyclic flow variations (CCFV) is a platelet-related phenomenon that occurred following reperfusion. Although CCFV predicts acute complications following thrombolytic therapy, its impact following percutaneous coronary interventions (PCI) has not been evaluated yet. METHODS AND RESULTS: One hundred and thirty-one patients with ST-segment Elevation Myocardial Infarction (STEMI) who underwent PCI were included in the analysis. All patients have 24-hour ST-segment monitoring. The development of CCFV was defined as > or = 3 ST-segment transitions (> or =150 microV). We divided the population in two groups according to the presence (n=14, 10.6%) or absence (n=117) of CCFV. The relation between CCFV and 30-day major adverse cardiac events (MACE) was analyzed using a multivariate logistic regression model adjusting for age, sex, diabetes, smoking, anterior infarct, Killip class, and final TIMI flow grade. Clinical and angiographic characteristics were similar between the two groups. Higher 30-day mortality (21.4 vs. 3.8%, p=0.022) and MACE rates (42.9 vs. 10.7%, p=0.005) were seen in the CCFV group. Multivariate regression analysis revealed that patients with CCFV were at increased risk of 30-day MACE (adjusted RR 5.09; 95% CI 1.3 19.1; p=0.0016). CONCLUSION: The presence of CCFV altered primary PCI may provide an early indication of insufficient myocardial perfusion and impending catastrophic outcome. PMID- 18164499 TI - Risks for bovine tuberculosis in British cattle farms restocked after the foot and mouth disease epidemic of 2001. AB - The foot and mouth disease (FMD) epidemic of 2001 was used to investigate herd breakdown (HBD) with bovine tuberculosis (bTB) in totally restocked herds of cattle. By August 2004, 2941 restocked cattle herds, with cattle movements from before and after 2001, had been tested for bTB for the first time since restocking. A total of 6% (177) of these herds broke down at the first bTB test. A binomial logistic regression model with HBD (at least one reactor bovine) at the first test after restocking as the outcome was used to investigate risks associated with HBD. The final model contained three risk factors. There was an increased risk for HBD in restocked herds with every log increase in herd size with an OR=1.38 (CI 1.16-1.64) to a maximum OR of 10.75. When there was a history of bTB on the restocked farm before 2001 the OR, with CI not including unity, were 5.92, 4.63, 3.8 and 2.9 for last HBD in 2000, 1999, 1998 and 1997, respectively, indicating a persistence in increased risk for restocked herds from farms with a history of HBD in the previous herd before restocking, i.e. a different population of cattle. Finally, for every log increase in the number of cattle purchased from herds with a greater than biennial frequency of testing for bTB in the previous 8 years (i.e. perceived high risk herds for bTB) there was an OR=1.35 (95% CI 1.22-1.49). The maximum OR was 9.27. These results indicate that both introduction of bTB through the purchase of cattle from farms with a high perceived risk of bTB infection and persistence of bTB on the restocked farm, (not the farm's original herd), were associated with an increased risk of HBD in the newly formed herds after restocking. PMID- 18164500 TI - Parameter estimation and simulations of a mathematical model of Corynebacterium pseudotuberculosis transmission in sheep. AB - Caseous lymphadenitis (CLA) is an infectious disease of sheep caused by Corynebacterium pseudotuberculosis. It is prevalent in most sheep producing countries and was introduced into the UK sheep population in 1991. The pathogen invades the host through epithelium and forms an abscess in the local draining lymph node. Typically, disease presents as clinical, with overt (externally visible) swollen lymph nodes (the parotid, submandibular, prefemoral, prescapular, popliteal or mammary) or sub-clinical, with abscesses in the lungs and associated thoracic (bronchial and mediastinal) lymph nodes. We present a mathematical model in which disease is categorised as overt and/or respiratory (sub-clinical), using the above groupings. In both situations sheep may be infected and may or may not be infectious. In the model, overt abscesses may resolve and respiratory abscesses are considered to be present for life. Using the location of the abscesses, three routes of transmission are postulated: overt to overt, respiratory to overt and respiratory to respiratory. Data from four naturally infected flocks were used to describe populations of sheep with epidemic CLA and to estimate transmission coefficients for each of the postulated transmission routes. The infection process parameters were derived from literature where possible. Parameters were estimated using maximum likelihood methods and compared to the data using a multinomial distribution. The distribution of abscesses in the flocks was similar to endemic data reported in other studies. In the model most infected sheep developed abscesses, and approximately 36% of sheep with overt abscesses recovered from infection. The average time for respiratory abscesses to become infectious was 41 days. In these data, overt to overt transmission was the most frequent route of transmission since it had the highest coefficient in the model compared with respiratory to overt and respiratory to respiratory transmission. Transmission coefficients specific for each flock significantly (P<0.05) improved the model fit to the data. In simulations using values of best-fitting parameter combinations, the proportion of sheep infected was between 0.39 and 0.60 at equilibrium. This is the first mathematical model of C. pseudotuberculosis infection, the parameter estimates indicate that aspects of the infection process could be utilised to design control strategies. PMID- 18164501 TI - Cortisol administration enhances the coupling of midfrontal delta and beta oscillations. AB - There is increasing evidence that the strength of the relation between slow (SW) and fast (FW) wave activity in the EEG is associated with specific motivational states and their corresponding neuroendocrine patterns. Enhanced correlations between SW and FW have been related to anxiety, behavioral inhibition and high basal cortisol levels. However, the direct effects of cortisol on SW-FW coupling have not been experimentally studied yet. The present study investigated whether cortisol administration increases SW-FW coupling. Resting state EEG recordings were obtained from 40 right-handed healthy male subjects with extreme low or high scores on a behavioral inhibition scale, after placebo and cortisol (50 mg) administration. As expected, cortisol resulted in a significant increase in correlation between SW (delta) and FW (beta) activity compared to placebo. In addition, delta-beta correlation was significantly higher in high compared to low behaviorally inhibited subjects in both conditions. These results suggest that cortisol can modify brain activity, increasing a pattern associated with anxiety and behavioral inhibition. This is in line with findings associating cortisol with behavioral inhibition and anxiety. PMID- 18164502 TI - Analysis of gene expression profiles in rat hippocampus following treatment with nicotine and an alpha7 nAChR selective agonist. AB - The nicotinic acetylcholine receptors (nAChRs) play critical roles in neuronal transmission and modulation. Among the diverse nAChRs, the alpha7 subtype has been considered as a potential therapeutic target for treating cognitive deficits associated with neuropsychiatric and neurodegenerative diseases. Although a number of mechanisms including neurotransmitter and biochemical effects linking alpha7 nAChR activation and cognitive function are beginning to be described, the underlying molecular processes especially following repeated administration remain unclear. To address this, we have performed gene expression analysis in rats treated with nicotine and a selective alpha7 nAChR agonist, PNU-282987. Our results showed significant overlap in gene expression changes induced by PNU 282987 and nicotine, suggesting convergent pathways triggered by these compounds. Treatment with nicotine also resulted in regulation of a number of genes that were not regulated by PNU-282987, consistent with the interaction of nicotine with other nAChRs beyond the alpha7 subtype. Interestingly, these gene expression changes were observed 24 h post-dose, suggesting that both nicotine and PNU 282987 cause protracted changes in gene expression. Overall, our results identify gene expression changes that may contribute to further defining the roles of nAChR activation in cognitive function. PMID- 18164503 TI - Upregulated expression of neuropeptide Y in hypothalamic-pituitary system of rats by chronic dexamethasone administration. AB - To study the effect of adrenal steroids on neuropeptide Y (NPY) synthesis in the hypothalamic-pituitary system, we examined NPY expression in rats treated with dexamethasone (a synthetic glucocorticoid) by in situ hybridization and immunohistochemistry. Rats were injected daily with dexamethasone (0.2mg/100g/day for 10 days, sc) or sesame oil (vehicle control), or non-injected (intact control). Relative staining area for corticotropin-releasing hormone or neurophysin II, a vasopressin carrier protein, was increased in the external zone of the median eminence in vehicle control, but was equivalent to that of intact control in the dexamethasone-injected group. Density of NPY-stained fiber varicosities was drastically increased in the external, but not the internal, zone of dexamethasone-injected group, coinciding with the increased NPY hybridization signal level in the arcuate nucleus. Dual-labeling experiments revealed no colocalization of NPY with hypophysiotropic or other peptides examined in single fibers of the median eminence. In the dexamethasone-injected group, expressions of NPY mRNA and peptide were detectable in a few pituitary cells, with some being corticotropes. These results suggest that NPY plays hormonal roles in the hypothalamic-pituitary-adrenal axis. PMID- 18164504 TI - IQ-ArfGEF/BRAG1 is a guanine nucleotide exchange factor for Arf6 that interacts with PSD-95 at postsynaptic density of excitatory synapses. AB - ADP ribosylation factor 6 (Arf6) is a small GTPase that regulates dendritic differentiation possibly through the organization of actin cytoskeleton and membrane traffic. Here, we characterized IQ-ArfGEF/BRAG1, a guanine nucleotide exchange factor (GEF) for Arf6, in the mouse brain. In vivo Arf pull down assay demonstrated that IQ-ArfGEF/BRAG1 activated Arf6 more potently than Arf1. IQ ArfGEF/BRAG1 mRNA was abundantly expressed in the brain with higher levels in forebrain structures and cerebellar granule cells. In hippocampal neurons, IQ ArfGEF/BRAG1 mRNA was localized not only at neuronal cell bodies but also at dendritic processes, indicating its dendritic transport and localization. Immunoprecipitation and in vitro binding experiments revealed that IQ ArfGEF/BRAG1 formed a protein complex with N-methyl-d-aspartate (NMDA)-type glutamate receptors through the interaction with a postsynaptic density (PSD) scaffold protein, PSD-95. Immunohistochemical analysis demonstrated that IQ ArfGEF/BRAG1 was localized preferentially at the postsynaptic density of asymmetrical synapses on dendritic spines, but was lacking at GABAa receptor carrying inhibitory synapses. Taken together, IQ-ArfGEF/BRAG1 forms a postsynaptic protein complex containing PSD-95 and NMDA receptors at excitatory synapses, where it may function as a GEF for Arf6. PMID- 18164505 TI - Edible alginate-based coating as carrier of antimicrobials to improve shelf-life and safety of fresh-cut melon. AB - The effect of malic acid and essential oils (EOs) of cinnamon, palmarosa and lemongrass and their main active compounds as natural antimicrobial substances incorporated into an alginate-based edible coating on the shelf-life and safety of fresh-cut "Piel de Sapo" melon (Cucumis melo L.) was investigated. Melon pieces (50 g) were coated with alginate-based edible coating containing malic acid (EC) and EOs or their active compounds before to be packed in air filled polypropylene trays and stored at 5 degrees C for shelf-life and sensory studies. On the other hand, melon pieces were inoculated with a Salmonella Enteritidis (10(8) CFU/ml) culture before applying the coatings containing malic acid and EOs or their active compounds to safety study. Controls of fresh-cut melon non-coated or coated with EC without EOs were also prepared. EC was effective to improve shelf-life of fresh-cut melon from microbiological (up to 9.6 days) and physicochemical (>14 days) points of view in comparison with non-coated fresh-cut melon, where microbiological and physicochemical shelf-life was up to 3.6 days and lower than 14 days, respectively. In addition, the incorporation of EOs or their active compounds into the edible coating prolonged the microbiological shelf-life by more than 21 days in some cases due probably to an enhanced antimicrobial effect of malic acid+EOs; however, some fresh-cut melon characteristics were affected such as firmness and color causing a reduction of physicochemical shelf-life. Significant reductions (p<0.05) of S. Enteritidis population in inoculated coated fresh-cut melon were achieved, varying the effectiveness of the coatings depending on the EOs or the active compound and their concentrations. According to the results, palmarosa oil incorporated at 0.3% into the coating appear to be a promising preservation alternative for fresh cut melon, since it had a good acceptation by panellists, maintained the fruit quality parameters, inhibited the native flora growth and reduced S. Enteritidis population. PMID- 18164506 TI - Tubulovesicular structures are a consistent (and unexplained) finding in the brains of humans with prion diseases. AB - Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler-Scheinker disease (GSS) and Fatal Familial Insomnia (FFI) are slow neurodegenerative disorders classified as transmissible spongiform encephalopathies (TSEs) or prion diseases, which appear in sporadic, hereditary or environmentally acquired forms. Tubulovesicular structures (TVS) are ultrastructural particles of unknown origin and chemical composition found in the brains of both animal and human forms of transmissible spongiform encephalopathies or prion diseases. In this paper, we report the results of a search for TVS in a total of 13 cases of sporadic Creutzfeldt-Jakob disease, three cases of Gerstmann-Straussler-Scheinker disease, two cases of Fatal Familial Insomnia, and individual cases of familial, iatrogenic, and variant CJD (vCJD). TVS were found in all but one sporadic and one familial case of CJD. As controls, we examined 15 cases of Alzheimer's disease (AD), two cases of Pick's disease, and one case of multiple system atrophy. TVS were not present in any of these cases. This study confirms the TSE-specificity of TVS, the morphology of which suggests a possible pathogenetic role and relationship to recently described virion-like arrays of 25nm particles in scrapie-infected tissue cultures. PMID- 18164507 TI - Interplay between oxidative stress and hepatic steatosis in the progression of chronic hepatitis C. AB - BACKGROUND/AIMS: The contribution of oxidative stress to the pathogenesis of chronic hepatitis C (CHC) is still poorly elucidated. This study investigated the relationship between oxidative stress, insulin resistance, steatosis and fibrosis in CHC. METHODS: IgG against malondialdehyde-albumin adducts and HOMA-IR were measured as markers of oxidative stress and insulin resistance, respectively, in 107 consecutive CHC patients. RESULTS: Oxidative stress was present in 61% of the patients, irrespective of age, gender, viral load, BMI, aminotransferase level, histology activity index (HAI) and HCV genotype. Insulin resistance and steatosis were evident in 80% and 70% of the patients, respectively. In the patients infected by HCV genotype non-3, but not in those with genotype 3 infection HOMA IR (p<0.03), steatosis (p=0.02) and fibrosis (p<0.05) were higher in the subjects with oxidative stress than in those without. Multiple regression analysis revealed that, HOMA-IR (p<0.01), fibrosis (p<0.01) and oxidative stress (p<0.05) were independently associated with steatosis, whereas steatosis was independently associated with oxidative stress (p<0.03) and HOMA-IR (p<0.02). Steatosis (p<0.02) and HAI (p=0.007) were also independent predictors of fibrosis. CONCLUSIONS: In patients infected by HCV genotype non-3, oxidative stress and insulin resistance contribute to steatosis, which in turn exacerbates both insulin resistance and oxidative stress and accelerates the progression of fibrosis. PMID- 18164508 TI - Early switch to pentoxifylline in patients with severe alcoholic hepatitis is inefficient in non-responders to corticosteroids. AB - BACKGROUND/AIMS: In severe alcoholic hepatitis (AH), 40% of patients will obtain no benefit from corticosteroids. Improvement in management of non-responders is warranted and only pentoxifylline can be considered an alternative. A two-step strategy was evaluated consisting of early withdrawal of corticosteroids and a switch to pentoxifylline for 28 additional days in non-responders identified using early change in bilirubin level. METHODS: One hundred and twenty-one patients with AH were treated prospectively with corticosteroids, and the two step strategy was proposed to 29 non-responders treated according to a two-step strategy who were compared to 58 matched non-responders treated with corticosteroids only. RESULTS: Clinical and biological features of the two groups were similar. There was no survival improvement at 2 months in patients treated with the two-step strategy compared to controls: 35.5+/-6.3% vs 31+/-8.6%. After 21 days, biological evolution was similar for prothrombin time (-0.25s vs +0.2s), bilirubin (0.8 mg/dl vs 2.03 mg/dl) and creatinine (+0.16 mg/dl vs -0.7 mg/dl). In multivariate analysis, only age, evolution of bilirubin during the first week, creatinine and DF were associated with 2-month survival. CONCLUSIONS: Non responders to corticosteroids do not obtain any benefit from an early switch to pentoxifylline. Thus, the issue of management of non-responders remains unresolved. PMID- 18164509 TI - Entry of new pharmacies in the deregulated Norwegian pharmaceuticals market- consequences for costs and availability. AB - OBJECTIVES: The objective of this study is to analyze the impact of the new regulation concerning entry of pharmacies into the Norwegian pharmaceuticals market in 2001 on cost and availability of pharmaceutical products. METHODS: In order to study costs, a translog cost function is estimated using data from the annual reports of a sample of Norwegian pharmacies before and after the deregulation of the market. Linear regression models for the number of pharmacies in each region in Norway are also estimated. RESULTS: The results show that the costs of the individual pharmacies have not decreased as a consequence of the deregulation of the Norwegian pharmaceuticals market. The deregulation of the market did, however, increase the availability to pharmacy services substantially. CONCLUSIONS: Increased availability of pharmacy services can be achieved by deregulating pharmaceutical markets as in Norway, but at the expense of increased costs for the pharmacies. PMID- 18164510 TI - Trastuzumab in early stage breast cancer: a cost-effectiveness analysis for Belgium. AB - OBJECTIVES: Although trastuzumab is traditionally used in metastatic breast cancer treatment, studies reported on the efficacy and safety of trastuzumab in adjuvant setting for the treatment of early stage breast cancer in HER2+ tumors. We estimated the cost-effectiveness and budget impact of reimbursing trastuzumab in this indication from a payer's perspective. METHODS: We constructed a health economic model. Long-term consequences of preventing patients to progress to metastatic breast cancer and side effects such as congestive heart failure were taken into account. Uncertainty was handled applying probabilistic modeling and through probabilistic sensitivity analyses. RESULTS: In the HERA scenario, applying an arbitrary threshold of euro30000 per life-year gained, early stage breast cancer treatment with trastuzumab is cost-effective for 9 out of 15 analyzed subgroups (according to age and stage). In contrast, treatment according to the FinHer scenario is cost-effective in 14 subgroups. Furthermore, the FinHer regimen is most of the times cost saving with an average incremental cost of euro668, euro-1045, and euro-6869 for respectively stages I, II and III breast cancer patients whereas the HERA regimen is never cost saving due to the higher initial treatment costs. CONCLUSIONS: The model shows better cost-effectiveness for the 9-week initial treatment (FinHer) compared to no trastuzumab treatment than for the 1-year post-chemotherapy treatment (HERA). Both from a medical and an economic point of view, the 9-week initial treatment regimen with trastuzumab shows promising results and justifies the initiation of a large comparative trial with a 1-year regimen. PMID- 18164511 TI - Development and pilot testing of the Neuromuscular Blockade Advisory System. AB - The neuromuscular blocker advisory system (NMBAS) is a computer program developed to provide advisory guidance to anesthesiologists on the timing and dose of rocuronium to paralyze patients during surgery. It is believed that the use of such a system will administer the minimally effective amount of drug, maintaining the patient in a state of paralysis that is useful for surgery yet easily reversible. This will improve patient safety and result in more efficient care. In this paper we present the NMBAS, its basic methodology, and its development though a pilot study. Novel methods of handling neuromuscular response data are presented, including relaxation measurement and the enhanced-train-of-four sensing modality. New methods of handling nonlinearities at the neuromuscular junction to allow application of adaptive control techniques are presented. A novel form of modelling combining model swapping and RLSE adaptation to accommodate the patient variation seen with NMB drugs is introduced. A pilot study testing the NMBAS was undergone to prepare the NMBAS for application in a full clinical trial, in which patients undergoing prostate brachytherapy surgeries using rocuronium for intubation were admitted. PMID- 18164512 TI - On tests of rate ratio under standard inverse sampling. AB - Inverse sampling suggests one continues to sample subjects until a pre-specified number of rare events of interest is observed. It is generally considered to be more appropriate than the usual binomial sampling when the subjects come sequentially, when the response probability is rare, and when maximum likelihood estimators of some epidemiological measures are undefined under binomial sampling. Reliable but conservative exact conditional procedure for the ratio of the response probabilities of subject without the attribute of interest has been studied. However, such a procedure is inapplicable to the risk ratio (i.e., ratio of the response probabilities of subject with the attribute of interest). In this paper, we investigate various test statistics (namely Wald-type, score and likelihood ratio test statistics) for testing non-unity risk ratio under standard inverse sampling scheme, which suggests one continue to sample until the predetermined number of index subjects with the attributes of interest is observed. Both asymptotic and numerical approximate unconditional methods are considered for P-value calculation. Performance of these test procedures are evaluated under different settings by means of Monte Carlo simulation. In general, the Wald-type test statistic is preferable for its satisfactory and stable performance with approximate unconditional procedures. The methodologies are illustrated with a real example from a heart disease study. PMID- 18164513 TI - Hydrogeochemical comparison and effects of overlapping redox zones on groundwater arsenic near the Western (Bhagirathi sub-basin, India) and Eastern (Meghna sub basin, Bangladesh) margins of the Bengal Basin. AB - Although arsenic (As) contamination of groundwater in the Bengal Basin has received wide attention over the past decade, comparative studies of hydrogeochemistry in geologically different sub-basins within the basin have been lacking. Groundwater samples were collected from sub-basins in the western margin (River Bhagirathi sub-basin, Nadia, India; 90 samples) and eastern margin (River Meghna sub-basin; Brahmanbaria, Bangladesh; 35 samples) of the Bengal Basin. Groundwater in the western site (Nadia) has mostly Ca-HCO(3) water while that in the eastern site (Brahmanbaria) is much more variable consisting of at least six different facies. The two sites show differences in major and minor solute trends indicating varying pathways of hydrogeochemical evolution However, both sites have similar reducing, postoxic environments (p(e): +5 to -2) with high concentrations of dissolved organic carbon, indicating dominantly metal-reducing processes and similarity in As mobilization mechanism. The trends of various redox-sensitive solutes (e.g. As, CH(4), Fe, Mn, NO(3)(-), NH(4)(+), SO(4)(2-)) indicate overlapping redox zones, leading to partial redox equilibrium conditions where As, once liberated from source minerals, would tend to remain in solution because of the complex interplay among the electron acceptors. PMID- 18164514 TI - Skin bradykinin-related peptides (BRPs) and their biosynthetic precursors (kininogens): comparisons between various taxa of Chinese and North American ranid frogs. AB - Bradykinins and related peptides (BRPs) occur in the defensive skin secretions of many amphibians. Here we report the structures of BRPs and their corresponding biosynthetic precursor cDNAs from the Chinese brown frog, Rana chensinensis, and the North American leopard frog, Lithobates pipiens. R. chensinensis skin contained four transcripts each encoding a different kininogen whose organizations and spectrum of encoded BRPs were similar to those reported for the pickerel frog, Lithobates palustris. In contrast, from L. pipiens, a single skin kininogen was cloned whose structural organization and spectrum of mature BRPs were similar to those reported for the Chinese piebald odorous frog, Huia schmackeri. These data also implied that the endogenous precursor processing proteases in each species pair have identical site-directed specificities, which in part may be dictated by the primary structures of encoded BRPs. Thus the spectra of skin BRPs and the organization of their biosynthetic precursors are not consistent with recent taxonomy. The natural selective pressures that mould the primary structures of amphibian skin secretion peptides are thought to be related to the spectrum of predators encountered within their habitats. Thus similarities and differences in skin bradykinins may be reflective of predator spectra rather than indicative of species relatedness. PMID- 18164515 TI - Upregulation of EMMPRIN after permanent focal cerebral ischemia. AB - Elevated activities of matrix metalloproteinases (MMPs) following ischemic stroke have been shown to mediate ischemic injury as well as neurovascular remodeling. The extracellular MMP inducer (EMMPRIN) is a 58-kDa cell surface glycoprotein, which has been known to play a key regulatory role for MMP activities. The roles of EMMPRIN in stroke injury are not clearly understood. In this study, we investigated changes of EMMPRIN in a mouse model of permanent focal cerebral ischemia, and examined potential association between EMMPRIN and MMP-9 expression. Adult male CD-1 mice were subjected to permanent focal ischemia by intraluminal occlusion of the left middle cerebral artery (MCAO) under anesthesia. EMMPRIN expression was markedly upregulated in the peri-infarct area at 2-7 days after ischemia compared to the contralateral non-ischemic hemisphere by Western blot analysis. Immunofluorescent double staining demonstrated that EMMPRIN signals co-localized with vwF-positive endothelial cells and GFAP positive peri-vascular astrocytes. In contrast, EMMPRIN signal did not co localize with NeuN-positive neurons, or MPO-positive neutrophils. Dual fluorescent staining revealed that EMMPRIN co-localized with MMP-9. Our data also demonstrated that increased EMMPRIN expression correlated with increased MMP-9 levels in a temporal manner. In summary, we report for the first time that EMMPRIN expression was significantly increased in a mouse model of permanent focal cerebral ischemia. The spatial and temporal association between increased EMMPRIN expression and elevated MMP-9 levels suggest that EMMPRIN may modulate MMP-9 activity, and participate in neurovascular remodeling after ischemic stroke. PMID- 18164516 TI - In vitro and in vivo evidences that antioxidant action contributes to the neuroprotective effects of the neuronal nitric oxide synthase and monoamine oxidase-B inhibitor, 7-nitroindazole. AB - The neuronal nitric oxide synthase (nNOS) inhibitor, 7-nitroindazole (7-NI) is neuroprotective against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced parkinsonism. Monoamine oxidase (MAO)-B inhibitory action partially contributes to this effect. We tested the hypothesis that 7-NI could be a powerful hydroxyl radical (OH) scavenger, and interferes with oxidative stress caused by MPTP. We measured OH, reduced glutathione (GSH), as well as superoxide dismutase (SOD) and catalase activities in the nucleus caudatus putamen and substantia nigra of Balb/c mice following MPTP and/or 7-NI administration. The nNOS inhibitor caused dose-dependent inhibition in the production of OH in (i) Fenton-like reaction employing ferrous citrate in a cell-free system in test tubes, (ii) in isolated mitochondrial preparation in presence of MPP+, and (iii) in the striatum of mice systemically treated with MPTP. An MPTP-induced depletion of GSH in both the nuclei was blocked by 7-NI, which was dose-dependent (10 50mg/kg), but independent of MAO-B inhibition. The nNOS-mediated recovery of GSH paralleled attenuation of MPTP-induced depletion of striatal dopamine. MPTP induced increase in the activities of striatal or nigral SOD and catalase were significantly attenuated by 7-NI treatment. These results suggest potent antioxidant action of 7-NI in its neuroprotective effects against MPTP-induced neurotoxicity. PMID- 18164517 TI - Synergistic Ca2+ and Cu2+ requirements of the FGF1-S100A13 interaction measured by quartz crystal microbalance: an initial step in amlexanox-reversible non classical release of FGF1. AB - It is known that fibroblast growth factor-1 (FGF1) lacking a conventional signal peptide sequence shows non-classical release independent of the endoplasmic reticulum-Golgi system. Recent studies reveal that FGF1 is co-released with S100A13, a Ca2+-binding protein that acts as an extracellular cargo molecule. Although both FGF1 and S100A13 are Cu2+-binding proteins, the role of Cu2+, as well as that of Ca2+, in non-classical release, remains to be clarified. In the present study we examined the requirements of both metal ions for the interaction between these two proteins. The addition of Ca2+ significantly increased the ka value, while decreasing the KD value, for the interaction between Strep-tagII S100A13 and GST-FGF1; both values were obtained by use of a quartz crystal microbalance, a real-time mass-measuring device. The EC50 of Ca2+ to enhance the interaction was 10.11 microM. Although the addition of Cu2+ alone had no effect, it caused a marked potentiation of the Ca2+-enhanced interaction. The EC50 of Cu2+ for the potentiation was 50.45 nM. On the other hand, the EC50 of Ca2+ and the KD values were decreased from 11.69 to 2.07 microM and 0.75 to 0.38 x 10( 7)M, respectively, by the addition of 200 nM Cu2+. The Cu2+-induced potentiation of this interaction was abolished by amlexanox, which inhibits non-classical release of FGF1. All of these findings suggest that synergistic effects of Ca2+ and Cu2+ play a key role in the interaction between FGF1 and S100A13, which is the initial step in non-classical release of FGF1. PMID- 18164518 TI - Synthesis of potential Rho-kinase inhibitors based on the chemistry of an original heterocycle: 4,4-dimethyl-3,4-dihydro-1H-quinolin-2-one. AB - A new series of substituted 4,4-dimethyl-3,4-dihydro-1H-quinolin-2-one have been prepared via condensation of 3,3-dimethylacryloyl chloride with aniline. Details of synthetic procedures are shown. Our aim was to investigate the potency of our original heterocycle in the inhibition of the Rho-kinase enzyme, known to be of major importance in the cascade reactions leading to arterial hypertension. Biological activity for the seven compounds has been investigated and is presented. PMID- 18164519 TI - The potential of a protease activation mutant of a highly pathogenic avian influenza virus for a pandemic live vaccine. AB - The most effective countermeasure against a pandemic originating from a highly pathogenic avian influenza virus (HPAIV) is immunoprophylaxis of the human population. We present here a new approach for the development of a pandemic HPAIV live vaccine. Using reverse genetics, we replaced the polybasic hemagglutinin cleavage site of an H7N7 HPAIV with an elastase motif. This mutant was strictly elastase-dependent, grew equally well as the wild-type in cell culture and was attenuated in mice unlike the lethal wild-type. Immunization at 10(6)pfu dosage protected mice against disease and induced sterile immunity; vaccination with homosubtypic or heterosubtypic reassortants led to cross protection. These observations demonstrate that a mutated hemagglutinin requiring elastase cleavage can serve as an attenuating component of a live vaccine against HPAIV. PMID- 18164520 TI - B cell immunopathology during HIV-1 infection: lessons to learn for HIV-1 vaccine design. AB - Induction of broad HIV-1 neutralizing antibodies should be a major goal of an effective HIV-1 vaccine. However, B cells are severely damaged during HIV-1 infection with loss of memory B cells and decline of serological memory. The molecular events leading to B cell damage must be further characterized with the aim of selecting vaccine components allowing preservation of B cell functions. This review focuses on B cell damage and antibody responses in HIV-1-infected patients during vaccination studies with viral and bacterial antigens. In addition novel data indicate that B cell activation may be at the basis of impaired immune responses. PMID- 18164521 TI - AIDS vaccine: efficacy, safety and ethics. AB - Human immunodeficiency virus (HIV) has infected 50 million people worldwide and killed 16 million so far, and the epidemic is still spreading with 16,000 new cases of HIV infection daily and a projection of 100 million infected individuals by the end of the next decade. There is no question that a safe and effective acquired immunodeficiency syndrome (AIDS) vaccine is urgently needed to bring the current AIDS pandemic under control. But, is preventive AIDS vaccine an attainable goal? Unfortunately, the results of many laboratory and clinical studies over the past two decades are not encouraging. We comment on the efficacy, safety and ethics of AIDS vaccine, and the urgent need for a new strategy for AIDS vaccine development. PMID- 18164522 TI - Distribution coefficients of tin in Japanese agricultural soils and the factors affecting tin sorption behavior. AB - Sorption behavior of tin (Sn) in Japanese agricultural soils was studied. Soil soil solution distribution coefficient (K(d)) of Sn (K(d)-Sn) for 142 soil samples ranged between 128 and 1,590,000 L kg(-1) with the geometric mean of 12 400 L kg(-1). The K(d)-Sn values for Andosol tended to be higher than those of the other soil groups. Among the relationships between K(d)-Sn values and soil properties, a high correlation was observed for soil active-Al (Al-(hydr)oxide and Al-humus complex) amount and K(d)-Sn. The pH effect on Sn sorption was also investigated. The results suggested that the low pH condition enhanced the Sn sorption in soils. The soil-sorbed Sn fractions in each type of soil material were also evaluated with selective extraction methods. The results showed that most of the soil-sorbed Sn was as organic matter bound or Al/Fe-(hydr)oxide-bound forms. PMID- 18164523 TI - Content, distribution and fate of 33 elements in sediments of rivers receiving wastewater in Hanoi, Vietnam. AB - Untreated industrial and domestic wastewater from Hanoi city is discharged into rivers that supply water for various agricultural and aqua-cultural food production systems. The aim of this study was to assess the content, distribution and fate of 33 elements in the sediment and pore water of the main wastewater receiving rivers. The sediment was polluted with potentially toxic elements (PTEs) with maximum concentrations of 73 As, 427 Cd, 281 Cr, 240 Cu, 218 Ni, 363 Pb, 12.5 Sb and 1240 Zn mg kg(-1) d.w. Observed distribution coefficients (log(10) K(d,obs)) were calculated as the ratio between sediment (mg kg(-1) d.w.) and pore water (mg L(-1)) concentrations. Maxima log(10) K(d,obs) were >4.26 Cd, >6.60 Cu, 4.78 Ni, 7.01 Pb and 6.62 Zn. The high values show a strong PTE retention and indicate the importance of both sorption and precipitation as retention mechanisms. Sulphide precipitation was a likely mechanism due to highly reduced conditions. PMID- 18164524 TI - Tobacco industry sociological programs to influence public beliefs about smoking. AB - The multinational tobacco companies responded to arguments about the social costs of smoking and hazards of secondhand smoke by quietly implementing the Social Costs/Social Values project (1979-1989), which relied upon the knowledge and authoritative power of social scientists to construct an alternate cultural repertoire of smoking. Social scientists created and disseminated non-health based, pro-tobacco arguments without fully acknowledging their relationship with the industry. After the US Surgeon General concluded that nicotine was addictive in 1988, the industry responded by forming "Associates for Research in the Science of Enjoyment" (c.1988-1999), whose members toured the world promoting the health benefits of the use of legal substances, including tobacco, for stress relief and relaxation, without acknowledging the industry's role. In this paper we draw on previously secret tobacco industry documents, now available on the Internet to show how both of these programs utilized academic sociologists, political scientists, anthropologists, psychologists, philosophers and economists, and allowed the industry to develop and widely disseminate friendly research through credible channels. Strategies included creating favorable surveys and opinions, infusing them into the lay press and media through press releases, articles and conferences, publishing, promoting and disseminating books, commissioning and placing favorable book reviews, providing media training for book authors and organizing media tours. These programs allowed the tobacco industry to affect public and academic discourse on the social acceptability of smoking. PMID- 18164525 TI - Socioeconomic context, distance to primary care and detection of hepatitis C: a French population-based study. AB - Previous research suggests that hepatitis C detection is lower in rural than in urban areas and depends on geographic accessibility to care. To what extent differences in socioeconomic context could explain these disparities remains unknown. This study assessed the respective influence of the socioeconomic context and of the distance to primary care on the hepatitis C detection rates in a well-defined population of 1,005,817 inhabitants covered by a universal health insurance system in the Cote d'Or and the Doubs areas of France. The 1938 cases of hepatitis C newly diagnosed in the survey area between 1994 and 2001 were included. The patients' 'cantons' of residence were classified into six socioeconomic clusters according to 19 quantitative indicators. Age and sex standardized hepatitis C detection rate ratios (DRR) for the six clusters were estimated and a multilevel Poisson model investigated whether detection rates varied across clusters after adjusting for distance to the nearest general practitioner. Standardized DRR (95% CI) were higher in both "lower-income urban" and "upper-income urban" clusters and lower in the following clusters: "outer suburb", "industrial rural" and economically "dynamic rural". Adjusting for distance to general practitioner reduced the rate difference between socioeconomic clusters: for a distance to general practitioner equalling 0km (practice in the municipality) and compared to the "upper-income urban" cluster, only the "lower-income urban" cluster had higher DRR. In the six clusters, detection rates decreased as the distance to general practitioner increased (DRR 0.89 95% CI 0.84-0.96 for a 1-km change) but the decrease was more marked in both urban clusters. In this population with good primary care affordability, geographic proximity to general practitioner, rather than socioeconomic context of neighborhood, appeared to be the main factor limiting hepatitis C detection. PMID- 18164526 TI - Childhood social class and cancer incidence: results of the globe study. AB - Despite increased recognition of the importance of investigating socio-economic inequalities in health from a life course perspective, little is known about the influence of childhood socio-economic position (SEP) on cancer incidence. The authors studied the association between father's occupation and adult cancer incidence by linking information from the longitudinal GLOBE study with the regional population-based Eindhoven Cancer Registry (the Netherlands) over a period of 14 years. In 1991, 18,973 participants (response rate 70.1%) of this study responded to a postal questionnaire, including questions on SEP in youth and adulthood. Respondents above the age of 24 were included (N=12,978). Cox regression was used to calculate hazard ratios (HR) for all cancers as well as for the five most frequently occurring cancers by respondent's educational level or occupational class, and by father's occupational class (adjusted for respondent's education and occupation). Respondents with a low educational level showed an increased risk of all cancers, lung and breast cancer (in women). Respondents with a low adult occupational level showed an increased risk of lung cancer and a reduced risk of basal cell carcinoma. After adjustment for adult education and occupation, respondents whose father was in a lower occupational class showed an increased risk of colorectal cancer as compared to those with a father in the highest social class. In contrast, respondents whose father was in a lower occupational class, showed a decreased risk of basal cell carcinoma as compared to those with a father in the highest occupational class. The association between childhood SEP and cancer incidence is less consistent than the association between adult SEP and cancer incidence, but may exist for colorectal cancer and basal cell carcinoma. PMID- 18164527 TI - Proverb interpretation changes in aging. AB - Recent investigations have emphasized the involvement of fronto-subcortical networks to proverb comprehension. Although the prefrontal cortex is thought to be affected by normal aging, relatively little work has been carried out to investigate potential effects of aging on proverb comprehension. In the present investigation participants in three age groups were assessed on a proverb comprehension task and a range of executive function tasks. The older group showed impairment in selecting correct interpretations from alternatives. They also showed executive function deficits, as reflected by reduced working memory and deficient set shifting and inhibition abilities. The findings of the present investigation showed proverb comprehension deficits in normal aging which appeared to be related to reduced executive skills. PMID- 18164528 TI - Aripiprazole as an adjunctive treatment for refractory unipolar depression. AB - INTRODUCTION: Aripiprazole may be an effective adjunctive treatment in outpatients with unipolar depression that has been refractory to treatment with SSRI or SNRI medication. METHODS: Fifteen subjects with a current DSM-IV diagnosis of MDD which had not responded to SSRI or SNRI treatment were enrolled in a 12 week open-label study of aripiprazole with a maximum dose of 30 mg/day. Patients' current episode averaged 10.4+/-16.6 years, with a range of 3 months to 54 years. Baseline severity averaged 30.1+/-7.1 on HDRS-24, and 19.7+/-8.4 on BDI. Patients had been treated with a mean dose of 79.2+/-28.2 mg/day of fluoxetine equivalents for an average of 1 year prior to starting the study. Five subjects were on SNRI medications and 10 on SSRIs. RESULTS: Seven of 14 (50.0%) subjects were classified as treatment responders, as defined by at least 50% reduction in the HDRS-24 at week 12. Four subjects (28.6%) achieved remission, based on STAR D criteria (HDRS-17 score